ANALELE STIINTIFICE biologie 2 2008

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ANALELE ŞTIINŢIFICE ALE

UNIVERSITĂŢII „Alexandru Ioan Cuza” DIN IAŞI

(SERIE NOUĂ)

SSEECCŢŢIIUUNNEEAA IIII aa.. BBIIOOLLOOGGIIEE VVEEGGEETTAALLĂĂ

Editura Universităţii „Alexandru Ioan Cuza” Iaşi

TOMUL LIV, FASCICULA 1 2008

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Editura Universităţii „Alexandru Ioan Cuza” din Iaşi

ANALELE ŞTIINŢIFICE ALE UNIVERSITĂŢII „Alexandru Ioan Cuza” DIN IAŞI (SERIE NOUĂ), SECŢIUNEA II a. BIOLOGIE VEGETALĂ

Comitetul de redacţie:

Profesor Dr. Constantin TOMA – Universitatea „Alexandru Ioan Cuza” din Iaşi Profesor Dr. Toader CHIFU – Universitatea „Alexandru Ioan Cuza” din Iaşi Profesor Dr. Mihai MITITIUC – Universitatea „Alexandru Ioan Cuza” din Iaşi Profesor Dr. Maria Magdalena ZAMFIRACHE - Universitatea „Alexandru Ioan Cuza” din

Iaşi Conferenţiar Dr. Cătălin TĂNASE – Universitatea „Alexandru Ioan Cuza” din Iaşi Conferenţiar Dr. Lăcrămioara IVĂNESCU - Universitatea „Alexandru Ioan Cuza” din Iaşi

Comisia de referenţi ştiinţifici: Academician Nicolae BOŞCAIU – Academia Română, Filiala Cluj-Napoca Academician Valeriu COTEA – Academia Română, Filiala Iaşi Profesor Dr. Constantin TOMA – Universitatea „Alexandru Ioan Cuza” din Iaşi, membru

corespondent al Academiei Române, Filiala Iaşi Profesor Dr. Leontin Ştefan PÉTERFI – Universitatea „Babeş-Bolyai” din Cluj-Napoca,

membru corespondent al Academiei Române, Filiala Cluj-Napoca Profesor Dr. Jean Pierre AUQUIÈRE – Universitatea Catolică din Louvain la Neuve, Belgia Profesor Dr. Maria COULADIS – Universitatea din Atena, Grecia Profesor Dr. Cvetomir DENCHEV – Academia de Ştiinţe din Bulgaria Profesor Dr. Franco PEDROTTI – Universitatea din Camerino, Italia Profesor Dr. Andrei MARIN – Universitatea din Bucureşti Profesor Dr. Ioan BURZO – Universitatea Agronomică şi de Medicină Veterinară din

Bucureşti Profesor Dr. Toader CHIFU – Universitatea „Alexandru Ioan Cuza” din Iaşi Profesor Dr. Mihai MITITIUC – Universitatea „Alexandru Ioan Cuza” din Iaşi Profesor Dr. Ursula STĂNESCU– Universitatea de Medicină şi Farmacie „Gr. T. Popa” din Iaşi Conferenţiar Dr. Cătălin TĂNASE – Universitatea „Alexandru Ioan Cuza” din Iaşi Redactor responsabil: Profesor Dr. Constantin TOMA,

membru corespondent al Academiei Române

Secretar de redacţie: Şef lucrări Dr. Naela COSTICĂ Tehnoredactare computerizată: Dr. Ramona Crina GALEŞ

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CONTENTS

IRINA STĂNESCU, C. TOMA – The leaf structure of some Nepenthes Danserspecies 5

IRINA BERCIU, C. TOMA – Histo-anatomical aspects reffering to thevegetative organs of two subspecies of Thymus pannonicus All. 16

LUMINIŢA HUŢANU-BASHTAWI, C. TOMA – Contributions to the histo-anatomical study of the Calendula officinalis L. leaves treated with thiophanatemethyl (topsin M) 22

IOANA MARCELA PĂDURE, LILIANA BĂDULESCU, TEODORA DEDIU,I. BURZO – Morpho-anatomical and phytochemical researches regardingPseudotsuga menziesii (Mirbel) Franco (Pinaceae) 33

ALEXANDRINA MURARIU, CORINA GRĂDINARIU, ANIŞOARASTRATU – The ecophysiological reaction of some varieties of apple tree, peartree and quince tree to the pathogenic agents attack 40

ELENA CRISTINA ROŞU, MARIA MAGDALENA ZAMFIRACHE, I. I.BĂRA - Physiological effects induced by purinic substances at Capsicumannuum L. 46

M. RÎŞCA, L. FĂRTĂIŞ, ANA LEAHU – The influence on the Mn2+ ionseffects on the wheat (Triticum aestivum L. ) seed germination 50

NICOLETA IANOVICI, I. E. JUHÁSZ, P. RADISIC, M. JUHÁSZ, B.SIKOPARIJA - Plantago atmospheric pollinic season in the Danube-Kris-Mures-Tisza euroregion (2000-2004) 54

L. POP, DORINA CACHIŢĂ – Araucaria excelsa L. vitrocultures initiation 64

ADRIANA PETRUŞ – VANCEA, C. F. BLIDAR, ANCA BACIU – Africanviolet (Saintpaulia ionantha L.) exvitroplantlets acclimatization, in differenttypes of substratum 71

TATIANA EUGENIA ŞESAN, J. KÖHL, WILMA M. L. MOLHOECK -Ulocladium atrum preuss - biological control agent of grey mould (Botrytiscinerea Pers.) of cropped plants 78

M. COSTICĂ, NAELA COSTICĂ - A new site in Romania for Spirulina(Arthrospira) platensis 92

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T. CHIFU, C. MÂNZU, OANA ZAMFIRESCU – Contribution to the study ofgrassy vegetation in the Ceahlău Mountain 94

J. HANGANU, M. DOROFTEI, N. ŞTEFAN - Assesment of ecologicalstatus of Danube Delta lakes using indicator macrophytes species 103

OANA ZAMFIRESCU – The plant communities with Phragmites australisfrom “The hayfields of Valea lui David” natural reservation (Iasi county) 109

IRINA BLAJ-IRIMIA – Associations of the Molinio-Arrhenatheretea R. Tx.1937 class in Vaslui river basin 113

LOREDANA ASOLTANI – Contributions to the study of paludal vegetationfrom Neagra Şarului river’s basin (Suceava county) 121

CARMEN AONCIOAIE –Protected taxa from the Bistriţa river basin betweenPiatra Neamţ and Bacău 129

MIHAELA AURELIA DANU, T. CHIFU – Contribution to the study of theclass Molinio-Arrhenatheretea R. Tx. 1937 in the upper basin of river Dorna(Suceava county) (I) 136

I. M. CIUMAŞU, NAELA COSTICĂ - Environmental education: education fortransition to sustainable development 146

C.TOMA, MARIA MAGDALENA ZAMFIRACHE - Review 153

C. DRĂGULESCU - Review 154

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Analele ştiinţifice ale Universităţii “Al. I. Cuza” Iaşi Tomul LIV, fasc. 1, s. II a. Biologie vegetală, 2008

THE LEAF STRUCTURE OF SOME NEPENTHES DANSER SPECIES

IRINA STĂNESCU∗, C. TOMA∗∗

Abstract: The authors analyze a few aspects referring to the modified leaf of four Nepenthes species, at different levels, stress being laid on the structure of the vascular bundles, digestive and nectariferous glands. Key words: Nepenthes, digestive glands, nectariferous glands, hydathodes.

Introduction

The Nepenthes genus consists of more than 80 tropical species [8], spread around S-

E Africa, Sri-Lanka and Madagascar. Etienne de Flancourt described it in 1658 for the first time [3] and in 1753 Linné called it Nepenthes. The plant is a climbing, weakly branched liana. It presents a basal rosette of leaves with short internodes when young; on sexual maturity, the internodes become elongated and the plant starts being climbing or prostrate, according to the species.

The plant creates a special impression by its bizarre leaves, consisting of a basal assimilatory part, a tendril which rolls up around different supports and a trap [2]. This trap is like a pitcher with a lid, which covers the trap, avoiding the dilution of the liquid from inside the trap by the rainwater. Some authors believe that the assimilatory part, the tendril and the pitcher belong to the petiole of an archaic leaf, while the lid represents the limb. Others consider that the pitcher and the lid form the limb, and the assimilatory part and the tendril form the petiole.

Darwin stated that a carnivorous plant attracts, captures and digests the prey; the supplementary nutritive elements brought by the prey are necessary in developing and blooming. The capturing system in Nepenthes is passive; the plant does not need to move to capture the prey, unlike those which have active traps, such as Dionaea muscipula.

The plants bear flowers with shiny colours and abundant nectar to attract the pollinating insects; on the other hand, the plants use different traps with different attraction elements: shiny colours or the reflection of the UV radiation, attracting odours or nectar secreted by the nectariferous extrafloral glands; all these characteristics belong to the leaf. Some authors [5, 7] evidenced the structure of the digestive glands; others [1, 6] considered that the digestive glands are closely associated with the vascular bundles. Some histo-anatomical aspects were evidenced in a previous work [9] devoted to Nepenthes maxima.

∗ Botanical Gardens of Iasi, Dumbrava Roşie Street, no. 7-9, Romania ∗∗ “Al. I. Cuza” University, Faculty of Biology, Carol I. Bd., no. 20A, 700506, Iaşi, Romania

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Materials and methods

The material under study, coming from the collection of the “Alexandru Borza” Botanical Gardens of Cluj-Napoca, belongs to four taxa: N. x coccinea Mast, N. distillatoria L., N. maxima Reinw. ex Nees and N. northiana Hook f.

The material subjected to analysis (the modified leaves of the plants) has been fixed and preserved in 70% ethylic alcohol. The sections (from the assimilatory part, tendril and pitcher) were cut with a microtome, then coloured with iodine green and alaun-carmine, mounted in gel and analyzed on a Novex (Holland) light microscope. The light micrographs were performed by means of Novex (Holland) microscope, using a Canon A95 camera.

Results and discussions

As already mentioned, the leaf of Nepenthes consists of three parts: a basal,

assimilatory one, a tendril and a pitcher which represents the trap of the plant. In front side view, the upper epidermis of the assimilatory part appears as formed of

polygonal cells (Fig. 1); here and there, a few hydathodes are present. The lower epidermis consists of small cells, bearing weakly waved walls (Fig. 2). Here and there, stomata of the anomocytic type and hydathodes are present. A hydathode bears a short pedicel formed of a few cells and a stellate part, formed of 4-10 cells. Another author [4] suggests that the hydathodes do not only secrete water, but even absorb it from time to time.

In cross section, the upper epidermis evidences small cells covered by a thick cuticle. Just beneath the epidermis, a few isodiametric-celled layers are present, forming an acviferous tissue (Fig. 3); some authors [5] call it an acviferous hypodermis. Then, a 2-3 layered palisade tissue, with short cells, in which chloroplasts can be observed, is present. The lacunary tissue is multi-layered, with small aeriferous spaces between the component cells. A lot of isolated mechanical cells (idioblasts) with spiral thickenings can be observed in the mesophyll; these were evidenced by other authors [5], too.

The lower epidermis consists of small, isodiametric cells, covered by a cuticle thinner than the one covering the upper epidermis. Numerous stomata are present, as well as numerous calcium oxalate crystals in the mesophyll.

The midvein is very prominent at the lower side of the assimilatory part (Fig. 4). A large number of vascular bundles is present (8 big bundles, one of its being situated in the centre or 6 bundles and a central one at N. northiana); most of them are implanted in a thick sclerenchyma ring, formed of sclerenchymatic fibres with thickened and lignified walls (or unlignified at N. coccinea). The vascular bundles have different orientation in the sclerenchymatic ring. A vascular bundle (Fig. 5) consists of a phloem (sieved tubes and companion cells) and a xylem (xylem vessels separated by celulosic parenchyma). Sometimes, the sclerenchyma sheath bears very small vascular bundles, consisting of a few phloem elements or of phloem and 1-2 xylem vessels.

In the fundamental, external parenchyma, idioblasts and small, isolated vascular bundles are present, often consisting of a few phloem elements, surrounded by a thin sclerenchymatic sheath.

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The tendril In cross section, the tendril shows a circular shape, with 7-8 ribs at N. maxima (Fig.

6). Small cells, covered by a thick cuticle, form the epidermis. Here an there, hydathodes, short, sometimes branched tector hairs and stomata prominig above the epidermis are present. The cortical parenchyma is formed of 5-6 layers of large cells. Most of the vascular bundles are implanted in a strong sclerenchyma ring. There is a high variability regarding the number of bundles and on their position (N. coccinea and N. maxima present a lot of vascular bundles of different size in the sclerenchyma ring and a central one in the fundamental parenchyma; N. distillatoria presents 2 big vascular bundles in the center of the parenchyma and a smaller one, close to them, while N. northiana shows the largest number of vascular bundles, implanted in the sclerenchyma ring, and also two smaller ones in the fundamental parenchyma, but close to the sclerenchyma. The tendril has an homogenous parenchyma, formed of big, turgescent cells and a few idioblasts.

Near the pitcher, the cross section of the tendril is quite circular. The vascular bundles form 2-3 rings (the internal bundles are bigger than the external ones, in the fundamental parenchyma). A sclerenchymatic sheath surrounds each vascular bundle. Numerous calcium oxalates are present in the fundamental parenchyma.

At the inferior level of the pitcher, a typical limb structure is present. In front side view, the internal epidermis presents polygonal elongated cells, with

thick walls. Here and there, a lot of multicellular digestive glands are present (Fig. 7); a small epidermal prolongation can be observed near each gland, yet without touching it. The external epidermis (Fig. 8) consists of small polygonal cells with thin walls, anomocytic stomata, hydathodes and nectariferous glands, which appear like multicellular, massive structures, communicating with the exterior through a short channel.

In cross section, the wall of the pitcher is quite thick. The internal epidermis shows elongated cells, covered by a thick cuticle. Numerous big digestive glands are present in small epidermal cavities (Fig. 9); the epidermal cells form a small fold, without touching the gland. A digestive gland shows 2-3 layers of oblate cells, 1-2 layers of isodiametrical cells and an external layer of columnar-shaped cells. Each digestive gland is associated with small vascular bundles (tracheids with ringed and spiral thickenings). In longitudinal section, the small fold can be better observed (Fig. 10).

The external epidermis consists of small cells, covered by a thin cuticle. The nectariferous glands are formed of three layers of cells delimiting a cavity which opens towards the exterior through a short channel (Fig. 11). The nectariferous glands attract the insects (the prey) to the trap and make them climb the wall of the pitcher to reach the slippery peristome. The assimilatory parenchyma is thick, homogenous, formed of small cells outside and bigger inside. A lot of calcium oxalates are present all over the parenchyma.

The vascular bundles have different sizes, the biggest ones occupying the external part of the parenchyma, while the smallest ones occupy the centre of it. Each vascular bundle consists in phloem facing the exterior part of the pitcher and a xylem facing the internal one, so that the internal epidermis represents the old upper epidermis of an archaic leaf and the external epidermis represents the old lower epidermis. All the studied species show mechanical sheaths surrounding the vascular bundles, consisting of fibres with

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moderately thickened and lignified walls. The assimilatory parenchyma also presents a few idioblasts.

The middle level of the pitcher presents a quite similar structure to that of the anterior level.

In front side view, the internal epidermis consists of polygonal cells with thick walls and secretory glands, smaller than those occurring at the inferior level; the integumentary fold do not touch the gland (Fig. 12). The external epidermis is formed of polygonal cells, anomocytic stomata, multicellular tector hairs, nectariferous glands and hydathodes (Fig. 13).

In cross section, the pitcher shows a thinner wall. The internal epidermis presents digestive glands which communicate with the tracheids (Fig. 14). In longitudinal section, the fold can be better observed (Fig. 15). The external epidermis consists of small cells covered by a thin cuticle. Anomocytic stomata, hydathodes, nectariferous glands (Fig. 16) and multicellular tector hairs, often branched, are present. In the assimilatory parenchyma, numerous calcium oxalates can be observed. Each vascular is bounded by a sclerenchymatic sheath (Fig. 17). There are vascular bundles consisting only of a few phloem elements. The assimilatory parenchyma presents idioblasts, too.

The superior level of the pitcher shows the same structure as the other levels. In front side view, the internal epidermis presents large polygonal cells and digestive glands (Fig. 18) smaller than those occurring at the other levels. The epidermal fold covers more than half of the digestive glands. The external epidermis (Fig. 19) consists of small polygonal cells, with waved lateral walls, tector hairs, hydathodes, nectariferous glands and anomocytic stomata.

Cross section through the superior level of the pitcher shows the digestive glands in their incipient stage of development (Fig. 20), consisting of a small number of cells. Almost half of the gland is covered by the integumentary fold (Fig. 21); the developing stages of the glands during ontogenesis were previously presented [9]. The external epidermis shows similar structures to those of the anterior levels (Figs. 22 and 23). The assimilatory parenchyma is thinner; the vascular bundles are bounded by a very thin mechanical sheath, consisting of fibres with thickened walls, but weakly lignified; some of the bundles present only a few phloem elements; calcium oxalates are not present.

The lid In front side view, both the upper and the lower epidermis present polygonal cells,

with waved lateral walls, anomocytic stomata, secretory glands surrounded by an integumentary fold (Fig. 24) and hydathodes (Fig. 25); tector hairs, sometimes branched, are present only in the lower epidermis. The cross section of the lid is similar to that occurring in the pitcher’s wall. Numerous digestive glands (Fig. 26) are present in both epidermis. The fundamental parenchyma presents vascular bundles of different sizes (Fig. 27), the largest occupying the centre of the parenchyma. All vascular bundles are surrounded by a thin sclerenchyma sheath formed of thin-walled and weakly lignified fibres.

The peristome (Fig. 28) is a common characteristic of the pitcher plants. All four investigated Nepenthes species have a ridged peristome. The epidermal cells are small, covered by a very thick cuticle. Stomata and hydathodes are present in the lower epidermis.

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The peristome has an homogenous parenchyma, with large cells, a few idioblasts and small vascular bundles, most of them consisting of phloem elements surrounded by sclerenchyma fibres, with cellulosed unthickened walls.

Conclusions

In spite of the high variability of the pitcher size recorded from one species to

another, their histo-anatomy is quite similar. There have been observed mostly quantitative differences (number of the vascular

bundles, size of the digestive glands at different levels, thickness of the sclerenchymatic sheath, length of the integumentary fold which covers each digestive gland) and not qualitative differences.

REFERENCES

1. ANDERSON A. N., 1994 - Secretion and absorbtion in glands of the carnivorous plant Nepenthes alata. B. A. honors thesis, Connecticut College, New London, C. T.

2. DALTON M. JOS., 1859 - Note sur l’ origin et le développement des urnes dans les plantes du genre Nepenthes. Ann. des Sci. Nat.; sér.Bot., 12: 125-129

3. LLOYD F. E., 1942 - The Carnivorous Plants. Chronica Botanica, 9. Ronald Press, New York 4. MACFARLANE J. M., 1889 - Observations on pitchered insectivorous plants. Part I. Ann. of Bot., 3: 253-

265 5. METCALFE C. R., CHALK L., 1972 - Nepenthaceae in Anatomy of the Dicotyledons. 1: 1105-1111,

Clarendon Press, Oxford 6. STERN K., 1917 - Contribution to the knowledge of Nepenthes. Flora, 109: 213 – 283 7. PARKES D. M., 1980 - Adaptive mechanisms of surfaces and glands in some carnivorous plants. MSc

Thesis, Monash University, Clayton, Victoria, Australia 8. STAROSTA P., LABAT J.-J., 1993. - L’univers des plantes carnivores. Éd. Du May, Paris 9. TOMA I., TOMA C., STĂNESCU I., 2002 - Histo-anatomical aspects of the Nepenthes maxima Reinw. ex

Nees metamorphosed leaf. Rev. Roum. Biol., sér. Biol. végét., 47, 1-2: 3-7

Acknowledgments

The authors gratefully thank to Felician Micle PhD, Ex-Director of the Botanical Garden of Cluj-Napoca and to Elena Rânba for supplying the material for the present investigation.

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Explanation of plates: Plate I 1. The upper epidermis of the assimilatory part of N. maxima, in front side view 2. The lower epidermis of the assimilatory part of N. distillatoria, in front side view 3. The mesophyll of the assimilatory part of N. distillatoria 4. Cross section through the midvein of the assimilatory part of N. maxima 5. The biggest vascular bundle of the midvein belonging to the assimilatory part of

N. maxima 6. Cross section through the tendril of N. maxima Plate II 7. The internal epidermis of the inferior level of N. distillatoria pitcher, in front side view 8. The external epidermis of the inferior level of N. distillatoria pitcher, in front side view 9. Cross section through the inferior level of N. distillatoria pitcher 10. Longitudinal section of the inferior level of N. maxima pitcher 11. Cross section through the inferior level of N. distillatoria pitcher 12. The internal epidermis of the middle level of N. northiana pitcher, in front side view Plate III 13. The internal epidermis of the middle level of N. maxima pitcher, in front side view 14. Cross section through the middle level of N. northiana pitcher 15. Longitudinal section through the middle level of N. maxima pitcher 16. Cross section through the middle level of N. distillatoria pitcher 17. Cross section through the middle level of N. maxima pitcher 18. The internal epidermis of the superior level of N. coccinea pitcher, in front side view Plate IV 19. The internal epidermis of the superior level of N. distillatoria pitcher, in front side

view 20. Cross section through the superior level of N. distillatoria pitcher 21. Longitudinal section through the superior level of N. maxima pitcher 22. Cross section through the superior level of N. distillatoria pitcher 23. Cross section through the superior level of N. northiana pitcher 24. The upper epidermis of the lid belonging to N. northiana pitcher, in front side view Plate V 25. The lower epidermis of the lid belonging to N. northiana pitcher, in front side view 26. Cross section through the lid of N. coccinea pitcher 27. Cross section through the lid of N. northiana pitcher 28. Cross section through the peristome of N. northiana pitcher

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IRINA STĂNESCU, C.TOMA PLATE I

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IRINA STĂNESCU, C.TOMA PLATE II

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IRINA STĂNESCU, C.TOMA PLATE III

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17 18

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IRINA STĂNESCU, C.TOMA PLATE IV

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23 24

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IRINA STĂNESCU, C.TOMA PLATE V

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HISTO-ANATOMICAL ASPECTS REFFERING TO THE VEGETATIVE

ORGANS OF TWO SUBSPECIES OF THYMUS PANNONICUS ALL.

IRINA BERCIU*, C. TOMA*

Abstract: The authors analyze comparatively two Thymus pannonicus subspecies (Thymus pannonicus ssp. auctus and Thymus pannonicus ssp. pannonicus). The vegetative organs (root, stem and leaf) of the two studied taxa were histo-anatomically investigated, evidencing the structure, localization and frequency of the secretory structures of essential oils. The secretory structures of essential oils are always multicellular, consisting of a basal cell, a unicellular pedicel and a gland which bears 1, 2 or 8 cells. Key words: Thymus, anatomy, vegetative organs, trichomes.

Introduction1

Thymus pannonicus is wide spread in our country, living in meadows, bushes, forest

glades, sands, heated rocks, from the steppe region to the holm level [2]. The plant presents vigorous, ascending at the basis or until the inflorescence,

branched stems, the entire surface being covered by trichomes measuring the same length as does the diameter of the stem. The plant presents a capitulum inflorescence, the calyx of the flowers reaching 3-4 mm; the corolla is red to mauve, reaching 6-7 mm [2].

In our country, T. pannonicus presents two subspecies which are focused in the present paper:

- T. pannonicus ssp. pannonicus - a villous-hairy plant, which presents a lot of trichomes on both sides of the leaves;

- T. pannonicus ssp. auctus (Lyka) Soó - a plant which bears glabrous leaves. Most of the Lamiaceae species have been investigated from anatomical point of

view; their general structure characteristics were presented in the fundamental tractates regarding the anatomy of the dicotyledons [3] or of the angiosperms (Napp-Zinn, 1973, 1974). Regarding the Thymus genera, in our country, Toma and Rugină have investigated only the anatomy of T. vulgaris.

We have publicized until now a paper referring to the seedling structure of Thymus vulgaris.

Material and methods

The material under study is represented by two subspecies: T. pannonicus ssp.

pannonicus, collected on 12.06.2007 from Potoci (Neamţ district) and T. Pannonicus ssp. auctus, collected on 16.07.2007 from Fălticeni (Suceava district).

∗„Al. I.Cuza” University, Faculty of Biology, Carol I Bd., no. 20A, 700506, Iasi, Romania

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The vegetal material has been fixed and preserved in 70% ethylic alcohol. The sections were cut with a microtome and a botanical razor. The vegetative organs, subterranean and overhead, were cross sectioned, on different levels, from the top to the basis. First of all, the sections were submitted to a discoloration process, using sodium hypochlorite (20-25’), then coloured (with iodine green and with alaun-carmine) and mounted in gel.

The drawings were performed by means of a Romanian (Projektionszeichenspiegel) microscope and the micrographs were performed by means of a Novex (Holland) microscope, using a Canon A95 camera.


The root (Plate I. 1, 2). There have not been observed major differences at the two subspecies; both of them

present a root with an early secondary structure generated by the activity of both lateral meristems: the cambium and the phellogen.

The phellogen is generated by a profound cortical layer. The cambium forms a thin phloem ring and 2-3 secondary xylem rings (the vessels are spread in the fundamentally massive of libriform). All the rings present a lot of early xylem vessels of large diameter and less lately xylem vessels of smaller diameter, resulting very easy to establish the limit between different rings.

The stem (Plate I. 3, 4, 5, 6; Plate II. 1, 2). The superior level of the stem belonging to T. pannonicus ssp. pannonicus has a

quadratic shape in cross section, with less prominent ribs where sheaths of angular collenchyma are present. The epidermis consists in isodiametric cells, having a bellied external wall thicker than the others and covered by a thick cuticle. T. pannonicus ssp. auctus presents tangentially elongated epidermal cells. Here and there both species present stomata (promining above the external part of the epidermis) and trichomes of two types: tector trichomes (almost all the trichomes are bi- or three-celled, with obtuse point) and short, multicellular secretory trichomes. T. pannonicus ssp. pannonicus presents a thicker cortex (5-6 layers) than T. pannonicus ssp. auctus (3-4 layers). The cortex does not present a casparian endodermis, opposite to other Lamiaceae species.

Both analyzed species show a central cylinder consisting in annular vascular tissues: an external, thin phloem ring and an internal, thick xylem ring (formed by xylem vessels and cells belonging to the xylem cellulosed parenchyma); a large meristematic region is present between them; the tracheogenesis is in process.

The middle level of the stem shows a quadratic shape in cross section, too, with very prominent ribs. T. pannonicus ssp. pannonicus presents longer tector trichomes and more numerous on unit area; the epidermal cells are covered by a thick cuticle (resulting a ridged profile); the internal layer of the cortex may be considered as playing the role of a casparian endodermis. T. pannonicus ssp. auctus does not present a typical endodermis yet.

The central cylinder shows a typical secondary structure at both taxa: a continuous, thin phloem ring and a thick xylem ring where the libriform is prevalent; the libriform fibers present thick and intense lignified walls.

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The meristematic region (which is present between the phloem and the xylem) is very thick. The tracheogenesis is still in process. Some of the cells belonging to the central part of the pith start the disarrangement process.

At the inferior level, the stem still shows a quadratic shape, but presenting less prominent ribs at both species. The epidermal cells are covered by a thicker cuticle; the cortex is thinner that the one belonging to the others levels because the most internal layers consist in strong oblate cells (with hardly noticeable lumina), the external ones are moderately collenchymatized and the collenchyma bands are present in the ribs.

Only at this level, a casparian endodermis can be distinguished at T. pannonicus ssp. auctus.

The structure of the central cylinder is quite the same, but the xylem ring is thicker and stronger lignified.

Most of the pith is disorganized, appearing a large aeriferous cavity, of irregular shape at T. pannonicus ssp. pannonicus and more narrowed at T. pannonicus ssp. auctus.

The foliar limb (Plate II. 3) In front side view, the epidermis consists in elongated cells, with uncurved lateral

walls, at both analyzed subspecies. A lot of tector trichomes, of different size, are present at the edges of the limb

belonging to T. pannonicus ssp. pannonicus; their structure varies from unicellular to multicellular, having an obtuse or narrow point; some of them are inflected toward the epidermis, bearing a large basal cell and a thick wall; they are frequently present in the lower epidermis.

The leaves are glabrous at T. pannonicus ssp. auctus; only the edges of the limb present some short aculeiform trichomes which are not mentioned in the literature.

The vascular bundles are big and present a very thick sclerenchymatic sheath at the phloemic pole; the sclerenchymatic fibers have a thick and lignified wall, at both analyzed subspecies.

The mesophyll is formed by palisade tissue and spongy tissue; the former is bi-layered, but the cells belonging to the hypodermic layer are higher than the others.

Both taxa shows multicellular secretory trichomes, bearing a basal cell, smaller than the adjacent epidermic cells, a short unicellular pedicel and a gland formed by 1, 2 or 8 secretory cells, all of them being covered by a common bellied cuticle.

The stem presents shorter secretory trichomes, their frequency rises from the basis to the apex of the organ. In front side view, the epidermic cells of the limb, which surrounds the secretory trichomes, are radialy elongated towards the cuticle and bear uncurved walls.

19

Conclusions

The superior level of the stem does not present a casparian endodermis at both subspecies; it appears at the middle level of the stem belonging to T. pannonicus ssp. pannonicus or at the inferior level of the stem belonging to T. pannonicus ssp. auctus. The xylem vessels are spread all over the libriform at T. pannonicus ssp. auctus or only in its internal part at T. pannonicus ssp. pannonicus. The aeriferous cavity appears earlier at T. pannonicus ssp. pannonicus than at T. pannonicus ssp. auctus. T. pannonicus ssp. pannonicus presents the largest number of secondary xylem vessels on unit area. The frequency of the tector trichomes decreases toward the basis of the stem at both subspecies.

REFERENCES

1. CIOCÂRLAN V., 2000 - Flora ilustrată a României. Pteridophyta et Spermatophyta. Ed. Ceres, Bucureşti:

670-675 2. GUŞULEAC M., 1961 - Thymus, In Flora Republicii Populare Române, VIII, Ed. Acad. RPR, Bucureşti:

301-334 3. METCALFE C.R., CHALK L., 1950 - Anatomy of Dicotyledons, vol II, Clarendon Press, Oxford: 1041-

1053 4. TOMA C., BERCIU I., 2007 - Morphological peculiaries of germination and structure of seedling in Thymus

vulgaris L.; Rom. Biol. Sci., 5, 1-2: 136-137 5. TOMA C., RUGINĂ R., 1998 - Anatomia plantelor medicinale. Atlas. Ed.Acad. Rom., Bucureşti: 169-172 Explanation of the plates: Plate I 1. Cross section through the root of T. pannonicus ssp. pannonicus (x400) 2. Cross section through the root of T. pannonicus ssp. auctus (x400) 3. Cross section through the inferior level of the stem of T. pannonicus ssp. pannonicus

(x400) 4. Cross section through the inferior level of the stem of T. pannonicus ssp. auctus

(x400) 5. Cross section through the middle level of the stem of T. pannonicus ssp. pannonicus

(x400) 6. Cross section through the middle level of the stem of T. pannonicus ssp. auctus (x400) Plate II 1. Cross section through the superior level of the stem of T. pannonicus ssp. pannonicus

(x 200) 2. Cross section through the superior level of the stem of T. pannonicus ssp. auctus

(x 200) 3. Cross section through the limb of T. pannonicus ssp. pannonicus (x400) 4. Glandular trichome of T. pannonicus ssp. auctus (x400) 5. Tector trichome of T. pannonicus ssp. pannonicus (scale bars=50µm) 6. Aculeiform trichome of T. pannonicus ssp. auctus (scale bars=50µm) 7. Multicellular secretory trichome of T. pannonicus ssp. auctus (x400) 8. Tector and glandular trichome of T. pannonicus ssp. pannonicus (x400)

20

IRINA BERCIU, C. TOMA PLATE I 1.

1 2

34

5 6

21

IRINA BERCIU, C. TOMA PLATE II

1 2

3

4

7 8

56

22


CONTRIBUTIONS TO THE HISTO-ANATOMICAL STUDY OF THE

CALENDULA OFFICINALIS L. LEAVES TREATED WITH THIOPHANATE METHYL (TOPSIN M)

LUMINIŢA HUŢANU-BASHTAWI∗, C. TOMA∗

Abstract. The study analyzes the histo-anatomical modifications of the Calendula officinalis leaf, caused by the treatment with thiophanate methyl, applied 3 times, in two different concentrations, of 0.1% and, respectively, 0.4%. The cross-sections made at the three levels of the leaf, as well as the surface ones, evidenced some quantitative differences between the two variants of treatment and the reference, while the differences of qualitative type are minimum, referring to the different distribution of the pallisadic tissue on the two sides of the foliar limb; consequently, the leaf structure is different: bifacial unequally equifacial in the reference and bifacial heterofacial, respectively, in the treated samples. The quantitative type modifications are related to the prominence extent of the median nervure, thickness of the meristematic area, size and number of the conducting fascicles and the xylem vessels (which are intensely stimulating parameters in the two treatments), the presence of secretory hairs, width and thickness of the foliar limb which, at a concentration of 0.4%, are slightly inhibited, in spite of the fact that the median nervure is much more prominent, even comparatively with the 0.1% concentration treatment. Keywords: Calendula, Topsin M, cytokinin hormone-type action, foliar limb, histo-anatomical modifications.

Introduction

The influence of fungicides on plants productivity is usually attributed to the primary fungicide/fungistatic effect of such substances, although, quite frequently, they may show secondary physiological effects, which may be either toxic or, on the contrary, beneficial to the plants subjected to such treatments [3]. The thiophanate methyl, a systemic fungicide belonging to the benzimidazole class, is largely utilized, as due to its large spectrum of action, as a curative and protecting substance for the cultivation of alimentary, industrial as well as medicinal plants [9]. Involvement of benzimidazoles and of some of their derivatives in the regulation of certain physiological processes developed at plant level has been extensively studied, being usually defined as a “cytokynin hormone-type action” [6, 10]. In the case of both thiophanate methyl and carbendazime, the main metabolite and the fungicide’s active substance, respectively, cytokinin-like effects have been demonstrated in some culture plants, being manifested by the inhibition of leaves senescence, lower degradation of chlorophyll, proteins and AND; more than that, synthesis of the photosynthetic pigments is stimulated, so that the treated leaves maintain their green colour over a longer period of time [1, 4, 5, 6]. The (20 g ml-1) carbendazime solution applied on the leaves of wheat

∗ “Al. I. Cuza” University, Faculty of Biology, 20A Carol I Bd., 700506, Iaşi , Romania

23

prevents the loss of electrolytes and amino acids, as well as disorganization of the cellular organites, the main mechanism of the antisenescence activity exercised by carbendazime being its protecting effect upon the membranary system. At higher concentrations (100 g ml-1), the fungicide losses its cytokinin-like activity; more than that, it even stimulates this loss of electrolytes and amino acids at the level of the membranary system of the treated leaves [7].

It is expected that the action exercised by carbendazime on the treated plants will be possibly extrapolated to the thiophanate methyl, in spite of the fact that, according to some authors [6], the cytokinin hormone-type action might be partially caused by carbendazime, in certain cases fungicides being more active, in this context, then pure carbendazim; on the other hand, the results of the experiments performed with various commercial formulae of the benzimidazolic fungicides are quite controversial as to their secondary effects [8]. Considering all these observations, the present paper analyzes the histo-anatomical modifications induced by thiophanate methyl and/or its main metabolite (MBC) upon the Calendula officinalis leaf, along their correlation with the cytokinin hormone-type effect of the fungicide, anatomically evidenced on the Cynara scolymus leaves [2], comparatively with the non-treated reference sample.


The experimental material, cultivated in the „Anastasie Fătu” Botanical Gardens of

Iasi, was obtained from seeds of the Petrana kind, provided by the Research Station for Medicinal and Aromatic Plants of Fundulea. Besides the treated plants (TM70 0.1%, a concentration, applied in agriculture and, respectively, a TM70 0.4%, a concentration value recommended for fungicides similar to thiophanate methyl), a sample batch, formed of nontreated plants, was prepared for comparative purposes. The administration of fungicide, as a moisty powder, was made three times (at intervals of 7 and 10 days), in the moment of branching or of the first anthodium formation, the plants possessing 30-35 nomophyles. The vegetal material, harvested 10 days after the last treatment, was fixed and conserved in 70% ethanol, then processed according to the methods commonly applied in studies of vegetal anatomy. Measurements were performed on a photonic microscope, by means of a micrometer (ocular and objective), while the light micrographs were performed on a Novex (Holland) microscope, using a Cannon A95 camera.

In this paper we used the following abbreviations: Ca. of. M - Calendula officinalis, control (untreated plants); Ca. of. TM 0,1% - Calendula officinalis, treated with Topsin M 0,1%; Ca. of. TM 0,4% - Calendula officinalis, treated with Topsin M 0,4%.


Cross-sections - the basal leaf, the middle third In the reference – the median nervure is visibly prominent on the inner side of the

limb, evidencing only one conducting fascicle (Fig. 1, 2). The mesophyl shows 2 layers of low pallisadic cells, with sinuous lateral walls on the upper side, and lacunary tissue (6-7

24

layers of cells) on the inferior side, the structure of the limb being bifacial heterofacial (dorsi-ventrally) (Fig. 9, 10).

In the TM 0.1% treated sample – the median nervure strongly prominent on the inferior side of the limb, evidencing a semi-circular contour and including 3 large conducting fascicles which, in the case of Asteraceae, is an exception (Fig. 1, 2). The mesophyl includes 2 layers of law pallisadic cells, on the upper side, and 6 layers of lacunary tissue, so that the structure of the limb is bifacial heterofacial (dorsi-ventrally) (Fig. 9, 10).

In the TM 0.4% treated sample – the median nervure strongly prominent on the inferior side of the limb, with a semi-elliptical contour, forming 3 ribs: a large one in the middle and two, smaller, lateral ones (Fig. 1, 2). The secretory hairs are more numerous on the surface unit, similarly with the tectory ones, which form large bunches on the edge of the limb (Fig. 14 a). The mesophyl evidences 2 layers of low pallisadic cells on the upper side, and 5 layers of lacunary tissue, with isodiametric cells. The cells of the abaxial layer do not form a typical palisade, being rather square-shaped, their ratios being of 1/1.5, while the mesophyl is visibly thinner at this sample (Fig. 9, 10; Tab. IV).

Cross-sections - the leaf from the mid strain, the middle third In the reference – the median nervure is pronouncedly prominent on the inferior

side of the limb and moderately prominent, respectively, on the upper side (Fig. 4). The mesophyl is of the pallisdic type under both epidermes, yet the cells from the adaxial side are taller; between the two pallisades, the cells of the assimilatory parenchyma are isodiametric, the structure of the leaf being therefore bifacial unequally equifacial (Fig. 11, 12). The median conducting fascicle is collaterally open, having a collenchyma girdle at each of the two poles (Fig. 5). All the other conducting fascicles are small, the latter ones possessing only phloem elements.

In the TM 0.1% treated sample – the median nervure, very thick and highly prominent on the inferior side of the limb, includes 3 conducting fascicles (of which, the median one is thicker), all of the open collateral type, each with a collenchyma girdle at both poles (Fig. 4, 5). The mesophyl from the adaxial side is of the pallisadic type, yet with lower cells, similar to those of the reference, while the one from the inferior side is mostly of lacunary type, which explains the different bifacial heterofacial structure of the limb, comparatively with the reference (Fig. 11, 12). The frequency of the secretory hairs is approximately equal to that recorded in the standard sample.

In the TM 0.4% treated sample – the median nervure is highly prominent on both sides of the limb, the hypodermic layer being of collenchymatic type (Fig. 4). The fundamental parenchyma of the median nervure evidences a single conducting fascicle of open collateral type (Fig. 4, 5); the lateral nervures of the first order show, too, relatively large conducting fascicles. The mesophyl is more lax, with the pallisadic tissue at its adaxial side (with visibly lower cells, as actually in the case of the untreated sample) and lacunary tissue on its inferior side, the limb’s structure being bifacial heterofacial, as in the 0.1% treatment (Fig. 11, 12).

25

Cross-sections - the upper leaf, the middle third In the reference – the mesophyl is almost wholly of the pallisadic type, yet with

visibly higher cells in the hypodermic cells; the cells in the middle of the mesophyl are not always perpendicularly elongated on the epidermis, yet oblique; the structure of the limb is, at this level, too, bifacial equifacial (Fig. 13).

In the TM 0.1% treated sample – the median nervure is prominent on both sides of the limb, evidencing isodiametric epidermal cells, with an extremely thick, almost wholly cutinized wall (Fig. 6). The fundamental parenchyma of the median nervure includes a single conducting fascicle, of open collateral type, its wooden vessels occurring in parallel radial rows (8-10) as well as a thin girdle of mechanical fibers, in the course of formation in periphloemic position (Fig. 7); the generating area between the xylem and the phloem has 4 cell layers, comparatively with the reference which evidences only 2 layers (Fig. 8). The mesophyl is homogeneous, formed of isodiametric cells with large aeriferous spaces among them; it is only the cells of the adaxial hypodermal layer that appear slightly higher, reminding of the pallisadic form, with sinuous lateral walls; here and there, up to two layers of “pallisadic” cells may be observed (Fig. 13). The secretory hairs are thicker than in the reference.

In the TM 0.4% treated sample – the secretory hairs are more frequent on the median nervure, yet fewer then in the untreated and 0.1% treated samples. The mesophyl is almost homogeneous, of lacunary type, only the cells of the adaxial hypodermal layer being slightly taller (Fig. 13). In front of the median nervure, the epidermis has isodiametric cells, with their internal and external wall thicker than the others, the external one being covered by a thin cuticle. Long tectory hairs are visible at the edges of the limb (Fig. 14 a); where, too, the mesophyl appears typically lacunary within the whole thickness of the limb. The fascicle of the median nervure evidences numerous (10-12) parallel rows of woody vessels (Fig. 7); the generating area between the xylem and the phloem is thicker (5-6 cell layer) (Fig. 8), while the phloemic pole has a girdle of sclerenchymatic fibers with moderately thickened and lignified walls.

Epidermis in front side view (the upper leaf) In the reference – the upper epidermis is formed of polygonal cells with straight

lateral walls. Here and there, stomatae of anomocytic type and secretory hairs may be observed. In the TM 0.1% treated sample, more stomatae occur on the unit of the surface. Besides the secretory hairs, very long tectory hairs, with a very long filiform terminal cell, have been also noticed. In the TM 0.4% treated sample, the epidermal cells are more numerous on the unit of surface, therefore they are smaller, yet the hairs have the same frequency as in the previous sample (Fig. 15 a).

In the reference – the inferior epidermis evidences some epidermal cells with slightly waved lateral walls. The stomatae and the secretory hairs have the same frequency on both sides of the limb, the gland showing secretory cells arranged on 3-4 levels, the ones situated at superior levels having a convex wall. In the TM 0.1% treated sample, the inferior side shows some cells with slightly waved walls. The frequency of hairs is the same on both sides, yet more numerous than in the untreated sample (3-4 in a microscopic field). In the TM 0.4% treated sample, all epidermal cells show slightly waved lateral walls.

26

The secretory hairs are more rare (1-2 in a microscopic field), yet the stomatae show the same frequency as in the TM 0.1% treatment (Fig.15 b).

Conclusions

The histo-anatomical modifications induced by the treatment with Topsin M depend

on both the administered dose and the leaves development stage in the moment of fungicide’s application. The response reactions of the tissues are more frequently of quantitative order, although mention should be also made of certain structural aspects that might influence the physiology and biochemistry of the plant and, consequently, the active principles synthesized by Calendula officinalis.

The first and most evident effect of thiophanate methyl involves an increase of the foliar surface, a process to be decompensated by a weaker development of the pallisadic tissue, along with a more lax texture of the lacunary one (especially in the 0.4% treatments). At higher concentrations, the fungicide visibly reduces the thickness of the foliar limb, by reducing the sizes of the pallisadic cells (Tab. IV, V, VI), the ratio of which become 1.5/1 on the upper side (mainly in the terminal leaves which, in the moment of spraying, appear in an incipient stage of development), and 1/1.5, respectively, on the inferior side (almost quadratic cells); consequently, the structure of the foliar limb gets modified, from bifacial equifacial in the reference, to bifacial heterofacial in the leaves collected from the middle of the stem, and to an almost wholly lacunary (isofacial) structure in those from the top of the Calendula officinalis stem (Fig. 9-14). However, the median nervure is much thicker, with a modified (either triangular or semi-circular) contour in cross-section, the growth being intensely stimulated by the fungicide action, through an increased number of conducting fascicles (Tab. I, II, III).

In Asteraceae, the presence of more numerous conducting fascicles in the median nervure constitutes an exception, to be possibly explained by inclusion of the first order nervures, as a result of the stimulating action exercised by the thiophanate methyl (the cytokinin hormone-type action of this fungicide being acknowledged). The woody conducting tissue shows a visible reaction to the treatment, by increasing the number of its vessels and, equally, of their diameter (TM 0.1%) (Tab. I, II, III); between the xylem and the phloem, the generating zone is thicker in the treated samples, including several cell layers (TM 0.4%) (Fig. 3, 5, 8).

The epidermal cells are more numerous and smaller, with more waved lateral walls than in the reference (Fig. 15), while the stomatae are more numerous on the unit surface, the stomatic index recording higher values in the treated materials (Tab. 7); the frequency of secretory hairs increases in inverse ratio to the concentration of Topsin M and with the development stage of leaves in the moment of fungicide’s application so that, in the terminal leaves, it records higher values in the TM 0.1% treatment (stimulating dose) and lower values, respectively, in the TM 0.4% one (inhibiting dose).

27

REFERENCES

1. EL MASHAD A.A.A., 2002 - Effect of thiophanate methyl on the growth and some metabolic activities of soybean plant. Egyptian J. Physiol. Sci., 24, 1: 83-102

2. HUŢANU-BASTAWI LUMINIŢA, TOMA C., 2006 - Considerations on the histo-anatomical study of the leaves of Cynara scolymus L. treated with thiophanate methyl (Topsin M). 4th Conference on Medicinal and Aromatic Plants of South-East European Countries - Iaşi, România, 28th – 31st of May 2006 :126-132

3. PETRÓCZI, M.I., MATUZ, J., KÓTAI C., 2002 - Study of pesticide side-effects in winter wheat trials. Acta Biologica Szegediensis, 46, 3-4: 207-208

4. PRIESTELY, R.H., BAYLES, A. ROSEMARY, 1982 - Effect of fungicide treatment on yield of winter wheat and spring barley cultivars. Plant Pathology, 31, 1: 31–37

5. STASKAWICZ B., KAUR-SAWHNEY R., SLAYBAUGH R., ADAMS W., GALSTON A.W., 1978 - The cytokinin-like action of methyl-2-benzimidazolecarbamate on oat leaves and protoplasts. Pesticide Biochemistry Physiology, 8, 1:106-110

6. THOMAS T.H., 1974 - Investigations into the cytokinin-like properties of benzimidazole-derived fungicides. Ann. Appl. Biol., 76: 237-241

7. TRIPATHI R.K., TANDON, K., SCHLÖSSER E., HESS W.M., 1981 - Effect of fungicides on the physiology of plants. Part IV: Protection of cellular organelles of senescent wheat leaves by carbendazim. Pesticide Science, 13, 4: 395 – 400

8. VAN IERSEL, M.W., BUGBEE, B., 1996 - Phytotoxic effects of benzimidazole fungicides on bedding plants. J. Am. Soc. Hort. Sci., 121, 6: 1095-1102

9. YEOUNG-SEUK, B., BYEONG-YONG P., TAE-JIN A., BYEONG-YEON Y., SUNG-WOO L., NAK-SUL S., 2006 - Selection of potential fungicides for control of Ginseng seedling damping-off and research on fungicide application for disease control in farms. Treat. Crop Sci., 7: 679-698

10. YOSHIDA Y., 1970 - Effect of benzimidazole on the senescence of wheat chloroplasts and their boat shape transformation. Plant Cell Physiology, 11, 3: 435-444

Table I. Numerical values - basal leaves (median nervure, middle level)

Variant Thickness

Length/width (μm Oc.10 x Ob.4)

No. of fascicles

No. woody vessels

Diameter of vessels

(μm Oc.10 x Ob.40)

Ca. of. M 1200/1075–1500/1400 1 78-89 15-22,5 Ca. of. TM 0.1 % 1850/2350–1925/2475 3 81-94 22,5-32,5 Ca. of. TM 0.4 % 2075/3325–2250/3500 3 111-115 22,5-30

Table II. Numerical values - middle leaves (median nervure, middle level)

Variant Thickness


No. of fascicles

No. woody vessels

Diameter of vessels

(μm Oc.10 x Ob.40)

Ca. of. M 1375/750–1500/1000 1 80-88 22,5-27,5 Ca. of. TM 0.1 % 1700/2250–1800/2600 3 -5 95-109 25-32,5 Ca. of. TM 0.4 % 2000/1900–2200/2075 2-3 132-139 22,5-27,5

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Table III. Numerical values - superior leaves (median nervure, middle level)

Variant Thickness


No. of fascicles

No. woody vessels

Diameter of vessels

(μm Oc.10 x Ob.40)

Ca. of. M 950/975-1000/1000 1 50-56 20-22,5 Ca. of. TM 0.1 % 1075/1050–1250/975 1 61-72 25 Ca. of. TM 0.4 % 1375/1200–1600/1450 2 68-88 25-30

Table IV. Numerical values - basal leaves (limb, middle level) (Oc.10 x Ob.20)

Variant Thickness of the limb (μm)

No. of layers in the limb

Thickness of the pallisade (μm)

Ca. of. M 375-450 7-8 135-160 Ca. of. TM 0.1 % 350-435 6-7 125-150 Ca. of. TM 0.4 % 325-375 7-8 115-130

Table V. Numerical values - middle leaves (limb, middle level) (Oc.10 x Ob.20)




Ca. of. M 350-400 6-7 150-175 Ca. of. TM 0.1 % 375-410 7-8 140-170 Ca. of. TM 0.4 % 300-350 6-7 110-120

Table VI. Numerical values - superior leaves (limb, middle level) (Oc.10 x Ob.20)




Ca. of. M 335-375 6 125-145 Ca. of. TM 0.1 % 325-375 7-8 125-135 Ca. of. TM 0.4 % 275-300 7 100-120

Table VII. Numerical values – epidermis of the upper leaves, middle level (Oc.10 x Ob.20) Upper epidermis Inferior epidermis

Samples No. of cells

No. of stomatae

Stomatic index

No. of cells

No. of stomatae

Stomatic index

Ca. of. M 63 9 11,11 76 13 12,74 Ca. of. TM 0.1 % 71 12 12,63 78 15 13,88 Ca. of. TM 0.4 % 75 15 14,28 88 17 13,93

29

Ca. of. M Ca. of. TM 0.1% Ca. of. TM 0.4%

Fig. 1. Cross-sections – median nervure of the limb, basal leaf, middle third

Ca. of. M Ca. of. TM 0.1% Ca. of. TM 0.4% Fig. 2. Cross-sections – median conducting fascicle, basal leaf, middle level


Fig. 3. Cross-sections – median conducting fascicle, basal leaf, middle level


Fig. 4. Cross-sections – median nervure of the limb, middle leaf, middle level

30


Fig. 5. Cross-sections – median conducting fascicle, middle leaf, middle level


Fig. 6. Cross-sections – median nervure of the limb, superior leaf, middle level


Fig. 7. Cross-sections – median conducting fascicle, superior leaf, middle level


Fig. 8. Cross-sections – median conducting fascicle, superior leaf, middle level

31


Fig. 9. Cross-sections – limb, basal leaf, middle level


Fig. 10. Cross-sections – mesophyl, basal leaf, middle level


Fig. 11. Cross-sections – limb, middle leaf, middle level


Fig. 12. Cross-sections – mesophyl, middle leaf, middle level

32


Fig. 13. Cross-sections – mesophyl, superior leaf, middle level

a

b Fig. 14. Calendula officinalis: tectory hairs (a), uni- and biseriated secretory hairs (b)

a

b Ca. of. M Ca. of. TM 0.1% Ca. of. TM 0.4%

Fig. 15. Epidermis in front side view - upper (a) and inferior epidermis (b), superior leaf, middle level

33


MORPHO-ANATOMICAL AND PHYTOCHEMICAL RESEARCHES REGARDING PSEUDOTSUGA MENZIESII (MIRBEL) FRANCO (PINACEAE)

IOANA MARCELA PĂDURE*, LILIANA BĂDULESCU, TEODORA DEDIU,

I. BURZO

Abstract: The paper presents morpho-anatomic and phytochemical researches of needles and young shoots of Pseudotsuga menziesii (Mirbel) Franco in Romania. The composition of needle and young shoots essential oils in Douglas-fir needles from different population are studied for the first time. Essential oil extraction of P. menziesii was performed using a neo-Clevenger-type apparatus and the analysis procedures with a GC-MS system. The major constituents of the oils were found to be sabinene, terpinolene, terpinene 4-ol, β-pinene, α-terpinen, β-phellandrene and α-pinene in various chemical concentrations due to analyzed vegetative organs (needles / young shoots), ecological conditions (chemotypes), analytical methods or plant phenophases. Keywords: Pseudotsuga menziesii, Pinaceae, needles, chemotype, essential oil, GC-MS, chromatogram.

Introduction

Pseudotsuga menziesii [syn. P. douglasii (Lindl.) Carrière] is an evergreen trees;

bark initially smooth with transverse resin blisters, with age becoming reddish brown, thick and corky, deeply fissured into scaly ridges or flaking; branches often pendulous, irregularly whorled; short shoots absent; leaf scars transversely elliptic, slightly raised proximally but essentially flush with twig distally. Buds elongate, not or slightly resinous, apex acute. Leaves borne singly, persisting 6-8 years, alternate, short-stalked, linear, flat, green and grooved above, with 2 white stomatal bands containing 5-8 lines of stomata beneath; 2 marginal resin ducts and 1 vascular bundle. Seed cones maturing first season, terminal on short branchlets, consisting of numerous spirally arranged scales, each scale 2-ovulated. Mature cones shed whole, deflexed or pendent from a 2-10 mm long peduncle, ellipsoid, ovoid, or cylindric, lacking apophysis and umbo; scales persistent, apex rounded; bracts ± exserted, apex 3-lobed with the middle lobe long and narrow. Seeds winged; cotyledons 2-12 [6]. It is in leaf all year, in flower from March to May, the seeds ripen from September to November. The scented flowers are monoecious and are pollinated by wind.

The plant prefers light (sandy), medium (loamy) and heavy (clay), acid to neutral soils. It cannot grow in the shade. It requires moist or wet soil. The plant can tolerate strong winds but not maritime exposure. Very slow growing for its first few years, growth soon becomes extremely fast with new shoots of up to 1.2 metres a year. New growth takes place from May to July. The genus Pseudotsuga contains about 7 species native to North America, and eastern Asia. Douglas-fir is named for Henry Douglas (1798-1834), a Scottish botanist who traveled in North America. The word Pseudotsuga means ‘false

* University of Agronomic Sciences and Veterinary Medicine (U.S.A.M.V.), Faculty of Horticulture, Department of Botany and Plant Physiology, Mărăşti 59 Bd., 011464, Bucharest, Romania

34

hemlock", while “menziesii” is used in recognition of Archibald Menzies (1754-1842), a Scotch physician and naturalist, who discovered Douglas-fir in 1793 on Vancouver Island.

Douglas fir was often employed medicinally by various native North American Indian tribes who used it to treat a variety of complaints: an antiseptic resin is obtained from the trunk; it is used as a poultice to treat cuts, burns, wounds and other skin ailments. The resin is used in the treatment of coughs and can be chewed as a treatment for sore throats. Infusions: the green bark has been used in the treatment of excessive menstruation, bleeding bowels and stomach problems; the leaves has been used as a wash and a sweat bath for rheumatic and paralyzed joints; the young sprouts has been used in the treatment of colds; the shoots has been used in the treatment of kidney and bladder problems. A decoction of the buds has been used in the treatment of venereal disease. Young shoots have been placed in the tips of shoes to keep the feet from perspiring. A mouthwash is made by soaking the shoots in cold water [5], [7].

Bogar et al. (1965), Spicer et al. (2000) and Apple et al. (2002) made anatomical investigations of seedling roots, needles and stem growth in P. menziesii [1]. The chemical composition of P. menziesii has been reported in several studies. Snajberk et al. (1974), Wagner et al. (1989), Gambliel & Cates (1995) determined that the essential oil of needles and shoots in Douglas-fir consists of terpenes (monoterpene, sesquiterpene, oxygenated monoterpene) [10] and oleoresins (linalool, methylsalicylate, bornyl acetate citronellol, geranyl acetate, methylthymol, citronellyl acetate, terpinen 4-ol, borneol, isopulegol, anethole, terpinen 4-ol acetate, camphor, geraniol, neryl acetate, and nerol) [8]. The oil of P. menziesii from different Bulgarian populations was constituted as main compounds the monoterpenes: β-pinene, sabinene, β-ocimene, α-terpinolene, α-terpineol, citronellyl acetate, α-terpinene, limonene and γ-terpinene [4].


The mature shoots with needles were collected from two different populations with

P. menziesii from Bucharest: chemotype 1: “I. Todor” Botanic Garden and chemotype 2: Faculty of Biotechnology in U.S.A.M.V. in 2005-2006. The fresh vegetal organs were free hand sectioned. The material were cleared with chloral hydrate, stained in carmine red and green iodine and mounted in gelatinized glycerin. Numerical characteristics were undertaken at ML-4M IOR microscope. The prepared material was viewed and photographed with a Nikon camera; anatomical photos are presented in Fig. 1-3.

Fresh needles of the collected plant were subjected to hydrodistillation for three hours using a neo-Clevenger-type apparatus to produce essential oils. Analysis conditions: FISIONS gas chromatograph with DB 5 column 25 m length and 0.25 mm internal diameter. Carrier gas has been helium; work temperatures by injector 250°C, interface 280°C and ionization source 200°C; EI mode 70eV; mass range 41-550 amu. Chemical compounds identification has been performed by library search represented by NIST and ESO 85. Kovats indices were used as a confirmation for the chromatographic peak position. These determinations were performed in Research Center for the Study of Food Quality and Useful Plant Compounds from U.S.A.M.V. Bucharest.

35


The needle and young shoots anatomy in Pseudotsuga menziesii: The two adaxial resin ducts of Pseudotsuga needle are located in direct contact with

epidermis. The needles present a central midvein with variable diameter (31,25-68,7 μm) surrounded by a thin endodermis. Each canal is sometimes partial surrounded by sclerenchyma fibers with lignified walls (fig. 1, 2). The resin ducts of young shoots with different diameter are located in cortical zone of stem. The stem presents secondary structure with three annual rings in the moment of analysis. In the cortex, each canal with different diameters (25-137,5 μm) is surrounded by secretory cells with thin walls (fig. 3).

The anatomical characteristics of needle and young shoots are presented in Tab. I.

Table I Anatomical characteristics of needle and young shoot in Pseudotsuga menziesii

Anatomical characteristics Minimum value [µm]

Maximum value [µm]

Average value [µm]

Needle Ecotype Ecotype Ecotype 1 2 1 2 1 2

1. Stomata length (40x) 19,4 17,8 32,4 32,4 24,3 18,5 2. Stomata wide (40x) 16,2 13 19,4 24,3 17,8 18,5 3. Number of stomata / mm2 266 207 414 335 - - 4. Stomatiferous zone wide (10x) 206,2 200 281,2 312,5 239 257,8 5. Number of stomata strings / on leaf wide (10x)

4 4 6 7 5 6

6. Midvein height, including endodermis (10x)

187,5 268,7 243,5 293,7 200 282,6

7. Vascular bundle height (10x) 125 218,7 168,7 231,2 143,8 220,8 8. Vascular bundle wide (10x) 162,5 185,3 218,7 220 184,4 203,5 9. Resin ducts diameter (10x) 31,3 31,25 62,5 68,7 45 51,3 10. Mesophyllum height (10x) 331,3 500 375 562,5 347,9 534,4 11. Palisade tissue height (10x) 93,75 143,7 156,2 218,7 120,6 172,2 12. Number of palisade cell layers 1 2 3 3 2 2 13. Septate cells height 187,5 281,2 250 406,2 226,2 348,2 14. Number of septate cell layers 4 3 7 6 5 5 15. Xylem thickness (40x) 19,4 40,5 43,74 65 30,29 48,3 16. Sclerenchyma thickness (40x) 12,9 29,1 32,4 42,1 25,5 33,7 17. Endodermis thickness (40x) 8,1 9,7 27,5 39 15,4 22 18. Phloem thickness (40x) 19,44 23,4 59,9 40,5 42,28 32,6 19. Adaxial epidermis thick (40x) 6,4 9,7 8,1 24,3 7,2 14 20. Abaxial epidermis thick (40x) 4,8 6,5 6,4 11,34 5,6 9 21. Cuticle thickness (40x) 1,62 6,5 8,1 11,3 5,7 9

Young shoot 22. Resin ducts number / leaf section 7 6 10 11 8 8 23. Secretory hair length (10x) 19 31,2 125 156 62,5 106,2 24. Xylem thickness (primary and secondary xylem) (10x)

375 75 419 156 400 106,2

25. Resin ducts diameter (10x) 37,5 25 137,5 56,2 70,3 42,5 26. Cortex length (10x) 162,5 375 394 500 255 411 27. Number of cell layers in cortex 5 7 7 12 6 10 28. Phloem thickness (primary and secondary phloem) (10x)

125 12,5 187,5 25 157 20

29. Annual ring 2004 thickness (10x) 56,2 125 87,5 187,5 70 144,6

36

30. Annual ring 2003 thickness (10x) 125 - 150 - 135,4 - 31. Annual ring 2002 thickness (10x) 75 - 125 - 99 - 32. Primary xylem thickness (10x) 31,2 6,25 50 31,2 40,6 18,7 33. Pith diameter (10x) 375 437,5 500 500 441,6 465,6 34. Epidermis thickness (40x) 8,1 4,8 13 9,7 10 7,2 35. Subepidermic cell layers number 2 - 2 - 2 - 36. Bark thickness (40x) 11,3 4,8 24,3 9,7 17,4 7,2 37. Number of bark cell layers 4 1 4 3 4 2 38. Pheloderm thickness (40x) 11,3 48,6 21 64,8 18,3 58 39. Number of pheloderm cell layers 2 3 3 4 3 4

The needle and young shoots phytochemistry in P. menziesii: The yield and composition of the essential oils of P. mensiesii chemotypes can be

seen in Table II, III. Twenty eight components representing between 91.08% and 99.56% of the total oil composition were identified for the needles, and twenty six for young shoots oil representing between 95.5 to 98.97% of the total oil composition. The remaining percentage consists of traces or remained unidentified by chemical searched library.

The major components of essential oils in needles and young shoots of P. menziesii in different chemotypes and phenophases are represented by sabinene (14.34-31.73%), terpinolene (12.71-23.23%), terpinen-4ol (7.53-14.82%), α-pinene (3.88-9.23 %) in needle oil and α-pinene (3.88-9.23%), β-pinene (12.6-15.22%), α-terpinene (21.05-32.87%), terpinolene (13.7-15.4%) in young shoots.

The most significant quantities of different compounds are noticed as follows: in needle oils- α-pinene 9.23% (chemotype 1, April), sabinene 31.73% (chemotype 1, March), β-pinene 30.28% (chemotype 2, April), α-terpinene 5.64% (chemotype 1, October), terpinen-4ol 14.82% (chemotype 1, October) and terpinolene 23.23% (chemotype 2, April) (Tab. II). In the oils of young shoots the most important components are represented by α-pinene 10.6 % and sabinene 12.3% (chemotype 2, April), α-terpinene 32.87% (chemotype 1, January), terpinolene 15.4% and terpinen-4ol 7.12% (chemotype 2, January) (Tab. III).

Table II. Chemical composition of essential oils in needle of Pseudotsuga menziesii

Chemical components CHEMOTYPE 1 CHEMOTYPE 2

October 2005

January 2006

March 2006

April 2006

January 2006

April 2006

α-thujen 1.84 1.95 2.00 1.82 1.97 1.81 α-pinene 3.88 7.31 4.72 3.9 4.44 9.23 Camphene 0.15 0.32 0.14 0.15 0.13 0.48 Sabinene 19.16 24.36 31.73 29.99 29.57 14.34 β-pinene 7.23 21.22 9.22 7.22 7.94 30.28 β-myrcene 1.23 1.64 1.43 1.37 1.39 1.85 α-phellandrene 0.64 0.3 0.41 0.42 0.48 0.37 p-cymene 1.01 0.69 1.21 1.17 1.15 0.64 α-terpinene 5.64 3.07 4.09 4.62 4.74 3.31 Cymol 0.93 0.33 0.32 0.5 0.51 0.29 Limonene 1.93 2.12 1.68 1.7 1.73 2.49 Eucalyptol 0.18 0.24 0.17 0.25 0.19 0.29 Cis β-ocimene 0.102 0.07 0.13 0.14 0.11 0.09 β-phellandrene 8.09 4.79 6.4 7.17 7.14 5.11

37

Trans β-ocimene 0.16 0.41 0.3 0.35 0.19 0.24 Terpinolene 20.31 16.05 22.99 23.23 23.06 12.71 Linalool 0.23 0.34 0.27 0.31 0.21 0.3 Fenchol 0.41 0.37 0.41 0.52 0.44 0.37 Citronellal 0.24 0.25 0.25 0.33 0.28 0.23 Isoborneol 0.11 0.23 0.11 0.12 0.18 0.18 Terpinen-4ol 14.82 7.53 9.03 11.49 10.68 9.16 α-terpineol 0.80 0.71 0.47 0.61 0.55 1.13 Cis-piperitol 0.26 0.29 0.17 0.23 0.25 0.27 L-terpinen 4-ol 0.53 0.26 0.47 0.38 0.49 0.25 propenil-anisol 0.17 0.24 0.07 0.14 0.09 0.32 Trans-piperitol 0.95 2.27 1.24 1.03 1.14 2.12 Citronellol-acetate 0 0.81 0 0.07 0 0.8 Selinenol 0.08 0.24 0.13 0.1 0.08 0.15

Total 91.08 % 98.41 % 99.56 % 99.33 % 99.13 % 98.81 %

Table III Chemical composition of essential oils in young shoot of Pseudotsuga menziesii Chemical components CHEMOTYPE 1 CHEMOTYPE 2

January 2006 January 2006 April 2006 α-thujen 1.16 1.38 1.29 α -pinene 7.89 10.47 10.6 Camphen 0.25 0 0 Sabinene 9.46 5.65 12.3 β-pinene 12.6 14.89 15.22 β-myrcene 2.52 2.45 2.66 α-phellandrene 0.39 0.304 0.27 α-terpinene 21.05 32.87 24.56 P-cymene 2.39 2.7 1.74 Cymol 0.68 0.59 0.71 Limonene 3.15 3.78 3.13 Cis α-ocimene 0.29 0.77 0.25 Trans β-ocimene 3.98 4.48 2.85 Terpinolene 15.4 14.61 13.7 Linalool 0.84 0 0.13 α- terpineol 0.08 0.22 0.36 terpinen-4ol 7.12 2.77 1.96 α-terpineol 1.07 0.4 0.22 Propenil-anisol 0.61 0.27 0.74 β-cubebene 1.32 0.12 1.76 Isoeugenol 1.05 0 0.09 β-cadinene 0.22 0 0.205 Selinenol 0.42 0.25 0.39 Cembrene 0.27 0 0.28 Kauren 0.34 0 0.36 Norkauren 1 0 1.023 Total 95.55 % 98.97 % 96.80 %

Conclusions

The major components of essential oils in needles and young shoots of P. menziesii

in different chemotypes and phenophases are represented by sabinene (14.34-31.73%), terpinolene (12.71-23.23%), terpinen-4ol (7.53-14.82%), α-pinene (3.88-9.23 %) in needle

38

oil and α-pinene (3.88-9.23%), β-pinene (12.6-15.22%), α-terpinene (21.05-32.87%), terpinolene (13.7-15.4%) in young shoots.

The most significant quantities of different compounds are noticed as follows: in needle oils- sabinene 31.73%, β-pinene 30.28% terpinen-4ol 14.82% and terpinolene 23.23%. In the oils of young shoots the most important components are represented by α-pinene 10.6 % and sabinene 12.3%, α-terpinene 32.87% and terpinolene 15.4%.

The each chemotype or populations are represented by several compounds varying from month to month in the investigated periods or type of analyzed organs. The chemical composition of essential oils in needle of P. menziesii are represented in Chemotype 1 by 27 compounds in October 2005 and March 2006, 28 compounds in January and April 2006; in Chemotype 2 there are 27 compounds in January 2006 and 28 compounds in April 2006.

The chemical composition of essential oils in young shoots of P. menziesii are represented in Chemotype 1 by 26 compounds in January 2006 and Chemotype 2 by 19 compounds in January 2006, respectively 25 in April 2006.

Several anatomical and morphological characters were found significant for an analysis of the relationship of Pseudotsuga populations such as resin ducts diameter in stem or needle or number of resin ducts in cortical zone of stem.

This original study bring new information in the general knowledge on the morphology and anatomy of cultivated plants from Pinaceae family and fill the gaps regarding the phytochemical composition of oil in Pseudotsuga menziesii, studied for the first time in Romania.

REFERENCES

1. APPLE M., TIEKOTTER K., SNOW M., YOUNG J., SOELDNER A., PHILLIPS D., TINGEY D. &

BOND B.J., 2002 - Needle anatomy changes with increasing tree age in Douglas-fir. Tree Physiol., 22:129–136

2. BOGAR G.D. & SMITH F.H., 1965 - Anatomy of seedling roots of Pseudotsuga menziesii. Am. J. Bot. 52, 7: 720-729

3. GAMBLIEL H.A. & CATES R.G., 1995. Terpene changes due to maturation and canopy levels in Douglas-fir (Pseudotsuga menziesii) flush needle oil. Bioch. Syst. Ecol. 23, 5: 469-476

4. JIROVETZ L., PUSCHMANN C., STOJANOVA A., METODIEV S. & BUCHBAUER G., 2000 - Analysis of the essential oil volatiles of Douglas fir (Pseudotsuga menziesii) from Bulgaria. Flavour Fragrance J. 15: 434-437

5. JOHNSTON W.H., KARCHESY J.J., CONSTANTINE G.H. & CRAIG A.M., 2001 - Antimicrobial activity of some Pacific Northwest Woods against Anaerobic Bacteria and Yeast. Phytotheraphy Res. 15: 586–588.

6. LIPSCOMB B., 1993 - Flora of North America North of Mexico, vol II, University Press Oxford 7. MOERMAN D., 1998 - Pseudotsuga in Native American Ethnobotany. Timber Press, Oregon 8. SNAJBERK K., LEE C.J. & ZAVARIN E., 1974 - Chemical composition of volatiles from cortical oleoresin

of Pseudotsuga menziesii. Phytochemistry 74, 13: 185-188 9. SPICER R., GARTNER B.L. & DARBYSHIRE R.L., 2000 - Sinuous stem growth in a Douglas-fir

(Pseudotsuga menziesii) plantation: growth patterns and wood-quality effects. Canad. J. For. Res. 30, 5: 761-768

10. WAGNER M.R., CLANCY K.M. & TINUS R.W., 1989 - Maturational variation in needle essential oils from Pseudotsuga menziesii, Abies concolor and Picea engelmannii. Phytochemistry 28, 3: 765-770

http://heronpublishing.com/tphome.html

39

Fig. 1 The anatomy of needle (A) and median vein (B) in P. menziesii [ob. 6x / 25x, oc. 12,5x, orig.]

Fig. 2 The anatomy of shoots in P. menziesii: cortical cells (A) and central cylinder zone (B) [ob. 6x / oc. 12,5x, orig.]

Fig. 3 Pseudotsuga mensiesii needle (A) and cortical resin ducts of stem (B) [ob. 40x, oc. 12,5x, orig.]

A B epidermis

endodermis

midvein

secretory duct

A B

A B phloem

cambium

xylem II

xylem I

pith

cortex

pith

xylem

medullary rays

xylem II (I II III: annual rings)

resin ducts from stem

epidermis phelogen

bark

epidermis

secretory cells

chlorenchyma cortex

40


THE ECOPHYSIOLOGICAL REACTION OF SOME VARIETIES OF APPLE TREE, PEAR TREE AND QUINCE TREE TO THE PATHOGENIC AGENTS

ATTACK

ALEXANDRINA MURARIU*, CORINA GRĂDINARIU*, ANIŞOARA STRATU*

Abstract: The research was carried out in 2004, on material from horticultural collections of the Didactical Station “Vasile Adamachi” within the University of Agricultural Sciences and Veterinarian Medicine of Iaşi. The investigations were performed on three fruit tree species: Cydonia oblonga Mill., Pyrus communis L., and Malus domestica Borkh. and consisted in revealing the morphological symptoms induced by the attack of the pathogen Erwinia amylovora (Burrill 1882) Winslow et al. The biochemical (contents of water, assimilatory pigments and total mineral) and physiological analyses (intensity of photosynthesis, respiration and transpiration) were carried out on healthy and with different infection degree material of two varieties for each species: “Jonathan” and “Generos” (apple tree), “Williams” and “Argesis” (pear tree), “Moşna” and “De Huşi” (quince tree). The results revealed the correlation between the physiological alterations and the severity of the pathogenic attack: in weakly attacked (5 – 8% per tree) “Jonathan” (apple), “Argesis” (pear) and “De Huşi” (quince) samples we recorded high contents of water and minerals and an increase of the net photosynthesis intensity compared to the “Williams” (pear) variety and relatively strong bacterial attack (12%). Large differences between healthy and infected leaves were recorded in “Moşna” (quince) at a bacterial attack of 38%, which induced a decrease in pigment content (20%), net photosynthesis (50%), water content (76%), transpiration (38%), and an increase in respiration (117%). Key words: Cydonia oblonga Mill., Pyrus communis L., and Malus domestica Borkh., Erwinia amylovora, physiological indicators

Introduction

Fruit trees can be attacked by a large number of pathogens (viruses, mycoplasmas,

bacteria, and fungi), nematodes and insects, which results in important economical losses in orchards and storehouses. Just for the apple tree, the scientific literature describes over 150 diseases and pests, 50 of which are presumably worldwide spread [ 1 ]. In Romania, there are cited three bacterial diseases in apple tree and pear tree and one in quince tree [ 8 ].

With regard to the attack of the bacteria Erwinia amylovora on species of the Family Rosaceae, the first investigation was completed by T. Săvulescu (1938). In 1992, two contagion centers were identified in Brăila and Mărăcineni, and after 1993, the disease spread into almost all the Romanian fruit growing areas [ 4 ]. The most susceptible species belong to the following genera: Cydonia, Pyrus, Malus, followed by Cotoneaster, Pyracantha, and Sorbus.

* “Al. I. Cuza” University, Faculty of Biology, Carol I Bd., no. 20A, 700506, Iaşi, Romania

41

The plant reaction to the pathogen attack represents the expression of some genetically induced characteristics, and hence, the understanding of the genomic interaction implies biochemical and physiological investigations.


The research was carried out in 2004, on material from horticultural collections of

the Didactical Station “Vasile Adamachi” within the University of Agricultural Sciences and Veterinarian Medicine of Iaşi.

The research material consisting of fresh leaves was sampled from healthy and infected trees of two varieties of each species: “Jonathan” and “Generos” (apple tree), “Williams” and “Argesis” (pear tree), “Moşna” and “De Huşi” (quince tree).

The material was sampled during the second decade of June, when the trees were at the beginning of fruit growth phenophase, given that the pathogen optimum period comes after blooming.

The fresh material was used to determine the water and assimilatory pigments contents, the intensity of photosynthesis, respiration and transpiration. After the inactivation of the enzymes (60°C exposure) the material was dried with air and was used to determine the total mineral content, through the analysis of the ash.

Additionally, a visual assessment of the Erwinia amylovora attack was made. The strength of the attack was expressed as percentage (Tab. I) and varied with the climatic conditions of the year (extreme heat and drought).

Table I. The strength of Erwinia amylovora attack in 2004 Species Variety Strength (%) Resistance

category* Jonathan 5.0 Susceptible Malus domestica Generos 1.0 Resistant Williams 12.0 Susceptible Pyrus communis Argensis 7.0 Very susceptible Moşna 38.0 Very susceptible Cydonia oblonga De Huşi 8.0 Susceptible

* after Cimpoeş, 2001 and Grădinaru, 2002

42


Even though the primary infection occurs in flowers, it spreads out to leaves, braches and trunk, causing ulcerations [5]. Metabolic dysfunctions appear in leaves due to profound biochemical and physiological alterations such as imbalances of the water content, photosynthesis and respiration, and biosynthesis of complex organic substances.

1. Imbalance of water content The general effect of the bacterial attack was the reduction of the leaf water content

in all the varieties (Tab. II). Leaf dehydration in infected varieties compared to the healthy leaves varied from 2% in “Generos” resistant variety (apple tree) to 76% in “Moşna” very susceptible variety (quince tree).

Total mineral content of mature leaves, at fructification, varies as a function of the strength of the bacterial attack. In apple tree and pear tree the bacterial attack induced a 10 – 45% increase, whereas in quince tree (“Moşna”) the infected leaves contained 27% less mineral elements.

The reduction of the leaf area because of necroses and irregularities in stomatal movements occurs concurrently with transpiration decrease in all analysed varieties. The bacterial attack favoured the transpiration reduction in susceptible varieties with 6% in apple tree (“Jonathan”), 45% in pear tree (“Argesis”), and 43% in quince tree (“De Huşi”).

2. Imbalance of photosynthesis and respiration During the pathological process, the chloroplasts suffer alterations that lead to

fragmentation and loss of chlorophyll due to increased quantities of chlorophyllase. Our research on the content of the assimilatory pigments (total and by forms –

chlorophylls and carotenoids) revealed the same correlation of the strength of bacterial attack and the quantitative fluctuations in infected and healthy leaves, comparatively.

Only in “De Huşi” variety (quince tree), the infected leaves were richer in pigments, which entails the activation of a certain defensive mechanism against the pathogen.

The quantity of chlorophylls (a + b) in leaves stays high during fructification, varying between 1.78 – 3.85 mg in healthy leaves and between 1.43 – 2.14 mg in infected leaves. The carotenoids showed little differences in all the analysed varieties.

Fruit presence represents a stimulating factor for the photosynthesis intensity in healthy leaves (1.24 – 5.14 mg CO2/g/h). In the infected leaves the simulative effect is weaker (1%) because of the resistance to the bacterial attack (“Generos” variety – pear tree).

The obvious water deficit of quince tree infected leaves (“Moşna” variety) diminishes the photosynthesis with 50%.

The bacterial attack induces an intensification of the respiration in the host plant because of the redox enzymes and toxins or represents a tendency to counteract the very intense respiration of the pathogenic agent.

Our results showed that in all the cases an increase in respiration intensity with approximately 44% in apple tree and pear tree, and with 117% in quince tree (“Moşna” variety).

43

Regarding the total organic substances content, the bacterial attack generally reduces the synthesis of complex organic substances, but intensifies their hydrolysis, which increases the content of soluble forms of sugars and proteins with approximately 121% (“Moşna” variety).

All the mentioned physiological modifications could be disease indicators, through their accumulation in cells, tissues and finally organs (Fig.1).

Conclusions

By correlating the obtained results with the strength of the bacterial attack, we

revealed the existence of a powerful relation between the physiological and biochemical modifications and the gravity of the attack of the studied pathogen.

In the varieties “Jonathan” (apple tree), “Argesis” (pear tree) and “De Huşi” (quince tree) in which the pathogen attack was weak (5 – 8%) there is a higher content of water and minerals and an increase in the net photosynthesis, in comparison to the variety “Williams” (pear tree) and situations of relatively strong bacterial attack (12%).

Large differences between healthy and infected leaves were recorded in “Moşna” (quince) at a bacterial attack of 38%, which induced a decrease in pigment content (20%), net photosynthesis (50%), water content (76%), transpiration (38%), and an increase in respiration (117%).

REFERENCES

1. BRANIŞTE N., ANDRIEŞ N., 1990 - Soiuri rezistente la boli şi dăunători în pomicultură. Ed. Ceres.

Bucureşti 2. CIMPOEŞ GH., BACARCIUC V., CAIMACAN I., 2001 - Soiuri de măr. Ed. I. E.P. Ştiinţa, Chişinău. 3. GRĂDINARU G., 2002. Pomicultura specială. Ed. “Ion Ionescu de la Brad”, Iaşi 4. MAC HARDY, W.E. 1996 - Apple scab - Biology, Epidemiology and Management, APS Press, Sant Paul,

Minnesota 5. MITITIUC M., 1994 - Fitopatologie. Ed. Universităţii “ Al. I. Cuza”, Iaşi 6. OKTEM, Y.E.; BENLIOGLU, K. , 1988 - Studies on fireblight [Erwinia amylovora (Burr.) Winsl. et al.] of

pome fruits. Journal of Turkish Phytopathology, 17, 106 7. SEVERIN V., 1996 - Focul bacterian al rozaceelor (Erwinia amylovora). Ed. Ceres, Bucureşti 8. VAN DER ZWET T., KEIL H. L., 1979 - Fire Blight: A Bacterial Disease of Rosaceous Plants. United

States Department Agriculture Handbook, 510, Washington DC 9. YOSHKAWA M., 1983 - Macromolecules, recognition and the traggerin of resistance. In :Biochemical Plant

Pathology (edited by: Callow J. A.), Plant Physiology, 73: 497- 506

44

Table II. The ecophysiological reaction of some varieties of apple tree, pear tree and quince tree to Erwinia amylovora in 2004

Assimilatory Pigments (mg/g fresh substance)

Photosynthesis (mg CO2/g/h) Sp

ecie

s

Var

iety

Stre

ngth

of

A

ttack

(%

)

Wat

er (%

)

Tran

spira

tion

(mg/

h)

Min

eral

s

Chl

orop

hyll

a

Chl

orop

hyll

b

Car

oten

oids

Tota

l

Net

Ph

otos

ynth

esis

Res

pira

tion

Gro

ss

Phot

osyn

thes

is

Dry

subs

tanc

e (%

)

Org

anic

su

bsta

nce

(%)

Healthy 61.33 62 5.53 3.27 0.58 0.30 4.15 5.148 1.072 6.22 38.67 33.14 Jonathan 5% 57.99 58 6.11 1.36 0.26 0.18 1.80 2.624 1.552 4.176 42.01 35.90 Healthy 60.16 25 6.43 1..60 0.29 0.25 2.14 2.496 0.374 2.87 39.84 33.41

Malus domestica

Generos

1% 59.29 14 7.86 1.22 0.21 0.16 1.59 3.120 0.561 3.681 40.71 32.85 Healthy 56.44 22 4.50 1.97 0.45 0.21 2.63 2.496 0.963 3.432 43.56 39.06 Williams 12% 51.28 12 5.22 1.45 0.25 0.18 1.88 0.624 1.360 1.984 48.72 43.50 Healthy 61.53 198 4.27 1.53 0.25 0.16 1.94 3.744 1.248 4.992 38.47 34.20

Pyrus communis

Argensis 7% 58.98 144 6.40 1.42 0.24 0.14 1.80 0.624 0.936 1.560 41.02 34.62 Healthy 54.64 71 9.33 1.53 0.25 0.20 1.98 1.248 0.744 1.992 45.36 36.03 Moşna 38% 13.24 44 6.83 1.24 0.19 0.18 1.61 0.624 1.616 2.240 86.76 79.93

Healthy 58.23 79 6.80 1.65 0.24 0.20 2.09 1.872 0.744 2.616 41.77 34.97

Cydonia oblonga

De Huşi 8% 56.63 45 8.42 1.81 0.33 0.23 2.37 2.496 0.936 3.432 43.37 34.95

45

0

10

20

30

40

50

60

70

80

90

Healthy Strengthof Attack

5 %


1 %


12 %


7 %


38 %


8 %

Jonathan Generos Williams Argensis Moşna De Huşi

Malus domestica Pyrus communis Cydonia oblonga

Water content ( %) Total mineral elements content ( %) Organic substance content ( %)

Fig.1. The physiological modifications in leaves of studied species

46


PHYSIOLOGICAL EFFECTS INDUCED BY PURINIC SUBSTANCES AT CAPSICUM ANNUUM L

ELENA CRISTINA ROŞU∗, MARIA MAGDALENA ZAMFIRACHE∗, I. I. BĂRA

Abstract: The paper present the physiological effects after the treatment with two purinic substances, at Capsicum annuum L. plantles in seedling degree. For the treatment was using 1, 3, 7 trimetil-xanthine (coffeine, theine) and 1,3-dimethil-xanthine (theophyline). As physiological parameters was using quantity of assimilatory pygments, water percentage and dry substance percentage. The treatment has determinated the decrease of quantity assimilatory pygments and the dry substance percentage, comparatively with the control variant. Keywords: purinic compounds, coffeine, theophylline, assimilatory pigments.

Introduction

Purinic compounds are substances with purinic nucleus, wich besides physicals

characteristic and chemicals properties, may replaced the nitrate bases on DNA and causing different mutations. For these properties, the substances were used in plant amelioration and for the study of aberations in mitotic divisions.

The Capsicum genre, is an important genre in Solanaceae family, including more than 245 species, initial in Central America. This plantes containes: capsaicinoids: (0.05 - 1.5 %), pungent phenolic amides including mostly capsaicin, dihydrocapsaicin and derivatives; carotenoids: carotene, capsanthin; volatile oil: trace; proteins, vitamins A, C, coumarins, steroidal alkaloids including solanidine, flavonoids.

Medicamentary properties: stimulant to heart and circulation; peripheral of circulatory insufficiency, intermittent claudication. Low vitality, cold, weak, debility syndromes. Stimulant and tonic to the gastro-intestinal tract; warms digestion, poor appetite, atonic conditions; membranes pale, relaxed, or flabby; impaired secretion; chronic gastric catarrh in absence of inflammation; atonic dyspepsia; gastric flatulence; migraines and cluster headaches.

Materials and methods

The biological material was represented by leafs grow from plantles of Capsicum

annuum L. three varietys: grossum, longum and tetragonum, in seedling degree. The treatment was used on seeds until germination, after that the seeds was hotbed planting.

Methods: The treatment – The seeds was treated with four concentrations, coffeine and

theophylline solutions: V1 (1 variant) = 0.025%, V2=0.05%, V3=0.1% , V4 =0.25% and the control variant with distilled water.

∗ “Al. I. Cuza” University, Faculty of Biology, Carol I Bd., no. 20A, 700506, Iaşi, Romania

47

Physiological determinations: The assimilatory pygments dosage according to spectrophotometrical method after

acetone extraction; The determination of humidity and dry substance through gravimetrical method

bringing to constant weight, at 1000C.


The quantity of assimilatory pigments: The substances applied on germination seeds, had establish the modifications of

physiological aspects trhough decrease of assimilatory pygments quantity comparatively with control variant.

In theophylline treatment case, the 0,1% concentration has determinated the decrease of pygments quantity (exception tetragonum variety), obtaining lesser values comparatively even with maxim concentration used (0,25%).

The 0,25% concentration had a different effect only in longum variety case, where it determinated the increasing of pygments quantity comparatively with those treatement variants, without outrun the control variant value (fig.1).

The coffeine has determinated the decrease of pygments quantity, the minim value obtaned at 0,25% concentration, with exception of grossum variety, where in this case the pygment quantity breede outruning the control variant value (fig. 1).

The quantity of carothenoids pygment, is generally decreased in all treatment variants inclusively at control variant. The treatment with coffeine has a strongly effect in tetragonum variety case, where the quantity of carothenoid pygments decreasee in proportions with the increasing of substance conentration (fig.2). The other varietys, inregistrated an variation of pygments quantity, with decreasing values at 0,025% and 0,1% concentrations, with the difference as whether longum variety case, the minim value is obtained at 0,25% concentration, and at grossum variety the same concentration determinated maxim value of pygments quantity, outruning even the control value .

Comparatively, between this two substances applied, the theophylline has a strongly effect as decreasing of pigments quantity comparatively with coffeine effect, and relation proportion doze-effect, with litlle exceptions, the quantity of assimilatory pygments decrease in proportion with increasing the substance concentration applied in treatment.

The percent of water and the drying substance: The treatment with both substance, has not determineted very large variation of

water percentage, only at tetragonum variety was observed a decrease of water percent at control variant comparatively with the percentages obtained at treated variant.

In case of theophylline using, we remarked a decreas of the drying substance percentage comparatively with control variant at longum and tetragonum varieties, unlike of grossum variety were with exception 0,25% concentration, obtained higer percentage comparatively with the control variant (fig. 3).

48

The report of chlorophyll a/b: The analysis of diagrams, showe, that at the treatment with theophylline, the report

chlorophyll a/b, varies much comparatively with control variant in longum variety case, were decrease unexpectedly at a minim value at 0,025% concentration, following an easye increasing at 0,25% without outrun the control variant value.

At grossum and tetragonum variety the minim values obtaned at V1 and V3 and maxim at V2 and V4 without outrun the control variant value.

The coffeine has determinated at grossum and longum varietyes case an increasing of report in proportion with increasing the substance concentration, outrun the control variant. At tetragonum variety, obtained minim values at V2, the value increasing until 0,25% were the control variant value is equalised (fig.4).

Fig.1. The quantity of assimilatory pygments after de treatment with coffeine (left) and theoffylline (right) at Capsicum annuum L

Fig 2. The quantity of carothenoids pygment after the tretment with coffeine (left) and theoffylline (right) at Capsicum annuum L

49

Fig 3. The percent of water and the dry substance after the tretment with coffeine (left) and theoffylline (right) at Capsicum annuum L

Fig 4. The report of chlorophyll a/b after the tretment with coffeine (left) and theoffylline (right) at Capsicum annuum L

Conclusions

Comparatively, between those two substances used, the theophylline has un strongly effect by decreasing of pygments quantity up to coffeine and abaut the relation of doze-effect report, with little exceptions, the pygments quantity decrease in proportion with increasing of substance concentration applied in treatment . The report of chlorophyll a/b, in the both substance case, had a general tendency by decreasing of the report at minim treatement variant, then (with exception longum variety), the report increased sometimes outgruning the control variant value.

REFERENCES

1. BĂRA I. I., CÂMPEANU MIRELA, 2003 - Genetica, Edit. Corson, Iaşi: 208-209 2. DIACONU P., 1971 - Ereditatea şi factorii mutageni, Edit. Ceres, Bucureşti

50


THE INFLUENCE OF THE Mn2+ IONS EFFECTS ON THE WHEAT (TRITICUM AESTIVUM L.) SEED GERMINATION

I. M. RÎŞCA∗, L. FĂRTĂIŞ∗, ANA LEAHU∗

Abstract: The testing of the Mn2+ ions on wheat seed were conducted in a growth chamber with controlled parameters and the results showed that germination rate and shoot length varies according to the ions concentration. Significant increases of the germination rate at high concentrations were observed as a probable consequence of the seed’s enzymatic system activation. The probable biochemical action mechanisms are discussed. Keywords: manganese ions, wheat, germination rate

Introduction

The microelements have a complex role in the living structures, with both negative

and positive effects, depending – inter alias – on the nature of the elements, their acting form and also their concentration. The essential role of some elements like Fe, Mg, Mn, Zn, Cu, B or Mo in the plant kingdom or Co, Se, Fe and I in the animal one, is well-known [2]. The role of other microelements is not yet well known.

The role of manganese in the unfolding of the oxidative processes at the cellular level and in the functioning of some enzymatic systems [2, 3, and 5] is also a matter of common knowledge; its action unfolds in direct connection with those of the iron [1]. Thus, as bivalent ion, the manganese is part of the superoxyd-dismutase (SOD) from the prokaryotes, an enzyme that annihilates – at the mitochondrial level – the super oxide anion which induces multiple negative biological effects, due to the formation of hydrogen peroxide, aggressive towards the cells [5]:

2O- ●2 + 2H+ → H2O2 + O2

Further, the hydrogen peroxide is removed by the metal enzyme: 2H2O2 → 2 H2O + O2

Among the biological effects of the superoxyd against animals and humans we can enumerate: destruction of the endothelial cells, increase of the micro vascular permeability, formation of some chemotactic factors like leukotriene B4, peroxidation and oxidation of lipids, deterioration of AND singlet chains and formation of peroxynitrite anion (ONOO-), a strong cytotoxic and pro-inflammation agent, according the reaction [5]:

O- ●2 + NO → NOO-

2 In the green plants, the photosystem II uses another manganoenzyme that is involved

in the water dissociation and the production of molecular oxygen, of protons and neutrons. On the other side, manganese is cytotoxic in high concentrations, those effects were studied especially on the animal kingdom were it produces Parkinson-like effects (rhythmical ∗ “Ştefan cel Mare “ University, Universităţii 13 Street, Suceava, Romania

51

trembling and muscular rigidity) but also effects at the psychical level like behavioural aggressiveness, probably due to the neurotoxic accumulations of manganese in globus pallidus. In fact it is well known that the professional exposure to manganese is a risk factor for the Parkinson disease.

The paper studies first of all the manganese effects on the wheat seed germination but also the effects on the wheat growth after the germination.


Apparatus. The germination was accomplished in a CONVIRON MP4030 - G30

growth chamber with the parameters settled as follows: temperature 200C, humidity 90%, without illumination.

Biological material. The wheat samples (Triticum aestivum) we used came from the Magistral variety, 37.5 g/1000 seeds, harvested in 2005 at S.C.D.A Suceava. We measured the germination (FG), according to the standards [4] and also the hypocotyls length (LH) of the germinated plants.

Reagents. We used MnCl2, analytical reagent (Chimopar) and bidistilled water. Applied treatments. The wheat seeds were treated with MnCl2 solutions;

7 concentrations were used: 1M, 0,5 M, 0,1 M, 5 x 10-2 M, 10-2 M, 5 x 10-3 M, 10-3 M, 5 x 10-4 M and a blank with distilled water, 3 x 50 seeds for each concentration, the witness included, in Petri dishes on filter paper.

Two treatment schemes were used: 1.The seeds were immersed for one hour in the treatment solutions, washed thereafter and placed in Petri dishes with distilled water; 2. The seeds were maintained throughout the germination period in the treatment solutions.

After 7 days the number of germinated seeds and the hypocotyls length for the germinated plants were measured. The data obtained was statistically analysed with an application that makes a multiple variance analysis.

Results

The experiments were fulfilled in order to establish the biological answer of the

wheat seed under the influence of mn2+ ions; the results are synthetically showed below (table I and figures 1 and 2).

Table I. Germination and hypocotyl length values under the influence of the

treatment with mncl2 solutions

concentration of

Mn2+

measured parameter (average values)

witness

1 m

0,5 m

0,1 m

0,05 m

0,01 m

0,005 m

0,001 m

germination 1h (%) 88,00 90,67 78,00 91,33 90,67 94,67 93,33 96,67

germination 7 d (%) 88,00 100,00 100,00 98,00 88,00 92,66 91,33 94,66

hypocotyls length 1h

(mm) 60,29 8,78 20,35 51,72 50,02 51,02 53,09 62,50

hypocotyls length 7 d

(mm) 60,29 1,00 1,00 8,69 26,71 56,82 65,27 64,60

52

Fig. 1: Germination analysis

Fig. 2: Hypocotyl length analysis

0

20

40

60

80

100

120

1 M 0,5 M 0,1 M 0,05 M 0,01 M 0,005 M 0,001 M

7 days1 hour

0

10

20

30

40

50

60

70

1 M 0,5 M 0,1 M 0,05 M 0,01 M 0,005 M 0,001 M

7 days1 hour

53

Conclusions and discussions

A first finding is that the stimulation effects of the manganese becomes manifest at high dilution. Thus, beginning with the dilution of 10-1M we observed an approach to the witness of the value of the hypocotyls length, especially for the short-term treatment (fig. 2). The drastically inhibition of the hypocotyls, especially at high concentrations and long-term treatment, could be generated by the secondary toxicity of the manganese ions on the plantlets.

On the other hand, analysing the influence of the manganese ions on the germination, an obvious positive reaction at high concentrations and long-term treatments (fig. 1) comes out, so that, for concentrations of 1M and 0.5 M, the germination is – practically – 100% and for the concentration of 0,1M – 98%. At lower values of the concentration the effect of manganese ions on the germination is not so obvious; the obtained differences are not so significant (table I).

The probable action mechanism is as follows: the high concentrations and the long action times of the manganese ions on the seed permits the diffusion of the ions through the seed tegument in a sufficient high concentration to permits the activation of the enzymatic systems controlled by the Mn2+ ions. At low concentrations and/or short action times this process does not take place any longer, according to the obtained results.

In the case of the action of manganese ions on the hypocotyls, this mechanism is no longer valuable (the plantlet has not a protection teguments like the seed) so that the higher Mn2+ ions concentrations act aggressively and therefore the growth of the wheat embryos is inhibited (fig. 2).

REFERENCES

1. CRICHTON, R., 2001 - Inorganic Biochemistry of Iron Metabolism. John Wiley & Sons, Ltd. Chichester -

New York – Weinheim – Brisbane – Singapore – Toronto 2. DAVIDESCU D., DAVIDESCU VELICICA, LĂCĂTUŞU R., 1988 - Microelementele în agricultură. Edit.

Acad. Rom., Bucureşti, 280 p. 3. Institutul Român de Standardizare, 1999 - Seminţe pentru însămânţare. Determinarea germinaţiei. SR1634:

iunie 1999, Bucuresti 4. KHAN A. A. EDITOR, 1980 - The physiology and biochemistry of seed dormancy and germination. New

York, Oxford 5. ROAT-MALONE, ROSETTE M., 2002 - Bioinorganic Chemistry – A short course. John Wiley & Sons,

Inc., Hoboken, New Jersey

54


PLANTAGO ATMOSPHERIC POLLINIC SEASON IN THE DANUBE-KRIS-MURES-TISZA EUROREGION (2000-2004)

NICOLETA IANOVICI∗, I. E. JUHÁSZ∗∗, P. RADISIC∗∗∗, M. JUHÁSZ∗∗,

B. SIKOPARIJA∗∗∗

Abstract: The aim of the present study is to compare the pollinic season of Plantago in four aerobiological stations within the Danube-Kris-Mures-Tisza euroregion. The Plantago pollen has generally been considered a minor cause of pollinosis. It is difficult to evaluate the role of Plantago airborne pollen in the pollinosis symptomatology because of the low rate of monosensitive patients. moreover, the allergy to Plantago airpollen is connected with the allergy to Poaceae pollen because of the simultaneous presence of the two types of pollen in the air. Since the onset of the allergy also depends on the airpollen concentration, in this study we intend to present the concentrations of the Plantago pollinic type within the Danube-Kris-Mures-Tisza euroregion. The analysis uses the data obtained during five years of monitoring with the help of four volumetric traps located in timişoara (Romania), Szeged (Hungary), Novi Sad and Ruma (Serbia). The Plantago pollen is present in the air from may until august. The highest total annual concentration was recorded at novi sad in 2001 (1326 pg/m3). The highest daily airpollen concentrations seldom exceed 30 pg/m3. The highest daily concentration was 64 pg/m3 (recorded in Novi Sad, 2001). In Szeged and Novi Sad the annual concentrations are on the decrease, while in Timişoara they are slightly on the increase. The presence of the pollen in the airplankton was considerably long in Timişoara, Novi Sad and Ruma in 2004. Key words: airborne pollen, airplankton of cities, Plantago

Introduction

Several authors have already reported that Plantago lanceolata (English plantain,

ribwort) belong to the most important pollens and should therefore be included in the test spectrum for allergological examinations. Allergic sensitization to Plantago pollen is fairly common. It was first reported by Bernton (1925) and there is a relatively large bibliography available on the subject; numerous researchers have carried out studies on this pollen type, including Tuft & Blumstein (1937), Serafini (1957), Duchaine & Spapen (1961), Charpin et al. (1962), Izco et al. (1972), Lewis (1977), Saenz de Rivas (1978), Bousquet et al. (1984), Subiza Martin et al. (1986), Watson & Constable (1991).

Few studies have tried to identify the allergens in Plantago pollen. In 1980, Baldo et al. detected at least six IgE-binding allergens in the molecular weight range of 10–300 kDa. More recently, Dreborg et al. found at least 13 allergens by immunoblotting with one component of 15 kDa with a high degree of specific IgE binding. Only a 30 kDa plantain allergen cross-reactive with the grass group 5 allergens has been identified to date, yet this cross-reactivity shows little or no clinical relevance, as suggested by Asero et al. (2004).

∗ Department of Biology, Faculty of Chemistry-Biology-Geography, West University of Timisoara, Romania ∗∗Department of Botany, University of Szeged, Hungary ∗∗∗Laboratory of Palynology, Institute of Biology, Faculty of Sciences, University of Novi Sad, Serbia

55

These authors also reported in the same article that IgE from monosensitized plantain-allergic patients mainly reacted with 17, 19 and 40 kDa allergens. Calabozo et al. found that the 17 and 20 kDa allergens are the unglycosylated and glycosylated forms of the same protein (Pla l 1) respectively, and that the 32–36 kDa protein is a dimeric form of the same Pla l 1 allergen. Moreover, the major complex N-glycan of Pla l 1 might be a potential source of cross-reactivity with other glycosylated pollen allergens that could be misleading in terms of false positive diagnoses of allergy to plantain when using natural extracts. Therefore the generation of a recombinant Pla l 1 without the complex N-glycan as part of its structure would be a very useful tool to diagnose patients specifically sensitized to plantain. The glycoprotein Pla l 1 is the major allergen from Plantago lanceolata pollen, which is a common cause of pollinosis in temperate areas.The major allergen of Olea europaea pollen has been found to share sequence similarity to Pla l 1[10].


The aerobiological monitoring concerning the pollen content of the airplankton was carried out by using the volumetric method of collecting data. This is the method that most researchers use for qualitative and quantitative studies of airborne pollen and fungi. It implies the repeated succession of two phenomena: the absorption of a constant volume of air and the immediate trapping of the airborne particles as they impact a trapping surface.

The traps used were of the Hirst type, model VPPS 2000, Lanzoni. The trap allows for an evaluation of the dynamics of the allergenic atmospheric pollen in the town/ city and its surroundings. In order to regularly collect the data and to get correct statistics, the traps are placed at locations higher than 20 meters, far from industrial areas and barriers which might prevent the circulation of the air currents. The bands inside the volumetric traps were changed weekly.

Pollen identification was carried out according to morphological criteria [23, 46]. The pollen concentration is expressed in number of pollen grains per m³ of air. Our bulletins were published weekly, from February until October on the following websites: Euroregional Polleninformation Service Danube-Kris-Mures-Tisza Euroregion (www.pollinfo.ini.hu) and www.nspolen.com. The DKMT Euroregion includes the following counties: Bács Kiskun, Csongrád, Jász-Nagykun-Szolnok, Békés (Hungary), Arad, Timiş, Hunedoara, Caraş Severin (Romania), and Vojvodina (Serbia).

56

Fig.1. The map of The Danube-Kris-Mures-Tisza Euroregion


The main pollen types with a role in allergic sensitization come from anemophilous plants. The pollination duration matches the symptomatology of clinic manifestations. Spieksma (1991) included Plantago pollen into sporomorphs revealing high level of allergenicity, and these taxa were placed on the list comprising the most important plants which should be considered in pollen monitoring in European research centres.

In 2000, the highest total annual concentration was recorded in Szeged (472 PG/m3). In Timişoara the concentration reached only half of the result recorded in Szeged, representing 1.72% of the pollinic range of the year 2000.

In 2001, the Novi Sad station reported a total concentration of 1326 PG/m3. In Timişoara, the concentration of Plantago pollen (148 PG/m3) diminished as compared to the concentration recorded the previous year and represented a mere 0.94% of the annual pollinic range. In 2002, the airpollen concentrations recorded in Novi Sad and Timişoara were relatively similar: 797 PG/m3 and 669 PG/m3. In Timişoara, the concentration represented 3.28% of the total annual of 20068 PG/m3. In 2003, the Ruma monitoring station started recording data in Serbia, alongside the Novi Sad station. The total annual concentration was highest in Timişoara, representing 2.67% of the total annual of 24557 PG/m3. In 2004, the highest concentration (486 PG/m3) was recorded in Timişoara. As to the interannual variation (fig.2), the linear regression model shows the decreasing trend in the annual concentrations for Szeged and Novi Sad, while an increasing trend can be noticed for Timişoara. We can state that in the DKMT Euroregion the Plantago airpollen is a constant presence in the pollinic range, yet its quantity is moderate. This situation is also present in other parts of Europe, where the annual concentrations do not exceed 10% [20]. In Europe, this pollinic type proved to be dominant in Montpellier, France [43], Athens [1], Bitlis, Turkey [12], London, Leiden, Brussels, Munich, Marseille [41], Madrid [44], Salamanca [22], Northwestern Spain [36], Estepona, Southern Spain [35], Belgium [42, 43].

57

The longest period (8 days) when the number of pollen grains exceeded 30 PG/m3 was reported in Novi Sad in 2001. Another two values over 30 PG/m3 were also registered in Novi Sad: one in 2000 and the second, in 2002. In Szeged and Ruma the threshold value was never exceeded. In Timişoara, the sensitization threshold value was exceeded for three days in 2003 and one day in 2002. The stations in the Euroregion did not register excessive values in 2004. The highest daily concentration (fig.3), 64 PG/m3, was registered in Novi Sad in 2001. For Szeged the highest value (22 PG/m3) was registered in 2000; for Timişoara the highest value (42 PG/m3) was registered in 2003. These concentrations are exceptions from the usual values throughout the pollinic season. Similar situations were recorded in Spain [20], Poland [50], Turkey [7, 21], Hungary [26], and Croatia [32].

The Plantago species pollinate from May until the end of August. By considering the data which refer to the number of days when the pollen was present in the airplankton, very wide variations were noted: 97 to 107 days in Szeged, 108 to 137 days in Novi Sad, 102 to 138 days in Ruma, and 76 to 136 days in Timişoara. In 2004, the presence of the Plantago airpollen until the first decade of September did not correlate with an increase in the daily or annual concentrations. In this paper, we determined the APS (Atmospheric Pollen Season) in accordance with the criteria used by the following authors: Nilsson and Persson (corresponding to 90% of the total pollen catch -the 90% method), Andersen and Torben (corresponding to 95% of the total pollen catch- the 95% method) [25]. The longest pollinic season was that in Ruma and the shortest, in Szeged (tab.I). The duration of the pollinic season and the total annual airpollen concentration were relatively close in Timişoara and Novi Sad. Similar variations were recorded in Poland: 95 to 105 days in Rzeszów, 69 to 92 days in Krasne [28], 62 to 98 days in Lublin [50]. In Ankara, Plantago was included in the pollinic group with a maximum pollinating period longer than 15 weeks [27].

Fig.2. Dynamics of annual concentrations of Plantago airpollen (2000-2004)

58

Tabel I. The duration of the atmospheric pollinic season in 2004

Fig.3. Number of Plantago airpollen on the peak day

Several authors report that between 3%-36% of patients are allergic to Plantago, most being polysensitized and, therefore, also allergic to the pollen of other plants, mainly Poaceae. Most patients positive to the skin prick test (SPT) with plantain-pollen extracts are hypersensitive to Poaceae [6, 49; 38], a fact which suggests that at least one allergen in grass and plantain pollens cross-reacts. In a series of 242 consecutive grass-pollen-allergic patients, 71 (29%) were positive in the SPT with plantain-pollen extract.

The results suggest the existence of common antigenic epitopes in melon and Plantago pollen, and in melon and grass pollen [18]. The pollen contains epitopes made up of major and minor determining (antigenic, allergenic) groups of amino acids. An individual may be sensitized either to several major and minor epitopes or to a single epitope, be it a minor one. Cross-reactivity phenomena may occur especially because of homology, but also because of the structural mimocrimy of some epitopes, both within the

2004 Novi Sad Ruma Szeged Timisoara First identification of the airpollen 7 V 4 V 15 V 1 V Last identification of the airpollen 21 IX 19 IX 28 VIII 13 IX Duration of the atmospheric pollinic season (Nilsson & Persson, 1981); 90%

89 days 108 days 73 days 95 days

Duration of the atmospheric pollinic season (Andersen, 1991; Torben, 1991); 95%

107 days 128 days 85 days 109 days

59

same species and between different species. The cross-reactivity phenomenon may be produced not only by various pollen types, but also by food [34]. In other countries, a relationship between changes in crops and variation in pollen sensitization has been observed [38].

Subiza et al. (1995) reported an average airborne Plantago pollen count in Madrid of 3.6%, with positive skin tests of 53% to P. lagopus pollen, 32% to P. lanceolata and 55% to P. lagopus and/or P. lanceolata. Garcia Gonzales (1995) in Málaga reported that 8% of patients were allergic to P. lancelata. Recently, several articles have yielded clinical results from tests carried out in several Spanish cities, with sensitization percentages varying between 15% for Málaga [47]) and 78.24% for Toledo [30]. Regarding patient sensitization, sensitivity was detected in Thessaloniki - Greece to plantain in 194 patients (14.6%) [19].

In the period of plantain pollination, Bryant et al. reported from Sydney that the patients developing asthma symptoms were simultaneously allergic to the plantain allergens. The allergy to plantain allergens was noted in 84% patients with asthma [8]. Of the 629 patients, 459 gave positive SPT results to at least one pollen. No statistical differences were found with respect to gender, habitat (rural or urban) and age.

Sensitizations to the different botanical families were as follows: 384 patients were sensitized to Poaceae family pollen, 348 patients to the Oleaceae family, 249 patients to the Plantaginaceae family, 211 patients to the Chenopodiaceae family, 153 patients to the Cupressaceae family, 94 patients to the Platanaceae family, 94 patients to the Compositae family, 80 patients to the Betulaceae family and 27 patients to Urticaceae pollen. Multiple Correspondence Analysis proved the existence of associations among pollen sensitizations, showing that they clustered into two groups: Group I which included Poaceae, Oleaceae, Cupressaceae, Chenopodiaceae and Plantaginaceae and Group II, which included Betulaceae, Platanaceae and Compositae. Pollens of the association Group I do not coincide with those collected in largest numbers in the Madrid atmosphere, since the total annual pollen grain count is highest for Poaceae, followed by Cupressaceae, Platanus, Olea and Plantago [44, 4]. Two distinct behaviors could be observed in Milan: a) a high propensity to develop new respiratory allergies characterized patients allergic to Poaceae (46%), Parietaria (35%), and Betula pollen (37%) whereas b) patients allergic to house dust mite (15%), Ambrosia (15%), Alternaria (11%), Artemisia (22%), and Plantago (20%) showed a much lower propensity to develop new allergies. The “new” allergens (Ambrosia and Betula) caused 228/256 (89%) new sensitizations detected in the whole study group, included patients allergic to Plantago [2].

It has been observed that is not a clear relationship between the amount of pollen collected in the air and the incidence in allergy people [38]. Percentage of patients displaying reactions to Plantago pollen type according to SPT was 13.33% in Cordoba and 21.42 % in Evora. Sum of daily pollen counts during the study period was 606 in Cordoba and 184 in Evora. Result of correlation between Plantago pollen counts and symptoms suggest a lower incidence of allergic diseases related to pollen in the city of Evora. This fact could be explained by two main causes.

Firstly, 73.34 % of patients in the city of Cordoba were aged between 11 and 30 years old, whereas only 50% of patients were included in group. A study performed across

60

Spain reported that the 14-25 age-group was the most affected by pollinosis. Secondly, the population in Evora has maintained a rural lifestyle for a long time whereas in Cordoba people have changed to an urban lifestyle along the last decades.

Some studies suggest that acquisition of certain infections or exposure to naturally occurring microbial exposures as encountered in the rural environment could confer protection from allergic diseases. The atmosphere in Evora is by far less contaminated than in Cordoba where the main source of solid material emissions into the air is road traffic, since the city lies on the route for goods transported from the southern to the central regions of the country. Córdoba has recently seen a reduction in the number of ecosystems where grasses are able to grow, due to the expansion of the city and to town-planning changes [38, 11]. Prevalence of allergic reactions to Plantago pollen type in each area was 9.24 in North, 10.15 in West, 11.21 in South, 6.05 in Centre, 12.52 in East. Percentage of patients displaying reactions to Plantago pollen type was: 29.16% (between 1984 -1990) and 33.87% (between 1999 -2001). A positive and significant correlation was observed between monthly pollen indices and antihistamine sales for Plantago [38].

In the sample of population of Sarajevo region during the 2002 year has been investigated on the pollen of weed plant species and Poaceae pollen. In the mixture of pollen weed plant species have been following plants: Plantago lanceolata, Chenopodium album, Solidago gigantea, Artemisia vulgaris and Urtica dioica. 589 have been tested patients by mixture of pollen mentioned plant species and found 115 as a sensitive on pollen alergy; 65 male and 50 females. Even 61 are children to 14 years, or 53% of total sick patients [39].

In Romania, hypersensitization to the mixture of pollen coming from grasses (Artemisia vulgaris, Plantago lanceolata, Rumex acetosella, Urtica dioica) was found in 13.13% of the cases [33] and 2.77% of the patients [16].

A study carried out in France points out that some of the children suffering from atopic dermatitis (9.8%) are also sensitive to Plantago aeroallergens [9].

The number of people allergic to plant aeroallergens has substantially increased in big cities and industrial areas [31]. Thus, monitoring of the pollen counts in the airplankton of cities is of relevant medical importance.

The concentration of pollen grains in the air over a city is determined by the individual rhythm of plant pollination, meteorological conditions, composition of local flora, geographic location and kind of urban structure (loose or compact housing, areas with many gardens or with scarce vegetation, industrial areas, agricultural areas or forests) [51]. The higher temperatures in a town can cause a longer vegetative period.

The microclimate of towns is characterized by reduced levels of relative air humidity, specific winds, an increased content of aerosols in the air, and a greater frequency of fogs. The generally accepted conclusion is that the participation of arboreal pollen in the pollen fall reflects regional conditions, while the content of pollen of herbaceous plants reflects local ones [20].

Results of this study demonstrate that Plantago seasons occurred at regular intervals between May and August each year; however, individual daily and seasonal Plantago counts were heterogeneous. The start of APS (Atmospheric Pollen Season) was relatively constant for Plantago, while the end of APS showed significant variations. Registered data

61

confirm the fact that at least quantitatively Plantago pollen is not an important allergenic factor. Overall, the pollen shedding course of the Plantago in Danube-Kris-Mures-Tisza Euroregion corresponds to that already described during the pollen season in other European areas [23].

Conclusions

In the DKMT Euroregion the Plantago airpollen is a constant presence in the

pollinic range, but it is moderately represented from a quantitative point of view. The highest annual concentration (1326 PG/m3) was recorded in Novi Sad in 2001,

while the lowest annual concentration (134 PG/m3) was recorded in Ruma in 2004. In Szeged and Novi Sad the annual concentrations are on the decrease, while in

Timişoara they are slightly on the increase. The highest daily concentration (64 PG/m3) was recorded in Novi Sad in 2001. The threshold value (30 PG/m3/day) was seldom exceeded. There is no correlation between the longer presence in the airplankton and an

increase in the concentrations.

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30. MORAL DE GREGORIO A., SENENT SANCHEZ C., CABANES HIGUERO N., GARCIA VILLAMUZA Y., GOMEZ-SERRANILLOS R., 1998 - Pólenes alergénicos y polinosis en Toledo durante 1995-96. Rev. Esp. Alergol. Inmunol. Clín. 13, 2: 126-134

31. NILSSON S., PERSSON S., 1981 - Tree pollen spectra in the Stockholm region (Sweden), 1973-1980, Grana, 20: 179-182

32. PETERNEL R., ČULIG J., MITIĆ B., VUKUŠIĆ I., ŠOSTAR Z., 2003 - Analysis of airborne pollen concentrations in Zagreb, Croatia, Ann Agric Environ Med, 10: 107–112

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34. RADU J.R., 1998. Alergiile reaginice. Imunoterapia specifică cu vaccinuri alergenice. Ed. Medicală Amaltea, Bucureşti

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36. RODRIGUEZ-RAJO F.J., JATO V., AIRA M.J., 2003 - Pollen content in the atmosphere of Lugo (NW Spain) with reference to meteorological factors (1999–2001). Aerobiologia, 19: 213–225

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http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Search&itool=pubmed_AbstractPlus&term=%22Saracevic+E%22%5BAuthor%5D

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64


ARAUCARIA EXCELSA L. VITROCULTURES INITIATION

L. POP∗; DORINA CACHIŢĂ*

Abstract: Araucaria excelsa L. is a well-known conifer, mostly used as an indoor ornamental plant. For the initiation of Araucaria excelsa L. vitrocultures, we have studied the reactions of explants, in the presence of different growth regulators, added in the aseptic nutritive media. We have prelevated apexes from a unique plant and used them as biological material. The explants were sterilized and inoculated on BM media, with and without growth regulators. This experiment, which lasted for 90 days, has brought forth the following conclusions: On V0 (control variant– BM without growth regulators), the inoculs have presented a very week regenerative capacity; the best medium for elongation of Araucaria excelsa L. was V2 (BM with 2 mg/l BA + 2 mg/l NAA); the most ramifications and buds can be obtained if using V4

experimental variant (BM with 0.5 mg/l NAA + 0.5 mg/l KIN); at any experimental variant the rhysogenesis wasn’t observed. Keywords: Araucaria excelsa L., vitroculture, growth regulators, conifer

Introduction

Araucaria excelsa L. is a beautiful conifer from Canar and Mader Irelands, and

commonly cultured to decorate different indoor and outdoor places. The multiplication of this tree is problematical [4]; therefore the “in vitro” micropropagation remains a good alternative to be studied.

The purpose followed on this experiment was the Araucaria excelsa L. vitroculture initiation and its evolution during 90 days observation.


For this research we have collected 2 cm length apexes from a 2 m height adult Araucaria excelsa L. plant, which is founded in the glasshouse of University of Oradea. The apexes were prelevated from basal zone of crown and sterilized in 96° alcohol for 1 minute submersion, followed by a Natrium Hypochloride 0.8%, for 15 minutes resubmersion. After these were done, the biological material was washed, more times, in sterile water [1].

In aseptic environment, the resulted pieces were shorted at 1 cm length and so the inoculs were obtained. They were inoculated (fig.1) on 5 different variants of nutritive media. The control experimental variant have consisted in Araucaria excelsa L. apexes, placed on Murashige and Skoog (1962) nutritive standard medium [3], abbreviated here BM (basic medium). The other variants have contained, in addition, different growth regulators.

∗ University of Oradea, Faculty of Science, Universităţii Street, no.1, 410087, Oradea, Romania

65

The Murashige&Skoog (1962) [4] mineral medium, which have consisted in

macroellements, FeEDTA, Heller microellements, vitamins (B6, B1 and PP), m-inositol, sucrose and agar, was used as basic medium (BM). In this mixture, growth regulators were added, as following:

- V0 –(control variant) – BM without growth regulators; - V1 –BM with 2 mg/l BA + 2.5 mg/l IBA; - V2 – BM with 2 mg/l BA + 2 mg/l NAA; - V3 – BM with 0.5 mg/l KIN + 2.5 mg/l IBA; - V4 – BM with 0.5 mg/l NAA + 0.5 mg/l KIN;

The growth media were sterilized at 121°C, during 30 minutes [2]. After their cooling, in the sterile room, we proceeded to inoculate the minicuttings, one piece per culture recipient, and place them on shelves, at 20-22°C, under fluorescent white light, at 1700 lux, with a 16h light/24h photoperiod.


The Araucaria excelsa L. vitroplantlets evolution has been observed during 90 days, and the watched parameters were noted and compared.

At 30 days after inoculation, the Araucaria excelsa L. vitroplantlets have presented stagnation, their height modification being mostly insignificant (fig.2, fig.4). The highest elongation was founded on V2 medium (BM with 2 mg/l BA + 2 mg/l NAA). No buds or ramifications were observed. The rhysogenesis was missing, too.

We didn’t find any infection on cultures, but some necroses have occurred in a few growth recipients, at all experimental variant, excepting V3 (BM with 0.5 mg/l KIN + 2.5 mg/l IBA), where all plantlets have survived. The lowest survival level (84.61%) was observed on V0 (control variant), where the growth regulators were missing (fig.3).

The general survival percents were pretty good, the best being founded on V3 (BM with 0.5 mg/l KIN + 2.5 mg/l IBA) (fig.3)

1 cm

Fig. 1. Making minicuttings and the inoculation process

inoculation Murashige and Skoog medium

Original plant

4 cm 1 cm

Explant

66

Fig. 2 Araucaria excelsa L. cultures after 30 days from inoculation (V0 (control

variant) – BM without growth regulators, V1 –BM with 2 mg/l BA + 2.5 mg/l IBA, V2 – BM with 2 mg/l BA + 2 mg/l NAA, V3 – BM with 0.5 mg/l KIN + 2.5 mg/l IBA, V4 – BM with 0.5 mg/l NAA + 0.5 mg/l KIN)

93,1

84,61

95,2

91,66

100

60

80

100

V0 V1 V2 V3 V4variants

%

Fig. 3. The survival percent of Araucaria excelsa L. plantlets, at 30 days after

inoculation

67

Fig. 4. The vitroplantlets elongation at 30 days after inoculation on aseptic media

At 60 days after inoculation the best elongation was observed also on V2 medium

(BM with 2 mg/l BA + 2 mg/l NAA) (fig.5, fig.6), but the ramifications were more on V4 medium (BM with 0.5 mg/l NAA + 0.5 mg/l KIN), (fig.5, fig.7).

Fig. 5. Araucaria excelsa L. cultures after 60 days from inoculation (V0 (control variant) – BM without growth regulators, V1 –BM with 2 mg/l BA + 2.5 mg/l IBA, V2 – BM with 2 mg/l BA + 2 mg/l NAA, V3 – BM with 0.5 mg/l KIN + 2.5 mg/l IBA, V4 – BM with 0.5 mg/l NAA + 0.5 mg/l KIN)

1.041.061.081.1

1.121.141.161.181.2

V0 V1 V2 V3 V4

Stalk length

68

0

0,5

1

1,5

2

2,5

V0 V1 V2 V3 V4

Stalk length


0

1

2

3

4

5

6

V0 V1 V2 V3 V4

Ram ifications

Fig. 7. The vitroplantlets stalk ramification at 60 days from inoculation The 90th day of this experiment has revealed us that in the V2 medium (BM with 2

mg/l BA + 2 mg/l NAA) the vitroplantlets were 30% taller than those from the control variant (BM without growth regulators) (fig.8, fig.9). The most ramifications were found again at V4 experimental variant (BM with 0.5 mg/l NAA + 0.5 mg/l KIN) (fig8, fig.10)

The control experimental variant V0 (BM without growth regulators) hasn’t manifested any ramification (fig.8, fig.10).

No one of experimental variants has manifested rhysogenesis.

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Fig. 8. Araucaria excelsa L. cultures after 90 days from inoculation (V0 (control

variant) – BM without growth regulators, V1 –BM with 2 mg/l BA + 2.5 mg/l IBA, V2 – BM with 2 mg/l BA + 2 mg/l NAA, V3 – BM with 0.5 mg/l KIN + 2.5 mg/l IBA, V4 – BM with 0.5 mg/l NAA + 0.5 mg/l KIN)

0

0,5

1

1,5

2

2,5

3

V0 V1 V2 V3 V4

Stalk length


70

0

1

2

3

4

5

6

V0 V1 V2 V3 V4

Ramifications

Fig. 10. The vitroplantlets stalk ramification at 90 days from inoculation

Conclusions

According to this research, the initiation of Araucaria excelsa L. vitoculture is possible, and this is a useful tool for micropropagation of this ornamental conifer specie.

On V0 (control variant– BM without growth regulators), the inoculs have presented a very week regenerative capacity.

No ramifications were observed on standard MS medium (V0) The best medium for elongation of Araucaria excelsa L. was V2 (BM with 2 mg/l

BA + 2 mg/l NAA). The most ramifications and buds can be obtained if using V4 experimental variant

(BM with 0.5 mg/l NAA + 0.5 mg/l KIN). 90 days seems to be a to short vitroculture period, for rhysogenesis occurrence. These positive results stimulate us to go forward with the experiments concerning

Araucaria excelsa L. “in vitro” micropropagation.

Abreviations: MS – Murashige&Skoog (1962), BM – basic medium, BA – benzyl-adenine; IBA – β–indolilbutilic acid; NAA – α-naftilacetic acid; KIN – kinetin

REFERENCES

1. CACHIŢĂ, C.D., 1987 - Metode „in vitro” la plantele de cultură. Bazele teoretice şi practice, Ed. Ceres, Bucureşti

2. CACHIŢĂ, C. D., SAND, C., 2000 - Biotehnologie vegetală. Baze teoretice şi practice. Vol.1, Ed. Mira Design, Sibiu: 272-276

3. MURASHIGE, T., SKOOG, F., 1962 - A revised medium for rapid growth and bioassays with tobacco tissues cultures. Physiologia Plantarum, 15: 155-159

4. PREDA, M, 1979 - Floricultura (ed. a II-a), Edit. Ceres, Bucureşti: 231-232

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AFRICAN VIOLET (SAINTPAULIA IONANTHA L.) EXVITROPLANTLETS

ACCLIMATIZATION, IN DIFFERENT TYPES OF SUBSTRATUM

ADRIANA PETRUŞ – VANCEA∗, C. F. BLIDAR∗, ANCA BACIU∗∗

Abstract: In micropropagation, the success of cloning depends on the surviving process of the exvitroplantlets after the acclimatization and on the quality of the resulting planting material. For the planting material, coming from vitroculture, to be competitive with the one obtained through classic vegetative multiplying, especially for African violets, it has to be cheaper and better quality. So, the „ex vitro” planting of the African violet plantlets, directly into greenhouse conditions, is possible and favorable if the substratum in which the acclimatization is being made is efficient and low cost. From the seven types of substratum we tested for exvitroplantlets acclimatization, the most efficient, regarding the post acclimatization survival percent, was 100%, the „Top soil” substratum, a worm compost, the exvitroplantlets growth spores set on this type of substratum having a very good rooting, these data being meaningful statistically speaking. Key words: Saintpaulia ionantha L., „ex vitro” acclimatization, substratum

Introduction

The proper characteristics for the „ex vitro” planting substratum for cultivating

African violets are the following [4]: consistence 0,75g/cm3, porosity 75,10%, ventilation 19,53%, available water 26,03% and easy available water 23,71%. Analysing different types of substratum mixtures, that had the previously described characteristics, namely: common peat : eucalyptus saw : sand; earthworm soil: vermiculite : sand; pine saw : earthworm soil; peat : vermiculite or other commercial substratum („Eucatex”, made of peat and „Vida Verde”, a commercial substratum especially produced for African violets), in equal proportion each, the authors [4] demonstrated that all variants proved themselves to be proper for cultivating African violets.

African violet exvitroplantlets (after removing them from „vitro”) can be acclimatized successfully to a septic medium, in different types of unconventional substratum [5]. In this sense, it was recommended the use of poplar saw, but never the beech one and no additional thermic treatments [2]. Also, glassy wool may be an „ex vitro” acclimatization substratum for African violets, as a cheap and good alternative in placing the African violet exvitroplantlets, for future planting in a septic medium [5], and adding the biogel [6], and also the zeolits [7], in the exvitroculture substratum, especially mixed with „Top soil”, proved to be favorable to this species, the post-acclimatization survival percent being of 100%, and the growth in the adapting to a septic medium period was suitable. In this last type of substratum, the peroxides activity from the rootlets of the African violet exvitroplantlets – as a marker of the rizogenesis process – determined after 30 days from

∗ University of Oradea, Faculty of Science, Universităţii Street, no. 1, 410087, Oradea, Romania ∗∗Potato Research & Development Station from Târgu –Secuiesc, Romania

72

their acclimatization to a septic medium, presented very significant positive differences statistically speaking, compared to the control (the rootlets of the plants grown in a natural medium, in a greenhouse) [3], which proves the presence of a intense process of rootlets forming.


The acclimatized vegetal material consisted by individual propaguls of African

violets (Saintpaulia ionantha) witch were detached from vitrobushes, having 3 – 4 rootlets, 1 – 1,5 cm waist and 4 – 5 leaflets with 1 cm foliar limb diameter, kept 60 days in vitroculture, in basal MURASHIGE-SKOOG (1962) medium [1], containing vitamins (HCL thiamine, HCL pyridoxine and nicotinic acid, 1 mg/l each), myo-inositol 100 mg/l, sucrose 20 g/l and agar – agar 7 g/l, without grown regulators; pH was adjusted to 5,7, prior autoclaving. The Saitpaulia propaguls were inoculated and cultivated “in vitro” in glass jar by 200 ml with 12 cm height and 7 cm diameter, in each recipient was shared 50 ml medium. The sterilization of recipients with culture medium was made by autoclaving at 121 ˚C for 25 minutes. The inoculation was operated after medium cooling.

After the inoculation, the jars were covered with colourless, transparent, polyethylene folia. The cultures were incubated at irradiance with white fluorescent light with 1700 lx intensity and 16/24 h light photoperiod, at 23 ˚C ± 2˚C in the light period and 20˚C ± 2˚C in the darkness period. After 60 days of vitroculture, the agarized medium was removed, by washing with tap water at lab temperature and the vitroplantlets were transferred to the septic medium, in greenhouse. In acclimatization process, was tested the efficiency of seven mixture types, as “ex vitro” culture substratum, with some characteristics (tab.1), namely: V0 – river sand, with fallow soil (3:1); V1 - white peat, with manure and river sand (1:1:1); V2 - perlite with white peat (1:1); V3 - river sand with white peat (1:1); V4 - manure with white peat (1:1); V5 – perlite; V6 – „Top soil”. “Top soil” is commercial name of a substratum made in a biobase (from Stei City, Bihor County), soil resulted from a vital activity of worm cultures growth on vermicompost. To ensure an optimum humidity in the atmosphere around plantlets and to avoid an excessive evapoperspiration, each plantlet was placed under a colorless plastic case, which was pierced in the upper part. This was made to ensure the evacuation of the excessive humidity from the interior vessel. During acclimatization period, all the substratum types were humidified with foul tap weather, and after 14 days from planting, those were weathered with Knop (1865) mineral solution, 100 ml/100 cm2; between 1100 – 1400 hours, exvitroplantlets were protected, by direct action of sunray, covering them with paper sheet.

Both in the moment of the “ex vitro” vitroplantlets transfer and also after 30 days from the beginning of the acclimatization process, were operated biometrics of the growth indices and was calculated the post-acclimatization survival process, the data being statistically insured. We have calculated the arithmetic average on each sample, at 30 days after their transfer into “ex vitro” culture substratum was related to values registered at the start moment of acclimatization, these being considered 100%.

73


The survival percent – registered at 30 days from “ex vitro” transfer of African violet vitroplantlets, given to initiation moment of acclimatization – has noted maximal values to the exvitrocultures that were made on perlite with white peat (1:1) (V2), river sand with white peat (1:1) (V3), perlite (V5) and Top soil” (V6), but higher values, meaning 98%, were recorded at the exvitroplantlets lot placed on the white peat with manure and river sand (1:1:1) mixture (V1) or on manure with white peat (1:1) (V4). A little less exvitroplantlets, 95%, were survived on river sand, with fallow soil (3:1) (V0) (fig. 1), but the percent is still a reasonable one.

During the acclimatization period, the rizogenesis was influenced by the exvitroculture substratum nature. So, the rootlets length increased given to the moment of planting the exvitroplantlets on soil, registered insignificant increased from statistically point of view (tab. 2), with 17% (fig. 1) percentage efficiency, in case of those placed on river sand with fallow soil (3:1) (V0), those size reaching only medium values of 1,35 cm (tab. 2), to the 130%. On the other hand, for the rootlets of exvitroplantlets cultivated on „Top soil” (V6), whose length was to the 2,95 cm, the efficiency was to the 130% and the difference against the control was statistically very significant. At exvitroplantlets placed on perlite with white peat (1:1) (V2) – to the 30 exvitroculture days – was reported a doubling of radicular system length, reaching values of 2,29 cm, related to rootlets size from the moment of vitroplantlets transfer on soil. At the end of acclimatization period, a little less rootlets, just in average of 4,5 samples was counted at the base of exvitroplantlets planted on river sand with fallow soil (3:1) (V0), the increased given to initiation moment being to 34%, and a lot of rootlets, 7,5 samples, was identified – similar to the his length – at exvitroplantlets planted on „Top soil” (V6), the efficiency given to control lot was 130%, all the values being statistically very significant (tab. 2).

The perlite substratum were identified to be optimal, alone or mixed 1:1 with peat (V2), according to variant V5, when the benefits, in comparison to the control lot, were 66%, respective 106% (fig. 1), same like in the case of rootlets length. We can observe in the figure 1 histograms that the highest benefits of exvitroplantlet growth indicators, in dependence of exvitroculture substratum, were registered to the indexes that express the rizogenesis, these ones touching highest values, 130%, referred to the plantlet rootlets size growth, in “Top soil”.

To those cultivated on mixture of river sand with fallow soil (3:1) (V0), white peat with manure and river sand (1:1:1) (V1), river sand with white peat (1:1)(V3) and manure with white peat (1:1)(V4), total leaflets number was represented by the summing of the leaflets with 0 – 0,4 cm diameter, those with 0,5 – 0,9 cm and 1,0 – 1,4 cm diameter, because exvitroplantlets did not disposed of higher diameter leaflets (tab. 2), those market medium values between 4,6 samples (V0), showing significant differences to the start moment of acclimatization and 6,05 samples (V4), witch represented statistically significant efficiency, percentage expressed by 12% values, respectively 41%, to the initiation of the acclimatization (fig. 1), though the leaflets number with reduced diameter (which is in class dimensions of the 0 - 0,4 cm and 0,5 – 0,9 cm), registered statistically significant minuses to 47% (V0), at the end of acclimatization period. Only those exvitroplantlets, cultivated on

74

the substratum containing perlite (V2 and V5) and those on the “Top soil” (V6) presented leaflets with an diameter between 1,5 – 1,9 cm (fig. 1), total leaflets number to these lots reaching values of 7,5 and 8,54 samples, to the first two lots, and to the final lot the efficiency was 59% and 92%, respectively 9,1 samples and the higher increase of 109%, the difference between those values, given to the acclimatization initiation, being very significantly, statistically point of view (tab. 2). The efficiency of the total leaflets number, registered to the end of the acclimatization period, given to the initiation moment, represent those leaflets which “ex vitro” new formed.

Conclusions

Analyzing the results of the present experiment, we can state that, in the case of

the African violets, all tested substratum led to a good or even very good post acclimatization survival, especially remarked was the mixture of perlite with peat, in a report of 1:1 (V2), or perlita (V5) and „Top soil”, as independent substratums from rooting, in this last case the growth parameters being the highest.

REFERENCES

1. MURASHIGE T., SKOOG F., 1962 - A revised medium for rapid growth bioassays with tobacco tissue

cultures. Physiol. Plant., 15: 473 - 497 2. PETRUŞ - VANCEA ADRIANA, 2005 - The acclimatization of African violet exvitroplantlets in substratum

consisting in sawdust. Analele Univ. Oradea, fasc. Biol., t. XII: 119 – 123 3. PETRUŞ – VANCEA ADRIANA, CACHIŢĂ C. DORINA, ŞIPOŞ MONICA, 2004 - Activitatea

peroxidazică în rădăciniţele vitro- şi exvitroplantulelor de crizanteme, violete africane şi de Cymbidium. An. SNBC, t. IX, 1., cap. III – Biologie celulară vegetală: 392 – 395

4. SALVADOR E.D., MINAMI K., JADOSKI S.O., 2003 - Evaluation of different substrates on African violets (Saintpaulia ionantha Wendl.) growth. International Symposium on Soilless Culture and Hydroponics. Acta Horticulturae, 697, www.actahort.org/books/697

5. VANCEA ADRIANA, CACHIŢĂ C. DORINA, 2002 - Aclimatizarea vitroplantulelor de Saintpaulia ionantha, prin plantarea acestora pe substraturi neconvenţionale. În: Lucrările celui de al X-lea Simpozion Naţional de Culturi de Ţesuturi şi Celule Vegetale. (eds. Cachiţă C. Dorina, Rakosy T. Lenuţa, Ardelean A.). Edit. Risoprint, Cluj- Napoca: 310 – 315

6. VANCEA ADRIANA, CACHIŢĂ C. DORINA, 2002 - Utilizarea biogelului în substratul destinat aclimatizării vitroplantulelor de saintpaulia ionantha la mediul septic de viaţă. în: Lucrările Simpozionului Ştiinţific „90 de ani de învăţământ agronomic universitar la Iaşi”. CD, Agrosoft – U.S.A.M.V., Iaşi

7. VANCEA ADRIANA, CACHIŢĂ C. DORINA, FENCE DANIELA, 2003 - Utilizarea zeolitului ca substrat de aclimatizare a vitroplantulelor la mediul septic de viaţă. în: Lucrările celui de al XI-lea Simpozion Naţional de culturi de ţesuturi şi celule vegetale. (eds. Cachiţă C. Dorina, Ardelean A. Edit. Daya, Satu-Mare: 212 – 224

http://www.actahort.org/books/697/index.htm

http://www.actahort.org/books/697/index.htm

Fig. 1. The survival percent and growing of the African violet (Saintpaulia ionantha L.) exvitroplantlets, at 30 days

from their “ex vitro” transfer and planting on different substratum types: V0 – river sand with fallow soil (3:1); V1 - white peat with manure and river sand (1:1:1); V2 - perlite with white peat (1:1); V3 - river sand with white peat (1:1); V4 - manure with white peat (1:1); V5 – perlite; V6 – „Top soil”, expressed in percentage values against the parameters registered at the level of exvitroplantlets biometred in first days of acclimatization, which were considered 100%

0%

50%

100%

150%

200%

250%

Survivalpercent

Rootletslength

Rootletsnumber

Propagulsnumber

Totalnumber of

leaflets

Leafletsnumber with

0-0,4 cmdiam.

Leafletsnumber with

0,5-0,9 cmdiam.

V0 V1 V2 V3 V4 V5 V6

I zi

76

Table I. Qualitative characteristics of some substratum type witch were used by us in African violet exvitroplantlets acclimatization, obtained from chemical analyses made at Chemical and Agrotechnical Research Center Oradea

Determinations Substratum type

eather

H 2 2O5 aO gO

mid.

g/100

Mineral residue

%

river sand + fallow soil, 3:1 ,1 0 1 8

0,13

white peat + manure + river

sand, 1:1:1 ,4 2 3 8 4 0

,14

perlite + white peat, 1:1 ,3 6 0 3 5 1

0,15

river sand + white peat, 1:1 ,2 9 1 2 9

0,18

manure + white peat, 1:1 ,1 6 3 7 2 0 5

0,25

„Top soil” ,9 2 1 8 5 0

0,14

Normal values 5 - 30 - 12 0 - 60 0 - 50 - 15

Table II. Statistic processing of biometric measurements done at the level of African violet extrovitroplantlets (Saintpaulia

ionantha L.), at 30 days of their “ex vitro” transfer and their plantation into different substratum type, namely: V0 – river sand with fallow soil (3:1); V1 - white peat with manure and river sand (1:1:1); V2 - perlite with white peat (1:1); V3 - river sand

with white peat (1:1); V4 - manure with white peat (1:1); V5 – perlite; V6 – „Top soil” Type V0 (control)

Rizogenesis Caulogenesis Biometrics

Statistic count

Rootlets length Rootlets number

Propaguls number

Total number of leaflets

Leaflets number with

0-0,4 cm diam.

Leaflets number with 0,5-0,9 cm

diam.


diam.


diam.

1 2 3 4 5 6 7 8 9

77

x ± S x 1,35 ± 0,08 4,50 ± 0,11 1,00 ± 0,00 4,60 ± 0,15 0,95 ± 0,05 1,90 ± 0,07 1,75 ± 0,10 0,00 ± 0,00

p ns *** ns * *** ** *** ns Type V1

1 2 3 4 5 6 7 8 9

x ± S x 1,59 ± 0,06 4,70 ± 0,10 1,00 ± 0,00 5,20 ± 0,09 1,20 ± 0,09 2,05 ± 0,09 1,95 ± 0,09 0,00 ± 0,00

p *** *** ns *** *** ns *** ns Type V2

x ± S x 2,29 ± 0,06 5,65 ± 0,11 1,00 ± 0,00 7,55 ± 0,22 1,90 ± 0,07 2,50 ± 0,11 2,30 ± 0,10 0,95 ± 0,05

p *** *** ns *** ** ns *** *** Type V3

x ± S x 1,95 ± 0,03 5,20 ± 0,09 1,00 ± 0,00 5,80 ± 0,09 1,40 ± 0,11 2,25 ± 0,12 2,15 ± 0,13 0,00 ± 0,00

p *** *** ns *** *** ns *** ns Type V4

x ± S x 1,81 ± 0,03 5,10 ± 0,12 1,00 ± 0,00 6,05 ± 0,05 1,75 ± 0,10 2,30 ± 0,10 2,00 ± 0,00 0,00 ± 0,00

p *** *** ns *** ** ns *** ns Type V5

x ± S x 2,46 ± 0,07 7,40 ± 0,11 1,00 ± 0,00 8,45 ± 0,11 2,10 ± 0,07 2,65 ± 0,11 2,55 ± 0,11 1,15 ± 0,08

p *** *** ns *** ns * *** *** Type V6

x ± S x 2,95 ± 0,05 7,50 ± 0,11 1,00 ± 0,00 9,10 ± 0,16 2,10 ± 0,07 2,85 ± 0,08 2,70 ± 0,10 1,45 ± 0,11

p *** *** ns *** ns *** *** ***

Note: x ± S x (average ± standard deviation of the average), s (standard deviation), S% (variability coefficient), p (significance limit of the difference against the control): ns – insignificant, * - significant, ** - distinctively significant, *** - very significant

78

Analele ştiinţifice ale Universităţii “Al. I. Cuza” Iaşi Tomul LIV, fasc. 1, s. II a. Biologie vegetală, 2008 ULOCLADIUM ATRUM PREUSS - BIOLOGICAL CONTROL AGENT OF GREY

MOULD (BOTRYTIS CINEREA PERS.) OF CROPPED PLANTS

TATIANA EUGENIA ŞESAN∗, J. KÖHL∗∗, WILMA M. L. MOLHOECK∗∗

Abstract. Ulocladium atrum Preuss is a promising candidate as biological control agent against Botrytis cinerea Pers. from different plants: onion, lily, geranium, cyclamen, kiwifruit, strawberry, grapevine, etc. During year 2000 the authors have been worked together in the Plant Research International Wageningen, The Netherlands, for the EU project 1898 - BIOSPORSUPPRESS - The biological control of air-borne necrotrophic plant pathogens by suppression of spore production -, leaded by PhD J. Köhl. The objectives of that research were: (1) Testing culture media for Ulocladium atrum (isolate 385 and isolates from soil organic fragments and necrotic leaf fragments) in order to select the most favourable one for detecting the fungus in soil organic material and necrotic leaf samples; (2) Detecting of U. atrum incidence in necrotic leaf and soil samples (soil organic fragments‚ soil suspensions) collected from U. atrum treated strawberry fields plated on ARSA medium and (3) Reisolation and purification of U. atrum from colonies developing on organic material or soil dilution plates from all the experimental plots, obtaining of U. atrum isolates for their characterization in comparaison with the isolate 385 used in the experimental field plots as a biocontrol agent against the grey mould of strawberry. Key words: biological control (biocontrol) of Botrytis-diseases, Ulocladium atrum biological control agent, strawberry, culture media.

Introduction

Ulocladium atrum Preuss (Mitosporic fungi/ Conidial Ascomycetes/ Ord.

Hyphomycetales/ Fam. Dematiaceae) (fig. 1) is an attractive candidate for applications as a biocontrol agent in the field as well as in greenhouses for suppressing sporulation of Botrytis cinerea and other Botrytis spp. of different crops, like: onion, lily, geranium, cyclamen, kiwifruit, strawberry, grapevine, etc. [1], [2], [4], [5], [6], [7], [8], [9], [10], [11], [12], [13], [14], [15], [17], [18], [19], [20], [21], [22]. For application of the antagonistic fungus as a bioproduct it is necessary to be studied his action on the environment.

∗ University of Bucharest, Faculty of Biology, Aleea Portocalilor, no. 1-3, 060101, Bucharest 35, Romania ∗ ∗Plant Research International, Bornesesteeg 65, 6708 Wageningen, The Netherlands

79

10μm

Fig. 1 – Ulocladium atrum Preuss: conidiofores and conidia (after David, 1995 [3])

The objectives of this research, in the frame of the international project 1898 -BIOSPORSUPPRESS - The biological control of air-borne necrotrophic plant pathogens by suppression of spore production - were:

1. Choice of culture media for Ulocladium atrum (isolate 385 and isolates from soil organic fragments and necrotic leaf fragments) in order to select the most favourable one for detecting the fungus in soil organic material and necrotic leaf samples;

2. Incidence of U. atrum in necrotic leaf and soil samples (soil organic fragments‚ soil suspensions) collected from U. atrum treated strawberry fields plated on ARSA medium.


Biological materials used for this investigation consisted of: strawberry necrotic

leaves only collected from the overwintering trial (1999-2000) and soil organic fragments obtained from soil samples collected on 16 February 2000 from 8 field experiments performed between 1997 and 2000 (table 1) in the biocontrol group of Plant Research International Wageningen.

80

Table I. Samples♦ from strawberry trials 1997-2000 year treatments plot numbers date of

collect-ing1)

1997-1 1. Control 2. Ua weekly during season 3. fungicides 4. Ua weekly during flowering

2,5,10,13,17 3,8,11,15,18 4,7,12,16,19 1,6,9,14,20

18.07.2000

1997-2 1. Control 2. Ua weekly during season 3. fungicides 4. Ua weekly during flowering

1,6,11,13,20 4,7,10,16,18 3,8,9,15,19 2,5,12,14,17

12.07.2000

1997/ 1998-a 1997- 1998-b

1. Control 2. Ua monthly sept.(IX)-april (IV) (a) 3. removal senescing/dead leaves 4. Ua planned during flowering; not performed 5. Ua monthly sept.(IX)-june (VI) (b)

1,8,15,19 3,7,12,17 4,10,11,18 2,6,14,16 5,9,13,20

28.06.2000

1998-1 1. Control 2. Ua at planting; weekly from bud to flowering;

twice weekly during flowering 3. fungicides 4. Ua twice weekly at flowering 5. removal senescing/dead leaves

1,8,15,19 3,9,12,17 4,6,11,18 2,10,14,16 5,7,13,20

27.06.2000

1998-2 1. Control 2. Ua at planting; weekly from bud to flowering;

twice weekly during flowering 3. fungicides 4. Ua twice weekly at flowering 5. removal senescing/dead leaves

1,6,13,20 4,8,15,19 3,9,14,18 5,10,11,17 2,7,12,16

26.06.2000

1998/ 1999

1. Control (Tween water) 2. fungicides 3. Ua fortnightly from winter to flowering; twice

weekly during flowering 4. Ua twice weekly during flowering 5. removal senescing/dead leaves

1,8,15,19 3,9,12,17 4,6,11,18 2,10,14,16 5,7,13,20

14.06.2000

1999

1. Control 2. Tween water every second day during

flowering 3. Ua 5x105 ml-1 every 4th day during flowering 4. Ua 5x105 ml-1 every 2nd day during flowering 5. fungicides

1,8,15,20 2,6,13,17 3,7,12,18 4,10,11,16 5,9,14,19

08.06.2000

81

1999/ 2000-a 2000-b 2000-c 2000-d

1. Control 2. Ua December 1999 (a) 3. Ua January 2000 (b) 4. Ua March 2000 (c) 5. Ua May 2000 (d)

3,10,14,16 2,6,12,18 5,9,13,20 1,7,15,19 4,8,11,17

30.05.2000

♦ Soil samples plus vegetation cover, 5 sub-samples per plot, collected in trays (40 x 30 x 8 cm) - left in field - on 16 February 2000, except for trial 8 (1999-2000); 1) Date of collecting material for laboratory work from the trays with soil put aside since 16 February. Samples of the 1999-2000 experiment were taken directly from the field. Legend: Performed laboratory tests

1. Choice of culture media for growth of Ulocladium atrum. Isolate 385 of U. atrum has been tested on different culture media: ARSA

(Alternaria radicina selective agar), WA (water agar), MA (malt agar) and PDA (potato-dextrose-agar).

ARSA medium was prepared in two parts: part A consisted of 16.0 g agar‚ 1.0 g KH2PO4, 1.0 g KNO3, 0.5 g KCl, 0.5 g MgSO4, and 500 ml H20; and part B consisted of 5.0 g sodium polypectate (Sigma P-1879) and 500 ml of H20. Parts A and B were autoclaved separately, cooled to 50°C, and combined. Subsequently 50 mg chlortetracycline HCl (Sigma C-4881), 50 mg streptomycin sulphate (Sigma S-6501), 4 mg dicloran (5 mg Botran 75PW), 100 mg triadimefon (200 mg Bayleton 50WP), 106 mg thiabendazole (0.25 g Mertect 340-F), and 10 mg 2,4-D (Sigma D-8407). The herbicide 2,4-D was added from a stock solution that consisted of 200 mg 2,4-D dissolved in 5 ml of hot ethanol and added slowly to 100 ml H20 [16].

WA (water-agar) medium was prepared from 15 g agar for 1 l of distilled water (Tuite, 1969, p. 75), 50 mg streptomycin sulphate (Sigma S-6501) was added to control bacteria.

MA (malt extract) medium was prepared from 25 g malt extract and 20 g agar for 1 l distilled water [23], 50 mg streptomycin sulphate (Sigma S-6501) was added to control bacteria. PDA (potato-dextrose-agar) was prepared from 39 g Oxoid PDA powder suspended in 1 l distilled water. Observations on U. atrum 385 have been performed by daily measuring the colony diameter 3 to 17 days after inoculation (table 2), till the whole surface of the culture medium has been covered by the fungal colony.

82

Soil organic material for plating has been obtained using the following method:

COLLECTING SOIL SAMPLES FROM THE FIELD (obtaining a composite sample consisting of 7 cores of soil, 15 mm φ,

5 cm deep, collected from different points of each plot/tray)

WEIGHING SOIL SAMPLES (70 - 100 g each)

WASHING SOIL SAMPLES (with running with tap water on two nematological sieves with mesh

0.5 and 1.0 mm diameter)

COLLECTING SOIL ORGANIC FRAGMENTS From the sieves in a tube containing 10 ml sterile water

FIRST WASHING with STERILE WATER (10 ml sterile water per tube)

SECOND WASHING with TWEEN 80 1% (9 ml sterile water + 1 ml Tween 80 per tube)

COLLECTING SOIL ORGANIC FRAGMENTS

PLATING SOIL ORGANIC FRAGMENTS ON CULTURE MEDIA (5 fragments/ plot/ 5 replications)

Incidence of U. atrum on leaf and soil organic fragments (0.5 and 1.0 mm sieves) has been established after plating the biological material in replicates on discs of the three media: MA‚ WA and ARSA (10 discs of 1.0 cm φ/ Petri dish/ replicate) [24], [25].

Observations have been performed after 14 days under the stereomicroscope.

2. For checking/estimating incidence of U. atrum on necrotic leaf and soil organic fragments the following methods were used:

Leaf fragments (about 1 mm diameter) were detached from necrotic leaves collected from the field of trial 8 (5 leaflets/plot). They have been plated directly on Petri plates with culture media, without washing or sterilizing.

83

Soil organic fragments have been plated in the same way (5 pieces / plot/ 5 replicates) (fig. 2).

Fig. 2 - Soil organic fragments plated in Petri plates with culture media

Soil suspensions have been prepared from each soil sample. One ml of a

suspension of 1 g soil in 9 ml sterile water has been plated on each of 3 Petri plates ( 9 cm φ) containing culture media (ARSA) (fig. 3).

Fig. 3 – Laboratory preparing of suspensions from soil samples collected from different strawberry experiments

84

3. Isolation/ reisolation and purification of Ulocladium atrum-like fungi from colonies developing on organic material or soil dilution plates has been attempted for all plots sampled (3-5 per plot). Isolates were randomly transferred to slants or Petri dishes (5 cm diameter) with PDA and incubated at 18oC. If necessary for purification a second or third transfer was done. Selection of obtained isolates has been performed by checking them under the stereomicroscope. Purified isolates are stored in a cold room at 4oC.


1. Growth of Ulocladium atrum (isolate 385 and isolates from soil organic

fragments and necrotic leaf fragments) on different culture media. As presented in table 2 and in fig. 3a and 3b, the development of U. atrum 385 was different on the three tested culture media. Table II. Colony diameter (cm) of Ulocladium atrum 385 colony on different culture media

after 3-17 days of inoculation

Days after inoculation: Culture media 3 4 5 6 7 9 12 13 14 17 MA 1.500 1.925 2.420 2.675 2.875 3.175 3.200 3.450 3.450 3.950 WA 1.150 1.500 2.800 3.625 5.000 6.900 7.900 8.050 8.275 8.320 ARSA 0.225 0.225 0.350 0.575 0.725 1.000 1.150 1.200 1.360 1.450

Fig. 3a - Diameter (cm) of Ulocladium atrum colony on different culture media

0

2

4

6

8

10

0 5 10 15 20

Incubation time (days)

Col

ony

diam

eter

(cm

)

MAWAARSA

85

The fastest growth of U. atrum mycelium has been noticed on WA medium‚ the maximal value of the colony diameter (8.050-8.320 cm) being recorded after 13-14 days of incubation. Contrary‚ on the same culture medium‚ a poor sporulation has been recorded.

3b1 3b2 3b3

Fig. 3b - Growth of Ulocladium atrum on different culture media

(after 70 days): 3b1. on ARSA medium; 3b2.on Malt Agar; 3b3. on Water agar media

On MA culture medium‚ the U. atrum isolate 385 had a moderate growth‚ as well as a moderate sporulation. Colony diameter recorded after 14 days of incubation was 3.450 cm. The lowest value of U. atrum growth has been recorded on ARSA medium with a colony diameter of 1.360 cm after 14 days of incubation. On this medium U. atrum sporulation was abundant.

Based on these first observations‚ necrotic leaf and soil organic fragments have been plated on the three culture media under test in order to check incidence of U. atrum (tables 3-4).

Table III. Incidence of Ulocladium atrum (%) in organic soil fragments (0.5 and 1.0 mm φ

sieves) from strawberry plots treated with U. atrum 1 month before soil sampling

Incidence of U. atrum % Experiment Culture media 0.5 mm φ 1.0 mm φ

MA 20.83a 17.50b WA 19.17a 5.83c

1999-2000 ( i )

ARSA 33.33a 31.67a MA 10.0a 6.67a WA 6.67a 1.33b

1999-2000 ( ii )

ARSA 8.00a 8.00a

The data on the incidence of U. atrum on soil organic fragments collected from the 0.5 and 1.0 mm diameter sieves an effect of culture media.

86

For the soil organic particles from the 1.0 mm sieve‚ the highest incidence of U. atrum was noticed on ARSA medium (31.67% in experiment i and 8.00% in experiment ii). The incidence on MA culture medium was second (17.50% in experiment i and 6.67% in experiment ii). The lowest incidence of U. atrum was on WA medium: 5.83% in experiment i and 1.33% in experiment ii.

No significant differences were found with particles from the 0.5 mm sieve. On necrotic leaf fragments plated on the three culture media no difference was found

(table 4).

Table IV. Incidence of Ulocladium atrum (%) in leaf fragments from strawberry plots treated with U. atrum 1 month before soil sampling

Experiment Culture media Incidence of U. atrum (%)

MA 38.23a WA 29.32a

1999-2000 ( i )

ARSA 38.23a

U. atrum grew and sporulated well on ARSA medium. On the other media - MA and WA - saprophytic fungi and bacteria quickly developed. This did not permit the estimation of U. atrum incidence on necrotic leaf pieces, soil organic fragments and in soil suspensions.

2. Incidence of U. atrum on necrotic leaf and soil samples (soil organic

fragments‚ soil suspensions) collected from U. atrum treated strawberry fields and plated on ARSA medium.

Incidence of U. atrum on organic soil fragments from strawberry plots treated with U. atrum 1-37 months before soil sampling was about twice as in the controls (table 5, fig. 4). The lowest incidence of U. atrum (30-45%) was recorded in the U. atrum treated plots from the experiment 1999-2000 and 1997-1. Higher incidence of U. atrum (67-91%) was recorded in the other experiments performed from 1997 until 1999.

Table V. Incidence of Ulocladium atrum (%) in organic soil fragments from strawberry

plots treated with U. atrum 1 – 37 months before soil sampling

Incidence of U. atrum (%) Experiment Time span between last U. atrum treatments

and sampling (months) untreated Ua treated

1997-1 35 21a1) 39a 1997-2 37 14a 44b 1997-1998 - a 22 38a 72b b 24 38a 87c 1998-1 23 74a 85b

87

1998-2 24 62a 91b 1998-1999 13 55a 86b 1999 11 41a 67b 1999-2000 - a 6 19a 36a b 4 19a 34a c 2 19a 45a d 1 19a 30a 1) Figures for U. atrum-treatment in the same time with same latter are not different from the control

Fig. 4 – Incidence of Ulocladium atrum (%) in organic soil fragments

Incidence of U. atrum in soil suspensions from strawberry plots (cfu/ml) treated with U. atrum 1-37 months before soil sampling was higher in the treated plots in comparison with the controls (table 6, fig. 5). The incidence of U. atrum was lowest in the 1999-2000 trial (227-326 cfu/ml soil suspension); the maximum was 1033 cfu/ml soil suspension in the 1998-1 trial. The difference between U. atrum treatment and control was not significant at low levels of incidence.

88

Table VI. Incidence of Ulocladium atrum (cfu/ml) in soil suspensions from strawberry plots treated with U. atrum 1 – 37 months before soil sampling

Incidence of U. atrum (cfu/ml soil

suspension) Experiment Time span between last

U. atrum treatments and sampling (months) untreated Ua treated

1997-1 35 316.7a1) 418.7a 1997-2 37 346.7a 834.7b 1997-1998 - a 22 669.5b 765.2b b 24 669.5 934.6c 1998-1 23 742.5a 1032.5b 1998-2 24 316.7a 725.0b 1998-1999 13 341.7a 570.0a 1999 11 548.3a 810.0b 1999-2000 - a 6 168.3a 226.7a b 4 168.3a 262.5a c 2 168.3a 327.5a d 1 168.3a 276.5a 1) See table 5

Fig. 5 - Incidence of Ulocladium atrum (cfu/ml) in soil suspensions

89

Incidence of U. atrum on necrotic leaf fragments from strawberry plots treated with U. atrum 1-6 months before sampling was twice to 3 times as high in the treated plots in comparison with the controls (table 7, fig. 6). Incidence tended to decrease with increasing time span between the last U. atrum treatments and sampling (1-6 months).

Table VII. Incidence of Ulocladium atrum (%) on necrotic leaves from strawberry plots treated with U. atrum 1 - 6 months before soil sampling

Incidence of U. atrum (cfu/ml soil

suspension) Experiment Time span between last

U. atrum treatments and sampling (months) untreated Ua treated

1999-2000 - a 6 27 a1) 50ab b 4 27a 62bc c 2 27a 72bc d 1 27a 85c

1) See table 5

Fig. 6 - Incidence of Ulocladium atrum (%) on necrotic leaves

Incidence tended to decrease with increasing time span between the last U. atrum

treatments and sampling (1-6 months).

90

Conclusions

1. From the three culture media (MA, WA, ARSA) tested for use to detecting Ulocladium atrum in necrotic leaf and soil organic fragments as well as in soil suspensions from U. atrum treated strawberry plots, ARSA medium (Pryor et al., 1994) proved to be the proper one for development of the fungus. MA and WA media were covered by other saprophytic fungi and bacteria, that did not permit the estimation of U. atrum incidence on the biological material collected from the field.

2. Incidence of U. atrum on all material (soil organic fragments, soil suspensions, necrotic leaf pieces) and in all experiments (1997-2000) was higher in the treated plots in comparison with untreated ones. U. atrum incidence was lower for semples from the oldest and the most recent field experiment than for the experiment in the intermediate period (1997-1998 to 1999).

3. A number of 168 isolates of U. atrum was purified from the biological material analyzed (leaf necrotic and soil organic fragments, soil suspensions) plated on ARSA medium.

REFERENCES

1. BOFF P., 2001 - Epidemiology and biological control of grey mould in annual strawberry crops. PhD

Thesis, WAU 2. BOFF P., KÖHL J., JANSEN M., HORSTEN P.J.F.M., LOMBAERS-VAN DER PLAAS KARIN,

GERLAGH M., 2002 - Biological control of grey mould with Ulocladium atrum in annual strawberry crops, Plant Disease 86, 3: 220-224

3. DAVID J.C. 1995. Ulocladium atrum, IMI Descriptions of Fungi and Bacteria, No. 1224 4. ELMER P. A. G.‚ KÖHL J., 1998 - The survival and saprophytic competitive ability of the Botrytis spp.

antagonist Ulocladium atrum in lily canopies. Eur. J. Plant Pathology, 104: 435-447 5. FRUIT LAETITIA, NICOT PH., 2007 - Use of Ulocladium atrum for biological control of Botrytis cinerea

stem infections in greenhouse tomatoes, INRA, http://www.ubourgogne.fr/UVV/P56.pdf. 6. GERLAGH T., AMSING J.J., MOELHOEK WILMA M.L., BOSKER-VAN ZESSEN A.I., LOMBAERS-

VAN DER PLAAS KARIN, KÖHL J., 2001 - The effect of treatment with Ulocladium atrum on Botrytis cinerea attack of geranium (Pelargonium zonale) stock plants and cutting. Eur. J. Plant Pathology, 107: 377-386

7. KESSEL G.J.T., KÖHL J., POWELL J.A., RABBINGE R., VAN DER WERF W., 2005 - Modeling spatialk characteristics in the biological control of fungi at the leaf scale: competiting substrate coloniyation by Botrytis cinerea and the saprophytic antagonists Ulocladium atrum. Phytopathology, 95: 439-448

8. KÖHL J., 1997-2000 - Contract FAIR-CT96-1898, BIOSPORSUPPRESS – The biological control of air-borne necrotrophic plant pathogens by suppression of spore production

9. KÖHL J.‚ MOELHOEK WILMA M.L., VAN DER PLAAS KARIN, FOKKEMA N. J., 1995 - Effect of Ulocladium atrum and other antagonist on sporulation of Botrytis cinerea on dead lily leaves exposed to field conditions. Phytopathology 85, 4: 383-403

10. KÖHL J.‚ FOKKEMA N. J. 1998. Strategies for biological control of necrotrophic fungal foliar pathogens, in BOLAND G. J. & KUYKENDALL L. D., 1998 - Plant-microbe interactions and biological control. Marcel Dekker Inc. New York-Basel-Hong Kong : 49-88

11. KÖHL J.‚ GERLAGH M.‚ DE HAAS B. H.‚ KRIJGER M. C., 1998 - Biological control of Botrytis cinerea in cyclamen with Ulocladium atrum and Gliocladium roseum under commercial growing conditions. Phytopathology, 88: 568-575

12. KÖHL J.‚ GERLAGH T., DE HAAS B.H., KRIEGER M.C., 1998 - Biological control of Botrytis cinerea in cyclamen with Ulocladium atrum and Gliocladium roseum under commercial growing conditions. Phytopathology, 88, 6: 568-575

http://www.ubourgogne.fr/UVV/P56.pdf

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13. KÖHL J.‚ LOMBAERS-VAN DER PLAAS KARIN, MOELHOEK WILMA M.L., KESSEL G.J.T., GOOSEN-VAN DER GEIJN HELEN M., 1999 - Competitive ability of the antagonists Ulocladium atrum and Gliocladium roseum at temperatures favourable for Botrytis spp. Development. BioControl, 44: 329-346

14. KÖHL J.‚ GERLAGH M.‚ GRIT G., 2000 - Biocontrol of Botrytis cinerea by Ulocladium atrum in different production systems of cyclamen. Plant Disease, 84: 569-573

15. KÖHL J.‚ KESSEL G.J.T., 2000 - Epidemiology of Botrytis spp. in different crops determines success of biocontrol by competitive substrate exclusion by Ulocladium atrum, p. 63, in Sixth Workshop of the IOBC/wprs Phytopathogens WG: Biocontrol agents modes of action and their interaction with other means of control, 30 Nov.-3 dec.200, Sevilla, Spania

16. PRYOR B. M., DAVIS R. M., GILBERTSON R. L., 1994 - Detection and eradication of Alternaria radicina on carrot seed, Plant Dis. 78 (5): 452-456

17. ROUDET J, DUBOS BERNADETTE., 2000 - Evaluation of a three year study of Ulocladium atrum (strain 385) as a biological control agent of vine grey rot in the Bordeaux region, 12th International Botrytis Symposium, Reims (France): 58

18. SCHOENE P.‚ KÖHL J., 1999 - Biologische Bekämpfung von Botrytis cinerea mit Ulocladium atrum in Reben und Cyclamen. Gesunde Pflanzen, 51, 3: 81-85

19. SCHOENE P., 2002 - Ulocladium atrum as an antagonist of grey mold (Botrytis cinerea) in grapevine. PhD Thesis Rheinische Friederich Wilhelms University, Germany

20. ŞESAN T. E., 2003 - Sustainable management of gray mold (Botrytis cinerea) on grapevine, strawberry and ornamentals, in H.-C. Huang & Acharya Suryia, Advances in plant diseases management. Research Signpost, Kerala, India: 121-152

21. ŞESAN T. E., 2005 - Bibliografia românească în domeniul combaterii biologice a micozelor plantelor. Sănătatea plantelor, ediţie specială: 15-22

22. ŞESAN T. E., 2006 - Integrated control of strawberry diseases, Phytopathologia Polonica, 39: 133-148 23. TUITE J., 1969 - Plant pathological methods. Fungi and bacteria, Burgess Publishing Company,

Minneapolis 55415: 44, 52, 75 24. WHIPPS J. M., 1987 - Behaviour of fungi antagonistic to Sclerotinia sclerotiorum on plant tissue segments.

Journal of General Microbiology, 133: 1495-1501 25. WHIPPS J. M., BUDGE S. P., 1990 - Screening for sclerotial mycoparasites of Sclerotinia sclerotiorum.

Mycological Research, 94, 5: 607-612

Acknoledgments

Acknoledgments for the Biocontrol Group in Plant Research International Wageningen, The Netherlands, leaded by PhD Jurgen Köhl, for the opportunity to work together for six monthes in 2000 for the project 1898, BIOSPORSUPPRESS.

Deep thanks to a special friend PhD Thijs Gerlagh from the same working group for his kindness, perenial help and cooperation in our research activity in the field of Botrytis and Sclerotinia biocontrol.

92


A NEW SITE IN ROMANIA FOR SPIRULINA (ARTHROSPIRA) PLATENSIS

M. COSTICĂ*, NAELA COSTICĂ*

Abstract: Spirulina (Arthrospira) platensis Geitl was found in the perimeter of the Tomesti village (district Iasi) for the first time in Moldavia (Romania). Its coenotic ambience and distribution in Romania are given. Key words: Spirulina (Arthrospira ) platensis, a new site in Romania

Introduction

In 1827, P.J.Turpin isolated Spirulina from a fresh water sample. In 1844, Wittrock and Nordstedt reported the presence of a green - blue microalgae named Spirulina jenneri f. platensis in the sample collected near the city of Motevideo. In 1852, Stizenberger gave the name Arthrospira for the septal form and multicellular structure, and Spirulina for without septal, but helical form. Geitler, in 1932 reunified the two genera under the designation Spirulina, considering only morphological similarity. [2]. The microalgae exploited as food belongs to the genus Arthrospira, but it has been called Spirulina for some time.

The taxonomy of the genus Arthrospira is quite confused, and at least 12 binomials are currently recognized: A. funiformis, A. fusiformis, A. geitleri, A. gomontiana, A. indica, A. jenneri, A. khannae, A. massartii, A. maxima, A. miniata, A. platensis, and A. tenuis. However, different interpretations were given to their descriptions, and these species are difficult to distinguish [4].


The samples were collected from small pools in the perimeter of the Tomesti village. and analysed at optical microscope. The morphological characterisation has been defined through measurement of microscopic details, such as: trichome wide, helix wide, coils of helix apart. The results were compared with the species description from speciality literature [5]. The identified species has been verified by phycologist, Prof.dr. Şalaru Vasile (University from .Kishinev , The Republic of Moldovia)


The basin of the Bahlui river is situated on North - Eastern of Romania with strong continental influence (9.6 Celsius degree; 518 mm precipitation; there is a large variation of these values in the years with low precipitations). There are many microhabitats, in this

* “Al. I. Cuza” University, Faculty of Biology, Carol I Bd., no.11, 700506, Iaşi, Romania

93

basin, where we could find some interesting species. In the perimeter of the Tomesti village we found a pool where Spirulina (Arthrospira) platensis grows.

Around this pool, there is the vegetal association (Cynodonto – Atriplicetum tataricae Morariu 1943), where we identified the species: Aster tripolium, Spergularia marina, Juncus gerardi. Cenotic ambience The characteristics of the water at the sampling site, at a depth of 20 – 25 cm are: pH= 8.9; temperature = 24 Celsius degree (in August 2004). The Spirulina grows together with species: Chlorella vulgaris, C. minutissima, Oscillatoria brevis, Synecoccocus sp., Chlamydomonas sp., Carteria sp., Sphaerellopsis sp.

Conclusions

Chorology in Romania Spirulina platensis was identified in basin of the Criş River (at Petea, Homorog), of the Argeş River (at Caldarusani, Chirnogi), of the Danube River (at Pardina and the Danube Delta) [1]. Site from Tomeşti is unique in the province of Moldovia.and the third quotation from Europe.

A short history of Spirulina in human consumption The first writing about this alga belonged to Bernal Diaz del Castillo (a member of

Hernan Cortez,s troops). He reported in 1521 that Spirulina was harvested from the Lake Texcocco, dried and consumed by people in a Tenochtitlan market (today Mexico City) [2].

In 1940 P. Dangeard (phycologist) mentioned that Spirulina has been consumed by people near the African Lake Chad (confirmed by Jean Leonard, 1966 cf. [2].).

In 1970, Germany supported studies on human consumption of Spirulina in Peru, Thailand and India. Spirulina is marketed in Germany, Brasil, Chile, Spain, France, Canada, Ireland, Argentina, India etc. [3]. Spirulina is one of the most extensively used microalgae for animal and human nutrition; its main interest is centered in its high protein content, 60-65% on a dry weight basis.

REFERENCES

1. CĂRĂUŞ I., 2002 - The Algae of Romania. St. Cerc., Univ. Bacău, (Biol.), 7: 1-694 2. CIFERRI O., 1983 - Spirulina, the edible microorganism. Microbiol. Rev., 47: 551- 578 3. LUCIA HELENA PELIZER et al ., 2003 - Influence of inoculum age and concentration in Spirulina

platensis cultivation. J. Food Engeneering, 56, 4: 371-375 4. KOMÁREK, J., LUND, J.W.G., 1990 - What is Spirulina platensis in fact ?. Algological Studies, 58: 1-13. 5. PASCHER A., 1925 - Cyanophyceae In: Pascher A.(ed) Suswasser- Flora Deutschlands, Osterreichs und der

Schweiz, Gustav Fischer, Jena.

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CONTRIBUTION TO THE STUDY OF GRASSY VEGETATION IN THE CEAHLĂU MOUNTAIN

T. CHIFU∗, C. MÂNZU*, OANA ZAMFIRESCU*

Abstract: This paper presents five types of grassland phytocoenoses, widely spread in the Ceahlău Mountain and classified by us, into five associations belonging to Cynosurion R. Tx. 1947 Alliance, Arrhenatheretalia R. Tx. 1931 Order, Molinio – Arrhenatheretea R. Tx. 1937 Class: Pastinaco – Arrhenatheretum elatioris Passarge 1964, Anthoxantho – Agrostietum capillaris Sillinger 1933, Festuco rubrae – Agrostietum capillaris Horvat 1951, Agrostio – Festucetum rupicolae Csürös – Kaptalan 1964 and Poo – Trisetetum Knapp ex Oberd. 1957. For these associations, we present the distribution in the studied territory, the ecology, the use, as well as the synthetic table. Key words: Arrhenatheretalia R. Tx. 1931 Order, grassland vegetation, Ceahlău Mountain.

Introduction

The phytocoenoses we analyzed are located at altitude between 530 m and 1270 m,

corresponding to the mixed beech and coniferous trees forests altitudinal horizon [6]. The primary vegetation of this altitudinal horizon consists in phytocoenoses of the Pulmonario rubrae – Fagetum (Soó 1969) Taüber 1987, Symphyto cordati – Fagetum Vida 1963, Leucanthemo waldsteinii – Fagetum (Soó 1964) Taüber 1987, Hieracio transsilvanico – Fagetum (Vida 1963) Taüber 1987, Geranio robertianae – Fagetum (Burduja et al. 1974) Chifu et Ştefan 1994, Galio schultesii – Fagetum (Burduja et al. 1973) Chifu et Ştefan 1994 associations. The main secondary vegetation is represented by the meadows of the Festuco rubrae – Agrostietum capillaris Horvat 1951 association, togheter with Pastinaco – Arrhenatheretum elatioris Passarge 1964, Anthoxantho – Agrostietum capillaris Sillinger 1933, Agrostio – Festucetum rupicolae Csürös – Kaptalan 1964 and Poo – Trisetetum Knapp ex Oberd. 1957 associations [6].


For the study of vegetation in the Ceahlău Mountain, we used the classical method of the phytosociological surveys, elaborated by the Zurich-Montpellier school [2].

To identify each association we carried out an appropriate number of surveys (between 5 and 16) on 25 to100 m2 sample areas.

The investigation lasted from June to August. For the identification of the species with special status in the analysed communities,

we used “The Red list of superior plants in Romania” [13], and whereas the ecological demands of the species was set according to “The Abstract of the spontaneous cormophytes in Romania” [14].

∗, „Al. I. Cuza” University, Faculty of Biology, Carol I Bd., no. 20A, 700506, Iaşi, Romania

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1. The communities of the Association Pastinaco – Arrhenatheretum elatioris Passarge 1964 grow on fresh, fertile soil, plane or less inclined slopes, generally southern orientated, at the base of the Ceahlău Mountain, between 600 m and 700 m of altitude.

The characteristic species, Arrhenatherum elatius, dominates in the communities together with Trisetum flavescens, Dactylis glomerata, Festuca rubra, Agrostis capillaris sau Medicago falcata. Over 70% of the component species are characteristic to the sintaxa of the Class Molinio-Arrhenatheretea, and many of them are constant: Campanula patula, Centaurea phrygia, Trifolium pratense, T. repens, Leucanthemum vulgare, Briza media, Tragopogon pratensis ssp. orientalis, Medicago lupulina, Taraxacum officinale etc. (Table I, column 1).

The plant communities display an obvious stratification: a superior layer, uniform and dense, consisted of Arrhenatherum elatius, Dactylis glomerata, Trisetum flavescens etc.; medium layer, consisted of Festuca rubra, Agrostis capillaris, Filipendula vulgaris, Achillea millefolium etc., and an inferior layer, consisted of Trifolium repens, Prunella vulgaris, Taraxacum officinale, Leontodon hispidus, L. autumnalis etc.

The communities of Arrhenatherum elatius are among the most productive and with good quality meadows, making excellent hayfields. Their floristic composition includes the endemit Primula elatior ssp. leucophylla, and the rarities Leontodon hispidus ssp. hyoseroides and Ranunculus acris ssp. friesianus.

2. The Association Anthoxantho – Agrostietum capillaris Sillinger 1933 is widely spread in hills and mountains, up to the limit of the common spruce woods. In the Ceahlău Mountain, the association occupies plane fields or gentle slopes, with sufficiently humid soil. The communities are rich in species. Many species are characteristic to the Alliance Cynosurion, Order Arrhenatheretalia and Class Molinio – Arrhenatheretea, and some of them are highly constant: Anthoxanthum odoratum, Leucanthemum vulgare, Rhinanthus minor, Leontodon hispidus, Festuca rubra, Plantago lanceolata, Prunella vulgaris etc. (Table 1, column 2). The floristic composition of the communities reflects the habitat conditions, 90% of the species are mesophilous and mesohygrophilous.

These meadows have mixed use: they are grazed in spring and autumn and mowed in summer. For this reason, the floristic composition is heterogeneous and the fields are often overgrazed.

3. The Association Festuco rubrae – Agrostietum capillaris Horvat 1951 is widely spread throughout the Carpathians, in mesophilous locations, mainly at the decidous forest level, descending in the common oak sub-level, and ascending in the common spruce level. In Ceahlău, the association is widely spread at the altitude of 550 – 1000 m, in the fields generally with gentle or medium inclined slopes, with varied orientation, and more rarely on flat fields. The phytocoenoses are mainly formed of Festuca rubra and Agrostis capillaris, which are in different relations of co-dominance. In other phytocoenoses, they associate also with other species with significant dominance, such as Leucanthemum vulgare, Trifolium montanum, T. alpestre, Filipendula vulgaris, Carex montana. To these species, several highly constant ones can be added: Campanula patula, Holcus lanatus, Briza media, Lotus corniculatus, Galium verum, Plantago media etc.

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In the structure of the association we distinguish three layers: the superior one, 40 - 50 cm high, where predominates Festuca rubra and Agrostis capillaries; a median layer, 20 - 30 cm high, formed of several species such as Anthoxanthum odoratum, Cynosurus cristatus, Trifolium pratense, Lotus corniculatus, Leucanthemum vulgare etc., and the inferior layer, up to 10 - 15 cm, which includes Plantago lanceolata, P. media, Trifolium repens, Carlina acaulis, Euphrasia stricta etc.

Two sub-associations represent the association: - typicum Coldea 1991, with a rich, more homogenous, floristic composition (Table I, column 3a); - nardetosum strictae (Csürös et Resm. 1960) Oroian 1998, which occupies fields, less rich in nutritive substances and more acid; the differential species are Nardus stricta, Antennaria dioica, Potentilla aurea and others characteristic to the Class Juncetea trifidi (Table 1, column 3b).

These meadows have a mixed use, however, the overgrazing and degradation let a series of weeds to appear. The evolution towards nardetosum strictae is the effect of grazing. The need to protect these fields is also supported by the presence of some endemic (Silene nutans ssp. dubia, Primula elatior ssp. leucophylla etc.) and rare species (Dactylorhiza maculata, Ranunculus oreophilus, Leontodon hispidus ssp. hyoseroides etc.).

4. Association Agrostio – Festucetum rupicolae Csürös – Kaptalan 1964 (Table I, column 4).

The phytocoenoses established by Festuca rupicola and Agrostis capillaris occurs on humid to dry soils in hill and mountain regions. The dominant of the two species is Festuca rupicola, which is accompanied in some phytocoenoses by Agrostis capillaris, Festuca rubra, Trifolium montanum, Thymus pulegioides, Leucanthemum vulgare etc.

Apart from the mesophyllous species characteristic to the class, a large group of xero-mesophyllous species from Class Festuco – Brometea is included in the floristic composition.

These fields are used for grazing, and therefore they are highly degraded and invaded by weeds.

5. Association Poo – Trisetetum Knapp ex Oberd. 1957 (Table I, column 5) The meadows of Trisetum flavescens are scarcely distributed in the Romanian

Carpathians, in the inferior and medium mountain level, on flat or gently inclined terrain, with moderately humid and poor in nutrients soils.

These meadows are distributed in patches, at altitudes between 600 m and 700 m, in the broad valleys of the Ceahlău Mountain.

Trisetum flavescens dominate the communities. In some phytocoenoses, this species is associated with Trifolium pratense, Festuca rubra, Taraxacum officinale etc. In addition, we must remark the presence of some constant species, such as Trifolium repens, Campanula patula, Centaurea phrygia, Carum carvi, Leucanthemum vulgare, Achillea millefolium, Medicago lupulina, Dactylis glomerata, Cerastium holosteoides, Ranunculus acris, Prunella vulgaris, Tragopogon pratensis ssp. orientalis etc.

The floristic composition is clearly dominated by the mesophilous species (approximately 85 %), in relation to the habitats of the communities belonging to this association.

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From the economic point of view, these meadows are valuable because they are used as hayfields.

Table I. Associations from the Arrhenatheretalia R. Tx. 1931 Order Association 1 2 3a 3b 4 5 Altitude (m. o. s.) 620 - 690 630 - 1230 600 - 1004 970 – 1270 530 – 780 570 – 820 Exposure S, SE NV, S NV, NE, S,

SE, V, E SV, S, NV, V

SE, S -

Slope (degrees) 0 - 5 0 - 20 0 – 15 0 – 10 0 - 25 - Vegetation coverage (%) 90 - 100 90 - 100 75 - 100 95 – 100 75 - 100 90 - 100 No. of surveys 5 8 16 8 6 10 Association’s characteristic species Pastinaca sativa I - - - - Anthoxanthum odotarum - IV IV IV III II Festuca rubra III IV V V V III Agrostis capillaris II V V V V II Poa pratensis III I - - II III Subassociation’s differential species Antennaria dioica - - I III - - Nardus stricta - - - V - - Potentilla aurea - - - III - - Arrhenatherion Arrhenatherum elatius V - - - - II Campanula patula V III IV II - IV Centaurea phrygia IV III IV II - IV Daucus carota I - I - - - Equisetum arvense - - I - - I Geranium pratense - - - - - II Pimpinella major - - I - I V Taraxacum officinale IV I - - - - Cynosurion Bellis perennis - - - - III I Cynosurus cristatus III III III V III II Leontodon autumnalis II III II II III III Lolium perenne - - I - - - Phleum pratense I II II I II - Plantago major - I - - - - Trifolium repens IV IV III V III IV Phyteumo – Trisetion Aegopodium podagraria - - I - - - Gladiolus imbricatus II I II - - II Hypericum maculatum I III III - - III Trisetum flavescens IV I I - - V Trollius europaeus - I III - - - Veratrum album I - - - - I Arrhenatheretalia Achillea millefolium III II V II IV V Ajuga reptans - - I - - - Avenula pubescens - - I - - - Briza media IV II V - IV II Bromus hordeaceus - - I - - -

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Campanula glomerata II III III III II II Carum carvi III III II III II IV Crepis biennis I I III - - II Dactylis glomerata IV II II - I II Heracleum sphondylium I - I - - III Holcus lanatus - I IV I - - Knautia arvensis I I III - - I Leontodon hispidus ssp. hispidus

- IV III - III I

Leucanthemum vulgare V V V II V V Luzula campestris - - II V - - Medicago lupulina IV I I - III IV Rhinanthus minor III V III - III III Stellaria graminea - - - I - - Thymus pulegioides III III V V V III Tragopogon pratensis ssp. orientalis

IV I III - III IV

Trifolium campestre - - I - - - Molinietalia (incl. Alopecurion, Filipendulion, Deschampsion, Molinion, Calthion) Agrostis canina - - I - - - Agrostis stolonifera - - I - - - Carex ovalis - - I I - - Carex pallescens - III III III I - Carex tomentosa - - I - - - Chaerophyllum hirsutum - I - - - - Colchicum autumnale III I III - - II Dactylorhiza maculata - - I - - - Deschampsia caespitosa I I I IV - - Dianthus superbus - - I - - - Festuca pratensis - III III - II II Filipendula ulmaria - - I - - - Gymnadenia conopsea I I III - - - Hypericum tetrapterum - - I - - - Juncus articulatus - - - I - - Laserpitium prutenicum - - I - - - Linum catharticum - III V - I I Lychnis flos-cuculi - - I - - - Lysimachia vulgaris - - I - - - Lythrum salicaria - - - - - I Myosotis scorpioides - I - I - - Molinia caerulea - - II - - - Platanthera bifolia - - I - - - Polygonum bistorta I - - - - - Serratula tinctoria - - I - - - Stachys officinalis - I V - II I Succisa pratensis - - II III - - Symphytum officinale - - - - - I Valeriana officinalis - I I - I - Potentillion anserinae et Potentillo – Polygonetalia Bromus commutatus II - - I I I Carex distans - - - - - I Elymus repens - I - - - -

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Inula britanica - - I - I - Ranunculus repens - - - - - I Molinio – Arrhenatheretea Alchemilla vulgaris agg. II II III V - II Centaurea jacea - - I - - - Cerastium holosteoides II I II II II IV Euphrasia officinalis ssp. pratensis

I I I - - I

Euphrasia stricta - III I IV - - Lathyrus pratensis - I III - - - Leontodon hispidus ssp. hyoseroides

I IV II I I -

Lotus corniculatus III III V V V III Plantago lanceolata II IV IV IV III II Polygala vulgaris II II III - III II Poa trivialis - - I - - I Primula veris - II III - II I Prunella vulgaris III III IV III III IV Ranunculus acris II III II IV - IV Rhinanthus angustifolius I - I - - - Rumex acetosa I II II - - III Stellaria graminea - III III III - - Trifolium alpestre - I III - III - Trifolium montanum III III V - IV I Trifolium pratense V V V V III V Vicia cracca II - II - II II Festuco – Brometea s. l. Ajuga genevensis - - I - I - Anthemis tinctoria - - I - - - Anthericum ramosum - - I - I - Anthyllis vulneraria - III III - III I Arenaria serpyllifolia - - I - II - Asperula cynanchica - - I - II - Astragalus cicer - - I - I - Astragalus onobrychis - - - - III - Bupleurum falcatum - - I - I - Carex montana - I III - III - Carlina vulgaris - - I - I - Centaurea stoebe - - - - IV - Cirsium pannonicum - - I - II - Dianthus carthusianorum - - III - - I Echium vulgare - - II - IV I Elymus hispidus - - - - I - Eryngium campestre - - - - I - Euphorbia cyparissias - - I - I - Festuca rupicola II - I - V I Filipendula vulgaris I I V - IV I Galium verum II III IV - II - Gentiana cruciata - I I - - - Helianthemum nummularium ssp. obscurum

- I III - III -

100

Hieracium bauhinii - I - - II I Hieracium pilosella - I II II III - Hypochoeris maculata - I III - I - Koeleria macrantha - - I - II - Linum austriacum - I I - II - Medicago falcata III I III - III III Onobrychis viciifolia II - - - - I Pimpinella saxifraga - - III II IV II Plantago media II V V III III III Potentilla argentea - - - - II - Potentilla arenaria - - - - III - Prunella grandiflora - I II - I - Ranunculus bulbosus - - - - II - Salvia nemorosa II - - - I I Salvia pratensis - - I - - - Scabiosa ochroleuca - I II - III - Teucrium chamaedrys - I I - III - Trifolium aureum - I II - I - Trifolium ochroleucon - - I - - - Veronica orchidea - - I - II - Juncetea trifidi s. l. Botrychium lunaria - - I - - - Campanula abietina - - I I - - Campanula serrata - - I - - - Carlina acaulis - III III II II II Genista tinctoria - - I - - - Gentianella austriaca - III II I - - Hieracium aurantiacum - - - I - - Hieracium umbellatum - - I - - - Hypochoeris radicata - I III II II I Ligusticum mutellina - - I - - - Potentilla erecta - I IV III II I Viola canina - - III II - - Festuca supina - - - I - - Variae syntaxa Achillea stricta - I I - - - Acinos arvensis - - - - I - Astragalus glycyphyllos - I - - - - Astrantia major - - III - - - Bunias orientalis II I I - II I Campanula persicifolia - - I - I - Campanula rapunculoides

- I - - I -

Campanula trachelium I - - - - - Carduus acanthoides - - - - I - Carex echinata - - - I - - Carex flava - I I I - - Cerastium arvense - - - - I - Cichorium intybus III I I - - III Cirsium arvense I I I - - - Cirsium decussatum - I - - - - Cirsium erisithales - I II - - I

101

Cirsium oleraceum I - - - - I Cirsium vulgare - - - - I - Clinopodium vulgare - I I - - - Convolvulus arvensis - - - - I - Coronilla varia - I - - II - Cruciata glabra III III I II I II Cruciata laevipes - I - - - - Dianthus deltoides - - I - - - Dianthus tenuifolius - - - - III - Erigeron acris - - I - IV - Festuca heterophylla - II - - II I Fragaria vesca - - I - III - Galium schultesii - - I - - - Gentiana asclepiadea - I I - - - Gentiana verna - - I - - - Gnaphalium sylvaticum - - - I - - Inula salicina - - I - I - Laserpitium latifolium - I III - I - Listera ovata - I - - - I Luzula luzuloides - II - - - - Luzula pilosa - - I - - - Matricaria recutita I - - - - - Melampyrum cristatum - - I - - - Myosotis alpestris - - I - - I Nepeta cataria - - - - I - Origanum vulgare - I I - - - Polygonatum verticillatum

- - II - - -

Primula elatior ssp. leucophylla

I - II I I -

Pteridium aquilinum - I III - II - Ranunculus acris ssp. friesianus

I - - - - -

Ranunculus oreophilus - - - I - - Ranunculus polyanthemos ssp. polyanthemoides

I I II - II II

Reseda lutea - - - - I - Rumex acetosella - - II - I - Sagina saginoides - - - I - - Salvia verticillata II - II - III I Senecio jacobea - - I - I - Senecio umbrosus - - - - - I Silene alba I - - - - I Silene nutans ssp. dubia - I I - III I Tanacetum corymbosum - - I - II - Telekia speciosa - I - - - - Thalictrum aquilegiifolium

- - II - - -

Thalictrum minus - - II - - - Trifolium medium - I II - - - Trifolium pannonicum - II V - - -

102

Veratrum album ssp. lobelianum

- - - III - -

Verbascum lychnytis - I - - - - Veronica chamaedrys III III I - - - Vicia sepium - - - - - I Vicia sylvatica - I I - - - 1. Pastinaco – Arrhenatheretum elatioris Passarge 1964 2. Anthoxantho – Agrostietum capillaris Sillinger 1933 3. Festuco rubrae – Agrostietum capillaris Horvat 1951

a. typicum b. nardetosum strictae (Csűrös et Resm. 1960) Oroian 1998

4. Agrostio – Festucetum rupicolae Csűrös – Kaptalan 1964 5. Poo – Trisetetum Knapp ex Oberd. 1957

REFERENCES

1. BELDIE AL., 1968 - Asociaţiile vegetale din Carpaţii României. Com. de Bot., Soc. Şt. Biol., Bucuresti, 6: 133-238

2. BORZA AL., BOŞCAIU N., 1965 - Introducere în studiul covorului vegetal, Edit. Acad. R. S. R., Bucureşti 3. BURDUJA C.,1968 - Muntele Ceahlău – flora şi vegetaţia, Ocrot. Nat., Bucureşti, 6: 63 – 92 4. BURDUJA C., DOBRESCU C., GRÂNEANU A., RĂVĂRUŢ M., CĂZĂCEANU I., BÂRCĂ C., RACLARU

P., TURENSCHI E., 1956 - Contribuţii la cunoaşterea pajiştilor naturale din Moldova sub raport geobotanic şi agroproductiv. St. cerc. şt. Acad. R. P. R. Fil. Iaşi, Biol. şt. Agric.7, 1: 1 – 37

5. CHIFU T., 1995 - Contribuţii la sintaxonomia vegetaţiei pajiştilor din clasele Molinio-Arrhenatheretea Tx. 1937 şi Agrostietea stoloniferae Oberd. in Oberd. et al. 1967 de pe teritoriul Moldovei. Bul Grăd. Bot. Iaşi, 5: 125 – 132

6. CHIFU T., MÂNZU C., ZAMFIRESCU O., 2006 - Flora şi vegetaţia Moldovei (România), vol. II, Edit. Univ. „Al. I. Cuza” Iaşi: 11 - 21, 211 – 303

7. CHIFU T., MITITELU D., DĂSCĂLESCU D., 1987 - Flora şi vegetaţia judeţului Neamţ. Mem. Secţ. Şt. Acad. Rom. , Seria IV, 10, 1: 281 – 302

8. COLDEA GH., 1991 - Prodrome des associations végétales des Carpates du sud-est (Carpates Roumaines), Docum. Phytosoc., Camerino, 13: 458 – 470

9. COLDEA GH., NEGREAN G., SÂRBU I., SÂRBU A.(coord.), 2001 - Ghid pentru identificarea şi inventarierea pajiştilor seminaturale din România. Edit. alo, Bucureşti!: 17 -58

10. ELLMAUER T., MUCINA L., 1993 - Molinio – Arrhenatheretea, in Mucina L., Grabherr G., Ellmauer Th. 1993. Die Pflanzengesellschaften Österreichs, I, Anthropogen Vegetation, Gustav Fischer Verlag Jena-Stuttgart-New York: 297 – 401

11. GRINTESCU I., 1924 - Considerations géobotaniques sur le Mont Ceahlău (Carpates Orientales), Bul. Soc. Şt. Cluj-Napoca, 2, 2: 104 - 112

12. NYÁRÁDY E., 1924 - Contribuţiuni la cunoaşterea vegetaţiei şi florei muntelui Ceahlău, Bul. Grăd. Bot. Cluj-Napoca, 4: 2 – 3

13. OLTEAN M., NEGREAN G., POPESCU A., ROMAN N., DIHORU G., SANDA V., MIHĂILESCU SIMONA, 1994 - Lista roşie a plantelor superioare din România, Stud., Sint., Docum. de Ecol., Acad. Rom., Bucureşti, I

14. POPESCU A., SANDA V., 1998 - Conspectul florei cormofitelor spontane din România, Acta Bot. Hort. Bucurestiensis

15. POPESCU A., SANDA V., DOLTU M. I. 1983 - Conspectul vegetaţiei ierboase din România. St. şi Com., Muz. Şt. Nat. Brukenthal, Sibiu, 25: 182 – 255

16. RESMERIŢĂ I., 1977 - La classe des Molinio - Arrhenatheretea Tx. 1937, dans les Carpathes Roumaines, Docum. Phytosoc., Lille, vol. I: 241 – 267

17. ZANOSCHI V., 1971 - Flora şi vegetaţia masivului Ceahlău. Teză de doctorat, Univ. Babeş- Bolyai, Cluj-Napoca

103


ASSESMENT OF ECOLOGICAL STATUS OF DANUBE DELTA LAKES USING INDICATOR MACROPHYTES SPECIES

J. HANGANU∗, M. DOROFTEI ∗, N. ŞTEFAN∗∗

Abstract: Implementation of European Water Framework Directive (WFD) legislation requires valuation of chemical and ecological status of surface water bodies. One of the biological indicators prescribes for such assessment is aquatic macrophytes taxonomic composition and abundance. In many member states the trophic status of the lakes is asses by calculating trophic index. This paper show the results of applying Schaumburg reference index for 39 water bodies in Danube Delta . Total P, secchi depth, connectivity and substrate type were the main environmental variables calculated versus index values. As lakes differs little in tot. P content, distribution of aquatic vegetation seems to be mainly determined by connectivity type, substrate and lake morphology. Key words: submerged macrophytes, lakes, classification, trophic index, hydromorphology.

Introduction

Danube Delta refers to the area between 3 main branches of the Danube river (from

north to south: Chilia, Sulina and Sfântu Gheorghe branch). This depresionary area is covered by over 200 km2 of reed beds and 140 km2 open water area. Total numbers of lakes is over 300 and lake size varies from 14 to 4530 ha. Lakes are supplied with fresh river water through vast networks of natural and artificial canals. Water level in lakes is variable and dependent of the river pulse. Higher water level is recorded in spring (May-June). With water depths of 1.5 - 4 meters and a chloride concentration below 60 mg/l, the Danube Delta lakes are characterized as shallow freshwater lakes and included in LCB2 GIG group. In the Danube River the median phosphorus concentration is 0.12 - 0.17 mg P/l. In the lakes the seasonal variation in phosphorus is more pronounced than in the river. In spring the concentration of P-total in the lakes is lower than in the river; in this season also P-ortho is lower in the lakes than in the river. Relative to spring the summer phosphorus concentration increases in all lakes to the level in the river; this increase occurs both in particulate and in dissolved phosphorus. Differences between lakes are small. Based on the classification scheme of Vollenweider [5] all lakes are eutrophic with respect to phosphorus. P-ortho concentrations in spring can be quite low, but are considerably higher in summer [3]. Some lakes in the Danube Delta are dominated by submerged vegetation, high water clarity and a high diversity of benthos and fish in contrast with other lakes dominated by phytoplankton, low water clarity and a low diversity of benthos and fish.

The man-made network of canals in the Delta has intensified the water circulation and input of river water into the lakes. Also nutrient content in river water has increase in

∗Danube Delta National Institute for Research & Development – Tulcea, 165 Babadag Street, 820112, Tulcea, Romania ∗ ∗ “Al. I. Cuza” University, Faculty of Biology, Carol I Bd., no. 20A, 700506, Iasi, Romania

104

the last century few times. As a results lakes reported before 1960 to be dominated by Charecee at present the dominate vegetation is Ceratophyllids and Potamides type.


691 vegetation surveys of the submerged vegetation from 39 water bodies in the

Danube Delta made between 1993 and 2002 were used to test Schaumburg reference index. All vegetation surveys were made in June at fully development of vegetation. Each vegetation surveys had an approximate diameter of 5 m, and were made from canoe. Submerged plants were collected using a rake, and the abundance of each species in the vegetation was visually estimated using a 7-point scale corresponding with the Braun-Blanquet scale. Only true aquatic macrophytes and characeae were included in the analyses. Additionally, measurements of water depth and transparency (secchi-depth: water depth) were made in each sampling point [2]. For each water body type dominant bottom substrate from the soil map was assigned. The substrate was classified in 4 classes according with the % of clay content; 1 = ≤ 5; 2 = 5 - 32, 3= 32-60, 4 = organic Type of connectivity for each lake has been use in the analyses. The connectivity type for each lake was calculated from modeling (SOBEK). Lake was classified in 3 connectivity classes: 1 = lakes with direct connectivity to the river and short residence time; 2= large lakes with long residence time; 3 = remote lakes, surrounded by reed beds.

For calculation of Schaumburg trophic index in Danube delta lakes we use the REBECCA reference list of species, slightly modified (tab. I) The quantity of species was estimated from the original data and transformed in 5 degree scale [2].

Calculation (1) is the same as Reference Index in Schaumburg et al. (2004):

100*Q

QQ)(

1

11

∑

∑∑

=

==

−= ng

igi

nB

iBi

nA

iAi

STI

Where: TI(S) = trophy-index based on quantity (identical to Reference index in

Schaumburg et al. 2004), QA = quantity of species i in group A (see table I), QB = quantity of species i in group B, QC = quantity of species i in all groups, nA = total number of species in group A, nB = total number of species in group B, nC = total number of species in all groups. Quantity = (semi quantitative score)

Results

Almost all TI values are negative (tab. II). The lower values are in lakes that are relatively deep (2 - 4 m) and large (> 200 ha), with sand-silt substrate and an intermediate inflow of river water with direct connectivity. Potamogeton trichoides, P. pectinatus, P. crispus are dominant species in those lakes and usually the cover is 100%.

105

Intermediate TI values are represented by lakes that are of medium size and water depth, loamy to clayey loamy top bottom substrate and are intensively flushed with river water and have a high seasonal dynamics in water level. Ceratophyllum demersum and Trapa natans may cover large part of the lakes.

Higher TI values are related to relatively small and shallow lakes, with peat bottom, surrounded by extensive reedbeds, hydrologically isolated from the river and dominated by Characeae.

Nymphaeids as Nuphar luteum, Nymphaea alba/candida can create large field at the border of large lakes or be dominant in small insulated lakes with peat bottom and surrounded by reed beds.

We could not found for Danube Delta lakes good relationship between P-total and Schaumburg trophic index (fig. 1). It confirmed previously detailed studies [3] on linking trophic statutes of the Danube delta lakes with biological elements as chlorophyll -a, aquatic vegetation and fish species composition in the lakes.

For the TP It was expected to be so as at present, the lakes differing little in nutrient concentrations. Transparency may be influence by other environmental factors and is quite variable during growing season and lakes types.

Fig.1. Trophic Reference Index (Schaumburg) vs. P-total However, hydro morphological parameters as soil substrate and connectivity type we

found to correlate” better” with above biological elements (fig. 2, 3). One explanation is that soil texture is dependent of lake hydrology (connectivity)

and geomorphology as shown in [3]. Lakes with direct connectivity are supplied with fresh sediments during flooding and large lake is former lagoon with sandy substrate. In the insulated lakes water is filtered by suspended solids by reed beds and more clear and accumulation of organic matter from decay of aquatic plant debris or dieback of peaces of floating reed beds is the dominant process. The input of toxic substances (e.g. humic acids

Total P vs. RI

R2 = 0.0011

-100

-50

0

50

100

0 100 200 300

P-tot

RI

106

or H2S) from the surrounding reed beds may give charophytes a competitive advantage over Potamogeton species in these lakes and low redox values in organic soils may favor

development of Nympaeides. Fig.2. Trophic Reference Index (Schaumburg) vs. Soil texture classes

Fig.3. Trophic Reference Index (Schaumburg) vs. Connectivity classes

RI vs. Soil texture classesR2 = 0.1273

-100-80-60-40-20

020406080

100

0 1 2 3 4

RI soil texture

Conectivity vs.RI

R2 = 0.228

-100

-50

0

50

100

0 1 2 3

RI Conectivity

107

Conclusions

Connectivity and residence time seems to play an important roll in distribution of aquatic macrophytes in the Danube delta lakes. In lakes with low residence time of the water the algae may be flush away and have not sufficient time to fully develop and so the transparency is higher giving a chance to some specific submerged aquatic vegetation to grow [3].

The use of Schaumburg TI for the Danube Delta lakes revealed the eutrophication stage of the lakes and the biological quality gradient between lakes as is perceived in the field. In the future a possible reduction in nutrients correlated with hydrological works to reduce the direct inflow of river water in lakes may lead to a switch of tolerant dominant vegetation species to sensitive dominant species as Characeae group.

REFERENCES 1. CARLSON B., 1995 - The Secchi Disk and the Volunteer Monitor. LakeLine, N. Am. Lake. Manage. Soc.,

15, 1: 28-29, 35-37 2. KOHLER A., 1978 - Methoden der Kartierung von Flora und Vegetation von Swasserbiotopen. Landschaft +

Stadt, 10: 23-85 3. OOSTERBERG W. et al., 2000 - Ecological gradients in the Danube Delta; present state and man-induced

changes. RIZA The Netherlands, Danube Delta National Institute Romania and Danube Delta Biosphere Reserve Authority, RIZA raport nr. 2000.015

4. SCHAUMBURG J., SCHRANZ C., HOFMANN G., STELZER D., SCHNEIDER S., SCHMEDTJE U., 2004 - Macrophytes and phytobenthos as indicators of ecological status in German lakes – a contribution to the implementation of the Water Framework Directive, Limnologica, 34: 302–314

5. VOLLENWEIDER R.A., KEREKES J., 1982 - Eutrophication of waters. Monitoring, assessment and control. OECD Cooperative programme on monitoring of inland waters (Eutrophication control). Environment Directorate, OECD, Paris

Acknowledgments

This study was funded by the European Commission under the 6th Framework

Program, Contract No.:SSP1-CT-2003-502158 – REBECCA.

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Table I. List of species groups with regard to eutrophication A Sensitive species B Tolerant species C Indifferent species Chara globularis Hydrocharis morsus-ranae Ceratophyllum demersum Elodea canadensis Nuphar luteum Elodea nuttallii Nitella flexilis Nymphaea alba Lemna gibba Nitellopsis obtusa Nymphaea candida Lemna minor Nitella mucronata Nymphoides peltata Lemna trisulca Potamogeton gramineus Potamogeton lucens Myriophyllum spicatum Potamogeton nodosus Potamogeton perfoliatus Myriophyllum verticillatum Potamogeton mucronatus Potamogeton pusillus Najas marina Tolypella glomerata Potamogeton natans Potamogeton crispus Ranunculus aquatilis Potamogeton berchtoldii Trapa natans Potamogeton compressus Zannichellia palustris Potamogeton trichoides Potamogeton pectinatus Salvinia natans Spirodella polyrrhiza Stratiotes aloides Utricularia vulgaris

Table II. TI values / lake Lake name TI Lake name TI Baclaneşti 1996 -99 Grl. Bratusca 1995 -52 Cuibul cu Lebede 2002 -64 Grl. Sireasa Veche1995 -73 Isac W 1998 -75 L. Corciovata 1994 -54 Plin 1997 -81 L. cu Cotete 1995 -58 Raducu 1997 -73 L. Gasca 1994 -53 Rosu 1998 -100 L. Lung 1995 -35 Rosulet 1998 -98 L. Merheiu Mic 1995 -87 Serbata 1997 -79 L. Mesteru 1995 -24 Uzlina 1998 -68 L. Parches 1994 -56 Iacub -20 L. Puiu 1993 -6 Miazazi -13 L. Rădăcinosu Mare 1995 -32 Nebunu -15 L. Rădăcinosu Mic 1995 -48 Oprio -8 L. Rotund 1994 -61 Raduculets 1997 -11 L. Somova 1994 -16 L. Erenciuc 1993 -14 L. Telincea 1994 -49 L. Merheiu Mare 1995 -39 Lopatna 1996 -93 Fortuna 1996 -98 M. Sacalin 1993 -52 Cuibul cu Lebede 1998 -93 O. Babina 1995 -62 Isac 2002 -82 Sf. Gheorghe 1995 -100 Uzlina 2002 -78 Sf. Gheorghe 1993 -6 Cn. Draghilea 1995 -58 Tataru 1998 -1 Cn. L.Tătaru 1995 -63 Chiril 2002 51 Cn. Mila 35 1995 -58 Ghearsim 2002 -41 Cn. Potcoava 1995 -68 Gorgova 2002 -92 Cn. Sireasa 1995 -92 Gorgova 1998 -76 Cn. Sulimanca 1995 -37 HO02_1996 -92 Grl. Somova 1994 -50 LacBabina 1997 -87 Grl. Şontea Veche 1995 -65 Pojarnia 2002 -35 J. Sulimanca 1995 -75 L. Casla 1994 -60 J. Urechea 1994 -39

109


THE PLANT COMMUNITIES WITH PHRAGMITES AUSTRALIS FROM “THE

HAYFIELDS OF VALEA LUI DAVID” NATURAL RESERVATION (IASI COUNTY)

OANA ZAMFIRESCU∗

Abstract: The association that comprises the arid reed communities was described based on an insufficient number of relevés. The plant communities with Phragmites australis from the arid and saline slopes of the natural reserve are significantly different from the typical reed beds that cover water banks. Such communities are phytosociologicaly classified in the Association Xero-Phragmitetum. These communities arise on the wide tolerance of reed to substrata water conditions. Keywords: reed beds, xerophylous vegetation, steppic meadow

Introduction

The natural reserve “The secular hayfields of Valea lui David”, located at 13 km

from Iaşi, comprises a patchy vegetation because of the rough relief and the large variation of humidity and salinity of the soil on relatively small areas.

The dominant vegetation of the protected area is mostly xerophilous and generally belongs to the Class Festuco-Brometea. Among the six associations of this class, we discuss the Association Xero-phragmitetum Şerbănescu 1955, which has not been studied from this zone. Additionally, the relevés from the areas where it was identified are not sufficiently numerous to justify the coenotaxonomical classification.


We used the phytocoenological relevé method created by the Zürich-Montpellier floristic-phytosociological school, and adapted by us to the local conditions. Therefore, we carried out many field observations, in optimal sampling periods, in order to obtain the relevés. For the vernal aspect, the observations were sampled in April, whereas for the aestival aspect, the relevés were sampled from May to July.

The sample area varied between 10m2 and 25m2, depending on area of the investigated habitat.

Due to the small areas occupied by these plant communities, we managed to draw just five relevés. Nevertheless, these relevés were very consistent with regard to the type of vegetation, and consequently they allowed the phytosociological classification.

∗ “Al. I. Cuza” University, Faculty of Biology, Carol I Bd., no. 20A, 700506, Iaşi, Romania

110


The communities formed by Phragmites australis grow on arid habitats, on flat areas or in small depressions where the water table riches the surface. The areas of these habitats are generally small, 10m2 to 20m2, with western orientation, and 100% vegetation covering.

The analysis of the relevés demonstrated the phytosociological classification through the high number of species fitting in the Class Festucion valesiacae, Order Festucetalia valesiacae, and in the Class Festuco-Brometea.

Table I – Ass. Xero-Phragmitetum Relevé No. 1 2 3 4 5 K Altitude (m) 103 105 123 98 94 Exposition NV V V V V V Slope (degree º) 20 10 15 5 10 Cover (%) 100 100 100 100 100 Area (m²) 10 10 15 10 25 Caract. Sp. Phragmites australis + 1 + 2 1 V Festucion valesiacae Festuca valesiaca 1 + 1 + + V Phlomis pungens + + - + + IV Adonis vernalis + + + + + V Agropyron cristatum 1 + 1 - + IV Linum austriacum + - + + - III Salvia nemorosa + + + - + IV Euphorbia glareosa + - - + + III Festucetalia valesiacae Achillea setacea + + - + - III Centaurea biebersteinii + - - - + II Elymus hispidus 2 + + + + V Inula hirta + - - + + III Erysimum odoratum + - - + + III Bromus inermis - - - + - I Festuco-Brometea Galium verum + - + + + IV Medicago falcata + - - + + III Dianthus carthusianorum + + + - - III Muscari racemosum + + + + - IV Thlaspi perfoliatum + - + + - III

111

Relevé No. 1 2 3 4 5 K Agrimonia eupatoria - + + - - II Thalictrum minus - - + + + III Salvia verticillata + + - - - II Melica ciliata - + - - - I Asperula cynanchica - - + + - II Molinio-Arrhenatheretea Dactylis glomerata + + - + + IV Elymus repens + + - - - II Knautia arvensis - + + - - II Rhinanthus rumelicus + + - + - III Variae syntaxa Melampyrum arvense - + + - - II Convolvulus arvensis + - - - - I Asparagus officinalis + + + - - III Peucedanum latifolium - - + - - IV Nepeta nuda - + + - - II Marrubium vulgare + - - - - I Centaurea orientalis - - + + - II Chenopodium album + - - - - I Conium maculatum - - + - - I Polygonum dumetorum - - - + - I Stachys annua - + - - - I Tribulus terestris + - + - - II

We compared our five relevés with other 4 from the literature, sampled from Frumoasa-Moara (Mititelu and Cojocaru, 1970) and Prut River Valley (Mititelu and Barabaş, 1975), and we noted their strong resemblance, given that it is the same type of vegetation.

On the other hand, our relevés differ very much from another type of community formed by Phragmites australis, i. e. the Association Phragmitetum vulgaris Soó 1927, both in floristic composition and in ecological characteristics. The latter occupies the banks of the lakes, ponds, and slow rivers, and its floristic composition is dominated by species characteristic of the Alliance Phragmition, Order Phragmitetalia, Class Pragmiti-Magnocaricetea, and other subordinated taxa of this class.

112

Conclusions

The plant communities with Phragmites australis from “The hayfields of Valea lui David” natural reserve differ significantly from those of the Association Phragmitetum vulgaris Soó 1927. Consequently, our observations support the classification of these types of communities in the Class Xero-Phragmitetum Şerbănescu 1955.

The characteristic species – Phragmites australis – is highly tolerant to soil moisture, given that it grows, not only in wet habitats but also in arid ones.

REFERENCES 1. BORKMAN J., MORAVEC J. & RAUSCHERT S., 1985 - Code de nomenclature phytosociologique Vegetatio.

Haga, 67, 3: 177-187 2. BURDUJA, C., 1959 - O rezervaţie ştiinţifică care trebuie înfiinţată "Fâneţele din Valea lui David" - Iaşi. Ocrot.

nat., Bucureşti, 4: 154 – 157 3. CHIFU T., MÂNZU C. & ZAMFIRESCU O., 2007 - Flora şi vegetaţia Moldovei. Edit. Univ "Al. I Cuza"

Iasi 4. MITITELU, D. & BARABAŞ N., 1972 - Răspândirea unor asociaţii ierboase în lunca Prutului. St. Com., Muz. Şt.

Nat. Bacău, 5: 189 – 196 5. MITITELU D. & BARABAŞ N., 1975 - Vegetaţia din lunca Prutului. St. Com. Muz. Şt. Nat. Bacău, 8: 219 – 285 6. MITITELU D. & COJOCARU V., 1970 - Flora şi vegetaţia rezervaţiei Frumoasa - Suceava. Ocrot. nat.,

Bucureşti, 14, 1: 35 – 40 7. MITITELU D. & COJOCARU V., 1981 - O nouă contribuţie la flora rezervaţiei botanice de la Frumoasa - Moara

( jud. Suceava ). St. Com. ocrot. nat. Suceava, 5: 394 – 395 8. MITITELU D., MOŢIU T., DĂSCĂLESCU D., TEŞU C. & VIŢALARIU C., 1969 - Flora şi vegetaţia

rezervaţiei "Valea lui David" - Iaşi. St. Com. Muz. Şt. Nat. Bacău, 2: 81 – 100 9. SANDA V., 2002 - Vademecum ceno - structural privind covorul vegetal din România. Ed. Vergiliu, Bucureşti 10. SANDA V., POPESCU & A. DOLTU M. I., 1980 - Cenotaxonomia şi corologia grupărilor vegetale din

România. St. Com. Şt. Nat. Muz. Brukenthal, Sibiu, 24

113

Analele ştiinţifice ale Universităţii “Al. I. Cuza” Iaşi Tomul LIV, fasc. 1, s. II a. Biologie vegetală, 2008 ASSOCIATIONS OF THE MOLINIO –ARRHENATHERETEA R. TX. 1937 CLASS

IN VASLUI RIVER BASIN

IRINA BLAJ - IRIMIA∗

Abstract: The paper presents 4 vegetal associations belonging to the Molinio – Arrhenatheretea R. Tx. 1937 class, associations found on the territory of Vaslui river basin. Each association is accompanied by a phytosociological table and an analysis of the bioforms, floristic elements and ecological indices. Key words: phytosociology, bioforms, floristic elements, ecological indices.

Introduction

The coenotaxonomic classification of the vegetal associations identified is the following:

MOLINIO – ARRHENATHERETEA R. Tx. 1937 Class MOLINIETALIA CAERULEA Koch 1926 Order

CALTHION R. Tx. 1937 Alliance Scirpetum sylvatici Ralski 1931 Association POTENTILLO – POLYGONETALIA R . Tx. 1947 Order

POTENTILLION ANSERINAE R. Tx. 1947 Alliance Ranunculetum repentis Knapp ex Oberd. 1957 Association Junco inflexi – Menthetum longifoliae Lohmeyer 1953 Association

PLANTAGINETALIA MAJORIS R. Tx. et Preising in R. Tx. 1950 Order

LOLIO-PLANTAGINION R. Tx. 1947 Alliance Schlerochloo – Polygonetum avicularis Soó ex Korneck 1969 Association


For the identification of plant associations, we used phytosociological research

methods according to the Central–European school. The establishment of the bioforms and floristic elements was made on the basis of Flora ilustrată a României – Pteridophyta et Spermatophyta, by V. Ciocârlan (2000). The ecological indices were noted having in mind the works of H. Ellenberg [4].


Ass. Scirpetum sylvatici Ralski 1931 (Syn.: Scirpetum sylvatici Schwickerath 1944, Scirpetum sylvatici Maloch 1935)

∗ “Al. I. Cuza” University, Faculty of Biology, Carol I Bd., no. 20A, 700506, Iasi, Romania

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Chorology: Codăeşti, Dobrovăţ Ecology: The coenoses represented by Scirpus sylvaticus are met on alluvial soils,

gleic and pseudogleic, having a large altitude distribution. The association was identified on plane surfaces, on soils with excessive humidity almost all the year. The phytocoenological composition: Together with the characteristic and dominant species, Scirpus sylvaticus, we can find numerous hygrophilic species, Lysimachia nummularia, Lythrum salicaria, Carex vulpina, but also some mesophilic ones, fact denoting the belonging to the classes Phragmiti-Magnocaricetea and Molinio-Arrhenatheretea (Table I). After the analysis of the surveys undertaken, the following was noticed: - from the spectrum of bioforms it is noticed the net predominance of the hemicryptophytes (70.83%), followed by geophytes (16.66%), hydrohelophytes (4.17%), hydrophytes (4.17%) and chamephytes (4.17%);

- from the phytogeographical spectrum we notice the dominance of the Euro–Asian elements (37.5%) and circumpolar ones (37.5%), followed by the cosmopolite ones (16.66%) and European (8.34%);

- from the spectrum of ecological indices we notice that the species bare weakly the shadow (47.82%), are amphitolerant to temperature (52.17%), with area of spreading in central Europe (34.78%), developing on humid–wet soils (usually not aerated) (26.08%), amphitolerant to the reaction of the soil (47.83%) and the quantity of mineral nitrogen in the soil.

Observations: This association is quoted for the first time in Vaslui river basin.

Ass. Ranunculetum repentis Knapp ex Oberd. 1957 (Syn.: Ranunculetum repentis Knapp 1946, Ranunculetum repentis Knapp 1948, Agrostio-Ranunculetum repentis (Knapp ex Oberd. 1957) Oberd. et al. 1967)

Chorology: Tăcuta (Mititelu D.,1975), Vaslui, Văleni (Mititelu D. and collab.,

1996), Soleşti Ecology: The association vegetates on plane fields, with excess of humidity met in

courtyards of houses. It stands well stagnant water for a period, after the withdrawal Ranunculus repens develops fast, covering by means of stolons, important surfaces. During the summer, it stands severe dryness of the soil at the surface. The phytocoenological composition: The floristic composition is not very rich because Ranunculus repens covers almost all the surface. Among the species frequently met in the association we mention: Potentilla reptans, Rumex crispus, Elymus repens etc. (Table II).

After the analysis of the surveys undertaken, the following was noticed: - the spectrum of bioforms indicates the predominance of the hemicryptophytes

(81.82%), followed by geophytes (18.18%); - the phytogeographical spectrum indicates the predominance of the Euro–Asian elements (63.63%), followed by the circumpolar ones (27.28%) and the cosmopolite ones (9.09%);

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- the spectrum of ecological indices indicates the presence of the species standing weakly the shadow (45.45%), developing on humid to wet soils (5-27.27%, 6-27.27%) and with a moderate content of mineral nitrogen (45.46%). They are amphitolerant to temperature (54.54%) and the reaction of the soil (63.64%).

Observations: The association was mentioned without floristic surveys by D. Mititelu (1975, 1996).

Ass. Junco inflexi– Menthetum longifoliae Lohmeyer 1953 (Syn: ass. Mentha longifolia-Juncus inflexus Passarge 1964)

Chorology: Popeşti, Satu Nou (Mititelu D., 1975), Vaslui (Mititelu D. and collab.,

1996), Ciorteşti, Codăeşti, Dobrovăţ, Fereşti Ecology: This association was frequently met in the river watersides, on soils with excess of humidity, where it develops isolated, in thick groupings. The phytocoenological characterization: The species characteristic and representative are Mentha longifolia and Juncus inflexus, together with which it also participate Agrostis stolonifera, Inula britannica, Potentilla reptans, Ranunculus repens etc. species characteristic to the class Molinio-Arrhenatheretea (Table III). The presence of the species in the classes Phragmiti-Magnocaricetea and Bidentetea is explained by the conditions of higher humidity characterizing this association. After the analysis of the surveys undertaken, the following was noticed:

- the spectrum of bioforms illustrates the dominance of the hemicryptophytes (62.49%), followed by geophytes (17.86%), terrophytes (12.50%), hemiterrophytes (5.36%) and fanerophytes (1.79%); - the phytogeographical spectrum indicates a predominance of the Euro-Asian elements (55.36%). Apart from them, there are also circumpolar elements (16.08%), cosmopolite ones (14.28%), continental Euro–Asian (5.35%), Mediterranean (3.56%), European (1.79%), Atlantic–European (1.79%) and pontic–Balkan elements (1.79%); - the spectrum of ecological indices indicates us the fact that the species stand weakly the shadow (45.28%), they are mesothermal (30.20%), with the area of spreading in central Europe (28.30%), mesohygrophilic (15.09%), amphitolerant to the reaction of the soil (54.72%) and develop on soils with a moderate content of mineral nitrogen (24.52%).

Observations: The association was mentioned in the Vaslui river basin by D. Mititelu (1975, 1996), but without presenting a table with floristic surveys.

As. Schlerochloo – Polygonetum avicularis Soó ex Korneck 1969 (Syn.: Polygonetum avicularis Gams 1927, Schlerochloo-Polygonetum avicularis Soó 1945)

Chorology: Bârnova (Mititelu D. and collab., 1995), Dăneşti, Tăcuta, Vaslui, Văleni

(Mititelu D. and collab., 1996), Deleni, Moara Grecilor, Popeşti Ecology: Both species characteristic Schlerochloa dura and Polygonum aviculare

are adapted to hardened fields, growing by the roads, paths, in courtyards, where the field is compact, but also with a content of nitrogen substances of organic nature.

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The phytocoenological composition: The association resists to human, animals or even light vehicles stepping over. The species Schlerochloa dura being an annual and vernal species, it develops in the first part of the season of vegetation, forming the aspect of spring (Sanda V. et al., 2001) (Tabel IV).

After the analysis of the surveys undertaken, the following was noticed: - the spectrum of bioforms indicates the predominance of the terrophytes

(58.62%) and hemicryptophytes (37.92%), followed by geophytes (3.46%); - the phytogeographical spectrum indicates a dominance of the Euro–Asian elements (48.27%) and cosmopolite (31.05%), followed by the Mediterranean Euro–Asian ones (6.88%), Mediterranean (3.45%), adventive (3.45%), continental Euro–Asian (3.45%) and pontic–Mediterranean–central European (3.45%);

- the spectrum of ecological indices indicates the fact that the species stand weakly the shadow (33.33%), they are to temperature (41.66%), develop on dry to moderately humid soils (41.66%), amphitolerant to the reaction of the soil (75%), with a high content of mineral nitrogen (29.16%).

Observations: The association was mentioned in Vaslui river basin by D. Mititelu (1996), but without presenting a table with floristic surveys.

REFERENCES

1. CIOCÂRLAN V., 2000 - Flora ilustrată a României – Pteridophyta et Spermatophyta. Ed. Ceres, Bucureşti 2. CHIFU T., 1995 - Contribuţii la sintaxonomia vegetaţiei pajiştilor din clasele Molinio – Arrhenatheretea

Tx.37 şi Agrostietea stoloniferae Oberd. in Oberd. et al.67 de pe teritoriul Moldovei. Bul. Grăd. Bot. Iaşi, 5: 125-132

3. CHIFU T., MÂNZU C., ZAMFIRESCU O., 2006 - Flora & vegetaţia Moldovei (România), vol. II, Ed. Univ. „Al. I. Cuza” Iaşi: 211-303

4. ELLENBERG H., 1974 - Indicator values of vascular plants in Central Europe. Scripta Geobotanica, vol. IX, Verlag Erich Goltze K.G., Göttingen: 1-97

5. ELLMAUER T., MUCINA L., 1993 - Molinio – Arrhenatheretea In: MUCINA L., GRABHERR G., ELLMAUER T., 1993 – Die pflanzengesellschaften Österreichs, Gustav Fischer Verlag Jena – Stuttgart – New York, vol. I: 297-401

6. MITITELU D., 1975 - Flora şi vegetaţia judeţului Vaslui. St. şi Com. Muz. Şt. Nat. Bacău, Biol. veget.: 67-162

7. MITITELU D., CHIFU T., SCARLAT A., ANIŢEI L., 1995 - Flora şi vegetaţia judeţului Iaşi. Bul. Grăd. Bot. Iaşi, 5: 99-124

8. MITITELU D., HUŢANU M., 1996 - Noi contribuţii la flora şi vegetaţia judeţului Vaslui. St. şi Cerc. Muz. Şt. Nat., Piatra-Neamţ, 8: 193-211

9. MUCINA L., 1997 - Conspectus of classes of European vegetation. Folia Geobot. Phytotax., Praha, 32, 2: 117-172

10. SANDA V., 2002 - Vademecum ceno-structural privind covorul vegetal din România. Bucureşti, Ed. Vergiliu

11. SANDA V., POPESCU A., 1991 - Studiul fitocenozelor clasei Molinio-Arrhenatheretea Tx. 37 din România. Acta Bot. Horti Bucurestiensis: 49-59

12. SANDA V., POPESCU A., BARABAŞ N., 1997 - Cenotaxonomia şi caracterizarea grupărilor vegetale din România. St. şi Comun. Muz. Şt. Nat. Bacău, Biol. veget., 14: 2-365

13. SANDA V., POPESCU A., STANCU D. I., 2001 - Structura cenotică şi caracterizarea ecologică a fitocenozelor din România. Ed. Conphis, Bucureşti

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Table I. Ass. Scirpetum sylvatici Ralski 1931

Number of survey 1 2 3 4 5 Altitude (m.s.m.) 210 121 121 210 210 Cover of the vegetation (%) 100 95 90 100 100 Surface of survey (m²) 10 25 25 10 10 Number of species 13 14 11 10 8 K Association’s characteristics Scirpus sylvaticus 5 5 5 5 5 V

Calthion Poa palustris + - + + + IV Epilobium hirsutum + - - + + III Myosotis scorpioides + - - - + II

Molinietalia Galium palustre + - + + + IV Lysimachia nummularia - + + + + IV Lythrum salicaria + - + + - III Juncus effusus - + - - - I Symphytum officinale - - + - - I

Potentillo-Polygonetalia Ranunculus repens 1 + + 1 + V Juncus inflexus + + - - 1 III Mentha aquatica - + + - - II Mentha longifolia - + + - - II Agrostis stolonifera - + - + - II

Molinio-Arrhenatheretea Ajuga reptans - + - - - I Poa pratensis - - + - - I

Phragmiti-Magnocaricetea Veronica beccabunga + + - + - III Veronica anagallis-aquatica + + - - - II Phragmites australis + - - + - II Alisma plantago-aquatica + - - - - I Carex vulpina - + - - - I

Variae syntaxa Juncus gerardii + - - + - II Equisetum arvense + - - - - I Lemna minor - + - - - I

Place and date of the surveys: 1,4,5. Dobrovăţ, 1.07.2004, 18.07.2004; 2,3. Codăeşti, 1.07.2004, 18.07.2004

Table II. Ass. Ranunculetum repentis Knapp ex. Oberd. 1957

Number of survey 1 2 3 4 5 Altitude (m.s.m.) 120 120 120 120 120 Cover of the vegetation (%) 90 95 100 85 100 Surface of survey (m²) 20 20 20 20 20 Number of species 5 4 6 6 5 K Association’s characteristics Ranunculus repens 5 5 5 4 5 V

Potentillion anserinae Potentilla reptans + 1 + 2 + V

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Elymus repens + - - + + III Rumex crispus - + + - + III Agrostis stolonifera + - - + - II

Potentillo-Polygonetalia Rorippa sylvestris ssp. sylvestris + - - + - II Plantago major - - + + - II Carex vulpina - - + - - I

Molinio-Arrhenatheretea Taraxacum officinale - - + - - I

Phragmiti-Magnocaricetea Phragmites australis - + - - - I Poa palustris - - - - + I

Place and date of the surveys: 1-5. Soleşti, 6.06.2004

Table III. Ass. Junco inflexi– Menthetum longifoliae Lohmeyer 1953

Number of survey 1 2 3 4 5 6 7 8 9 10 Altitude (m.s.m.) 170 210 94 110 110 110 120 110 110 210

Exposition SV V SE N N SV NV V NV loc plan

Slope (º) - - - - - - - - - - Cover of the vegetation (%) 60 100 70 70 85 60 70 100 85

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Surface of survey (m²) 25 20 20 25 25 15 25 20 50 20 Number of species 8 7 8 7 9 7 10 17 11 18 K Association’s characteristics Juncus inflexus 3 5 4 3 3 3 4 4 4 3 V Potentillion anserinae et Potentillo-Polygonetalia Mentha longifolia 2 1 + - 2 1 - 2 1 2 IV Mentha pulegium - 1 - - - + + - + - II Agrostis stolonifera - - - 2 - + + - - - II Ranunculus repens - - - - 2 1 - - - + II Trifolium repens - - - - + + - - - 2 II Inula britannica - - + - - - - - + - I Verbena officinalis - - - - + - - - + - I Myosoton aquaticum - - - - - - + - - - I Potentilla reptans - - - - - - - + - + I Molinietalia Galium palustre - - - - - - - - - + I Trifolium hybridum - - - - - - - - - + I Molinio-Arrhenatheretea Daucus carota + - - - - - - + + - II Lotus corniculatus - - + - - - - + - + II Trifolium pratense - - + - - - - + - - I Cichorium intybus - - - + - - - - - - I Plantago major - - - - - - - 1 - + I Medicago lupulina - - - - - - - + - - I Centaurea jacea - - - - - - - + - - I Ranunculus acris ssp. acris - - - - - - - - -

+

I

Festuca pratensis - - - - - - - - - + I

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Festuco-Brometea Prunella vulgaris - + - - - - - + - - I Galium humifusum - + - - - - - - - - I Achillea setacea - - - + - - - - + - I Salvia nemorosa - - - - - - - + + - I Plantago media - - - - - - - + - + I Artemisietea vulgaris Artemisia absinthium + - - - - - - - - - I Tanacetum vulgare + - - - - - + - - - I Setaria viridis - - - - + - - + - - I Arctium lappa - - - - - - - + - - I Equisetum arvense - - - - - - - - - + I Stellarietea mediae Cannabis sativa ssp. ruderalis - - + - - - - - - -

I

Lathyrus tuberosus - - - + - - - - - - I Chenopodium vulvaria - - - - + - - - - - I Linaria vulgaris - - - - - - + - - - I Sonchus arvensis - - - - - - - + - - I Anagallis arvensis - - - - - - - + - - I Phragmiti-Magnocaricetea Lythrum salicaria + - - - - - 1 + 1 - II Typha angustifolia + - - - - - - - - - I Lycopus europaeus - - 1 - - - - + - - I Equisetum palustre - + - - - - - - - + I Carex riparia - - - + - - - - - - I Typha latifolia - - - - - - + - - - I Phragmites australis - - - - - - + - - - I Carex vulpina - - - - - - - - - + I Bidentetea Bidens tripartita - + + 1 1 + - - - - III Polygonum hydropiper - - - - - - - - 1 - I Rumex conglomeratus - - - - - - - - - + I Rumex crispus - - - - - - - - - + I Variae syntaxa Schoenoplectus tabernaemontani + - - - - - - - -

-

I

Polygonum aviculare - - - - + - - - - - I Salix alba - - - - - - + - - - I Chaemerion angustifolium - - - - - - - - +

- I

Trifolium fragiferum - - - - - - - - - + I Juncus gerardi - - - - - - - - - + I

Place and date of the surveys: 1. Ciorteşti, 27.07.2003; 2, 10. Dobrovăţ, 23.08.2003, 1.07.2004; 3. Vaslui, 12.08.2003; 4-6,8,9. Codăeşti, 24.08.2003, 6.08.2003; 7. between Codăeşti and Dobrovăţ, 24.08.2003

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Table IV. Ass. Schlerochloo-Polygonetum avicularis Soó ex Korneck 1969

Number of survey 1 2 3 4 5 Altitude (m.s.m.) 220 220 270 95 190 Exposition E E - - NE Slope (º) 1-2 1-2 - - 2-3 Cover of the vegetation (%) 95 100 70 80 75 Surface of survey (m²) 50 25 25 50 25 Number of species 14 14 11 16 14 K Association’s characteristics Schlerochloa dura + - - + - II Lolio-Plantaginion et Plantaginetalia Polygonum aviculare 5 5 3 4 4 V Poa annua + 1 2 + + V Lolium perenne + - + + - III Lepidium ruderale + + - + - III Matricaria perforata + - - - + II Hordeum murinum + - + - - II Cynodon dactylon + - - - - I Malva pusilla - + - - - I Cichorium intybus - - - - + I Molinio-Arrhenatheretea Plantago major + + - 1 + IV Trifolium repens + + - - + III Plantago lanceolata + - + - + III Verbena officinalis + - - - + II Stellarietea mediae Capsella bursa-pastoris + - 1 + 1 IV Cardaria draba - - + + 1 III Malva neglecta - - + + + III Convolvulus arvensis - + - + + III Thlaspi arvensis - + - + - II Bromus arvensis - + - + - II Salvia nemorosa - - - + + II Chenopodium album - + - - - I Atriplex tatarica - + - - - I Amaranthus retroflexus - - + - - I Portulaca oleracea - - + - - I Artemisietea vulgaris Xanthium strumarium + + - + - III Taraxacum officinale - + - + + III Artemisia absinthium - - + + - II Artemisia annua - + - - - I

Place and date of the surveys: 1,2. Micleşti, 11.08.2004; 3. Deleni, 10.08.2004; 4. Moara Grecilor, 11.08.2004; 5. Popeşti, 11.08.2004

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CONTRIBUTIONS TO THE STUDY OF PALUDAL VEGETATION FROM THE NEAGRA ŞARULUI RIVER’S BASIN

(SUCEAVA COUNTY)

LOREDANA ASOLTANI∗

Abstract: This paper presents three vegetal paludal associations identified in Neagra Şarului river’s basin: Epilobio – Juncetum effusi Oberd. 1957, Scirpetum sylvatici Ralski 1931 şi Filipendulo – Geranietum palustris W. Koch 1926., described in a phytocoenological table and analysed from the point of view of bioforms, floristic elements and ecological indices.

Key words: paludal vegetation, phytocoenology, Neagra Şarului river basin.

Introduction

Neagra Şarului river basin is situated in the north-east side of central groupe from Oriental Carpathians, in the Dornelor Depression, ascending the higher peaks of the Călimani Mountains in its south side. The Neagra Şarului river springs on the northern slope of Călimani Mountains, at an absolute altitude of 1832 m, and flows into Bistriţa river downstream to Vatra Dornei town [9]. Regarding the paludal vegetation existing in the investigated area, we present in this paper three associations, included in the following phytocoeno-system [1, 4, 6, 7]: MOLINIO – ARRHENATHERETEA R. Tx. 1937: MOLINIETALIA CAERULEAE Koch 1926 CALTHION PALUSTRIS R. Tx. 1937 Scirpetum sylvatici Ralski 1931 Epilobio – Juncetum effusi Oberd. 1957 FILIPENDULION Segal 1966 Filipendulo – Geranietum palustris W. Koch 1926 Ass. Scirpetum sylvatici Ralski 1931 (Syn.: Scirpetum sylvatici Schwickerath 1944, Scirpetum sylvatici Maloch 1935)


The association has a large spreading in Neagra Şarului river basin, populating the rivers valleys on alluvial soils, at 824 – 1250 m altitude, on plane or easy sloping soils. Phytocoenosis of this association have been identified in Plaiul Şarului area, on Sărişoru Mic river valley, in Coverca area on Negru and Deluganu rivers valley, in Păltiniş area, in Gura Haitii area, on Tamău and Haita rivers valley. ∗ „Al. I. Cuza” University, Faculty of Biology, Carol I Bd., no. 20A, 700506, Iasi, Romania

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As an observations, the association was first recorded in 1975, by M. Toma in his doctor’s theses, in Şaru Dornei with two relevées, but we have to underline that this results didn’t find again into a speciality publication. In 1989, it was also mentioned by D. Mititelu, without attaching a table of floristic relevées. The floristic composition of the assocition is relatival richly in species, respective 45 species. Sirpus sylvaticus is the main species in this association, realizing coverings of 90-100%. In the analysed phytocoenosis were identified 5 species characteristic to Calthion alliance (11,11%), 8 species characteristic to Molinietalia caeruleae order (17,78%) and 10 species characteristic to Molinio – Arrhenatheretea class (22,22%). The excess of humidiy favours the presence of some species characteristic to Filipendulion alliance and Phragmiti – Magnocaricetea and Scheuchzerio – Caricetea fuscae classes (tab. I). After the analysis of the relevées, we notice the following: - the bioform spectrum shows the dominance of hemicryptophytes (75,56%) followed by geophyte (11,11%), hydro-helophytes (6,67%) and chamaephytes, phanerophytes and the therophytes in equal proportions (2,22% each); - the phytogeographic spectrum shows the predominance of Eurasian elements (42,22%), followed by circumpolar elements (28,89%), European (8,89%) and central element – European (6,67%), cosmopolitan (11,11%) and alpine ones (2,22%); - within the spectrum of ecological indices, there is a predominance of species which have a low lever of tolerance of shade (51,11%), amphitolerant towards the temperature indice (44,45%), with a spreading area in oceanic climate (46,67%), adapted to excessive humidity (48,89%), amphitolerant to the soil reaction (48,49%) and to the content of mineral nitrogen in soil (20%).

Ass. Epilobio – Juncetum effusi Oberd. 1957 (Syn.: Ranunculus repens – Juncus effusus Paucă 1941)

The phytocoenosis enlightened by juncus effusus have been found on plane surfaces, with a humidity surplus of soil and a low content of nutritive substance, at an altitude of 900-1370 m, in Neagra Şarului, Sărişor, Coverca, Panaci and Şaru dornei area.

The association was mentioned before in Neagra Şarului river basin by M. Toma, in 1975 (Neagra ŞaruluI), presenting just a floristic list, and also by T. Seghedin in 1986 (Coverca) and D. Mititelu in 1989 (Panaci, Păltiniş), without attaching a table of floristic relevées.

the restrictive conditions from stations populated by this phytocoenosis are reflected by the presence of species characteristic Epilobium palustre and Juncus articulatus and dominant species Juncus effusus which is realizing coverings of average 95%, and also by the presence of species Deschampsia caespitosa in most of the analised phytocoenosis, sometimes realizing important covering. alongside of this, there are other species that are part of the Calthion alliance and Molinietalia order, but also of the Phragmiti – Magnocaricetea and Scheuchzerio – Caricetea fuscae classes, characteristics to the biotopes with a hight humidity of soil (tab. II).

After the analysis of the relevées, we notice the following:

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- the bioform spectrum shows the dominance of hemicryptophytes (80%) followed by geophyte (12,72%), chamaephytes (4,25%), therophytes (2,13%) and hydro-helophytes (1,82%); - within the phytogeographic spectrum, one may notice the presence of a large number of Eurasian (40%) and circumpolar (36,37%) elements, followed by cosmopolitan and central European in equal proportions (7,27% each), European (5,45%) and alpine elements (3,64%); - within the spectrum of ecological indices, there is a predominance of species which have a low lever of tolerance of shade (50,91%), amphitolerant towards the temperature indice (52,73%), with a spreading area in oceanic climate (47,27%), adapted to excessive humidity (41,81%), amphitolerant to the soil reaction (49,09%) and in equal proportions, amphitolerant to the content of mineral nitrogen in soil and adapted to a low content of mineral nitrogen in soil (18,18% each).

Ass. Filipendulo – Geranietum palustris W. KOCH 1926

The association has been found on humid fields, with a high level of nitrates owing to a high antropo-zoological influence, at an altitude of 825-1200 m, in Plaiul ŞaruluI, Şaru Dornei, Şaru Bucovinei, Sărişor, Coverca and Tarniţa river valley.

The association was not quoted before in the investigated area; it only was mentioned by D. Mititelu, in 1989, in Dornelor depression, but with no precise location.

The main species in this association, Filipendula ulmaria, realize an average covering degree of 75-100%; in some of the analized phytocoenosis, the other dominant species, Geranium palustre, realize an important covering of 25% average. the hight humidity of soils populated by the investigated phytocoenosis, induces the presence of some species characteristic to the Filipendulion and Calthion alliances and Molinietalia order, also to the Phragmiti – Magnocaricetea AND Scheuchzerio – Caricetea fuscae classes (tab. III).

After the analysis of the relevées, we notice the following: - within the bioform spectrum, one may notice the net dominance of the hemicryptophytes (81,36%), followed by geophyte (8,48%), hydro-helophytes (5,08%), hemitherophytes (3,39%) and therophytes (1,69%); - within the phytogeographic spectrum, one may notice the presence of a large number of Eurasian elements (47,46%), followed by circumpolar (23,73%), cosmopolitan (11,87%), European (10,17%), central European (5,08%) and alpine elements (1,69%); - within the spectrum of ecological indices, there is a predominance of species which have a low lever of tolerance of shade (50,84%), amphitolerant towards the temperature indice (54,24%), with a spreading area in oceanic climate (44,07%), adapted to moderate to hight humidity (20,34%), amphitolerant to the soil reaction (55,93%) and to the content of mineral nitrogen in soil (23,73%).

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REFERENCES

1. CHIFU T., MÂNZU C., ZAMFIRESCU O., 2006 - Flora şi vegetaţia Moldovei (România), vol. I, II, Edit. Univ. “Al. I. Cuza” Iaşi

2. CIOCÂRLAN V., 2000 - Flora ilustrată a României – Pteridophyta et Spermatophyta. Ed. Ceres, Bucureşti

3. MITITELU D., CHIFU T., PASCAL P., 1989 - Flora şi vegetaţia judeţului Suceava, Anuar. Muz. Şt. Nat. Suceava, Şt. Nat., 10: 93-120

4. MUCINA L., 1997 - Conspectus of classes of European vegetation. Folia Geobot. Phytotax (Praha), 32, 2: 117-172

5. POPOVICI D., CHIFU T., MITITELU D., CIUBOTARIU C., LUPAŞCU GH., DAVIDESCU G., PASCAL P., 1996 - Pajiştile din Bucovina, Edit. Helios

6. SANDA V., 2002 - Vademecum ceno-structural privind covorul vegetal din România. Ed. Vergiliu, Bucureşti

7. SANDA V., POPESCU A., BARABAŞ N., 1997 - Cenotaxonomia şi caracterizarea grupărilor vegetale din România. St. şi Comun. Muz. Şt. Nat. Bacău, Biol. veget., 14

8. SEGHEDIN T. G., 1986 - Flora şi vegetaţia munţilor Bistriţei, Rezumatul tezei de doctorat, Inst. Agr. „Ion Ionescu de la Brad” Iaşi

9. STOICA D. L., 2007 - Cercetări de geografie fizică pe versantul nordic al Masivului Călimani. Rezumatul tezei de doctorat, Univ. „Al. I. Cuza” Iaşi

10. TOMA M., 1975 - Cercetări asupra florei şi vegetaţiei din Depresiunea Dornelor (jud. Suceava), Rezumatul tezei de doctorat, Univ. „Babeş - Bolyai” Cluj-Napoca

Table I – SCIRPETUM SYLVATICI RALSKI 1931

Number of relevé 1 2 3 4 5 6 7 8 9 10 11 12

Altitude (m.s.m.) 1209 920 910 920 1013 1016 1046 1100 1250 1069 831 824

Covering (%) 100 100 100 95 95 95 95 95 95 90 90 90

Surface (m2) 50 100 100 100 50 50 25 50 50 50 100 25

No. of species 15 15 17 16 13 19 21 15 9 12 11 21

K

Association’s characteristics Scirpus sylvaticus 5 5 5 5 5 5 4 5 5 4 5 5 V

Calthion palustris Caltha palustris + - - - - - 1 - + + - + III

Chaerophyllum hirsutum + - - - + - + - - + - - II

Cirsium rivulare - - - - - + - - - - - - I

Geum rivale + - - - - + + - + - - + III

Myosotis scorpioides + + + + - + + + + + - + V

Deschampsion Carex ovalis - + + + + + + + - - - - III

Deschampsia caespitosa - + + + + - + + + 1 - + IV

Juncus conglomeratus - - - + - + + - - - - - II

Filipendulion Equisetum arvense - - - - + + + - - - - + II

Filipendula ulmaria + + + - + - - + - - - + III

Lysimachia vulgaris - - - - - - + - - - + + II

Mentha longifolia + - - + + + - + - + - + III

Molinietalia caeruleae Cirsium palustre + + + + - + + + + - - + IV

Equisetum palustre + + + - + - - + - - + - III

Galium palustre + + + + - + + - - + - - III

Juncus effusus + + + + + + + + + + + + V

Lychnis flos-cuculi - + + - - - + - - + - - II

Lythrum salicaria - - + - - - - - - - + I

Symphytum officinale - - - - - - - - - - + - I

125

ssp. officinale Trifolium hybridum ssp. hybridum

+ - - - + - - + - - - II

Molinio – Arrhenatheretea Agrostis stolonifera ssp. stolonifera

- + + - - + - - - - + + III

Alchemilla vulgaris - - + + - + + - - - - - II

Alnus incana juv. - - - - + - - - - - - - I

Holchus lanatus - - - + - - + - - - + + II

Lysimachia nummularia - + + - - + - + - - - + III

Phleum pratense - - - + - - - - - - + - I

Prunella vulgaris - - - + - - + - - - - - I

Stellaria graminea - - - - - - + - - - - - I

Phragmiti - Magnocaricetea s. l. Carex riparia - + + - + - - - - - + - II

Epilobium palustre - - - - - + - - - + - - I

Glyceria notata - - - - - - - - - - - + I

Lycopus europaeus - - - - - - - - - - + + I

Ranunculus repens - + - + - + - + - - - + III

Veronica beccabunga + + - - - - + + - + - - III

Scheuchzerio – Caricetea fuscae s. l. Carex flava + - + + - - - - - + - - II

Carex nigra ssp. nigra - - - + + + - - - - - - II

Variae syntaxa Epilobium montanum - - - - - - + - - - - - I

Festuca pratensis - - - - - - - - - - - + I

Luzula campestris - - - - - + - - - - - - I

Potentilla erecta + - + - - - + - - - + - II

Rorripa sylvestris ssp. sylvestris

- - - - - - - - - - - + I

Rumex crispus - - - - - - - - - - - + I

Stellaria nemorum - - - - - - - + + - - - I

Valeriana tripteris - - - - - + - + + - - - II

Place and date of relevées: 1. Tamău river valley (16.08.2007); 2, 3, 4. Sărişoru Mic river valley (14.07.2007); 5. Gura Haitii (20.08.2006); 6. Coverca – Negru river valley (16.08.2006); 7. Coverca – Deluganu river valley (16.08.2006); 8, 9. Neagra river valley (21.08.2006); 10. Păltiniş (13.07.2007); 11, 12. Plaiul Şarului (8.08.2007).

Table II – EPILOBIO – JUNCETUM EFFUSI OBERD. 1957 Number of relevé 1 2 3 4 5 6 7 8 9 10

Altitude (m.s.m.) 914 920 998 1000 1124 1372 995 993 980 906

Covering (%) 95 95 95 95 95 90 90 90 90 90

Surface (m2) 100 50 50 100 50 50 100 50 30 100

No. of species 21 19 22 16 25 13 15 17 17 18

K

Association’s characteristics Epilobium palustre - + + + + - - + - + III

Juncus articulatus + + - + + + + + - - IV

Calthion palustris Caltha palustris - - + + + + + - - - III

Chaerophyllum hirsutum

- - - - - + - - - - I

Geum rivale - - + - + - - + - - II

Myosotis scorpioides + + + + + - + + + + V

Scirpus sylvaticus + + - + - - + - + - III

126

Deschampsion Carex ovalis + + - - + - - - + + III

Deschampsia caespitosa + + + + + 1 - - 2 1 IV

Juncus conglomerates + - + + + - - + + + IV

Filipendulion Cirsium erysithales - - - - - - - + - - I

Filipendula ulmaria + + - - - - - - + + II

Mentha longifolia + + - - - - - - + - II

Molinietalia caeruleae Cirsium palustre - + + - - + - - - - II

Equisetum palustre - + + - - - + + - - II

Galium palustre - + + + + - + + + + IV

Juncus effusus 5 5 5 5 5 4 5 5 4 4 V

Succisa pratensis + + - - + - - - - - II

Trifolium hybridum ssp. hybridum

- - - - - - - - - + I

Molinio – Arrhenatheretea Agrostis capillaris - - - - - - - - + - I

Agrostis stolonifera ssp. stolonifera

- - + + - - - - - 1 II

Alchemilla vulgaris + - + + + + + + - + IV

Anthoxanthum odoratum

- - + - + 1 - + - - II

Briza media - - - + - - - - + - I

Carex hirta + - + - - - - - - - I

Carex pallescens + + - + + - - - - - II

Cynosurus cristatus - - + - + - - - - - I

Dactylis glomerata - - + - - - - - - - I

Festuca rubra - - - - - - - - + - I

Holchus lanatus - - - - + + - - - - I

Luzula campestris - - - - - + + + - - II

Lysimachia nummularia + + - - - - + + + - III

Phleum pratense - - - + - - - - - + I

Prunella vulgaris + + - - + + + + - + IV

Ranunculus acris - - - - - - + - - + I

Rumex crispus + - - - - - + - - - I

Trifoiul repens ssp. repens

- - - - - - - - - + I

Phragmiti – Magnocaricetea s. l. Alisma plantago aquatica

- + - + + - - - - - II

Carex riparia - - + - - - - - - - I

Lycopus europaeus - - + - - - - - - - I

Ranunculus repens - + - - + - + + + - III

Scheuchzerio – Caricetea fuscae s. l. Carex flava + + + - + - - + - - III

Carex nigra ssp. nigra - - - - + - + - + + II

Eriophorum angustifolium

- - + - - - - + - - I

Variae syntaxa Carduus personatus ssp. personatus

- - - - + - - - - + I

Cruciata glabra + - - - + - - - - - I

Epilobium montanum - - - - - + + - + - II

127

Equisetum sylvaticum - - - - - + - - - - I

Homogyne alpina + - - - - - - - - - I

Hypericum maculatum + - - - - + - - - - I

Plantago media - - + - - - - - - - I

Potentilla erecta + + + + + - - + + + IV

Valeriana tripteris - - - + + - - - - - I

Veratrum album + - - - + - - - - - I

Place and date of relevées: 1. Neagra Şarului (14.08.2006); 2. Sărişoru Mic river valley (14.07.2007); 3. Coverca – Călimănel river valley (16.08.2006); 4. Coverca – Negru river valley (16.08.2006); 5. Păltiniş (13.07.2007); 6. Panaci (18.082006); 7. Coverca – Negru river valley (16.08.2006); 8. Coverca – Buciniş river valley (16.08.2006); 9. Panaci – Rusului peak (18.08.2006); 10. on the outskirts Tinovului Mare Şaru Dornei (15.07.2007).

Table III – Filipendulo – Geranietum palustris W. Koch 1926 Number of relevé 1 2 3 4 5 6 7 8 9 10 11 12

Altitude (m.s.m.) 999 851 825 1037 1035 1016 1210 998 920 830 830 830

Covering (%) 100 100 100 95 95 90 90 90 95 95 95 95

Surface (m2) 25 25 50 50 50 25 25 25 50 100 50 100

No. of species 12 10 18 10 15 18 19 13 19 12 14 12

K

Association’s characteristics Filipendula ulmaria 5 5 5 5 5 5 5 5 5 5 4 4 V

Filipendulion Chaerophyllum hirsutum

- - - + + + + - - - - - II

Equisetum arvense - - - - + - - - - - - - I

Geranium palustre - + + + - + - + + 1 2 1 IV

Lysimachia vulgaris - - + - - - - - + + + - II

Lythrum salicaria - - + - - - - - + + + + III

Mentha longifolia - + - - - - + + + - - - II

Calthion palustris Alchemilla vulgaris - - + + + + + - + - - - III

Briza media - + + - - - - + + - - - II

Caltha palustris - - - - - + + - - - - - I

Cirsium rivulare - - - - - - - + - - - - I

Geum rivale - - - - + + + - - - - - II

Myosotis scorpioides

+ + - + + + + + + + + + V

Scirpus sylvaticus - - - - + + + + - - + 1 III

Deschampsion Carex ovalis - - - - + - - - - - - - I

Deschampsia caespitosa

- + + - - - + + + + + + IV

Juncus conglomeratus

+ - - - + - - - + + - - II

Molinietalia caeruleae Cirsium oleraceum - + - - - + - + - - - - II

Cirsium palustre - - - - - - + - - + + + II

Equisetum palustre - - - - - - - - - - + + I

Galium palustre - - - - - + + - - - - - I

Juncus effusus + - - + + - + + - + + 1 IV

Lychnis flos-cuculi - - - - - + + + + - - - II

Succisa pratensis - + - - - - - - + - - + II

Molinio – Arrhenatheretea Achillea millefolium - - + - - - + - + - - - II

128

Agrostis stolonifera ssp. stolonifera

- - - - + - - + - - - - I

Ajuga reptans - - + - - - - - - - - - I

Angelica sylvestris - - - - - - - - - + + - I

Campanula glomerata

- - + - - - - - + - - - I

Centaurea jacea - - + - - - - - + - - - I

Cynosurus cristatus - - - + - - - - + - - - I

Festuca pratensis - - + - - - - - - - - - I

Luzula campestris + - - - - - - - - - - - I

Phleum pretense - - - - + - - - - + - + II

Prunella vulgaris - - - - + + + - + - - - II

Ranunculus acris - - - - + - - - - - - - I

Rumex acetosa - - + - - + - - - - - - I

Rumex crispus - - - + - - - - - - - - I

Trifolium repens ssp. repens

- + + - - + + - + - - - III

Phragmiti – Magnocaricetea s. l. Epilobium palustre - - - - - - + - - - - + I

Glyceria notata - - - + - - - - - - - - I

Ranunculus repens + + - - - + + + - - - - III

Veronica beccabunga

- - - - - + - - - - - - I

Scheuchzerio – Caricetea fuscae s. l. Carex nigra ssp. nigra

+ - - - - - - - - - - - I

Ligularia sibirica + - - - - - - - - - - - I

Variae syntaxa Agrimonia eupatoria - - + - - - - - - - - - I

Astragalus glycyphyllos

- - - - - - - - - - + - I

Athyrium filix-femina

- - - - - + - - - - - - I

Cirsium arvense - - + - - - - - - - - - I

Epilobium montanum

+ - - - - - - - - - - - I

Equisetum sylvaticum

+ - - - - - - - - - - - I

Galeopsis tetrahit - - - - - - - - - + + - I

Holchus lanatus - - - - - - - - - - + - I

Hypericum perforatum

- - + - - - - - - - - - I

Hypochoeris uniflora

+ - - - - - - - - - - - I

Mentha arvensis - - - + - - - - - - - - I

Plantago media - - + - - - - - + - - - I

Potentilla erecta + - + - + - - - - - - - II

Valeriana tripteris - - - - - + + - - - - - I

Place and date of relevées: 1. Sărişor – Sărişorul Mare river valley (14.08.2006); 2. Şaru Dornei (14.08.2006); 3. Coverca – Negru river valley (16.08.2006); 4. Coverca – Deluganu river valley (16.08.2006); 5. Coverca – Deluganu river valley (16.08.2006); 6. Coverca – Buciniş river valley (16.08.2006); 7. Tamău river valley (16.08.2007); 8. Plaiul Şarului (19.08.2006); 9. Şaru Bucovinei – Sărişoru Mic river valley (14.07.2007); 10, 11, 12. Plaiul Şarului (8.08.2007).

129

Analele ştiinţifice ale Universităţii “Al. I. Cuza” Iaşi Tomul LIV, fasc. 1, s. II a. Biologie vegetală, 2008 PROTECTED TAXA FROM THE BISTRIŢA RIVER BASIN BETWEEN PIATRA

NEAMŢ AND BACĂU

CARMEN AONCIOAIE∗

Abstract: The results presented in this article were obtained during a study made in 2005 and 2006 in the Bistriţa river inferior basin in the region between Piatra Neamţ and Bacău. As a result of this study were catalogued a number of 124 taxa of vascular plants in different categories of protection, using several specialty papers. Key words: Red List, Berne Convention, Bistriţa inferior basin, Bacău, Piatra Neamţ

Introduction

The studied area belongs to the lower course of the Bistriţa river, being situated on two counties from Moldova region – Neamţ and Bacău. Geomorphologicaly speaking, the studied region spreads on the following four natural units (1400 Km2): Oriental Carpathians (the Goşmanu Mountains), the Moldavian sub-Carpathians with two subdivisions (Bistriţa sub-Carpathians and Cracău – Bistriţa depression) and the Moldavian Plateau (a very small region between Racova and Bacău). The climate is temperate – continental with variations, depending on the altitude of the relief and its particularities. The climate has excessive nuances in East and moderated nuances in West, presenting noteworthy variations with the altitude, with cold winters and hot, often dry, summers.

The hydrographical network is represented by Bistriţa and its affluents. The most important affluent in his sector is Cracău, followed by a number of streams like Calu, Iapa, Nechit, Trebiş, Negel and so on. Building hydro-electric power stations on the river leaded to the appearance of some artificial lakes like Bacău, Buhuşi, Gârleni, Lilieci and Racova.


For analyzing the vascular flora of the region were used the classical methods and materials for this kind of research. The working stages begun with documentation and study of the bibliography, followed by a terrain research stage and then a herbarium stage, finally ended with a stage of data interpretation and a complete list of taxa.

Among the identification guides used are: Flora României (vol. I – XIII) 1952 – 1976; Ciocârlan V. – Flora ilustrată a României. Pteridophyta et Spermatophyta., 2000; Sârbu I. et al. – Flora ilustrată a plantelor vasculare din estul României (vol. I and II) 2001.

∗ „Al. I. Cuza” University, Iaşi, Faculty of Biology, Carol I Bd., no. 20A, 700506, Iasi, Romania

130

There were used two papers for including the taxa in different protection categories: Gh. Dihoru, Alexandrina Dihoru – Plante rare, periclitate şi endemice în flora României – Lista Roşie, 1993 – 1994 and M. Oltean, G. Negrean, A. Popescu et al. –Studii, sinteze, documentaţii de ecologie, I/1994.


At the moment, the floristic inventory (of the studied region) has a number of 1.436

taxa among witch there are species included in diverse categories of protection. These are presented below from the point of view of two well known scientific papers (from Romania) and also after the Berne Convention.

The first column of the table presents the species registered in „Studii, sinteze, documente de ecologie – Lista Roşie a plantelor superioare din România” by Oltean M., Negrean G., Popescu A., Roman N., Dihoru G., Sanda V., Mihăilescu S. (1994) and the second column presents the analysis made after “Plante rare, periclitate şi endemice în flora României – Lista Roşie” by Dihoru Gh., Dihoru Alexandrina (1993 – 1994). Both columns of the table gather a number of 124 taxa.

After the paper by Oltean M. et al. a number of 95 taxa are protected and after the paper by Dihoru Gh. only a number of 58.

There is a difference of 37 taxa between the two papers and different visions on many species. Only 29 taxa are common in both papers and even about the protection category there are different opinions. This problem may occur because of the great variety of the Romanian territory and a list of protected species should be made for each region of the country, knowing that stationary factors are different and in certain places certain species may not be endangered (tab. I).

Another analysis was made after the data presented by the “Berne Convention” (1979) to witch Romania adhered in 1993. The analysis showed that a number of 5 strictly protected species are present in the territory:

Cypripedium calceolus L. Eleocharis carniolica Koch. Salvinia natans (L.) All. Typha minima Funk in Hoppe Typha schuttlewortii Koch et Sonder.

Conclusions

This article presents a number of 124 taxa of vascular plants from diverse categories of protection, representing 8,63 % out of the total number of taxa identified in the studied region.

131

REFERENCES

1. BELDIE AL., 1977 - Flora României. Determinator ilustrat al plantelor vasculare. Vol. I, II, Ed. Acad. R.S.R., Bucureşti

2. CIOCÂRLAN V., 2000 - Flora ilustrată a României. Pteridophyta et Spermatophyta. Ed. Ceres, Bucureşti 3. DIHORU GH., DIHORU A., 1993 – 1994 - Plante rare, periclitate şi endemice în flora României – Lista

Roşie, Acta Botanica Horti Bucurestiensis, Bucureşti, 1994: 173-179 4. DIHORU G., PÂRVU C., 1987 – Plante endemice în Flora României. Ed. Ceres, Bucureşti 5. MITITELU DUM., BARABAŞ N., ŞTEFAN N. 1987. Contribuţii la corologia unor plante rare în Moldova

şi Muntenia. An. Şt. Univ. “Al. I. Cuza” Iaşi , s. II, a., Biol. veget., t. XXXIII: 20 – 24 6. OLTEAN M. et al., 1994 - Lista roşie a plantelor superioare din România. Studii., Sinteze, Documentaţii de

Ecologie, Acad. Rom. 7. POPESCU A., SANDA V., 1966 - Consideraţii corologice asupra plantelor endemice din România. St. Cerc.

Biol., ser. Bot., 18, 5;;437 – 466 8. RUGINĂ R., MITITIUC M., 2003 - Plante ocrotite în România. Ed. Univ. „Al. I. Cuza” Iaşi 9. SÂRBU I., ŞTEFAN N., IVĂNESCU LĂCRĂMIOARA, MÂNZU C., 2001 – Flora ilustrată a plantelor

vasculare din estul României. Vol. I, II, Ed. Univ. „Al. I. Cuza” Iaşi 10. ŢOPA E., 1979 - Ocrotirea naturii în Judeţul Neamţ. Reflexii, istoric, realizări şi sugestii. **Conventia de la Berna (1979) *** 1952 – 1976 Flora R. P. R-r. S. R., I – XIII. Ed. Acad. R.S.R., Bucureşti

Table I: Protected taxa

M. Oltean et al. Gh. Dihoru, Alexandrina Dihoru category specie category specie

E Abies alba Miller - - - - R Adonis flammea Jacq. - - V Adonis vernalis L. R Allium schoenoprasum L. ssp.

sibiricum (L.) Hartman - -

- - R Alopecurus arudinaceus Poiret R Anacamptys pyramidalis (L.)

L.C.M. Richards - -

K Bromus racemosus L. K Bromus racemosus L. - - K Bryonia dioica Jacq. R Campanula carpatica Jacq. - - I Carex brevicollis DC. I (R) Carex brevicollis DC. R Carex dioica L. R Carex dioica L. R Carex hallerana Asso - - R Centaurea melanocalathia

Borbás - -

VR Centaurium littorale (D. Turner) R Centaurium littorale (D. Turner)

132

Gilmour ssp. uliginossum (W et K) G. Beck

Gilmour ssp. uliginossum (W et K) G. Beck

V Cephalanthera damassonium (Miller) Druce

- -

V Cephalanthera longifolia (L.) Fritsch

- -

R Cephalanthera rubra (L.) M.L. C. Richards

- -

V Cirsium furiens Griseb. et Schenk

K Cirsium furiens Griseb. et Schenk

R Cirsium grecescui Griseb et Schenk

R Cirsium grecescui Griseb et Schenk

R Coeloglossum viride (L.) Hartman

- -

R Colutea arborescens L. - - VR Corynephorus canescens (L.) P.

Beauv. E Corynephorus canescens (L.) P.

Beauv. V Crocus reticulatus Steven - - R Cyperus serrotinus Rottb. - -

VR Cypripedium calceolus L. V Cypripedium calceolus L. R Dactylorhizza incarnata (L.)

Soó - -

R Dactylorhizza maculata (L.) Soó ssp. maculata

- -

R Dactylorhizza maculata (L.) Soó ssp. schurii (Klinge) Soó

- -

R Dactylorhizza sambucina (L.) Soó

- -

R Dianthus collinus Waldst et Kit ssp. collinus

- -

R Dianthus collinus Waldst et Kit ssp. glabriusculus (Kit) Thaisz

I Dianthus collinus Waldst et Kit ssp. glabriusculus (Kit) Thaisz

VR Dictamnus albus L. - - R Dryopteris cristata (L.) A.Gray R Dryopteris cristata (L.) A.Gray - - V Eleocharis carniolica Koch R Epipactis helleborine (L.)

Crantz - -

R Epipactis palustris (L.) Crantz - - R Epipactis purpurata Sm. - - R Eryssimum witmannii Zawadzki

ssp. transilvanica (Schur) P.W. Ball

- -

133

- - R Evonymus latifolius (L.) Miller K Fragaria moschata Duchesne - -

VR Fritillaria orientalis Adams - - nt Galanthus elwesii Hooker fil. E Galanthus elwesii Hooker fil. nt Galanthus nivalis L. - K Galium sylvaticum L. - - - - R Galium tenuissimum Bieb. - - R Gladiolus imbricatus L. - - R Goodyera repens (L.) R. Br. nt Hepatica transsilvanica Fuss. nt Hepatica transsilvanica Fuss. V Hippuris vulgaris L. - - R Gymnadenia conopsea (L.) R.

Br. - -

R Herniaria hirsuta L. - - - - R Iris aphylla L. - - R Iris sibirica L. - - R Isolepis setacea (L.) R. Br. R Koeleria macrantha (Ledeb)

Schultes ssp. transsilvanica (Schur) A. Nyàr.

nt Koeleria macrantha (Ledeb) Schultes ssp. transsilvanica (Schur) A. Nyàr.

R Lactuca virosa L. R Lactuca virosa L. R Lapulla deflexa (Lehm.) Cesati - -

VR Lepidium cartilagineum (J.Mayer) Thell ssp. crassifolium (W. et K.) Thell

V Lepidium cartilagineum (J.Mayer) Thell ssp. crassifolium (W. et K.) Thell

R Listera ovata (L.) R. Br. - - R Luzula pallescens Swartz. R Luzula pallescens Swartz. - - R Melampyrum nemorosum L. R Melmpyrum saxosum Baumg. - - R Monotropa hypopytis L. - - R Myosotis discolor Pers.

R Myosotis stenophylla Knaf. R Myosotis stenophylla Knaf. R Neottia nidus-avis (L.) L.C.M.

Richards - -

R Oenanthe peucedanifolia Pollisch

- -

R Orchis coriophora L. - - R Orchis laxiflora Lam. ssp.

elegans (Heuffel) Soó - -

R Orchis mascula (L.) L. ssp. signifera (Vest) Soó

- -

R Orchis militaris L. - -

134

R Orchis morio L. ssp. morio - - R Orchis purpurea Hudson - - R Orchis ustulata L. - - R Orobanche lucorum A. Braun - - R Pinus sylvestris L. - - - - V Plantago cornuti Gouan. R Plantago schwarzenbergiana

Schur - -

R Platanthera bifolia (L.) L.C.M. Richards

- -

R Platanthera clorantha - - R Pleurospermum austriacum (L.)

Hoffm. R Pleurospermum austriacum (L.)

Hoffm. R Potamogeton trichoides Cham.

et Schlecht. R Potamogeton trichoides Cham. et

Schlecht. R Primula elatior L. ssp.

leucophylla (Pax.) H. Harrison ex W.W. Sm. et Fletcher

nt Primula elatior L. ssp. leucophylla (Pax.) H. Harrison ex W.W. Sm. et Fletcher

R Pulsatilla grandis Wenderoth - - - - R Pyrola chlorantha Swartz R Rhynchospora alba (L.) Vahl. R Rhynchospora alba (L.) Vahl. R Ribes spicatum Robson R Ribes spicatum Robson R Rorippa islandica (Oeder)

Borbás -

- - R Rorippa prolifera (Heuffel) Weiche

R Rosa micrantha Sm. - - - R Rubus glandulosus Bellardi - - R Rumex aquaticus L. - - R Rumex longifolius DC. in Lam. et

DC. R Salix aurita L. - - R Salix daphnoides Vill. - - R Salvinia natans (L.) All - - R Scirpus radicans Schkuhr - - - - R Sedum caespitosum (Cav.) DC. R Sempervivum zeleborii Schott - - R Serratula radiata (Welk) Bieb. - - - - R Seseli tortuosum L.

VR Seseli hippomarathrum Jacq. R Seseli hippomarathrum Jacq. - - K Silene italica Retz. ssp. italica - - E Sisymbrium altissimum L.

135

- - E Sisymbrium irio L. - - E Sisymbrium loeselii L. - - E Sisymbrium officinale (L.) Scop. - - E Sisymbrium strictissimum L. R Sorbus aria (L.) Crantz. - - R Spirea crenata L. - - R Stellaria palustris Retz. - - R Symphytum tauricum Willd. - -

VR Taxus baccata L. R Taxus baccata L. - - nt Thymus comosus Heuffel K Thymus serpyllum L. V Thymus serpyllum L. V Traunsteinera globosa (L.)

Reichenb. - -

R Trisetum macrotrichum Hackel - - R Trollius europaeus L. ssp.

europaeus V Trollius europaeus L. ssp.

europaeus R Typha minima Funk in Hoppe R Typha minima Funk in Hoppe

VR Typha shuttlewortii Koch et Sonder

- -

R Vicia peregrina L. R Vicia peregrina L. K Vicia tenuifolia Roth. - - R Viola jooi Janka R Viola jooi Janka R Wolffia arrhiza (L.) Horkel ex

Wimmer - -

Abbreviations: I- indeterminate species and subspecies R- rare species and subspecies V- vulnerable species and subspecies Ex - extinct species and subspecies K – less known species and subspecies nt - not endangered endemic species and subspecies P- endangered species and subspecies

136


CONTRIBUTIONS TO THE STUDY OF THE CLASS MOLINIO-ARRHENATHERETEA R. TX. 1937

IN THE UPPER BASIN OF RIVER DORNA (SUCEAVA COUNTY) (I)

MIHAELA AURELIA DANU∗, T. CHIFU∗

Abstract: This paper represents an analysis of 2 vegetal associations (Agrostideto – Festucetum pratensis Soó 1949 and Festuco rubrae – Agrostietum capillaris Horvat 1951) classified from the coenotaxonomical point of view in the class Molinio-Arrhenatheretea R. Tx. 1937. The phytocoenoses of these mesophilic association, identified on the territory of the upper basin of the river Dorna (district of Suceava), are described from both the phytocoenological point of view, as well as from the point of view of the bioforms, floristic elements and ecological indices. Key words: class Molinio-Arrhenatheretea, mesophilic phytocoenoses, upper basin of Dorna.

Introduction

The upper basin of the river Dorna is located in the south-west part of the district of Suceava. Integrated in the central-northern part of the Oriental Carpathians, the basin is characterized by a temperate continental climate, the average annual temperatures being low (4.2ºC). The average value of precipitations is over 740 mm/an. In the area can be identified different types of soil belonging to 7 classes: non-evolved soils, truncated or cleaned, hydromorph soils, cambisoils, spodosoils, muddy luvisoils, shadowed soils, hysto soils.

The vegetal associations analysed in this paper have not been noticed so far in the upper basin of the river Dorna.


For the study of the vegetation we used the method of the phytocoenological School in Zurich-Montpellier, perfected by J. Braun-Blanquet and J. Pavillard. On taking into consideration few phytosociological papers of classification [7], [8], [9], [10], the associations were framed in the following coenosystem:

Cls. Molinio – Arrhenatheretea R. Tx. 1937 Ord. Molinietalia caeruleae Koch 1926

Al. Alopecurion pratensis Passarge 1964 As. Agrostideto – Festucetum pratensis Soó 1949

Ord. Arrhenatheretalia R. Tx. 1931 Al. Cynosurion R. Tx. 1947

As. Festuco rubrae-Agrostietum capillaris Horvat 1951

∗ “Al. I. Cuza” University, Faculty of Biology, Carol I Bd., no. 20A, 700506, Iasi, Romania

137

As. Agrostideto – Festucetum pratensis Soó 1949

Corollogy: Piatra Fântânele Ecology. The phytocoenoses of Festuca pratensis and Agrostis stolonifera were

identified in the upper basin of the river Dorna at altitudes between 1100 and 1130 m, on fields with slopes between 2° and 20°, having a cover of vegetation between 80 and 100%. The phytocoenoses of this association are developed on humid soils, with high humidity especially in the vernal season.

The floristic and phytocoenological characterization. The floristic composition of the association is rich (51 species), varied, with numerous mesophilic and meso-eutrophic species. The mesophilic character of the association is underlined also by the presence in big number of some species characteristic for the order Arrhenatheretalia: Achillea millefolium ssp. millefolium, Carum carvi, Dactylis glomerata, Leucanthemum vulgare ssp. vulgare, etc., species with a high constancy (tab. I).

As for the spectre of bioforms, the basic found of these meadows is represented by hemicryptophytes (H.-70.59%) and, in proportion of over 11%, by terrophytes (T.-11.76%) (fig. 1 a).

From the phytogeographical point of view, the important contribution in the flora composition is brought by the Euro-Asian and European elements (fig. 2 a).

The spectre of ecological indices shows that, due to high humidity of the under-layer, in these phytocoenoses are present many (over 45%) mesohygrophilic species (Agrostis stolonifera, Festuca pratensis ssp. pratensis, Lychnis flos-cuculi, Rhinanthus angustifolius ssp. angustifolius, Alchemilla vulgaris). The analysis of the spectre of ecological indexes underlines also the fact that in the structure of these phytocoenoses are dominant the species of light, which prefer weakly the shadow (over 90%); as for the temperature, many species are amphitolerant, but over 30% are plants characteristic to the chilly areas. Concerning the content of mineral nitrogen, half of the species identified in these phytocoenoses prefer the soils with low content till moderate (fig. 2 b).

As. Festuco rubrae – Agrostietum capillaris Horvat 1951 The meadows with Festuca rubra and Agrostis capillaris have a wide spread in the

Romanian Carpathians, on the coasts moderately sloped, with regime of moderate humidity, with brown rainy and brown acid soils, moderate-low acid and with a moderate content of nutritive substances.

The species characteristic and representative, Agrostis capillaris and Festuca rubra, are in a proportion of co-dominance, according to the content of nutritive substances in the soil and the degree of aeration of the soil. Thus, the species Agrostis capillaris is dominant on the fields recently covered with herbal vegetation and fertilized, while Festuca rubra dominates on the fields more beaten and less rich in nutritive substances.

Corollogy: Piatra Fântânele Ecology. The meadows with the phytocoenoses of this association were identified at

altitudes between 1100 and 1150 m, on soils slightly sloped (1°-20°), having a cover of vegetation between 80% and 100%.

138

The floristic and phytocoenological characterization. The floristic composition is rich (62 species) and varied, being noticed the predominance of the species characteristic for the alliances Cynosurion (Bellis perennis, Cynosurus cristatus, Phleum pratense, Trifolium repens ssp. repens, Prunella vulgaris), Arrhenatherion (Campanula patula, Centaurea phrygia, Taraxacum officinale), order Arrhenatheretalia (Achillea millefolium ssp. millefolium, Briza media, Carum carvi) şi clasei Molinio – Arrhenatheretea (Anthoxanthum odoratum, Cerastium holosteoides, Euphrasia officinalis ssp. pratensis, Rhinanthus minor, Stellaria graminea, Trifolium pratense), in proportion of 62,9%, which denotes the character mainly mesophilic of this association. In the frame of the phytocoenoses of this association, there were also identified, in reduced proportions, species characteristic to the class Festuco – Brometea (12.9%), Calluno – Ulicetea (9.67%) and Juncetea trifidi (6.45%) (tab. II).

The spectre of bioforms is dominated by hemicryptophytes (H.) situation underlined by the percentage of 74.19%. Follow the hemiterrophytes (Ht.) that realize 9.68%, and in lower quantities are represented the species terrophytes (T.) (9.68%), geophytes (G.) and camephytes (Ch.) being represented in equal proportion (each 4.84%) (fig. 1 b).

Among the elements of flora, 50% belong to the Euro-Asian element (Euras.); follow the European element (Eur.) represented by 30.65% and the cosmopolite species (Cosm.) with 8.06%. The circumpolar species (Circ.) are represented equally as the alpine elements (each 3.23%); the pontic, Carpathian – Balkan elements and endemic elements have one representative each in these phytocoenoses (fig. 3 a).

The spectre of ecological indices indicates the following preferences of the species to the ecological factors: from the point of view of the preferences to light, the heliophylic species, which bear weakly the shadow, are dominant (over 90%). From the point of view of the temperature, dominate the species amphitolerant (62.9%), and almost 30% are mesothermal plants. The association has a mesophylic character (demonstrated by the presence of the species developed on soils moderately humid), 33.87% being mesophylic plants. We notice that 37.15% among the species are xerophylic, which can be explained by the fact that some phytocoenoses are located on sloped surfaces, with reduced capacity of retaining water. As for the preference of plants for the pH of the soil, over 53% are euriionic, and 24.19% are acidophilic and moderate-slight acidophilic plants. Most of the species belonging to the phytocoenoses develop on soils poor in mineral nitrogen (51.61%), 22.58% among the species prefer the soils with content of mineral nitrogen varying from moderate to excessive, and almost 21% of the species are amphitolerant from this point of view (fig. 3 b).

REFERENCES

1. BELDIE AL., 1977 - Flora României – Determinator ilustrat al plantelor vasculare, vol. I-II, Edit. Acad. R.S.R., Bucureşti

2. CHIFU T., 1995 - Contribuţii la sintaxonomia vegetaţiei pajiştilor din clasele Molinio – Arrhenatheretea Tx. 37 şi Agrostietea stoloniferae Oberd. in Oberd. et al. 67 de pe teritoriul Moldovei. Bul. Grăd. Bot. Iaşi, 5: 125-132

139

3. CHIFU T., MÂNZU C., ZAMFIRESCU O., 2006 - Flora şi vegetaţia Moldovei, vol. II, Ed. Univ. „Al. I. Cuza” Iaşi

4. CHIRIŢĂ V., 2003 - Depresiunea Dornelor: studiu fizico – geografic. Edit. Univ., Suceava 5. CIOCÂRLAN V., 2000 - Flora ilustrată a României – Pteridophyta et Spermatophyta, Edit. Ceres,

Bucureşti 6. ELLENBERG H., 1974 - Indicator values of vascular plants in Central Europe, Scripta Geobotanica, vol. IX,

Verlag Erich Goltze K.G., Göttingen: 1-97 7. SANDA V., 2002 - Vademecum ceno-structural privind covorul vegetal din România, Edit. Vergiliu,

Bucureşti 8. SANDA V., POPESCU A., ARCUŞ M., 1999 - Revizia critică a comunităţilor de plante din România, Edit.

Tilia Press International1, Constanţa 9. SANDA V., POPESCU A., BARABAŞ N., 1997 - Cenotaxonomia şi caracterizarea grupărilor vegetale din

România. St. şi Com. Muz. Şt. Nat. Bacău, Biol. veget., 14: 2-365 10. SANDA V., POPESCU A., STANCU D. I., 2001 - Structura cenotică şi caracterizarea ecologică a

fitocenozelor din România, Edit. Conphis, Bucureşti

Table I - As. Agrostideto – Festucetum pratensis Soó 1949

Relevé number 1 2 3 4 5 6 Altitude (m) 1130 1125 1125 1100 1120 1120 Exposition E V NE E NV V Slope (°) 5 15 2 20 10 10 Cover of the vegetation (%) 90 95 100 90 80 85 Surface of the relevé (m²) 100 100 100 100 100 100 Number of species 43 36 28 34 24 27 K Caract. as. Agrostis stolonifera 3 3 2 3 3 3 V Alopecurion pratensis Festuca pratensis ssp. pratensis 1 2 3 2 2 1 V Phleum pratense + + + + - - IV Molinion caeruleae et Molinietalia caeruleae Briza media + + + - - + IV Gymnadenia conopsea ssp. conopsea + - - - + - II Lychnis flos-cuculi - + - - + - II Arrhenatherion Campanula patula - + + - + + IV Centaurea phrygia 1 + - + - + IV Cynosurion Cynosurus cristatus + 1 + + + + V Leontodon autumnalis ssp. autumnalis

+ + + - - - III

Trifolium repens ssp. repens - + + - - - II Arrhenatheretalia Achillea millefolium ssp. millefolium

+ + + + 1 + V

Carlina acaulis ssp. acaulis + + + + + 1 V Carum carvi + + - + - + IV Crepis biennis + - - - + - II Dactylis glomerata - + + - - + III Heracleum sphondylium ssp. sphondylium

+ - - + - + III

Knautia arvensis ssp. arvensis - + - + - - II

140

Leucanthemum vulgare ssp. vulgare + 1 + 1 + + V Tragopogon pratensis ssp. orientalis + + + + - - IV Thymus pulegioides + + + - + + V Veronica chamaedrys ssp. chamaedrys

- + + - - + III

Molinio – Arrhenatheretea Anthoxanthum odoratum + 1 1 1 + + V Campanula glomerata ssp. glomerata + - + - - - II Cerastium holosteoides + + - + - + IV Euphrasia officinalis ssp. pratensis + - + + + + V Lotus corniculatus + + + + + + V Plantago lanceolata ssp. lanceolata + + + + - - IV Polygala vulgaris - + + + - - III Ranunculus acris ssp. acris + + + + - - IV Rhinanthus angustifolius ssp. angustifolius

1 + 1 + + + V

Rumex acetosa + - - + - + III Trifolium pratense + + + + - - IV Vicia cracca - + - + - + III Potentillo – Nardion Arnica montana + - - - + - II Cruciata glabra + + - - + - III Scorzonera rosea + - - + - - II Juncetea trifidi Campanula serrata + - - - + - II Dianthus deltoides + - - + - - II Hypochaeris uniflora + + - + + 1 V Nardus stricta + + + + - + V Potentilla erecta + + + + + + V Festuco – Brometea Plantago media + + - - + - III Trifolium alpestre + + + + - + V Trifolium pannonicum + - - + - - II Variae syntaxa Alchemilla vulgaris 1 + + + + + V Galeopsis speciosa + - - - - + II Hypericum maculatum ssp. maculatum

+ - - + + - III

Stellaria media + + - - - - II Tanacetum corymbosum + - - - + - II Viola tricolor + - - + - - II

Place and date of the relevées: Piatra Fântânele 1 - 27.07.2006; 2, 3, 4 - 20.08.2006; 5, 6 - 1.09.2006

141

Table II - As. Festuco rubrae – Agrostietum capillaris Horvat 1951

Relevé number 1 2 3 4 5 6 7 8 9 10 11 Altitude (m) 1130 1140 1140 1100 1100 1125 1150 1148 1145 1150 1150 Exposition V E S NV NV NE E V S V E Slope (°) 5 5 5 1 3 20 10 10 2 2 10 Cover of the vegetation (%) 95 95 95 80 90 90 90 95 100 100 90 Surface of the relevé (m²) 100 100 100 100 100 100 100 100 100 100 100 Number of species 30 32 26 24 37 29 26 36 24 26 29 K Caract. as. Agrostis capillaris 1 2 3 4 + 1 3 3 2 1 1 V Festuca rubra 3 3 2 2 4 4 2 2 3 4 4 V Cynosurion Bellis perennis - + - + - - + - - - - II Cynosurus cristatus + + - + + + + + + 1 - V Leontodon autumnalis ssp. autumnalis

+ + - + - - - + - - - II

Phleum pratense + + - - - + - + - 1 + III Plantago major ssp. major - + - - - - - - + - + II Prunella vulgaris - + + + + + + + - - + IV Trifolium repens ssp. repens - - + + - - + + 1 + + IV Veronica serpyllifolia ssp. serpyllifolia

- + - + - - + - - - - II

Arrhenatherion Campanula patula - + + + + - - + + + + IV Centaurea phrygia - + + - + + - + - - + III Taraxacum officinale - + + + - - + + - - - III

142

Phyteumo – Trisetion Hypericum maculatum ssp. maculatum

- + - - + - - + + + + III

Luzula luzuloides ssp. luzuloides + - - - - - - - + - + II Arrhenatheretalia Achillea millefolium ssp. millefolium

- + - + + + + + - - + IV

Briza media + + - + + + - + + + + V Campanula glomerata ssp. glomerata

+ - - - - - - - - - - I

Carlina acaulis ssp. caulescens + + + - - + - + - + + IV Carum carvi - + + - + + - - - - - II Leucanthemum vulgare ssp. vulgare + + + - + + - + 1 + + V Knautia arvensis + - - - - - - + - - + II Thymus pulegioides - + + + 1 + + + + + + V Tragopogon pratensis ssp. orientalis - - + - - - + - + - + II Molinietalia Gymnadenia conopsea ssp. conopsea

+ - + - + - - - + - - II

Lychnis flos – cuculi - + - - - - + - - - - I Molinio – Arrhenatheretea Alchemilla vulgaris 1 - + + + + + + + + + V Anthoxanthum odoratum + + - - + + 1 1 1 + + V Centaurea jacea - - - - + - + + - + + III Cerastium holosteoides - + - - + - - - - - - I Euphrasia officinalis ssp. pratensis - - - - + - + + + + - III Lotus corniculatus - + + + + + - + - - + IV Plantago lanceolata ssp. lanceolata + + - + + + + + - - - IV

143

Polygala vulgaris - - + - + + - - + + + III Ranunculus acris ssp. acris + - + - - - + - - + - II Rhinanthus minor 1 1 1 + + + + + + + + V Rumex acetosa + - - - + + - + - - - II Stellaria graminea + - - - - - - + - - - I Trifolium pratense + - - + + + - - + - 1 III Festuco – Brometea Anthyllis vulneraria ssp. vulneraria - + + - + + - - - + - III Echium vulgare - - - - + - - - - - - I Euphrasia stricta ssp. stricta - + + - - - - - - - - I Galium verum - - - + - - + + - - - II Hieracium pilosella - - + 1 + + - - - + - III Ranunculus polyanthemos ssp. polyanthemoides

- + - - + + - + - - - II

Trifolium alpestre - + + - + - + + + + + IV Trifolium pannonicum + - - + - + - + - - - II Juncetea trifidi Campanula serrata + + - - + + - + - - - III Nardus stricta + - - - - - - - - - - I Potentilla ternata - + + - + - - + + + - III Scorzonera rosea + - - - - - + - + - + II Calluno – Ulicetea Antennaria dioica - - + - - - - - - + - I Arnica montana + - - - + + - - - + - II Dianthus deltoides + + - - - - - + - - + II Genista tinctoria ssp. tinctoria + - - + + - + - - - - II Gentianella austriaca + - - - + + - - - - - II Potentilla erecta - - + - 1 + 1 + + + + IV

144

Variae syntaxa Cirsium vulgare - - - + - - + - - + - II Cruciata glabra + - + - + - - + - - - II Pteridium aquilinum - - - + + + - - - - - II Rumex acetosella ssp. acetosella - - - - - - - + + - + II Tanacetum corymbosum ssp. corymbosum

+ - - - - - + - - - - I

Place and date of the relevées: 1 - Piatra Fântânele, 27.07.2006; 2 - 11 - Piatra Fântânele, 21, 22.08.2006

Fig. 1. The bioforms spectrum: a) as. Agrostideto-Festucetum pratensis Soó 1949; b) as. Festuco rubrae-Agrostietum capillaris Horvat 1951

b

H - 74,19%

G - 4,84%

Ch - 4,84%

Ht - 9,68%

T - 6,45%

a

T - 11,76%

Ht - 9,8%

Ch - 3,92%

G - 3,92%

H - 70,59%

Fig. 2. a) The floristic elements spectrum; b) The ecological indices spectrum (L-light; T-temperature; Ct-continent; U-humidity; R-soil reaction; N-nitrogen)

– as. Agrostideto-Festucetum pratensis Soó 1949

Fig. 3. a) The floristic elements spectrum; b) The ecological indices spectrum (L-light; T-temperature; Ct-continent; U-humidity; R-soil reaction; N-nitrogen)

– as. Festuco rubrae-Agrostietum capillaris Horvat 1951

b

01020304050607080

1 2 3 4 5 6 7 8 9 10 11 x ?

%LTCtURN

a

Euras. - 44,77%

Eur. - 12,11%

Centr. eur. - 9,91%

Circ. - 18,35%

Atl. - 0,92%

Dacic - 0,18%

Pont. - 1,47%

Medit. -0,37%

a

Euras. - 50%

Eur. - 30,65%

Circ. - 3,23%

Pont. - 1,61%

End. - 1,61%

Cosm - 8,06%

Carp. balc. - 1,61%

Alp. - 3,23%

b

010203040506070

1 2 3 4 5 6 7 8 9 x ?

%LTCtURN

146


ENVIRONMENTAL EDUCATION: EDUCATION FOR TRANSITION TO

SUSTAINABLE DEVELOPMENT

I. M. CIUMAŞU∗, NAELA COSTICĂ∗∗

Abstract: It is general accepted that the principles of sustainable development can’t be reached without education, public awareness and training. In this regard, the present paper presents rationales and approaches in Environmental Education, as well as the importance of institutional and curricular aspects in implementing this type of education. Key words: Environmental Education, institutional and curricular aspects.

Rationales and educational approaches

While civilization is being maintained through institutions, it is kept alive and

growing through education. As the current unsustainability of humanity resides the inharmonious (conflicting) relationship between nature and human society, we need environmental education. Therefore we need a coherent program to train environmental educators.

Chapter 36 of Agenda 21 calls each nation to bring together experts from various

disciplines to prepare a national strategy for environmental education (EE) and training [33, 28, 17].

Lucas [16, 7] identified three meanings/facets of environmental education: 1. education about the environment (concerned with cognitive understanding of

environmental issues); 2. education for the environment (concerned with environmental protection via

particular purposes and aims); 3. education in the environment (concerned with environmental experience as

educational mean outside the classroom). In EE institutions, the organisational strategy and the curricular strategy should be

complementary. The first has a greater effect on values, attitudes and behaviour, whereas the second influence more the conceptual/knowledge understanding. EE is not effective if the organisational strategy contradicts the curricula [30]. For example, an Integrated

∗ “Al. I. Cuza” University, Faculty of Biology, Carol I Bd., no.11, 700506, Iaşi, Romania ∗∗ Associated researcher at: “Al. I. Cuza” University, Faculty of Biology, Centre of expertise on sustainable exploitation of ecosystems (CEXDUREC), B-dul Carol I no.11, 700506, Iaşi, Romania. and Technische Universität München, Arcistrasse 21, 83000, München, Deutschland, Wissenschaftszentrum Weihenstephen für Ernährung, Landnutzung und Umwelt, Biowissenschaftlische Grundlagen, Alte Akademie 8, 85354, Freising, Deutschland, Lehrstuhl für Ökologische Chemie und Umweltanalytik Weiehnstephaner Steig 23, 85350, Freising-Weihenstephan, Deutschland.

147

Action Model [7] can be used to identify motivational profile of students (this also includes age categories). These can be used together with social and situational conditions to design adapted curricula for different categories of students (target groups). However, such precision must not be applied at the expense of community-based cooperation in EE, but in the same time with it (and integrating it).

There are four possible target groups in Environmental Education, as related to their

personality: 1. The Technical Group needs to know how to gauge environmental parameters; 2. The Subject Specialist Group needs to understand environmental systems; 3. The Management Group requires skills and abilities to resolve complex

environmental problems; 4. The Lay Group needs to have attitudes, philosophies and values about the

environment. The old-fashion one-direction model of teaching is already outdated (though it resists

in many countries). The teacher must accept that, in the Internet era, he cannot hold control of the learning as in former times (when he was the one possessing information).

On the contrary, the teacher must extend teaching beyond the walls of the classrooms, and capture students' attention in more subtle ways, on the basis of reciprocal respect of the other and of the common values. This is of course not to say that the teacher-student relationship should be loose. The do-it-yourself learning (a lessez-faire approach) based on technology-in-the-classroom must be backed by reciprocal assuming of responsibility for the learning process. Otherwise, internet-chatting and other free-time activities will replace learning. Thus, internet in the classroom does not automatically bring educational progress [10]. An effective education approach is the two-ways, dialogical lesson.

This dialogue requirement holds also when environmental teaching kits are being used, eventually in combination with web-based and other methods and materials. As such kits tend to be more employed at lower ages [4], they could be employed more heavily in kindergartens and progressively be replaced at older school ages with web-based lessons of appropriate difficulty.

Given the ever changing and enlarging context of the teacher-student relationship,

any curriculum must be rather elaborated for educating the teacher how to educate the student. Such "rehearsal curriculum" must be written in a way that motivates the teacher to learn and update and diversify its skills [11, 24].

Thus, in order to structure a problem-based EE, various types of questions can be identified and labeled [6]:

- encyclopaedic, - meaning-oriented, - relational, - value-oriented, - solution-oriented

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For example, web-based systems for environmental management [21, 29] can (and should) be introduced in classrooms with internet availability through questions and dialogue. There exist even international web-based, hands-on EE programs, e.g. Global Learning and Observations to Benefit the Environment – GLOBE is such a program which allows school children to be involved in a dynamic student-teacher-scientist partnership: they learn about the environment by taking carefully supervised scientific measurements of their natural surroundings (land cover, soil, hydrology, phenology, haze/aerosols, and atmosphere) and sharing information/data with scientists and other students in remote locations via Internet [34]. More, there have already been experimented internet-based, inter-institutional teaching systems, linking education institutions with with actors in the socio-economic environment (local authorties, companies, etc). In sucha case has a profoundly applied character, because it also includes the real world decision-taking processes [18]. The children's sense of participation and discovery is the best medium for EE and for nurturing responsible attitudes towards the environment. We will come back to this aspect when addressing "community-based learning".

Institutional aspects

We should no be left with the impression that there are no limits to this approach. For example, the idea of green schools looks shiny, but is dangerous. The big risk is that creating a new category of school out of the mainstream education will ascribe to EE a marginal importance in the collective perception of the citizens. In fact, EE is still largely without focus and side-lined [13, 27, 5]. I mean we must learn from the mistakes of politicization of the environmental issues, and not confound environmental education with political activism [8]. Thus the greens insist on the fact that only a green party can do the necessary change for environmental protection. While it is true that politic efforts must be focused, the focus should be on problems, not on political activism, and solutions must be found within mainstream politics: main parties, main governments. EE (and EE curricula) must be present at the core educational programs, not mere a specific but marginal one.

Such marginalizing risks exist with green schools, despite being a great concept. Surely, sustainability and earth conservation is an emergency, and we want to achieve and see fast progresses toward sustainability. However, short-term fast progresses are all too often false progresses, and a pathway to profound deceit (they are a dead-end road). The idea that all schools will follow the example of the green schools and turn green themselves is rather wishful thinking. Undoubtedly, the fact that green schools exist is a working idea is good think, itself a sign of a healthy democratic, diverse society. The fact that green school can exist is a victory for sustainability. But if green curriculum can only be implemented as segregated from the regular school, in green school and the like, this is no victory, but a defeat.

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Curricular aspects

At present, EE is part of various disciplines/curricula. Our project is a way to synthesize and update EE methodologies and strategies in the partner institutions; hence it offers a model path for others. In addition, the project acts for the development and proposing of a common EE framework in the European Union. This work is therefore one of the international, real life efforts to implement sustainability.

A common European Curriculum for training the trainers in Environmental Education cannot avoid overlapping some specialized programs and curricula. Our aim is not to propose some sort of imposition of a common European curriculum. Still, a European curriculum should exist, at least as an authoritative reference.

The first obstacle to overcome is the fact that there is no unified theory or scientific

body of knowledge regarding environmental knowledge. The same is true for the environmental education itself. In fact even local/institutional knowledge is both largely contingent and not monolithic [22]. But that's ok, this should be so. This variety of opinion may appear to hamper decision-making process.

The idea is to create the necessary conditions and working framework that allow professional and democratic involvement, which is the scope of what is known as the science of governance (not governing; governing pertains to governmental decisions, while governance pertains to multiplayer decision – governments, NGOs, scientists, and all stakeholders). For this, it is essential to establish the common grounds and the separate freedoms and responsibilities of each player. This is what we want to achieve with the current curriculum project.

The workshop (kick-off meeting) held between May 30 –June 01 in Iasi (Alexandru

Ioan Cuza University), with the participation of all partners within the Leonardo da Vinci project RO/05/B/P/PP175010, allowed intensive discussions on the background of EE. The debates allowed reaching the common position that an environmental education (EE) curriculum must include the following character features:

• interdisciplinary and holistic [1, 31], • value- and fairness driven, • critical thinking and problem solving orientation, • participatory, • applicable in real-life and local contexts, • favouring creativity, • acceptance of change.

The last point is particularly relevant because it is a condition for necessary reforms in Central Eastern European member countries of the European Union, but also in older members.

A two-step education project at Purdue University in the US, a "dual-level

professional development model for changing teacher practice" where Level I

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participants were trained by University staff and trained at their turn their colleagues - Level II participants) similar to our educating-educators project showed high effectiveness in changing classroom practice (83% in Level I participants and 68 % in Level II participants). Hands-on approaches were the most effective [26]. While these results suggests that peer-education can be employed as highly effective (68%), they also hint to a more effective education of educators through direct contact with University staff, probably because the later (besides being higher qualified; but being also research professionals) have a deeper hands-on experience.

For instance, writing is an integral part of the Environmental Education research [15]. Therefore, innovative methods in the formation of EE teachers should include writing EE texts: conducting literature reviews, interviewing decision-makers and scientists, as well as synthesizing and documenting management problems (with related science and other issues that might constrain or drive the solution (legislation, social pressures, politics, personalities, etc) [3]. This should also include in every EE institution writing and periodically updating a document describing the organizational strategy of "greening" the EE institution. EE educators should acquire basic training and some working knowledge on what it means to green a Centre; plus related documents / working knowledge on environmental issues, environmental management, alternative systems, etc. This document should explain how the centres work (decision-making bodies, budgets, etc). It is recommended that "greening" mechanisms be professional, transparent and democratic (democratic does not signify lack of hierarchical responsibilities) [30].

A diversity of approaches is needed – there is no single general valid method [25].

While EE methods are already very diverse, the EE outcomes in schools can be understood as both:

1. well-established evidence (EE outcomes: students' environmental knowledge, attitudes and behaviour) and

2. emerging evidence (EE processes: students' perceptions of nature, experience of learning and influence on adults).

While later aspects have received less attention from researchers than the former, they disserve more dedication: as EE is not once and for all (but a life-long process), understanding EE processes insures better adaptability to new concerns and foci in time. Currently, there is a need to restore equilibrium in this sense [23].

Having in mind the importance of community involvement in governance and

sustainable development, an interesting approach in EE is that of community-based schools, where parents and other community members are actively taking part in school-based EE curriculum and various indoor and outdoor EE activities [32]. Some authors even talk of a "school-family-community ecosystem" as approach in environmental education [2]. Local environmental knowledge is also influenced by active participation in land use practices and outdoor recreation [19]. This is valuable bottom-line experience. Learning about "places" associated with local cultures is a good way to do effective EE. This learning can be done via [20]:

• childhood experiences,

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• learning from elders and family, • action and observation, • comparisons between places, • via festivals and community events, • external sources, • seeing a place under different conditions (during summer, winter, conflict,

drought, floods, etc, • continuity in connection to a place. For example, in schools that are closed to significant water bodies, EE can be done

through water quality analyses, e.g. in the sea [12] or across watersheds [9].

REFERENCES 1. Barrett, G.W., 2001 - Closing the ecological cycle: the emergence of integrative science. Ecosystem Health,

7, 2:79-84 2. Bennett, S.K., Bennett, D.B., 2004 - Lecture: regarding the ecology of science education: connections to

environmental and science education. Journal of Science Education and Technology 13, 2:137-146 3. Bradley, M.P., Hanson, R., Walbeck, E.S., 2004 - Innovative environmental education contributes to

improved management practices in the Mid-Atlantic region of the United States. Environmental Monitoring and Assessment, 94: 205-215

4. Chan, K., 2000 - Use of environmental teaching kits in Hong Kong. The Environmentalist, 20:113-121 5. Chapman, D., Sharma, K., 2001 - Environmental attitudes and behaviour of primary and secondary students

in Asian cities: an overview strategy for implementing an eco-schools programme. The Environmentalist, 21:265-272

6. Dahlgren, M.A., Öberg, G., 2001 - Questioning to learn and learning to question: structure and function of problem-based learning scenarios in environmental science education. Higher Education, 41:263-282

7. Dempsey, R., Gresele, C., Bögeholtz, S., Martens, T., Mayer, J., Rode, H., Rost, J., 1997 - Empirical studies on environmental education in Germany: contributions by the Institute for Science Education. Research in Science Education, 28, 2: 259-279.

8. Disinger, J.F., 1997 - Environmental education research news. The Environmentalist, 17:153-156 9. Donahue, T.P., Lewis, L.B., Price, L.F., Schmidt, D.C., 1998 - Bringing science to life through community-

based watershed education. Journal of Science Education and Technology, 7, 1:15-23 10. Elstad, E., 2006 - Understanding the nature of accountability failure in a technology-filled, laissez-faire

classroom: disaffected students and teachers who give in. Journal of Curriculum Studies, 38, 4: 459-481 11. Fien, J., Poh, I. T.-C., Yencken, D., Sykes, H., Treagust, D., 2002 - Youth environmental attitudes in

Australia and Brunei: implications for education. The Environmentalist, 22:205-216 12. Gough, G.A., Robottom, I., 1993 - Towards a socially critical environmental education: water quality studies

in a coastal school. Journal of Curriculum Studies, 25, 4: 301-316 13. Harvey, T., 1995 - An education 21 programme: orienting environmental education towards sustainable

development and capacity building for Rio. The Environmentalist ,15, 3: 202-210 14. John, P.D., 2006 - Lesson planning and the student teacher: re-thinking the dominant model. Journal of

Curriculum Studies, 38, 4:483-498 15. Lotz-Sisitka, H. Burt, 2002 - Being brave: writing environmental education research texts. Canadian Journal

of Environmental Education 7, 1: 9 http://www.uleth.ca/edu/research/ictrd/cjee/volume_7.1/9.pdf 16. Lucas, A.M., 1980 - Science and environmental education: pious hopes, self praise and disciplinary

chauvinism. Studies in Science Education, 7: 1-26 17. Lucie, S., 1999 - Environmental Education between modernity and postmodernity: searching for an

integrating educational framework. Canadian Journal of Environmental Education, 4: 9-35 18. Manring, S. & Moore, S.B., 2006 - Creating and managing a virtual inter-organizational learning network for

greener production: a conceptual model and case study. Journal of Cleaner Production, 14: 891-899

http://www.uleth.ca/edu/research/ictrd/cjee/volume_7.1/9.pdf

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19. McDaniel, J., Alley, K.D., 2005 - Connecting local environmental knowledge and land use practices: a human ecosystem approach to urbanization in West Virginia. Urban Ecosystems, 8: 23-38.

20. Measham, T.G., 2006 - Learning about environments: the significance of primal landscapes. Environmental Management, 38, 3: 426-434

21. Okada, M., Tarumi, H., Yoshimura, T., Moriya, K., Sakai, T., 2002 - Realization of a digital environmental education – a future style of environmental education in dynamically changing virtual environment. Book chapter in: Digital cities II: second Kyoto Workshop on digital cities, Kyoto, Japan, October 18-20, 2001

22. Reid, A., Petocz, P., 2006 - University lecturers' understanding of sustainability. Higher Education , 51: 105-123

23. Rickinson, M., 2001 - Learners and learning in environmental education: a critical review of the evidence. Environmental Education Research, 7, 3: 207-320.

24. Schwartz, M., 2006 - For whom do we write the curriculum? Journal of Curriculum Studies, 38, 4: 449-457. 25. Scott, D., 2004 - Transforming the "market-model University": environmental philosophy, citizenship and

the recovery of the humanities. Worldviews 8, 2-3: 162-184 26. Shepardson, D.P., Harbor, J., 2004 - ENVISION: the effectiveness of a dual-level professional development

model for changing teacher practice. Environmental Education Research, 10, 4: 471-492 27. Simmons, B., 2000 - Towards excellence in environmental education. A view from the United States. Water,

Air and Soil Pollution, 123: 517-524 28. Smyth, J.C., 1996 - A national strategy for environmental education: an approach to a sustainable future?

The Environmentalist, 16, 1: 27-35. 29. Sugumaran, R., Meyer, J.C., Davis, J., 2004 - A web-based environmental decision support system

(WEDSS) for environmental planning and watershed management. Journal of Geographical Systems, 6: 307-322

30. Sureda, J., Calvo, A.M., 2001 - Organization of school centres and environmental education: in search of action models for the greening of school organization. The Environmentalist, 21: 287-296

31. Stables, A., Scott, W., 2002 - The quest for holism in education for sustainable Development. Environmental Education Research, 8, 1: 53-60

32. Tal, R.T., 2004 - Community-based environmental education – a case study of teacher-parent collaboration. Environmental Education Research , 10, 4: 523-543

33. UN Agenda 21. 2005 - United Nations Agenda 21. http://www.un.org/esa/sustdev/documents/agenda21/english/agenda21toc.htm

34. Wormstead, S.J., Becker, M.L., Congalton, R.G., 2002 - Tools for successful student-teacher-scientist partnerships. Journal of Science Education and Technology, 11, 3: 277-287

http://www.un.org/esa/sustdev/documents/agenda21/english/agenda21toc.htm

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REVIEW MARIA DUCA, 2006, Plant physiology, Stiinta Publishing House in Chisinau, 287 p. (ISBN: 978-9975-67-596-3)

„Plant physiology”, book issued in 2006 at Science Publishing House in Chisinau, is considered to be a very important editorial event, which helps the increase of the knowledge level in plant physiology field. The author, Mrs. Maria DUCA, PhD Habilitated Professor, Corresponding Member of the Moldavian Academy of Sciences and Dean of The Faculty of Biology and Pedology from University of State from Moldavia, is an important figure for the Moldavian education and research activity, which, by this book, remarked once again in the field of expertise that she has been serving for the past two decades.

As the professor stated, the volume does not only reflects the basic principles and directions of the plant physiology, using specialized literature filtered by own thinking, but also includes results of personal research, by synthesizing a large amount of experimental data, and offering concrete possibilities to direct application of obtained knowledge in practice.The theme is very ample - it is enough to prospect the content of each of the 11 chapters included in the volume – so it will be a mistake to persist in the idea to make an exhaustive analysis of the book; even more, we would diminish the possibilities to know and to get thoroughly into the real novelties, exposed in a form inciting to meditation by running through the text. In the almost 300 pages, highly illustrated with suggestive color schemes and figures, the author approaches in a very systematic manner top contemporary plant physiology themes such as: auto regulation, signal perception and transduction, physiologic basis to perform the genetic program, biorhythms, compound elimination by the plants, as well as the resistance of these organisms to improper environmental factors.

To ensure the possibility to evaluate the new acquired knowledge, each chapter ends with evaluation tests of different complexity degrees, a glossary for specific terms, as well as a short, but comprehensive and actualized bibliography, that comes to help the ones interested in further studies on the discussed phenomena. The data transmitted in this manner helps to form the biologists, by detailing the biochemical and molecular mechanisms of the vital processes and functions, firstly having an informative function, but also a pronounced formative character. The book tries to build a systematic and logical thinking on the plant organism’s vital functions, as well as to form the reader’s competence in the field. The volume is correctly dimensioned, according to the contemporary biology requirements, and also offers to the biologists the necessary tools that allow the manipulation of plant organism, tools that are situated at the foundation of contemporary biotechnologies. The present volume, constructed in accord with the intrinsic changes in the Moldavian education system - that is trying to adhere to the Bologna process - by reviewing the information presentation mode and by offering landmarks for the self training of students, recommends itself to everyone and especially to the ones preoccupied with the study of plant physiology and interested in self education and up to date information in this scientific field.

Prof. dr. C. TOMA, Prof. dr. MARIA MAGDALENA ZAMFIRACHE

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REVIEW TĂNASE C., ŞESAN TATIANA EUGENIA, 2006, Concepte actuale în taxonomia ciupercilor, Editura Universităţii „Alexandru Ioan Cuza”, Iaşi: 510 pp. (ISBN 973-703-144-X; 978-973-703-144-0)

Rod al unei minuţioase munci, Concepte actuale în taxonomia ciupercilor este cel mai complex şi mai complet manual universitar tratând atât clasificarea ciupercilor cât şi morfologia, citologia, ultrastructura, biologia, ecologia şi reproducerea lor, cu informaţii utile şi asupra utilizării multora dintre ele.

Această carte se adresează în primul rând specialiştilor, dar şi studenţilor de la facultăţile de Biologie (cu secţii de Biologie, Ecologie şi Protecţia Mediului, Biochimie, Biotehnologie), Silvicultură, Agricultură, Horticultură, Farmacie, Medicină veterinară şi umană, Industrii alimentare, precum şi celor interesaţi de cunoaşterea şi conservarea diversităţii ciupercilor, de protecţia patrimoniului documentar, de carte, a monumentelor istorice, a diverselor tipuri de materiale ş.a.

Autorii – cadre didactice din două prestigioase universităţi ale ţării, Iaşi şi Bucureşti – au proiectat această lucrare sub forma unui tratat pe baza sintezei informaţiilor teoretice selectate din literatura de specialitate, a rezultatelor personale şi a experienţei acumulate în domeniul micologiei. Această experienţă, este concretizată în tratarea minuţioasă a unor aspecte originale care vizează atât identificarea acestor organisme, cât şi analiza critică a unor aspecte actuale referitoare la biologia, ecologia, corologia, biochimia şi taxonomia ciupercilor.

Tratatul este structurat în 5 capitole, care reunesc atât aspecte teoretice cât şi practice din domeniul micologiei. Ciupercile sunt prezentate ca un grup de organisme polifiletice reunite în 11 încrengături, care aparţin la 3 regnuri: Chromista, Fungi şi Protozoa. În aceste categorii taxonomice sunt descrise 1170 specii de ciuperci grupate în 115 familii.

Lucrarea cuprinde 510 pagini, în care sunt inserate 366 de figuri, 120 fotografii color originale şi 30 de tabele. Tratatul a fost realizat de cei doi autori, pe baza celor 580 de lucrări de specialitate consultate (marea majoritate apărute după 1990), a rezultatelor personale şi a experienţei acumulate de-a lungul multor ani de activitate în domeniu.

Prin analiza critică a cercetărilor din domeniul micologiei generale şi aplicate, inserate în baze de date internaţionale sau în reviste şi volume ale unor manifestări ştiinţifice internaţionale, autorii evaluează reperele şi realizările obţinute până în prezent, precum şi direcţiile de perspectivă.

În acest context, evidenţiez experienţa şi rezultatele autorilor, racordate la realizările internaţionale, foarte multe comunicate şi publicate în reviste de specialitate din ţară şi din străinătate.

Prof. dr. C. DRĂGULESCU

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INSTRUCTIONS TO AUTHORS

The Journal Analele Ştiinţifice ale Universităţii „Al. I. Cuza” din Iaşi (serie nouă), Secţiunea II a. Biologie vegetală, includes original articles of cytology, morpho-anatomy, physiology, taxonomy, phytosociology, mycology, phytopathology, along with book reviews and anniversary announcements.

All the papers must be submitted to our redaction address (Dr. Naela COSTICĂ, „Al. I. Cuza” University, Faculty of Biology, Department of Biology, Bd. Carol I., no. 20A, 700506, Iasi) both as printed manuscripts and electronic format. For the graphic uniformity of the volume, please consider followings: PAGE FORMAT: 18 x 12,7 cm; margins settings: 5,8 cm top, 6 cm bottom, 4 cm left, 4 cm right. TEXT: • the papers will be printed in one of the following languages: English, French, German • the text must be typed (in a PC compatible text editor) with Times New Roman 10,

single-spaced, on A4 paper; • the Abstract and the Key words must be typed in English or French, using the font

Times New Roman 8 points; • the title must be typed in Times New Roman 10 bold capitals; • authors' names and surnames must be typed in caps; male surnames must be

abbreviated; • the address of each author must be provided in footnote; • the text will be partitioned as follows: Introduction, Material and methods, Results

and discussions, Conclusions, References. All subtitles must be centred typed in Times New Roman, bold 10

ex.: Introduction • the scientific names must be typed in italics; • the text references of tables and figures (included in plates) must be typed between

round brackets: ex: (fig. 2, Pl. I), (tab. II);

• the text references of the cited bibliography must be typed between square brackets: ex: [5];

• the References subtitle must be centred and typed in bold 10 points caps:. ex.: REFERENCES

• bibliography references must be alphabetically ordered and typed in Times New Roman 8 point font, as follows:

for books: 1. BELDIE Al., 1972 - Plantele din Munţii Bucegi. Edit. Acad. Rom., Bucureşti for articles: 2. DAUB E. M., 1981 - Cercosporin, a photosensitizing toxin from Cercospora species.

Phytopathology, 72: 370 – 374

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3. REDZIĆ S., TUKA M., PAJEVIĆ A., 2006 - Research into microscopic structure and essential oils of endemic medicinal plant species Satureja subspicata Bartl. ex. Vis. (Lamiaceae). Bosn. J. Basic Med. Sci., 6, 2: 25-31

4. RUGINĂ R., TOMA C., IVĂNESCU L., 2007 – Morphological and histo-anatomical aspects at some dicotyledonate seedlings related to the vascular transition. An. Şt. Univ. “Al. I. Cuza” Iaşi, s. II a., Biol. veget., 52: 133- 142

FIGURES (colour or black and white photos, drawings) • all figures must be grouped in plates on separated pages; the number of the plate (ex.

PLATE I) must appear in the top-right position and the names of the authors (Times New Roman 10, caps, bold) must appear in the top-left position of each plate;

• the Explanation of plates (including figures) must be typed on separated page (after References)

• the figures must be printed on tracing paper (photocopies are not acceptable, just original materials) and must not exceed 18 x 12,7 cm; all materials must be accompanied by graphical scale.

TABLES • the tables must be printed on separated pages and must be numbered with roman digits

(Table I, Table II,…). For Book Reviews:

- mention the followings: author (name, surname, in caps), coma, year, title with italic bold characters, coma, place of publishment, number of pages, ISBN. Leave a blanc line and write the text of the reviews (paragraphs as few as possible) single spaced, on A4 paper with Times New Roman 10 font. Recommendations:

The submitted paper must not exceed 10 pages (illustration included) and must have an even number of pages (including an even number of pages with colour plates). The papers will be further submitted to the reference comity and will be published for a charge in Analele Ştiinţifice ale Universităţii “Al. I. Cuza” din Iaşi, Secţ. II a. Biologie vegetală.

The editorial comity reserves the right to: • reject certain papers (one paper as first author and another one in collaboration would

be acceptable) • reduce the number of figures, in case they are to many

Only papers presented in the Plant Biology section of the scientific congress organised by our Faculty will be published.

Responsibility upon the articles content belongs to the author (s). Papers that do not meat these rules will be returned to the author (s).

Editorial comity

ANALELE STIINTIFICE biologie 2 2008

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