Analele Stiintifice Biologie 2 2008

ANALELE TIINIFICE
ALE
UNIVERSITII Alexandru Ioan Cuza

DIN IAI
(SERIE NOU)
SECIUNEA II
a. BIOLOGIE VEGETAL
TOMUL LIV, FASCICULA 1
Editura Universitii Alexandru Ioan Cuza Iai
2008
Editura Universitii Alexandru Ioan Cuza din Iai

ANALELE TIINIFICE ALE UNIVERSITII Alexandru Ioan Cuza DIN
IAI (SERIE NOU), SECIUNEA II a. BIOLOGIE VEGETAL
Comitetul de redacie:
Profesor
Profesor
Profesor
Profesor
Dr. Constantin TOMA Universitatea Alexandru Ioan Cuza din Iai

Dr. Toader CHIFU Universitatea Alexandru Ioan Cuza din Iai
Dr. Mihai MITITIUC Universitatea Alexandru Ioan Cuza din Iai
Dr. Maria Magdalena ZAMFIRACHE - Universitatea Alexandru Ioan Cuza din
Iai
Confereniar Dr. Ctlin TNASE Universitatea Alexandru Ioan Cuza din Iai
Confereniar Dr. Lcrmioara IVNESCU - Universitatea Alexandru Ioan Cuza din Iai
Comisia de refereni tiinifici:

Academician Nicolae BOCAIU Academia Romn, Filiala Cluj-Napoca
Academician Valeriu COTEA Academia Romn, Filiala Iai
Profesor Dr. Constantin TOMA Universitatea Alexandru Ioan Cuza din Iai, membru
corespondent al Academiei Romne, Filiala Iai
Profesor Dr. Leontin tefan PTERFI Universitatea Babe-Bolyai din Cluj-Napoca,
membru corespondent al Academiei Romne, Filiala Cluj-Napoca
Profesor Dr. Jean Pierre AUQUIRE Universitatea Catolic din Louvain la Neuve, Belgia
Profesor Dr. Maria COULADIS Universitatea din Atena, Grecia
Profesor Dr. Cvetomir DENCHEV Academia de tiine din Bulgaria
Profesor Dr. Franco PEDROTTI Universitatea din Camerino, Italia
Profesor Dr. Andrei MARIN Universitatea din Bucureti
Profesor Dr. Ioan BURZO Universitatea Agronomic i de Medicin Veterinar din
Bucureti
Profesor Dr. Toader CHIFU Universitatea Alexandru Ioan Cuza din Iai
Profesor Dr. Mihai MITITIUC Universitatea Alexandru Ioan Cuza din Iai
Profesor Dr. Ursula STNESCU Universitatea de Medicin i Farmacie Gr. T. Popa din Iai
Confereniar Dr. Ctlin TNASE Universitatea Alexandru Ioan Cuza din Iai
Redactor responsabil: Profesor Dr. Constantin TOMA,
membru corespondent al Academiei Romne
Secretar de redacie: ef lucrri Dr. Naela COSTIC
Tehnoredactare computerizat: Dr. Ramona Crina GALE
CONTENTS
IRINA STNESCU, C. TOMA The leaf structure of some Nepenthes Danser
species
IRINA BERCIU, C. TOMA Histo-anatomical aspects reffering to the

vegetative organs of two subspecies of Thymus pannonicus All.
16
LUMINIA HUANU-BASHTAWI, C. TOMA Contributions to the histoanatomical study of the Calendula officinalis L. leaves treated with thiophanate
methyl (topsin M)
22
IOANA MARCELA PDURE, LILIANA BDULESCU, TEODORA DEDIU,
I. BURZO Morpho-anatomical and phytochemical researches regarding
Pseudotsuga menziesii (Mirbel) Franco (Pinaceae)
33
ALEXANDRINA MURARIU, CORINA GRDINARIU, ANIOARA
STRATU The ecophysiological reaction of some varieties of apple tree, pear
tree and quince tree to the pathogenic agents attack
40
ELENA CRISTINA ROU, MARIA MAGDALENA ZAMFIRACHE, I. I.
BRA - Physiological effects induced by purinic substances at Capsicum
annuum L.
46
M. RCA, L. FRTI, ANA LEAHU The influence on the Mn2+ ions
effects on the wheat (Triticum aestivum L. ) seed germination
50
NICOLETA IANOVICI, I. E. JUHSZ, P. RADISIC, M. JUHSZ, B.
SIKOPARIJA - Plantago atmospheric pollinic season in the Danube-KrisMures-Tisza euroregion (2000-2004)
54
L. POP, DORINA CACHI Araucaria excelsa L. vitrocultures initiation
64
ADRIANA PETRU VANCEA, C. F. BLIDAR, ANCA BACIU African

violet (Saintpaulia ionantha L.) exvitroplantlets acclimatization, in different
types of substratum
71
TATIANA EUGENIA ESAN, J. KHL, WILMA M. L. MOLHOECK Ulocladium atrum preuss - biological control agent of grey mould (Botrytis
cinerea Pers.) of cropped plants
78
M. COSTIC, NAELA COSTIC - A new site in Romania for Spirulina
92
(Arthrospira) platensis
T. CHIFU, C. MNZU, OANA ZAMFIRESCU Contribution to the study of

grassy vegetation in the Ceahlu Mountain
94
J. HANGANU, M. DOROFTEI, N. TEFAN - Assesment of ecological
status of Danube Delta lakes using indicator macrophytes species
103
OANA ZAMFIRESCU The plant communities with Phragmites australis
from 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 vegetation
from Neagra arului rivers basin (Suceava county)
121
CARMEN AONCIOAIE Protected taxa from the Bistria river basin between
Piatra Neam and Bacu
129
MIHAELA AURELIA DANU, T. CHIFU Contribution to the study of the
class Molinio-Arrhenatheretea R. Tx. 1937 in the upper basin of river Dorna
(Suceava county) (I)
136
I. M. CIUMAU, NAELA COSTIC - Environmental education: education for
transition to sustainable development
146
C.TOMA, MARIA MAGDALENA ZAMFIRACHE - Review
153
C. DRGULESCU - Review
154
Analele tiinifice ale Universitii Al. I. Cuza Iai

Tomul LIV, fasc. 1, s. II a. Biologie vegetal, 2008
THE LEAF STRUCTURE OF SOME NEPENTHES DANSER SPECIES

IRINA STNESCU , 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 SE Africa, Sri-Lanka and Madagascar. Etienne de Flancourt described it in 1658 for the first
time [3] and in 1753 Linn 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 histoanatomical aspects were evidenced in a previous work [9] devoted to Nepenthes maxima.
Botanical Gardens of Iasi, Dumbrava Roie Street, no. 7-9, Romania
Al. I. Cuza University, Faculty of Biology, Carol I. Bd., no. 20A, 700506, Iai, Romania
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.
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
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 pitchers 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.
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.
2.
3.
4.
5.
6.
7.
8.
9.
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.
DALTON M. JOS., 1859 - Note sur l origin et le dveloppement des urnes dans les plantes du genre
Nepenthes. Ann. des Sci. Nat.; sr.Bot., 12: 125-129
LLOYD F. E., 1942 - The Carnivorous Plants. Chronica Botanica, 9. Ronald Press, New York
MACFARLANE J. M., 1889 - Observations on pitchered insectivorous plants. Part I. Ann. of Bot., 3: 253265
METCALFE C. R., CHALK L., 1972 - Nepenthaceae in Anatomy of the Dicotyledons. 1: 1105-1111,
Clarendon Press, Oxford
STERN K., 1917 - Contribution to the knowledge of Nepenthes. Flora, 109: 213 283
PARKES D. M., 1980 - Adaptive mechanisms of surfaces and glands in some carnivorous plants. MSc
Thesis, Monash University, Clayton, Victoria, Australia
STAROSTA P., LABAT J.-J., 1993. - Lunivers des plantes carnivores. d. Du May, Paris
TOMA I., TOMA C., STNESCU I., 2002 - Histo-anatomical aspects of the Nepenthes maxima Reinw. ex
Nees metamorphosed leaf. Rev. Roum. Biol., sr. Biol. vgt., 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 Rnba for supplying the material for the present
investigation.
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
10
IRINA STNESCU, C.TOMA
PLATE I
11
PLATE II
10
11
12
12
PLATE III
13
14
15
16
17
18
13
PLATE IV
19
20
21
22
23
24
14
PLATE V
25
26
27
28
15

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.
Introduction 1
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) So - 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 Rugin 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 (Neam district) and T. Pannonicus ssp.
auctus, collected on 16.07.2007 from Flticeni (Suceava district).
Al. I.Cuza University, Faculty of Biology, Carol I Bd., no. 20A, 700506, Iasi, Romania
16
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.
17
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 bilayered, 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.
18
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.
2.
3.
4.
5.
CIOCRLAN V., 2000 - Flora ilustrat a Romniei. Pteridophyta et Spermatophyta. Ed. Ceres, Bucureti:
670-675
GUULEAC M., 1961 - Thymus, In Flora Republicii Populare Romne, VIII, Ed. Acad. RPR, Bucureti:
301-334
METCALFE C.R., CHALK L., 1950 - Anatomy of Dicotyledons, vol II, Clarendon Press, Oxford: 10411053
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
TOMA C., RUGIN R., 1998 - Anatomia plantelor medicinale. Atlas. Ed.Acad. Rom., Bucureti: 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=50m)
6. Aculeiform trichome of T. pannonicus ssp. auctus (scale bars=50m)
7. Multicellular secretory trichome of T. pannonicus ssp. auctus (x400)
8. Tector and glandular trichome of T. pannonicus ssp. pannonicus (x400)
19
IRINA BERCIU, C. TOMA
PLATE I
1.
20
IRINA BERCIU, C. TOMA
PLATE II
3
6
21

CONTRIBUTIONS TO THE HISTO-ANATOMICAL STUDY OF THE

CALENDULA OFFICINALIS L. LEAVES TREATED WITH THIOPHANATE
METHYL (TOPSIN M)
LUMINIA HUANU-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 fungicides 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, Iai , Romania
22
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 Ftu 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
23
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 limbs structure being bifacial heterofacial, as in the
0.1% treatment (Fig. 11, 12).
24
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.
25
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
fungicides 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 fungicides 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).
26
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.
HUANU-BASTAWI LUMINIA, 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 - Iai, Romnia, 28th 31st of May 2006 :126-132
3.
PETRCZI, M.I., MATUZ, J., KTAI 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: 3137
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., SCHLSSER 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., NAKSUL 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)

Diameter
Thickness
No.
of vessels
No. of
Variant
Length/width (m
woody
fascicles
(m Oc.10
Oc.10 x Ob.4)
vessels
x Ob.40)
Ca. of. M
1200/10751500/1400
1
78-89
15-22,5
Ca. of. TM 0.1 %
1850/23501925/2475
3
81-94
22,5-32,5
Ca. of. TM 0.4 %
2075/33252250/3500
3
111-115
22,5-30
Table II. Numerical values - middle leaves (median nervure, middle level)
Diameter
Thickness
No.
of vessels
No. of
Variant
Length/width (m
woody
fascicles
(m Oc.10
Oc.10 x Ob.4)
vessels
x Ob.40)
Ca. of. M
1375/7501500/1000
1
80-88
22,5-27,5
Ca. of. TM 0.1 %
1700/22501800/2600
3 -5
95-109
25-32,5
Ca. of. TM 0.4 %
2000/19002200/2075
2-3
132-139
22,5-27,5
27
Table III. Numerical values - superior leaves (median nervure, middle level)
Diameter
Thickness
No.
No. of
of vessels
Variant
Length/width (m
woody
fascicles
(m Oc.10
Oc.10 x Ob.4)
vessels
x Ob.40)
Ca. of. M
950/975-1000/1000
1
50-56
20-22,5
Ca. of. TM 0.1 %
1075/10501250/975
1
61-72
25
Ca. of. TM 0.4 %
1375/12001600/1450
2
68-88
25-30
Table IV. Numerical values - basal leaves (limb, middle level) (Oc.10 x Ob.20)
Thickness of the
No. of layers in
Thickness of the
Variant
limb (m)
the limb
pallisade (m)
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)
Thickness of the
No. of layers in
Thickness of the
Variant
limb (m)
the limb
pallisade (m)
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)
Thickness of the
No. of layers in
Thickness of the
Variant
limb (m)
the limb
pallisade (m)
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
No. of
No. of
Stomatic
No. of
No. of
Stomatic
Samples
cells
stomatae
index
cells
stomatae
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
28
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
Ca. of. M
Ca. of. TM 0.1%
Ca. of. TM 0.4%
Fig. 3. Cross-sections median conducting fascicle, basal leaf, middle level
Ca. of. M
Ca. of. TM 0.1%
Ca. of. TM 0.4%
Fig. 4. Cross-sections median nervure of the limb, middle leaf, middle level
29
Ca. of. M
Ca. of. TM 0.1%
Ca. of. TM 0.4%
Fig. 5. Cross-sections median conducting fascicle, middle leaf, middle level
Ca. of. M
Ca. of. TM 0.1%
Ca. of. TM 0.4%
Fig. 6. Cross-sections median nervure of the limb, superior leaf, middle level
Ca. of. M
Ca. of. TM 0.1%
Ca. of. TM 0.4%
Fig. 7. Cross-sections median conducting fascicle, superior leaf, middle level
Ca. of. M
Ca. of. TM 0.1%
Ca. of. TM 0.4%
Fig. 8. Cross-sections median conducting fascicle, superior leaf, middle level
30
Ca. of. M
Ca. of. TM 0.1%
Ca. of. TM 0.4%
Fig. 9. Cross-sections limb, basal leaf, middle level
Ca. of. M
Ca. of. TM 0.1% Ca. of. TM 0.4%
Fig. 10. Cross-sections mesophyl, basal leaf, middle level
Ca. of. M
Ca. of. TM 0.1%
Ca. of. TM 0.4%
Fig. 11. Cross-sections limb, middle leaf, middle level
Ca. of. M
Ca. of. TM 0.1% Ca. of. TM 0.4%
Fig. 12. Cross-sections mesophyl, middle leaf, middle level
31
Ca. of. M
Ca. of. TM 0.1% Ca. of. TM 0.4%
Fig. 13. Cross-sections mesophyl, superior leaf, middle level
b
Fig. 14. Calendula officinalis: tectory hairs (a), uni- and biseriated secretory hairs (b)
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
32

MORPHO-ANATOMICAL AND PHYTOCHEMICAL RESEARCHES

REGARDING PSEUDOTSUGA MENZIESII (MIRBEL) FRANCO (PINACEAE)
IOANA MARCELA PDURE *, LILIANA BDULESCU, 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, -pinene, -terpinen, -phellandrene and -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.) Carrire] 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 2ovulated. 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, Mrti 59 Bd., 011464, Bucharest, Romania
33
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: -pinene, sabinene, -ocimene, -terpinolene, -terpineol, citronellyl
acetate, -terpinene, limonene and -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 250C, interface
280C and ionization source 200C; 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.
34

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 m)
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 m) 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
Needle
1. Stomata length (40x)
2. Stomata wide (40x)
3. Number of stomata / mm2
4. Stomatiferous zone wide (10x)
5. Number of stomata strings / on
leaf wide (10x)
6. Midvein height, including
endodermis (10x)
7. Vascular bundle height (10x)
8. Vascular bundle wide (10x)
9. Resin ducts diameter (10x)
10. Mesophyllum height (10x)
11. Palisade tissue height (10x)
12. Number of palisade cell layers
13. Septate cells height
14. Number of septate cell layers
15. Xylem thickness (40x)
16. Sclerenchyma thickness (40x)
17. Endodermis thickness (40x)
18. Phloem thickness (40x)
19. Adaxial epidermis thick (40x)
20. Abaxial epidermis thick (40x)
21. Cuticle thickness (40x)
Young shoot
22. Resin ducts number / leaf section
23. Secretory hair length (10x)
24. Xylem thickness (primary and
secondary xylem) (10x)
25. Resin ducts diameter (10x)
26. Cortex length (10x)
27. Number of cell layers in cortex
28. Phloem thickness (primary and
secondary phloem) (10x)
29. Annual ring 2004 thickness (10x)
Minimum value
[m]
Ecotype
1
2
19,4
17,8
16,2
13
266
207
206,2
200
4
4
Maximum value
[m]
Ecotype
1
2
32,4
32,4
19,4
24,3
414
335
281,2
312,5
6
7
Average value
[m]
Ecotype
1
2
24,3
18,5
17,8
18,5
239
257,8
5
6
187,5
268,7
243,5
293,7
200
282,6
125
162,5
31,3
331,3
93,75
1
187,5
4
19,4
12,9
8,1
19,44
6,4
4,8
1,62
218,7
185,3
31,25
500
143,7
2
281,2
3
40,5
29,1
9,7
23,4
9,7
6,5
6,5
168,7
218,7
62,5
375
156,2
3
250
7
43,74
32,4
27,5
59,9
8,1
6,4
8,1
231,2
220
68,7
562,5
218,7
3
406,2
6
65
42,1
39
40,5
24,3
11,34
11,3
143,8
184,4
45
347,9
120,6
2
226,2
5
30,29
25,5
15,4
42,28
7,2
5,6
5,7
220,8
203,5
51,3
534,4
172,2
2
348,2
5
48,3
33,7
22
32,6
14
9
9
7
19
375
6
31,2
75
10
125
419
11
156
156
8
62,5
400
8
106,2
106,2
37,5
162,5
5
125
25
375
7
12,5
137,5
394
7
187,5
56,2
500
12
25
70,3
255
6
157
42,5
411
10
20
56,2
125
87,5
187,5
70
144,6
35

32. Primary xylem thickness (10x)
33. Pith diameter (10x)
34. Epidermis thickness (40x)
35. Subepidermic cell layers number
36. Bark thickness (40x)
37. Number of bark cell layers
38. Pheloderm thickness (40x)
39. Number of pheloderm cell layers
125
75
31,2
375
8,1
2
11,3
4
11,3
2
6,25
437,5
4,8
4,8
1
48,6
3
150
125
50
500
13
2
24,3
4
21
3
31,2
500
9,7
9,7
3
64,8
4
135,4
99
40,6
441,6
10
2
17,4
4
18,3
3
18,7
465,6
7,2
7,2
2
58
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%), -pinene (3.88-9.23 %) in needle
oil and -pinene (3.88-9.23%), -pinene (12.6-15.22%), -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- -pinene 9.23% (chemotype 1, April), sabinene 31.73% (chemotype 1, March),
-pinene 30.28% (chemotype 2, April), -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 pinene 10.6 % and sabinene 12.3% (chemotype 2, April), -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
-thujen
-pinene
Camphene
Sabinene
-pinene
-myrcene
-phellandrene
p-cymene
-terpinene
Cymol
Limonene
Eucalyptol
Cis -ocimene
-phellandrene
CHEMOTYPE 1
October
2005
1.84
3.88
0.15
19.16
7.23
1.23
0.64
1.01
5.64
0.93
1.93
0.18
0.102
8.09
January
2006
1.95
7.31
0.32
24.36
21.22
1.64
0.3
0.69
3.07
0.33
2.12
0.24
0.07
4.79
March
2006
2.00
4.72
0.14
31.73
9.22
1.43
0.41
1.21
4.09
0.32
1.68
0.17
0.13
6.4
36
CHEMOTYPE 2
April
2006
1.82
3.9
0.15
29.99
7.22
1.37
0.42
1.17
4.62
0.5
1.7
0.25
0.14
7.17
January
2006
1.97
4.44
0.13
29.57
7.94
1.39
0.48
1.15
4.74
0.51
1.73
0.19
0.11
7.14
April
2006
1.81
9.23
0.48
14.34
30.28
1.85
0.37
0.64
3.31
0.29
2.49
0.29
0.09
5.11
Trans -ocimene
Terpinolene
Linalool
Fenchol
Citronellal
Isoborneol
Terpinen-4ol
-terpineol
Cis-piperitol
L-terpinen 4-ol
propenil-anisol
Trans-piperitol
Citronellol-acetate
Selinenol
Total
0.16
20.31
0.23
0.41
0.24
0.11
14.82
0.80
0.26
0.53
0.17
0.95
0
0.08
91.08 %
0.41
16.05
0.34
0.37
0.25
0.23
7.53
0.71
0.29
0.26
0.24
2.27
0.81
0.24
98.41 %
0.3
22.99
0.27
0.41
0.25
0.11
9.03
0.47
0.17
0.47
0.07
1.24
0
0.13
99.56 %
0.35
23.23
0.31
0.52
0.33
0.12
11.49
0.61
0.23
0.38
0.14
1.03
0.07
0.1
99.33 %
0.19
23.06
0.21
0.44
0.28
0.18
10.68
0.55
0.25
0.49
0.09
1.14
0
0.08
99.13 %
0.24
12.71
0.3
0.37
0.23
0.18
9.16
1.13
0.27
0.25
0.32
2.12
0.8
0.15
98.81 %
Table III Chemical composition of essential oils in young shoot of Pseudotsuga menziesii
Chemical components
-thujen
-pinene
Camphen
Sabinene
-pinene
-myrcene
-phellandrene
-terpinene
P-cymene
Cymol
Limonene
Cis -ocimene
Trans -ocimene
Terpinolene
Linalool
- terpineol
terpinen-4ol
-terpineol
Propenil-anisol
-cubebene
Isoeugenol
-cadinene
Selinenol
Cembrene
Kauren
Norkauren
Total
CHEMOTYPE 1
January 2006
1.16
7.89
0.25
9.46
12.6
2.52
0.39
21.05
2.39
0.68
3.15
0.29
3.98
15.4
0.84
0.08
7.12
1.07
0.61
1.32
1.05
0.22
0.42
0.27
0.34
1
95.55 %
CHEMOTYPE 2
January 2006
April 2006
1.38
1.29
10.47
10.6
0
0
5.65
12.3
14.89
15.22
2.45
2.66
0.304
0.27
24.56
32.87
2.7
1.74
0.59
0.71
3.78
3.13
0.77
0.25
4.48
2.85
14.61
13.7
0
0.13
0.22
0.36
2.77
1.96
0.4
0.22
0.27
0.74
0.12
1.76
0
0.09
0
0.205
0.25
0.39
0
0.28
0
0.36
0
1.023
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%), -pinene (3.88-9.23 %) in needle
37
oil and -pinene (3.88-9.23%), -pinene (12.6-15.22%), -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%, -pinene 30.28% terpinen-4ol 14.82% and terpinolene
23.23%. In the oils of young shoots the most important components are represented by pinene 10.6 % and sabinene 12.3%, -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 Douglasfir (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: 586588.
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: 761768
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
38
B
epidermis
endodermis
midvein
secretory duct
Fig. 1 The anatomy of needle (A) and median vein (B) in P. menziesii [ob. 6x / 25x, oc. 12,5x, orig.]
phloem
cortex
cambium
B
medullary rays
xylem II
xylem I
pith
pith
xylem
xylem II (I II III: annual rings)
Fig. 2 The anatomy of shoots in P. menziesii: cortical cells (A) and central cylinder zone (B)
[ob. 6x / oc. 12,5x, orig.]
epidermis
epidermis
phelogen
bark
secretory cells
resin ducts from stem
cortex
chlorenchyma
Fig. 3 Pseudotsuga mensiesii needle (A) and cortical resin ducts of stem (B)
[ob. 40x, oc. 12,5x, orig.]
39

THE ECOPHYSIOLOGICAL REACTION OF SOME VARIETIES OF APPLE

TREE, PEAR TREE AND QUINCE TREE TO THE PATHOGENIC AGENTS
ATTACK
ALEXANDRINA MURARIU *, CORINA GRDINARIU *, ANIOARA 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 Iai. 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), Mona and De Hui (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 Hui (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 Mona (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. Svulescu (1938). In 1992, two
contagion centers were identified in Brila and Mrcineni, 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, Iai, Romania
40
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 Iai.
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), Mona and De Hui (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 (60C 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 (%)
Malus domestica
Jonathan
Generos
Williams
Argensis
Mona
De Hui
5.0
1.0
12.0
7.0
38.0
8.0
Pyrus communis
Cydonia oblonga
* after Cimpoe, 2001 and Grdinaru, 2002
41
Resistance
category*
Susceptible
Resistant
Susceptible
Very susceptible
Very susceptible
Susceptible

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 Mona 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 (Mona) 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 Hui).
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 Hui 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 (Mona 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 (Mona
variety).
42
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%
(Mona 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 Hui (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 Mona
(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.
2.
3.
4.
5.
6.
7.
8.
9.
BRANITE N., ANDRIE N., 1990 - Soiuri rezistente la boli i duntori n pomicultur. Ed. Ceres.
Bucureti
CIMPOE GH., BACARCIUC V., CAIMACAN I., 2001 - Soiuri de mr. Ed. I. E.P. tiina, Chiinu.
GRDINARU G., 2002. Pomicultura special. Ed. Ion Ionescu de la Brad, Iai
MAC HARDY, W.E. 1996 - Apple scab - Biology, Epidemiology and Management, APS Press, Sant Paul,
Minnesota
MITITIUC M., 1994 - Fitopatologie. Ed. Universitii Al. I. Cuza, Iai
OKTEM, Y.E.; BENLIOGLU, K. , 1988 - Studies on fireblight [Erwinia amylovora (Burr.) Winsl. et al.] of
pome fruits. Journal of Turkish Phytopathology, 17, 106
SEVERIN V., 1996 - Focul bacterian al rozaceelor (Erwinia amylovora). Ed. Ceres, Bucureti
VAN DER ZWET T., KEIL H. L., 1979 - Fire Blight: A Bacterial Disease of Rosaceous Plants. United
States Department Agriculture Handbook, 510, Washington DC
YOSHKAWA M., 1983 - Macromolecules, recognition and the traggerin of resistance. In :Biochemical Plant
Pathology (edited by: Callow J. A.), Plant Physiology, 73: 497- 506
43
Gross
Photosynthesis
Dry substance (%)
Organic
(%)
substance
Respiration
De Hui
Net
Photosynthesis
Mona
Total
Cydonia
oblonga
Carotenoids
Argensis
Chlorophyll b
Williams
Chlorophyll a
Pyrus
communis
Minerals
Generos
Transpiration (mg/h)
Jonathan
Photosynthesis (mg CO2/g/h)
Water (%)
Variety
Malus
domestica
Assimilatory Pigments (mg/g fresh

substance)
Strength of Attack
(%)
Species
Table II. The ecophysiological reaction of some varieties of apple tree, pear tree and quince tree to Erwinia amylovora in 2004
Healthy
5%
Healthy
61.33
57.99
60.16
62
58
25
5.53
6.11
6.43
3.27
1.36
1..60
0.58
0.26
0.29
0.30
0.18
0.25
4.15
1.80
2.14
5.148
2.624
2.496
1.072
1.552
0.374
6.22
4.176
2.87
38.67
42.01
39.84
33.14
35.90
33.41
1%
Healthy
12%
Healthy
7%
Healthy
38%
59.29
56.44
51.28
61.53
58.98
54.64
13.24
14
22
12
198
144
71
44
7.86
4.50
5.22
4.27
6.40
9.33
6.83
1.22
1.97
1.45
1.53
1.42
1.53
1.24
0.21
0.45
0.25
0.25
0.24
0.25
0.19
0.16
0.21
0.18
0.16
0.14
0.20
0.18
1.59
2.63
1.88
1.94
1.80
1.98
1.61
3.120
2.496
0.624
3.744
0.624
1.248
0.624
0.561
0.963
1.360
1.248
0.936
0.744
1.616
3.681
3.432
1.984
4.992
1.560
1.992
2.240
40.71
43.56
48.72
38.47
41.02
45.36
86.76
32.85
39.06
43.50
34.20
34.62
36.03
79.93
Healthy
8%
58.23
56.63
79
45
6.80
8.42
1.65
1.81
0.24
0.33
0.20
0.23
2.09
2.37
1.872
2.496
0.744
0.936
2.616
3.432
41.77
43.37
34.97
34.95
44
90
80
70
60
50
40
30
20
10
0
Healthy
Strength Healthy Strength Healthy Strength Healthy Strength Healthy Strength Healthy Strength
of Attack
of Attack
of Attack
of Attack
of Attack
of Attack
5%
1%
12 %
7%
38 %
8%
Jonathan
Generos
Malus domestica
Water content ( %)
Williams
Argensis
Pyrus communis
Total mineral elements content ( %)
Fig.1. The physiological modifications in leaves of studied species
45
Mona
De Hui
Cydonia oblonga
Organic substance content ( %)

PHYSIOLOGICAL EFFECTS INDUCED BY PURINIC SUBSTANCES AT

CAPSICUM ANNUUM L
ELENA CRISTINA ROU , MARIA MAGDALENA ZAMFIRACHE, I. I. BRA
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.
46
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).
47
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
48
Fig 3. The percent of water and the dry substance after the tretment with coffeine (left) and
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 dozeeffect 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.
2.
BRA I. I., CMPEANU MIRELA, 2003 - Genetica, Edit. Corson, Iai: 208-209
DIACONU P., 1971 - Ereditatea i factorii mutageni, Edit. Ceres, Bucureti
49

THE INFLUENCE OF THE Mn2+ IONS EFFECTS ON THE WHEAT (TRITICUM

AESTIVUM L.) SEED GERMINATION
I. M. RCA , L. FRTI, 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 seeds 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
tefan cel Mare University, Universitii 13 Street, Suceava, Romania
50
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
2+
Mn
witness
1m
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
60,29
8,78
20,35
51,72
50,02
51,02
53,09
62,50
60,29
1,00
1,00
8,69
26,71
56,82
65,27
64,60
measured parameter
(average values)
hypocotyls length 1h
(mm)
hypocotyls length 7 d
(mm)
51
120
100
80
7 days
1 hour
60
40
20
0
1M
0,5 M
0,1 M
0,05 M
0,01 M
0,005 M
0,001 M
Fig. 1: Germination analysis

70
60
50
40
7 days
1 hour
30
20
10
0
1M
0,5 M
0,1 M
0,05 M
0,01 M
0,005 M
Fig. 2: Hypocotyl length analysis
52
0,001 M
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 longterm 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.
2.
3.
4.
5.
CRICHTON, R., 2001 - Inorganic Biochemistry of Iron Metabolism. John Wiley & Sons, Ltd. Chichester New York Weinheim Brisbane Singapore Toronto
DAVIDESCU D., DAVIDESCU VELICICA, LCTUU R., 1988 - Microelementele n agricultur. Edit.
Acad. Rom., Bucureti, 280 p.
Institutul Romn de Standardizare, 1999 - Semine pentru nsmnare. Determinarea germinaiei. SR1634:
iunie 1999, Bucuresti
KHAN A. A. EDITOR, 1980 - The physiology and biochemistry of seed dormancy and germination. New
York, Oxford
ROAT-MALONE, ROSETTE M., 2002 - Bioinorganic Chemistry A short course. John Wiley & Sons,
Inc., Hoboken, New Jersey
53

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

NICOLETA IANOVICI , I. E. JUHSZ, P. RADISIC, M. JUHSZ,
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 timioara (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
Timioara they are slightly on the increase. The presence of the pollen in the airplankton was considerably
long in Timioara, 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 10300 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
54
These authors also reported in the same article that IgE from monosensitized plantainallergic 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 3236 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 m 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: Bcs Kiskun, Csongrd, Jsz-Nagykun-Szolnok, Bks (Hungary),
Arad, Timi, Hunedoara, Cara Severin (Romania), and Vojvodina (Serbia).
55
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 Timioara 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
Timioara, 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 Timioara
were relatively similar: 797 PG/m3 and 669 PG/m3. In Timioara, 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 Timioara, representing 2.67% of the total annual of 24557
PG/m3. In 2004, the highest concentration (486 PG/m3) was recorded in Timioara. 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 Timioara. 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].
56
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 Timioara, 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
Timioara 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 Timioara. 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
Timioara and Novi Sad. Similar variations were recorded in Poland: 95 to 105 days in
Rzeszw, 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)
57
Tabel I. The duration of the atmospheric pollinic season in 2004

2004
First identification of the airpollen
Last identification of the airpollen
Duration of the atmospheric pollinic season
(Nilsson & Persson, 1981); 90%
Duration of the atmospheric pollinic season
(Andersen, 1991; Torben, 1991); 95%
Novi Sad
7V
21 IX
89 days
Ruma
4V
19 IX
108 days
Szeged
15 V
28 VIII
73 days
Timisoara
1V
13 IX
95 days
107 days
128 days
85 days
109 days
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
58
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 Mlaga 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 Mlaga [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
59
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. Crdoba 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
60
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
Timioara 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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63

ARAUCARIA EXCELSA L. VITROCULTURES INITIATION

L. POP ; DORINA CACHI*
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 wasnt 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, Universitii Street, no.1, 410087, Oradea, Romania
64
inoculation
Original plant
4 cm
Murashige and Skoog medium
1 cm
Explant
1 cm
Fig. 1. Making minicuttings and the inoculation process

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 121C, 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 20-22C, 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 didnt 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)
65
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)
%
100
100
95,2
93,1
91,66
84,61
80
60
variants
V0
V1
V2
V3
V4
Fig. 3. The survival percent of Araucaria excelsa L. plantlets, at 30 days after

inoculation
66
1.2
1.18
1.16
1.14
1.12
1.1
1.08
1.06
1.04
Stalk length
V0
V1
V2
V3
V4
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
67
2,5
Stalk le ngth
2
1,5
1
0,5
0
V0
V1
V2
V3
V4
Ram ifications
5
4
3
2
1
0
V0
V1
V2
V3
V4
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) hasnt
manifested any ramification (fig.8, fig.10).
No one of experimental variants has manifested rhysogenesis.
68
Fig. 8. Araucaria excelsa L. cultures after 90 days from inoculation (V0 (control
Stalk le ngth
2,5
2
1,5
1
0,5
0
V0
V1
V2
V3
V4
69
Ramifications
5
4
3
2
1
0
V0
V1
V2
V3
V4
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 benzyladenine; IBA indolilbutilic acid; NAA -naftilacetic acid; KIN kinetin
REFERENCES
1.
2.
3.
4.
CACHI, C.D., 1987 - Metode in vitro la plantele de cultur. Bazele teoretice i practice, Ed. Ceres,
Bucureti
CACHI, C. D., SAND, C., 2000 - Biotehnologie vegetal. Baze teoretice i practice. Vol.1, Ed. Mira
Design, Sibiu: 272-276
MURASHIGE, T., SKOOG, F., 1962 - A revised medium for rapid growth and bioassays with tobacco tissues
cultures. Physiologia Plantarum, 15: 155-159
PREDA, M, 1979 - Floricultura (ed. a II-a), Edit. Ceres, Bucureti: 231-232
70

AFRICAN VIOLET (SAINTPAULIA IONANTHA L.) EXVITROPLANTLETS

ACCLIMATIZATION, IN DIFFERENT TYPES OF SUBSTRATUM
ADRIANA PETRU 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, Universitii Street, no. 1, 410087, Oradea, Romania
Potato Research & Development Station from Trgu Secuiesc, Romania
71
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 2C in the light period and
20C 2C 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%.
72

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
73
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. PETRU - VANCEA ADRIANA, 2005 - The acclimatization of African violet exvitroplantlets in substratum
consisting in sawdust. Analele Univ. Oradea, fasc. Biol., t. XII: 119 123
3. PETRU VANCEA ADRIANA, CACHI C. DORINA, IPO MONICA, 2004 - Activitatea
peroxidazic n rdciniele vitro- i exvitroplantulelor de crizanteme, violete africane i de Cymbidium. An.
SNBC, t. IX, 1., cap. III Biologie celular vegetal: 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, CACHI C. DORINA, 2002 - Aclimatizarea vitroplantulelor de Saintpaulia
ionantha, prin plantarea acestora pe substraturi neconvenionale. n: Lucrrile celui de al X-lea Simpozion
Naional de Culturi de esuturi i Celule Vegetale. (eds. Cachi C. Dorina, Rakosy T. Lenua, Ardelean A.).
Edit. Risoprint, Cluj- Napoca: 310 315
6. VANCEA ADRIANA, CACHI C. DORINA, 2002 - Utilizarea biogelului n substratul destinat
aclimatizrii vitroplantulelor de saintpaulia ionantha la mediul septic de via. n: Lucrrile Simpozionului
tiinific 90 de ani de nvmnt agronomic universitar la Iai. CD, Agrosoft U.S.A.M.V., Iai
7. VANCEA ADRIANA, CACHI C. DORINA, FENCE DANIELA, 2003 - Utilizarea zeolitului ca substrat
de aclimatizare a vitroplantulelor la mediul septic de via. n: Lucrrile celui de al XI-lea Simpozion Naional
de culturi de esuturi i celule vegetale. (eds. Cachi C. Dorina, Ardelean A. Edit. Daya, Satu-Mare: 212 224
74
V0
V1
V2
V3
V4
V5
V6
250%
200%
150%
I zi
100%
50%
0%
Survival
percent
Rootlets
length
Rootlets
number
Propaguls
number
Total
Leaflets
Leaflets
number of number with number with
leaflets
0-0,4 cm
0,5-0,9 cm
diam.
diam.
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%
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
M
Determinations
eather
Substratum type
river sand
+ fallow soil, 3:1
white peat
+ manure + river
sand, 1:1:1
perlite +
white peat, 1:1
river sand
+ white peat, 1:1
manure + white peat,
1:1
Top soil
mid.
2
2O5
aO
gO
ineral
residue
%
g/100
0
1
,13
,14
,15
,18
,25
,14
,1
,4
,3
,2
,1
,9
Normal values
5 - 30
- 12
0 - 60
0 - 50
0
0
0
0
0
0
- 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
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.
Leaflets
number with
1,0-1,4 cm
diam.
Leaflets
number with
1,5-1,9 cm
diam.
Statistic
count
1
76
Sx
p
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
ns
***
ns
***
**
***
ns
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
***
***
ns
***
***
ns
***
ns
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
***
***
ns
***
**
ns
***
***
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
***
***
ns
***
***
ns
***
ns
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
***
***
ns
***
**
ns
***
ns
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
***
***
ns
***
ns
***
***
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
***
***
ns
***
ns
***
***
***
Type V1
1
Sx
p
Type V2
Sx
p
Type V3
Sx
p
Type V4
Sx
p
Type V5
Sx
p
Type V6
Sx
p
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
77

ULOCLADIUM ATRUM PREUSS - BIOLOGICAL CONTROL AGENT OF GREY

MOULD (BOTRYTIS CINEREA PERS.) OF CROPPED PLANTS
TATIANA EUGENIA ESAN , J. KHL , 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. Khl. 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
78
10m
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.
79
year
1997-1
1997-2
1997/
1998-a
19971998-b
1998-1
1998-2
1998/
1999
1999
Table I. Samples from strawberry trials 1997-2000

treatments
plot numbers
1.
2.
3.
4.
1.
2.
3.
4.
1.
2.
3.
4.
5.
1.
2.
3.
4.
5.
1.
2.
3.
4.
5.
1.
2.
3.
4.
5.
1.
2.
3.
4.
5.
Control
Ua weekly during season
fungicides
Ua weekly during flowering
Control
Ua weekly during season
fungicides
Ua weekly during flowering
Control
Ua monthly sept.(IX)-april (IV) (a)
removal senescing/dead leaves
Ua planned during flowering; not performed
Ua monthly sept.(IX)-june (VI) (b)
Control
Ua at planting; weekly from bud to flowering;
twice weekly during flowering
fungicides
Ua twice weekly at flowering
Control
Ua at planting; weekly from bud to flowering;
twice weekly during flowering
fungicides
Ua twice weekly at flowering
Control (Tween water)
fungicides
Ua fortnightly from winter to flowering; twice
weekly during flowering
Ua twice weekly during flowering
Control
Tween water every second day during
flowering
Ua 5x105 ml-1 every 4th day during flowering
Ua 5x105 ml-1 every 2nd day during flowering
fungicides
80
2,5,10,13,17
3,8,11,15,18
4,7,12,16,19
1,6,9,14,20
1,6,11,13,20
4,7,10,16,18
3,8,9,15,19
2,5,12,14,17
1,8,15,19
3,7,12,17
4,10,11,18
2,6,14,16
5,9,13,20
1,8,15,19
3,9,12,17
4,6,11,18
2,10,14,16
5,7,13,20
1,6,13,20
4,8,15,19
3,9,14,18
5,10,11,17
2,7,12,16
1,8,15,19
3,9,12,17
4,6,11,18
2,10,14,16
5,7,13,20
1,8,15,20
2,6,13,17
3,7,12,18
4,10,11,16
5,9,14,19
date of
collecting1)
18.07.2000
12.07.2000
28.06.2000
27.06.2000
26.06.2000
14.06.2000
08.06.2000
30.05.2000
1999/
1. Control
3,10,14,16
2. Ua December 1999 (a)
2,6,12,18
2000-a
3. Ua January 2000 (b)
5,9,13,20
2000-b
4. Ua March 2000
(c)
1,7,15,19
2000-c
5. Ua May 2000
(d)
4,8,11,17
2000-d
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 (potatodextrose-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 50C, 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.
81
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 ,
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 / 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.
82
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
) containing culture media (ARSA) (fig. 3).
Fig. 3 Laboratory preparing of suspensions from soil samples collected from

different strawberry experiments
83
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
Culture
media
MA
WA
ARSA
3
1.500
1.150
0.225
4
1.925
1.500
0.225
5
2.420
2.800
0.350
6
2.675
3.625
0.575
Days after inoculation:

7
9
12
2.875
3.175
3.200
5.000
6.900
7.900
0.725
1.000
1.150
13
3.450
8.050
1.200
Fig. 3a - Diameter (cm) of Ulocladium atrum colony on different culture media
10
MA
WA
ARSA
Colony diameter (cm)
8
6
4
2
0
0
10
15
Incubation time (days)
84
20
14
3.450
8.275
1.360
17
3.950
8.320
1.450
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
sieves) from strawberry plots treated with U. atrum 1 month before soil sampling
Experiment
1999-2000 ( i )
1999-2000 ( ii )
Culture media
MA
WA
ARSA
MA
WA
ARSA
Incidence of U. atrum %
0.5 mm
1.0 mm
20.83a
17.50b
a
19.17
5.83c
a
33.33
31.67a
a
10.0
6.67a
a
6.67
1.33b
a
8.00
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.
85
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
1999-2000 ( i )
Culture media
MA
WA
ARSA
Incidence of U. atrum (%)

38.23a
29.32a
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
Experiment
1997-1
1997-2
1997-1998 1998-1
a
b
Time span between last

U. atrum treatments
and sampling (months)
35
37
22
24
23
86
Incidence of U. atrum (%)

untreated
a1)
21
14a
38a
38a
74a
Ua treated
39a
44b
72b
87c
85b
24
62a
91b
a
13
55
86b
a
11
41
67b
a
a
6
19
36a
a
b
4
19
34a
a
c
2
19
45a
a
d
1
19
30a
1)
Figures for U. atrum-treatment in the same time with same latter are not different from
the control
1998-2
1998-1999
1999
1999-2000 -
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.
87
Table VI. Incidence of Ulocladium atrum (cfu/ml) in soil suspensions from strawberry plots
treated with U. atrum 1 37 months before soil sampling
Experiment

U. atrum treatments
1997-1
1997-2
1997-1998 -
35
37
22
24
23
24
13
11
6
4
2
1
1998-1
1998-2
1998-1999
1999
1999-2000 -
1)
a
b
a
b
c
d
Incidence of U. atrum (cfu/ml soil

suspension)
untreated
Ua treated
316.7a1)
346.7a
669.5b
669.5
742.5a
316.7a
341.7a
548.3a
168.3a
168.3a
168.3a
168.3a
418.7a
834.7b
765.2b
934.6c
1032.5b
725.0b
570.0a
810.0b
226.7a
262.5a
327.5a
276.5a
See table 5
Fig. 5 - Incidence of Ulocladium atrum (cfu/ml) in soil suspensions
88
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
Experiment

U. atrum treatments
1999-2000 -
6
4
2
1
a
b
c
d
1) See table 5
Incidence of U. atrum (cfu/ml soil

suspension)
untreated
Ua treated
27 a1)
27a
27a
27a
50ab
62bc
72bc
85c
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).
89
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., KHL 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. KHL 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., LOMBAERSVAN DER PLAAS KARIN, KHL 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: 377386
7.
KESSEL G.J.T., KHL 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.
KHL J., 1997-2000 - Contract FAIR-CT96-1898, BIOSPORSUPPRESS The biological control of airborne necrotrophic plant pathogens by suppression of spore production
9.
KHL 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. KHL 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. KHL 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. KHL 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
90
13. KHL 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. KHL 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. KHL 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. KHL J., 1999 - Biologische Bekmpfung 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. ESAN 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. ESAN T. E., 2005 - Bibliografia romneasc n domeniul combaterii biologice a micozelor plantelor.
Sntatea plantelor, ediie special: 15-22
22. ESAN 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 Khl, 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.
91

A NEW SITE IN ROMANIA FOR SPIRULINA (ARTHROSPIRA) PLATENSIS

M. COSTIC *, NAELA COSTIC *
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. alaru 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, Iai, Romania
92
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 Cri River (at Petea, Homorog), of
the Arge River (at Caldarusani, Chirnogi), of the Danube River (at Pardina and the Danube
Delta) [1].
Site from Tometi 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.
2.
3.
4.
5.
CRU I., 2002 - The Algae of Romania. St. Cerc., Univ. Bacu, (Biol.), 7: 1-694
CIFERRI O., 1983 - Spirulina, the edible microorganism. Microbiol. Rev., 47: 551- 578
LUCIA HELENA PELIZER et al ., 2003 - Influence of inoculum age and concentration in Spirulina
platensis cultivation. J. Food Engeneering, 56, 4: 371-375
KOMREK, J., LUND, J.W.G., 1990 - What is Spirulina platensis in fact ?. Algological Studies, 58: 1-13.
PASCHER A., 1925 - Cyanophyceae In: Pascher A.(ed) Suswasser- Flora Deutschlands, Osterreichs und der
Schweiz, Gustav Fischer, Jena.
93

CONTRIBUTION TO THE STUDY OF GRASSY VEGETATION IN THE

CEAHLU MOUNTAIN
T. CHIFU , C. MNZU*, OANA ZAMFIRESCU*
Abstract: This paper presents five types of grassland phytocoenoses, widely spread in the Ceahlu
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 Csrs 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, Ceahlu 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 (So 1969) Taber 1987, Symphyto cordati Fagetum Vida 1963,
Leucanthemo waldsteinii Fagetum (So 1964) Taber 1987, Hieracio transsilvanico
Fagetum (Vida 1963) Taber 1987, Geranio robertianae Fagetum (Burduja et al. 1974)
Chifu et tefan 1994, Galio schultesii Fagetum (Burduja et al. 1973) Chifu et tefan 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 Csrs Kaptalan 1964 and Poo Trisetetum
Knapp ex Oberd. 1957 associations [6].
For the study of vegetation in the Ceahlu 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, Iai, Romania
94

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 Ceahlu 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 Ceahlu
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 Ceahlu, 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.
95
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 (Csrs 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 Csrs 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 Ceahlu 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.
96
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
Altitude (m. o. s.)
Exposure
1
620 - 690
S, SE
Slope (degrees)
0-5
Vegetation coverage (%)
90 - 100
No. of surveys
5
Associations characteristic species
Pastinaca sativa
I
Anthoxanthum odotarum
Festuca rubra
III
Agrostis capillaris
II
Poa pratensis
III
Subassociations differential species
Antennaria dioica
Nardus stricta
Potentilla aurea
Arrhenatherion
Arrhenatherum elatius
V
Campanula patula
V
Centaurea phrygia
IV
Daucus carota
I
Equisetum arvense
Geranium pratense
Pimpinella major
Taraxacum officinale
IV
Cynosurion
Bellis perennis
Cynosurus cristatus
III
Leontodon autumnalis
II
Lolium perenne
Phleum pratense
I
Plantago major
Trifolium repens
IV
Phyteumo Trisetion
Aegopodium podagraria
Gladiolus imbricatus
II
Hypericum maculatum
I
Trisetum flavescens
IV
Trollius europaeus
Veratrum album
I
Arrhenatheretalia
Achillea millefolium
III
Ajuga reptans
Avenula pubescens
Briza media
IV
Bromus hordeaceus
-
2
630 - 1230
NV, S
3b
970 1270
SV, S, NV,
V
0 10
95 100
8
4
530 780
SE, S
5
570 820
-
0 - 20
90 - 100
8
3a
600 - 1004
NV, NE, S,
SE, V, E
0 15
75 - 100
16
0 - 25
75 - 100
6
90 - 100
10
IV
IV
V
I
IV
V
V
-
IV
V
V
-
III
V
V
II
II
III
II
III
I
-
III
V
III
III
III
I
IV
IV
I
I
I
-
II
II
-
I
-
II
IV
IV
I
II
V
-
III
III
II
I
IV
III
II
I
II
III
V
II
I
V
III
III
III
II
III
I
II
III
IV
I
III
I
I
-
I
II
III
I
III
-
II
III
V
I
II
II
-
V
I
I
V
I
II
-
IV
IV
-
V
II
-
97
Campanula glomerata
II
III
III
III
Carum carvi
III
III
II
III
Crepis biennis
I
I
III
Dactylis glomerata
IV
II
II
Heracleum sphondylium
I
I
Holcus lanatus
I
IV
I
Knautia arvensis
I
I
III
Leontodon hispidus ssp. IV
III
hispidus
Leucanthemum vulgare
V
V
V
II
Luzula campestris
II
V
Medicago lupulina
IV
I
I
Rhinanthus minor
III
V
III
Stellaria graminea
I
Thymus pulegioides
III
III
V
V
Tragopogon
pratensis IV
I
III
ssp. orientalis
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
Carex tomentosa
I
Chaerophyllum hirsutum
I
Colchicum autumnale
III
I
III
Dactylorhiza maculata
I
Deschampsia caespitosa
I
I
I
IV
Dianthus superbus
I
Festuca pratensis
III
III
Filipendula ulmaria
I
Gymnadenia conopsea
I
I
III
Hypericum tetrapterum
I
Juncus articulatus
I
Laserpitium prutenicum
I
Linum catharticum
III
V
Lychnis flos-cuculi
I
Lysimachia vulgaris
I
Lythrum salicaria
Myosotis scorpioides
I
I
Molinia caerulea
II
Platanthera bifolia
I
Polygonum bistorta
I
Serratula tinctoria
I
Stachys officinalis
I
V
Succisa pratensis
II
III
Symphytum officinale
Valeriana officinalis
I
I
Potentillion anserinae et Potentillo Polygonetalia
Bromus commutatus
II
I
Carex distans
Elymus repens
I
-
98
II
II
I
III
II
IV
II
II
III
I
I
V
III
III
V
III
V
IV
III
III
IV
I
II
I
II
I
II
II
I
I
I
I
-
I
-
I
I
-
Inula britanica
Ranunculus repens
Molinio Arrhenatheretea
Alchemilla vulgaris agg.
II
Centaurea jacea
Cerastium holosteoides
II
Euphrasia officinalis ssp. I
pratensis
Euphrasia stricta
Lathyrus pratensis
Leontodon hispidus ssp. I
hyoseroides
Lotus corniculatus
III
Plantago lanceolata
II
Polygala vulgaris
II
Poa trivialis
Primula veris
Prunella vulgaris
III
Ranunculus acris
II
Rhinanthus angustifolius
I
Rumex acetosa
I
Stellaria graminea
Trifolium alpestre
Trifolium montanum
III
Trifolium pratense
V
Vicia cracca
II
Festuco Brometea s. l.
Ajuga genevensis
Anthemis tinctoria
Anthericum ramosum
Anthyllis vulneraria
Arenaria serpyllifolia
Asperula cynanchica
Astragalus cicer
Astragalus onobrychis
Bupleurum falcatum
Carex montana
Carlina vulgaris
Centaurea stoebe
Cirsium pannonicum
Dianthus carthusianorum
Echium vulgare
Elymus hispidus
Eryngium campestre
Euphorbia cyparissias
Festuca rupicola
II
Filipendula vulgaris
I
Galium verum
II
Gentiana cruciata
Helianthemum
nummularium
ssp.
obscurum
I
-
I
-
II
I
I
III
I
II
I
V
II
-
II
-
II
IV
I
III
I
IV
I
III
II
IV
I
III
IV
II
II
III
III
II
III
I
III
V
-
V
IV
III
I
III
IV
II
I
II
III
III
V
V
II
V
IV
III
IV
III
V
-
V
III
III
II
III
III
IV
III
II
III
II
II
I
I
IV
IV
III
I
V
II
III
I
I
III
I
I
I
I
I
III
I
I
I
I
III
I
I
III
II
I
I
V
IV
I
III
I
I
III
II
II
I
III
I
III
I
IV
II
IV
I
I
I
V
IV
II
III
I
I
I
I
I
-
99
Hieracium bauhinii
Hieracium pilosella
Hypochoeris maculata
Koeleria macrantha
Linum austriacum
Medicago falcata
Onobrychis viciifolia
Pimpinella saxifraga
Plantago media
Potentilla argentea
Potentilla arenaria
Prunella grandiflora
Ranunculus bulbosus
Salvia nemorosa
Salvia pratensis
Scabiosa ochroleuca
Teucrium chamaedrys
Trifolium aureum
Trifolium ochroleucon
Veronica orchidea
Juncetea trifidi s. l.
Botrychium lunaria
Campanula abietina
Campanula serrata
Carlina acaulis
Genista tinctoria
Gentianella austriaca
Hieracium aurantiacum
Hieracium umbellatum
Hypochoeris radicata
Ligusticum mutellina
Potentilla erecta
Viola canina
Festuca supina
Variae syntaxa
Achillea stricta
Acinos arvensis
Astragalus glycyphyllos
Astrantia major
Bunias orientalis
Campanula persicifolia
Campanula
rapunculoides
Campanula trachelium
Carduus acanthoides
Carex echinata
Carex flava
Cerastium arvense
Cichorium intybus
Cirsium arvense
Cirsium decussatum
Cirsium erisithales
III
II
II
II
-
I
I
I
I
I
V
I
I
I
I
-
II
III
I
I
III
III
V
II
I
II
I
II
I
I
II
II
III
-
II
III
I
II
II
III
IV
III
II
III
I
II
I
III
III
I
II
I
III
I
II
III
I
-
III
III
I
I
-
I
I
I
III
I
II
I
III
I
IV
III
-
I
II
I
I
II
III
II
I
II
II
II
-
II
I
I
-
II
-
I
I
I
I
I
III
I
I
-
I
II
I
I
I
-
I
III
I
-
I
I
I
I
I
I
I
I
II
I
I
-
I
I
-
III
I
100
Cirsium oleraceum
Cirsium vulgare
Clinopodium vulgare
Convolvulus arvensis
Coronilla varia
Cruciata glabra
Cruciata laevipes
Dianthus deltoides
Dianthus tenuifolius
Erigeron acris
Festuca heterophylla
Fragaria vesca
Galium schultesii
Gentiana asclepiadea
Gentiana verna
Gnaphalium sylvaticum
Inula salicina
Laserpitium latifolium
Listera ovata
Luzula luzuloides
Luzula pilosa
Matricaria recutita
Melampyrum cristatum
Myosotis alpestris
Nepeta cataria
Origanum vulgare
Polygonatum
verticillatum
Primula
elatior
ssp.
leucophylla
Pteridium aquilinum
Ranunculus acris ssp.
friesianus
Ranunculus oreophilus
Ranunculus
polyanthemos
ssp.
polyanthemoides
Reseda lutea
Rumex acetosella
Sagina saginoides
Salvia verticillata
Senecio jacobea
Senecio umbrosus
Silene alba
Silene nutans ssp. dubia
Tanacetum corymbosum
Telekia speciosa
Thalictrum
aquilegiifolium
Thalictrum minus
Trifolium medium
Trifolium pannonicum
I
III
I
-
I
I
III
I
II
I
I
I
II
I
-
I
I
I
I
I
I
I
I
I
III
I
I
I
I
II
II
I
-
I
I
II
I
III
IV
II
III
I
I
I
-
I
II
I
I
I
-
II
I
-
III
-
II
-
II
I
-
II
II
II
I
-
I
I
-
II
II
I
I
I
II
I
-
I
I
III
I
III
II
-
I
I
I
I
-
I
II
II
II
V
101
Veratrum album ssp. lobelianum

Verbascum lychnytis
I
Veronica chamaedrys
III
III
I
Vicia sepium
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 (Csrs et Resm. 1960) Oroian 1998
4. Agrostio Festucetum rupicolae Csrs Kaptalan 1964
5. Poo Trisetetum Knapp ex Oberd. 1957
III
I
-
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102

ASSESMENT OF ECOLOGICAL STATUS OF DANUBE DELTA LAKES USING

INDICATOR MACROPHYTES SPECIES
J. HANGANU , M. DOROFTEI , N. TEFAN
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 Sfntu 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
103
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 BraunBlanquet 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):
nA
TI ( S ) =
nB
Q Ai Q Bi
i =1
i =1
ng
Q
i =1
*100
gi
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%.
104
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.
Total P vs. RI
R2 = 0.0011
100
RI
50
0
0
100
200
300
-50
-100
P-tot
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
105
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
RI
RI vs. Soil texture classes

100
80
60
40
20
0
-20 0
-40
-60
-80
-100
R2 = 0.1273
soil texture
1
development of Nympaeides.
Fig.2. Trophic Reference Index (Schaumburg) vs. Soil texture classes
Conectivity vs.RI
100
R2 = 0.228
RI
50
0
Conectivity
0
-50
-100
Fig.3. Trophic Reference Index (Schaumburg) vs. Connectivity classes
106
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.
2.
3.
4.
5.
CARLSON B., 1995 - The Secchi Disk and the Volunteer Monitor. LakeLine, N. Am. Lake. Manage. Soc.,
15, 1: 28-29, 35-37
KOHLER A., 1978 - Methoden der Kartierung von Flora und Vegetation von Swasserbiotopen. Landschaft +
Stadt, 10: 23-85
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
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: 302314
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.
107
Table I. List of species groups with regard to eutrophication

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

Lake name
Baclaneti 1996
Cuibul cu Lebede 2002
Isac W 1998
Plin 1997
Raducu 1997
Rosu 1998
Rosulet 1998
Serbata 1997
Uzlina 1998
Iacub
Miazazi
Nebunu
Oprio
Raduculets 1997
L. Erenciuc 1993
L. Merheiu Mare 1995
Fortuna 1996
Cuibul cu Lebede 1998
Isac 2002
Uzlina 2002
Cn. Draghilea 1995
Cn. L.Ttaru 1995
Cn. Mila 35 1995
Cn. Potcoava 1995
Cn. Sireasa 1995
Cn. Sulimanca 1995
Grl. Somova 1994
Grl. ontea Veche 1995
J. Sulimanca 1995
J. Urechea 1994
TI
-99
-64
-75
-81
-73
-100
-98
-79
-68
-20
-13
-15
-8
-11
-14
-39
-98
-93
-82
-78
-58
-63
-58
-68
-92
-37
-50
-65
-75
-39
108
Lake name
Grl. Bratusca 1995
Grl. Sireasa Veche1995
L. Corciovata 1994
L. cu Cotete 1995
L. Gasca 1994
L. Lung 1995
L. Merheiu Mic 1995
L. Mesteru 1995
L. Parches 1994
L. Puiu 1993
L. Rdcinosu Mare 1995
L. Rdcinosu Mic 1995
L. Rotund 1994
L. Somova 1994
L. Telincea 1994
Lopatna 1996
M. Sacalin 1993
O. Babina 1995
Sf. Gheorghe 1995
Sf. Gheorghe 1993
Tataru 1998
Chiril 2002
Ghearsim 2002
Gorgova 2002
Gorgova 1998
HO02_1996
LacBabina 1997
Pojarnia 2002
L. Casla 1994
TI
-52
-73
-54
-58
-53
-35
-87
-24
-56
-6
-32
-48
-61
-16
-49
-93
-52
-62
-100
-6
-1
51
-41
-92
-76
-92
-87
-35
-60

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 relevs. 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 Iai, 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 erbnescu 1955, which has not been studied
from this zone. Additionally, the relevs from the areas where it was identified are not
sufficiently numerous to justify the coenotaxonomical classification.
We used the phytocoenological relev method created by the Zrich-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
relevs. For the vernal aspect, the observations were sampled in April, whereas for the
aestival aspect, the relevs 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 relevs. Nevertheless, these relevs were very consistent with regard to the type of
vegetation, and consequently they allowed the phytosociological classification.
109

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 relevs 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

Relev No.
1
2
3
4
Altitude (m)
103 105 123 98
Exposition
NV V
V
V
Slope (degree )
20
10
15
5
Cover (%)
100 100 100 100
Area (m)
10
10
15
10
Caract. Sp.
Phragmites australis
+
1
+
2
Festucion valesiacae
Festuca valesiaca
1
+
1
+
Phlomis pungens
+
+
+
Adonis vernalis
+
+
+
+
Agropyron cristatum
1
+
1
Linum austriacum
+
+
+
Salvia nemorosa
+
+
+
Euphorbia glareosa
+
+
Festucetalia valesiacae
Achillea setacea
+
+
+
Centaurea biebersteinii
+
Elymus hispidus
2
+
+
+
Inula hirta
+
+
Erysimum odoratum
+
+
Bromus inermis
+
Festuco-Brometea
Galium verum
+
+
+
Medicago falcata
+
+
Dianthus carthusianorum +
+
+
Muscari racemosum
+
+
+
+
Thlaspi perfoliatum
+
+
+
110
5
94
V
10
100
25
+
+
+
+
+
+
V
IV
V
IV
III
IV
III
+
+
+
+
-
III
II
V
III
III
I
+
+
-
IV
III
III
IV
III
Relev No.
Agrimonia eupatoria
Thalictrum minus
Salvia verticillata
Melica ciliata
Asperula cynanchica
Molinio-Arrhenatheretea
Dactylis glomerata
Elymus repens
Knautia arvensis
Rhinanthus rumelicus
Variae syntaxa
Melampyrum arvense
Asparagus officinalis
Peucedanum latifolium
Nepeta nuda
Marrubium vulgare
Centaurea orientalis
Chenopodium album
Conium maculatum
Polygonum dumetorum
Stachys annua
Tribulus terestris
1
+
-
2
+
+
+
-
3
+
+
+
4
+
+
5
+
-
K
II
III
II
I
II
+
+
+
+
+
+
+
+
-
+
+
+
-
IV
II
II
III
+
+
+
+
+
+
+
+
+
-
+
+
+
+
+
+
+
+
+
-
II
I
III
IV
II
I
II
I
I
I
I
II
We compared our five relevs with other 4 from the literature, sampled from
Frumoasa-Moara (Mititelu and Cojocaru, 1970) and Prut River Valley (Mititelu and
Baraba, 1975), and we noted their strong resemblance, given that it is the same type of
vegetation.
On the other hand, our relevs differ very much from another type of community
formed by Phragmites australis, i. e. the Association Phragmitetum vulgaris So 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 PragmitiMagnocaricetea, and other subordinated taxa of this class.
111
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 So 1927. Consequently, our observations support the classification of these types
of communities in the Class Xero-Phragmitetum erbnescu 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.
2.
3.
4.
5.
6.
7.
8.
9.
10.
BORKMAN J., MORAVEC J. & RAUSCHERT S., 1985 - Code de nomenclature phytosociologique Vegetatio.
Haga, 67, 3: 177-187
BURDUJA, C., 1959 - O rezervaie tiinific care trebuie nfiinat "Fneele din Valea lui David" - Iai. Ocrot.
nat., Bucureti, 4: 154 157
CHIFU T., MNZU C. & ZAMFIRESCU O., 2007 - Flora i vegetaia Moldovei. Edit. Univ "Al. I Cuza"
Iasi
MITITELU, D. & BARABA N., 1972 - Rspndirea unor asociaii ierboase n lunca Prutului. St. Com., Muz. t.
Nat. Bacu, 5: 189 196
MITITELU D. & BARABA N., 1975 - Vegetaia din lunca Prutului. St. Com. Muz. t. Nat. Bacu, 8: 219 285
MITITELU D. & COJOCARU V., 1970 - Flora i vegetaia rezervaiei Frumoasa - Suceava. Ocrot. nat.,
Bucureti, 14, 1: 35 40
MITITELU D. & COJOCARU V., 1981 - O nou contribuie la flora rezervaiei botanice de la Frumoasa - Moara
( jud. Suceava ). St. Com. ocrot. nat. Suceava, 5: 394 395
MITITELU D., MOIU T., DSCLESCU D., TEU C. & VIALARIU C., 1969 - Flora i vegetaia
rezervaiei "Valea lui David" - Iai. St. Com. Muz. t. Nat. Bacu, 2: 81 100
SANDA V., 2002 - Vademecum ceno - structural privind covorul vegetal din Romnia. Ed. Vergiliu, Bucureti
SANDA V., POPESCU & A. DOLTU M. I., 1980 - Cenotaxonomia i corologia gruprilor vegetale din
Romnia. St. Com. t. Nat. Muz. Brukenthal, Sibiu, 24
112

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 So ex Korneck 1969 Association
For the identification of plant associations, we used phytosociological research
methods according to the CentralEuropean school. The establishment of the bioforms and
floristic elements was made on the basis of Flora ilustrat a Romniei Pteridophyta et
Spermatophyta, by V. Ciocrlan (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)
113
Chorology: Codeti, Dobrov

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 MolinioArrhenatheretea (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 humidwet 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: Tcuta (Mititelu D.,1975), Vaslui, Vleni (Mititelu D. and collab.,
1996), Soleti
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).
- 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 EuroAsian
elements (63.63%), followed by the circumpolar ones (27.28%) and the cosmopolite ones
(9.09%);
114
- 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: Popeti, Satu Nou (Mititelu D., 1975), Vaslui (Mititelu D. and collab.,
1996), Ciorteti, Codeti, Dobrov, Fereti
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.
- 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 EuroAsian (5.35%), Mediterranean (3.56%),
European (1.79%), AtlanticEuropean (1.79%) and ponticBalkan 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 So ex Korneck 1969
(Syn.: Polygonetum avicularis Gams 1927, Schlerochloo-Polygonetum avicularis
So 1945)
Chorology: Brnova (Mititelu D. and collab., 1995), Dneti, Tcuta, Vaslui, Vleni
(Mititelu D. and collab., 1996), Deleni, Moara Grecilor, Popeti
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.
115
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).
- 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 EuroAsian
elements (48.27%) and cosmopolite (31.05%), followed by the Mediterranean EuroAsian
ones (6.88%), Mediterranean (3.45%), adventive (3.45%), continental EuroAsian (3.45%)
and ponticMediterraneancentral 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.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
CIOCRLAN V., 2000 - Flora ilustrat a Romniei Pteridophyta et Spermatophyta. Ed. Ceres, Bucureti
CHIFU T., 1995 - Contribuii la sintaxonomia vegetaiei pajitilor din clasele Molinio Arrhenatheretea
Tx.37 i Agrostietea stoloniferae Oberd. in Oberd. et al.67 de pe teritoriul Moldovei. Bul. Grd. Bot. Iai, 5:
125-132
CHIFU T., MNZU C., ZAMFIRESCU O., 2006 - Flora & vegetaia Moldovei (Romnia), vol. II, Ed. Univ.
Al. I. Cuza Iai: 211-303
ELLENBERG H., 1974 - Indicator values of vascular plants in Central Europe. Scripta Geobotanica, vol.
IX, Verlag Erich Goltze K.G., Gttingen: 1-97
ELLMAUER T., MUCINA L., 1993 - Molinio Arrhenatheretea In: MUCINA L., GRABHERR G.,
ELLMAUER T., 1993 Die pflanzengesellschaften sterreichs, Gustav Fischer Verlag Jena Stuttgart
New York, vol. I: 297-401
MITITELU D., 1975 - Flora i vegetaia judeului Vaslui. St. i Com. Muz. t. Nat. Bacu, Biol. veget.: 67162
MITITELU D., CHIFU T., SCARLAT A., ANIEI L., 1995 - Flora i vegetaia judeului Iai. Bul. Grd.
Bot. Iai, 5: 99-124
MITITELU D., HUANU M., 1996 - Noi contribuii la flora i vegetaia judeului Vaslui. St. i Cerc. Muz.
t. Nat., Piatra-Neam, 8: 193-211
MUCINA L., 1997 - Conspectus of classes of European vegetation. Folia Geobot. Phytotax., Praha, 32, 2:
117-172
SANDA V., 2002 - Vademecum ceno-structural privind covorul vegetal din Romnia. Bucureti, Ed.
Vergiliu
SANDA V., POPESCU A., 1991 - Studiul fitocenozelor clasei Molinio-Arrhenatheretea Tx. 37 din Romnia.
Acta Bot. Horti Bucurestiensis: 49-59
SANDA V., POPESCU A., BARABA N., 1997 - Cenotaxonomia i caracterizarea gruprilor vegetale din
Romnia. St. i Comun. Muz. t. Nat. Bacu, Biol. veget., 14: 2-365
SANDA V., POPESCU A., STANCU D. I., 2001 - Structura cenotic i caracterizarea ecologic a
fitocenozelor din Romnia. Ed. Conphis, Bucureti
116
Table I. Ass. Scirpetum sylvatici Ralski 1931

Number of survey
Altitude (m.s.m.)
Cover of the vegetation (%)
Surface of survey (m)
Number of species
Associations characteristics
Scirpus sylvaticus
Poa palustris
Epilobium hirsutum
Galium palustre
Lysimachia nummularia
Lythrum salicaria
Juncus effusus
Ranunculus repens
Juncus inflexus
Mentha aquatica
Mentha longifolia
Ajuga reptans
Poa pratensis
Veronica beccabunga
Veronica anagallis-aquatica
Alisma plantago-aquatica
Carex vulpina
Juncus gerardii
Equisetum arvense
Lemna minor
1
210
100
10
13
2
121
95
25
14
3
121
90
25
11
5
5
5
Calthion
+
+
+
+
Molinietalia
+
+
+
+
+
+
+
+
Potentillo-Polygonetalia
1
+
+
+
+
+
+
+
+
+
+
+
Phragmiti-Magnocaricetea
+
+
+
+
+
+
+
Variae syntaxa
+
+
+
-
4
210
100
10
10
5
210
100
10
8
+
+
-
+
+
+
IV
III
II
+
+
+
-
+
+
-
IV
IV
III
I
I
1
+
+
1
-
V
III
II
II
II
I
I
+
+
-
III
II
II
I
I
+
-
II
I
I
Place and date of the surveys:

1,4,5. Dobrov, 1.07.2004, 18.07.2004; 2,3. Codeti, 1.07.2004, 18.07.2004
Table II. Ass. Ranunculetum repentis Knapp ex. Oberd. 1957
Number of survey
Altitude (m.s.m.)
Number of species
Ranunculus repens
Potentilla reptans
1
120
90
20
5
2
120
95
20
4
5
5
Potentillion anserinae
+
1
117
3
120
100
20
6
4
120
85
20
6
5
120
100
20
5
Elymus repens
Rumex crispus
+
+
+
Potentillo-Polygonetalia
Rorippa sylvestris ssp. sylvestris
+
Plantago major
Carex vulpina
+
Poa palustris
-
+
-
+
+
+
+
-
III
III
II
+
+
+
+
-
II
II
I
I
I

1-5. Soleti, 6.06.2004
Table III. Ass. Junco inflexi Menthetum longifoliae Lohmeyer 1953
Number of survey
Altitude (m.s.m.)
1
170
2
210
3
94
4
110
Exposition
SV
V
SE
N
Slope ()
Cover of the
60
100
70
70
vegetation (%)
25
20
20
25
Number of species
8
7
8
7
Juncus inflexus
3
5
4
3
Potentillion anserinae et Potentillo-Polygonetalia
Mentha longifolia
2
1
+
Mentha pulegium
1
2
Ranunculus repens
Trifolium repens
Inula britannica
+
Verbena officinalis
Myosoton aquaticum
Potentilla reptans
Molinietalia
Galium palustre
Trifolium hybridum
Daucus carota
+
Lotus corniculatus
+
Trifolium pratense
+
Cichorium intybus
+
Plantago major
Medicago lupulina
Centaurea jacea
Ranunculus acris ssp.
acris
Festuca pratensis
-
118
5
110
6
110
7
120
8
110
9
110
N
-
SV
-
NV
-
V
-
NV
-
10
210
loc
plan
-
85
25
9
60
15
7
70
25
10
100
20
17
85
50
11
85
20
18
2
2
+
+
-
1
+
+
1
+
-
+
+
+
-
2
+
1
+
+
+
-
2
+
2
+
IV
II
II
II
II
I
I
I
I
+
+
I
I
+
+
+
1
+
+
+
-
+
+
-
II
II
I
I
I
I
I
+
+
I
I
Festuco-Brometea
Prunella vulgaris
Galium humifusum
Achillea setacea
Salvia nemorosa
Plantago media
Artemisietea vulgaris
Artemisia absinthium
+
Tanacetum vulgare
+
Setaria viridis
Arctium lappa
Equisetum arvense
Stellarietea mediae
Cannabis sativa ssp.
ruderalis
Lathyrus tuberosus
Chenopodium vulvaria
Linaria vulgaris
Sonchus arvensis
Anagallis arvensis
Lythrum salicaria
+
Typha angustifolia
+
Lycopus europaeus
Equisetum palustre
Carex riparia
Typha latifolia
Carex vulpina
Bidentetea
Bidens tripartita
Polygonum hydropiper
Rumex conglomeratus
Rumex crispus
Variae syntaxa
Schoenoplectus
tabernaemontani
+
Polygonum aviculare
Salix alba
Chaemerion
angustifolium
Trifolium fragiferum
Juncus gerardi
-
+
+
-
+
-
+
+
+
+
+
-
I
I
I
I
I
+
-
+
-
+
+
-
I
I
I
I
I
+
-
+
-
+
-
+
-
+
+
I
I
I
I
I
I
+
-
1
-
+
-
1
+
+
-
+
+
-
1
-
+
+
II
I
I
I
I
I
I
I
+
-
+
-
1
-
1
-
+
-
1
-
+
+
III
I
I
I
+
-
I
I
I
I
+
-
+
+
I
I

1. Ciorteti, 27.07.2003; 2, 10. Dobrov, 23.08.2003, 1.07.2004; 3. Vaslui, 12.08.2003; 46,8,9. Codeti, 24.08.2003, 6.08.2003; 7. between Codeti and Dobrov, 24.08.2003
119
Table IV. Ass. Schlerochloo-Polygonetum avicularis So ex Korneck 1969

Number of survey
Altitude (m.s.m.)
Exposition
Slope ()
Number of species
Schlerochloa dura
Lolio-Plantaginion et Plantaginetalia
Polygonum aviculare
Poa annua
Lolium perenne
Lepidium ruderale
Matricaria perforata
Hordeum murinum
Cynodon dactylon
Malva pusilla
Cichorium intybus
Plantago major
Trifolium repens
Plantago lanceolata
Verbena officinalis
Stellarietea mediae
Capsella bursa-pastoris
Cardaria draba
Malva neglecta
Thlaspi arvensis
Bromus arvensis
Salvia nemorosa
Chenopodium album
Atriplex tatarica
Amaranthus retroflexus
Portulaca oleracea
Artemisietea vulgaris
Xanthium strumarium
Artemisia absinthium
Artemisia annua
1
220
E
1-2
95
50
14
2
220
E
1-2
100
25
14
3
270
70
25
11
4
95
80
50
16
5
190
NE
2-3
75
25
14
II
5
+
+
+
+
+
+
-
5
1
+
+
-
3
2
+
+
-
4
+
+
+
-
4
+
+
+
V
V
III
III
II
II
I
I
I
+
+
+
+
+
+
-
+
-
1
-
+
+
+
+
IV
III
III
II
+
-
+
+
+
+
+
-
1
+
+
+
+
+
+
+
+
+
+
+
-
1
1
+
+
+
-
IV
III
III
III
II
II
II
I
I
I
I
+
-
+
+
+
+
-
+
+
+
-
+
-
III
III
II
I

1,2. Micleti, 11.08.2004; 3. Deleni, 10.08.2004; 4. Moara Grecilor, 11.08.2004; 5.
Popeti, 11.08.2004
120

CONTRIBUTIONS TO THE STUDY OF PALUDAL VEGETATION

FROM THE NEAGRA ARULUI RIVERS BASIN
(SUCEAVA COUNTY)
LOREDANA ASOLTANI
Abstract: This paper presents three vegetal paludal associations identified in Neagra arului rivers
basin: Epilobio Juncetum effusi Oberd. 1957, Scirpetum sylvatici Ralski 1931 i 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 arului river basin.
Introduction
Neagra arului 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
Climani Mountains in its south side. The Neagra arului river springs on the northern
slope of Climani Mountains, at an absolute altitude of 1832 m, and flows into Bistria 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 arului 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 arului area, on Srioru
Mic river valley, in Coverca area on Negru and Deluganu rivers valley, in Pltini area, in
Gura Haitii area, on Tamu and Haita rivers valley.
121
As an observations, the association was first recorded in 1975, by M. Toma in his

doctors theses, in aru Dornei with two releves, but we have to underline that this results
didnt find again into a speciality publication. In 1989, it was also mentioned by D.
Mititelu, without attaching a table of floristic releves.
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 releves, 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 Pauc 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 arului, Srior, Coverca, Panaci and aru dornei area.
The association was mentioned before in Neagra arului river basin by M. Toma, in
1975 (Neagra aruluI), presenting just a floristic list, and also by T. Seghedin in 1986
(Coverca) and D. Mititelu in 1989 (Panaci, Pltini), without attaching a table of floristic
releves.
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).
122
- the bioform spectrum shows the dominance of hemicryptophytes (80%)

followed by geophyte (12,72%), chamaephytes (4,25%), therophytes (2,13%) and hydrohelophytes (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%);
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 aruluI, aru
Dornei, aru Bucovinei, Srior, Coverca and Tarnia 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).
- 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%);
moderate to hight humidity (20,34%), amphitolerant to the soil reaction (55,93%) and to the
content of mineral nitrogen in soil (23,73%).
123
REFERENCES
1.
CHIFU T., MNZU C., ZAMFIRESCU O., 2006 - Flora i vegetaia Moldovei (Romnia), vol. I, II,
Edit. Univ. Al. I. Cuza Iai
2.
CIOCRLAN V., 2000 - Flora ilustrat a Romniei Pteridophyta et Spermatophyta. Ed. Ceres,
Bucureti
3.
MITITELU D., CHIFU T., PASCAL P., 1989 - Flora i vegetaia judeului Suceava, Anuar. Muz. t.
Nat. Suceava, t. 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., LUPACU GH., DAVIDESCU G.,
PASCAL P., 1996 - Pajitile din Bucovina, Edit. Helios
6.
SANDA V., 2002 - Vademecum ceno-structural privind covorul vegetal din Romnia. Ed. Vergiliu,
Bucureti
7.
SANDA V., POPESCU A., BARABA N., 1997 - Cenotaxonomia i caracterizarea gruprilor
vegetale din Romnia. St. i Comun. Muz. t. Nat. Bacu, Biol. veget., 14
8.
SEGHEDIN T. G., 1986 - Flora i vegetaia munilor Bistriei, Rezumatul tezei de doctorat, Inst. Agr.
Ion Ionescu de la Brad Iai
9. STOICA D. L., 2007 - Cercetri de geografie fizic pe versantul nordic al Masivului Climani.
Rezumatul tezei de doctorat, Univ. Al. I. Cuza Iai
10. TOMA M., 1975 - Cercetri asupra florei i vegetaiei din Depresiunea Dornelor (jud. Suceava),
Rezumatul tezei de doctorat, Univ. Babe - Bolyai Cluj-Napoca
Table I SCIRPETUM SYLVATICI RALSKI 1931

1
Number of relev
1209
Altitude (m.s.m.)
100
Covering (%)
50
Surface (m2)
15
No. of species
5
Scirpus sylvaticus
Calthion palustris
+
Caltha palustris
+
Chaerophyllum hirsutum
Cirsium rivulare
+
Geum rivale
+
Deschampsion
Carex ovalis
Juncus conglomeratus
Filipendulion
Equisetum arvense
+
Filipendula ulmaria
Lysimachia vulgaris
+
Mentha longifolia
Molinietalia caeruleae
+
Cirsium palustre
+
Equisetum palustre
+
Galium palustre
+
Juncus effusus
Lychnis flos-cuculi
Lythrum salicaria
10
11
12
920
910
920
1013
1016
1046
1100
1250
1069
831
824
100
100
95
95
95
95
95
95
90
90
90
100
100
100
50
50
25
50
50
50
100
25
15
17
16
13
19
21
15
12
11
21
III
II
III
III
IV
II
II
III
II
III
IV
III
III
II
124
ssp. officinale
Trifolium hybridum
+
+
+
II
ssp. hybridum
Agrostis stolonifera ssp.
+
+
+
+
+
III
stolonifera
+
+
+
+
II
Alchemilla vulgaris
+
I
Alnus incana juv.
+
+
+
+
II
Holchus lanatus
+
+
+
+
+
III
+
+
I
Phleum pratense
+
+
I
Prunella vulgaris
+
I
Stellaria graminea
Phragmiti - Magnocaricetea s. l.
+
+
+
+
II
Carex riparia
+
+
I
Epilobium palustre
+
I
Glyceria notata
+
+
I
Lycopus europaeus
+
+
+
+
+
III
Ranunculus repens
+
+
+
+
+
III
Veronica beccabunga
Scheuchzerio Caricetea fuscae s. l.
+
+
+
+
II
Carex flava
+
+
+
II
Carex nigra ssp. nigra
Variae syntaxa
+
I
Epilobium montanum
+
I
Festuca pratensis
+
I
Luzula campestris
+
+
+
+
II
Potentilla erecta
Rorripa sylvestris ssp.
+
I
sylvestris
+
I
Rumex crispus
+
+
I
Stellaria nemorum
+
+
+
II
Valeriana tripteris
Place and date of releves: 1. Tamu river valley (16.08.2007); 2, 3, 4. Srioru 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. Pltini (13.07.2007); 11, 12. Plaiul arului (8.08.2007).
Table II EPILOBIO JUNCETUM EFFUSI OBERD. 1957

1
Number of relev
914
Altitude (m.s.m.)
95
Covering (%)
100
Surface (m2)
21
No. of species
Epilobium palustre
+
Juncus articulatus
Calthion palustris
Caltha palustris
Chaerophyllum
hirsutum
Geum rivale
+
+
Scirpus sylvaticus
10
920
998
1000
1124
1372
995
993
980
906
95
95
95
95
90
90
90
90
90
50
50
100
50
50
100
50
30
100
19
22
16
25
13
15
17
17
18
III
IV
III
II
III
125
Deschampsion
+
+
Carex ovalis
+
+
+
Juncus conglomerates
Filipendulion
Cirsium erysithales
+
+
Filipendula ulmaria
+
+
Mentha longifolia
+
Cirsium palustre
+
Equisetum palustre
+
Galium palustre
5
5
Juncus effusus
+
+
Succisa pratensis
Trifolium hybridum ssp.
hybridum
Agrostis capillaris
Agrostis stolonifera ssp.
stolonifera
+
Alchemilla vulgaris
Anthoxanthum
odoratum
Briza media
+
Carex hirta
+
+
Carex pallescens
Cynosurus cristatus
Dactylis glomerata
Festuca rubra
Holchus lanatus
Luzula campestris
+
+
Phleum pratense
+
+
Prunella vulgaris
Ranunculus acris
+
Rumex crispus
Trifoiul repens ssp.
repens
Phragmiti Magnocaricetea s. l.
Alisma plantago
+
aquatica
Carex riparia
Lycopus europaeus
+
Ranunculus repens
+
+
Carex flava
Carex nigra ssp. nigra
Eriophorum
angustifolium
Variae syntaxa
Carduus personatus ssp.
personatus
+
Cruciata glabra
Epilobium montanum
III
IV
IV
II
II
II
II
IV
II
II
IV
II
II
II
III
IV
II
III
III
II
II
126
+
I
Equisetum sylvaticum
+
I
Homogyne alpina
+
+
I
Hypericum maculatum
+
I
Plantago media
+
+
+
+
+
+
+
+
IV
Potentilla erecta
+
+
I
Valeriana tripteris
+
+
I
Veratrum album
Place and date of releves: 1. Neagra arului (14.08.2006); 2. Srioru Mic river valley (14.07.2007); 3. Coverca
Climnel river valley (16.08.2006); 4. Coverca Negru river valley (16.08.2006); 5. Pltini (13.07.2007); 6.
Panaci (18.082006); 7. Coverca Negru river valley (16.08.2006); 8. Coverca Bucini river valley (16.08.2006);
9. Panaci Rusului peak (18.08.2006); 10. on the outskirts Tinovului Mare aru Dornei (15.07.2007).
Table III Filipendulo Geranietum palustris W. Koch 1926

1
Number of relev
999
Altitude (m.s.m.)
100
Covering (%)
25
Surface (m2)
12
No. of species
5
Filipendula ulmaria
Filipendulion
Chaerophyllum
hirsutum
Equisetum arvense
Geranium palustre
Lysimachia vulgaris
Lythrum salicaria
Mentha longifolia
Calthion palustris
Alchemilla vulgaris
Briza media
Caltha palustris
Cirsium rivulare
Geum rivale
Myosotis
+
scorpioides
Scirpus sylvaticus
Deschampsion
Carex ovalis
Deschampsia
caespitosa
Juncus
+
conglomeratus
Cirsium oleraceum
Cirsium palustre
Equisetum palustre
Galium palustre
+
Juncus effusus
Lychnis flos-cuculi
Succisa pratensis
Achillea millefolium
10
11
12
851
825
1037
1035
1016
1210
998
920
830
830
830
100
100
95
95
90
90
90
95
95
95
95
25
50
50
50
25
25
25
50
100
50
100
10
18
10
15
18
19
13
19
12
14
12
II
IV
II
III
II
III
II
II
III
IV
II
II
II
IV
II
II
II
127
+
+
I
ssp. stolonifera
+
I
Ajuga reptans
+
+
I
Angelica sylvestris
Campanula
+
+
I
glomerata
+
+
I
Centaurea jacea
+
+
I
Cynosurus cristatus
+
I
Festuca pratensis
+
I
Luzula campestris
+
+
+
II
Phleum pretense
+
+
+
+
II
Prunella vulgaris
+
I
Ranunculus acris
+
+
I
Rumex acetosa
+
I
Rumex crispus
Trifolium repens
+
+
+
+
+
III
ssp. repens
Phragmiti Magnocaricetea s. l.
+
+
I
Epilobium palustre
+
I
Glyceria notata
+
+
+
+
+
III
Ranunculus repens
Veronica
+
I
beccabunga
Carex nigra ssp.
+
I
nigra
+
I
Ligularia sibirica
Variae syntaxa
+
I
Agrimonia eupatoria
Astragalus
+
I
glycyphyllos
Athyrium filix+
I
femina
+
I
Cirsium arvense
Epilobium
+
I
montanum
Equisetum
+
I
sylvaticum
+
+
I
Galeopsis tetrahit
+
I
Holchus lanatus
Hypericum
+
I
perforatum
Hypochoeris
+
I
uniflora
+
I
Mentha arvensis
+
+
I
Plantago media
+
+
+
II
Potentilla erecta
+
+
I
Valeriana tripteris
Place and date of releves: 1. Srior Sriorul Mare river valley (14.08.2006); 2. aru 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 Bucini river valley (16.08.2006); 7. Tamu river valley
(16.08.2007); 8. Plaiul arului (19.08.2006); 9. aru Bucovinei Srioru Mic river valley (14.07.2007); 10, 11,
12. Plaiul arului (8.08.2007).
128

PROTECTED TAXA FROM THE BISTRIA RIVER BASIN BETWEEN PIATRA

NEAM AND BACU
CARMEN AONCIOAIE
Abstract: The results presented in this article were obtained during a study made in 2005 and 2006 in the
Bistria river inferior basin in the region between Piatra Neam and Bacu. 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, Bistria inferior basin, Bacu, Piatra Neam
Introduction
The studied area belongs to the lower course of the Bistria river, being situated on
two counties from Moldova region Neam and Bacu. Geomorphologicaly speaking, the
studied region spreads on the following four natural units (1400 Km2): Oriental Carpathians
(the Gomanu Mountains), the Moldavian sub-Carpathians with two subdivisions (Bistria
sub-Carpathians and Cracu Bistria depression) and the Moldavian Plateau (a very small
region between Racova and Bacu).
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 Bistria and its affluents. The most
important affluent in his sector is Cracu, followed by a number of streams like Calu, Iapa,
Nechit, Trebi, Negel and so on. Building hydro-electric power stations on the river leaded
to the appearance of some artificial lakes like Bacu, Buhui, Grleni, 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 Romniei (vol. I XIII) 1952
1976; Ciocrlan V. Flora ilustrat a Romniei. Pteridophyta et Spermatophyta., 2000;
Srbu I. et al. Flora ilustrat a plantelor vasculare din estul Romniei (vol. I and II)
2001.
Al. I. Cuza University, Iai, Faculty of Biology, Carol I Bd., no. 20A, 700506, Iasi, Romania
129
There were used two papers for including the taxa in different protection categories:
Gh. Dihoru, Alexandrina Dihoru Plante rare, periclitate i endemice n flora Romniei
Lista Roie, 1993 1994 and M. Oltean, G. Negrean, A. Popescu et al. Studii, sinteze,
documentaii 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 Roie a plantelor superioare din Romnia by Oltean M.,
Negrean G., Popescu A., Roman N., Dihoru G., Sanda V., Mihilescu S. (1994) and the
second column presents the analysis made after Plante rare, periclitate i endemice n
flora Romniei Lista Roie 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.
130
REFERENCES
1.
BELDIE AL., 1977 - Flora Romniei. Determinator ilustrat al plantelor vasculare. Vol. I, II, Ed. Acad.
R.S.R., Bucureti
2.
CIOCRLAN V., 2000 - Flora ilustrat a Romniei. Pteridophyta et Spermatophyta. Ed. Ceres, Bucureti
3.
DIHORU GH., DIHORU A., 1993 1994 - Plante rare, periclitate i endemice n flora Romniei Lista
Roie, Acta Botanica Horti Bucurestiensis, Bucureti, 1994: 173-179
4.
DIHORU G., PRVU C., 1987 Plante endemice n Flora Romniei. Ed. Ceres, Bucureti
5.
MITITELU DUM., BARABA N., TEFAN N. 1987. Contribuii la corologia unor plante rare n Moldova
i Muntenia. An. t. Univ. Al. I. Cuza Iai , s. II, a., Biol. veget., t. XXXIII: 20 24
6.
OLTEAN M. et al., 1994 - Lista roie a plantelor superioare din Romnia. Studii., Sinteze, Documentaii de
Ecologie, Acad. Rom.
7.
POPESCU A., SANDA V., 1966 - Consideraii corologice asupra plantelor endemice din Romnia. St. Cerc.
Biol., ser. Bot., 18, 5;;437 466
8.
RUGIN R., MITITIUC M., 2003 - Plante ocrotite n Romnia. Ed. Univ. Al. I. Cuza Iai
9.
SRBU I., TEFAN N., IVNESCU LCRMIOARA, MNZU C., 2001 Flora ilustrat a plantelor
vasculare din estul Romniei. Vol. I, II, Ed. Univ. Al. I. Cuza Iai
10. OPA E., 1979 - Ocrotirea naturii n Judeul Neam. Reflexii, istoric, realizri i sugestii.
**Conventia de la Berna (1979)
*** 1952 1976 Flora R. P. R-r. S. R., I XIII. Ed. Acad. R.S.R., Bucureti
Table I: Protected taxa

category
E
R
R
K
R
I
R
R
R
VR
M. Oltean et al.
specie
Abies alba Miller
Allium schoenoprasum L. ssp.
sibiricum (L.) Hartman
Anacamptys pyramidalis (L.)
L.C.M. Richards
Bromus racemosus L.
Campanula carpatica Jacq.
Carex brevicollis DC.
Carex dioica L.
Carex hallerana Asso
Centaurea melanocalathia
Borbs
Centaurium littorale (D. Turner)
131
Gh. Dihoru, Alexandrina Dihoru

category
specie
R
Adonis flammea Jacq.
V
Adonis vernalis L.
R
K
K
I (R)
R
R
Alopecurus arudinaceus Poiret

Bromus racemosus L.
Bryonia dioica Jacq.
Carex brevicollis DC.
Carex dioica L.
Centaurium littorale (D. Turner)
V
V
R
V
R
R
R
VR
V
R
VR
R
R
R
R
R
R
VR
R
R
R
R
R
Gilmour ssp. uliginossum (W et

K) G. Beck
Cephalanthera damassonium
(Miller) Druce
Cephalanthera longifolia (L.)
Fritsch
Cephalanthera rubra (L.) M.L.
C. Richards
Cirsium furiens Griseb. et
Schenk
Cirsium grecescui Griseb et
Schenk
Coeloglossum viride (L.)
Hartman
Colutea arborescens L.
Corynephorus canescens (L.) P.
Beauv.
Crocus reticulatus Steven
Cyperus serrotinus Rottb.
Dactylorhizza incarnata (L.)
So
Dactylorhizza maculata (L.) So
ssp. maculata
Dactylorhizza maculata (L.) So
ssp. schurii (Klinge) So
Dactylorhizza sambucina (L.)
So
Dianthus collinus Waldst et Kit
ssp. collinus
ssp. glabriusculus (Kit) Thaisz
Dictamnus albus L.
Dryopteris cristata (L.) A.Gray
Epipactis helleborine (L.)
Crantz
Epipactis palustris (L.) Crantz
Epipactis purpurata Sm.
Eryssimum witmannii Zawadzki
ssp. transilvanica (Schur) P.W.
Ball
132
Gilmour ssp. uliginossum (W et

K) G. Beck
-
Cirsium furiens Griseb. et

Schenk
Cirsium grecescui Griseb et
Schenk
-
R
E
V
-
Corynephorus canescens (L.) P.

Beauv.
-
I
R
V
-

ssp. glabriusculus (Kit) Thaisz
Dryopteris cristata (L.) A.Gray
Eleocharis carniolica Koch
-
K
VR
nt
nt
K
nt
V
R
R
R
R
R
VR
R
R
R
R
R
R
R
R
R
R
R
Fragaria moschata Duchesne

Fritillaria orientalis Adams
Galanthus elwesii Hooker fil.
Galanthus nivalis L.
Galium sylvaticum L.
Hepatica transsilvanica Fuss.
Hippuris vulgaris L.
Gymnadenia conopsea (L.) R.
Br.
Herniaria hirsuta L.
Koeleria macrantha (Ledeb)
Schultes ssp. transsilvanica
(Schur) A. Nyr.
Lactuca virosa L.
Lapulla deflexa (Lehm.) Cesati
Lepidium cartilagineum
(J.Mayer) Thell ssp.
crassifolium (W. et K.) Thell
Listera ovata (L.) R. Br.
Luzula pallescens Swartz.
Melmpyrum saxosum Baumg.
Monotropa hypopytis L.
Myosotis stenophylla Knaf.
Neottia nidus-avis (L.) L.C.M.
Richards
Oenanthe peucedanifolia
Pollisch
Orchis coriophora L.
Orchis laxiflora Lam. ssp.
elegans (Heuffel) So
Orchis mascula (L.) L. ssp.
signifera (Vest) So
Orchis militaris L.
133
R
E
R
R
R
nt
-
Evonymus latifolius (L.) Miller

Galanthus elwesii Hooker fil.
R
R
R
nt
R
R
R
R
-
Iris aphylla L.
Iris sibirica L.
Isolepis setacea (L.) R. Br.
Koeleria macrantha (Ledeb)
Schultes ssp. transsilvanica
(Schur) A. Nyr.
Lactuca virosa L.
Lepidium cartilagineum
(J.Mayer) Thell ssp. crassifolium
(W. et K.) Thell
Luzula pallescens Swartz.
Melampyrum nemorosum L.
Myosotis discolor Pers.
Myosotis stenophylla Knaf.
-
R
V
Galium tenuissimum Bieb.

Gladiolus imbricatus L.
Goodyera repens (L.) R. Br.
Hepatica transsilvanica Fuss.
-
R
R
R
R
R
R
Orchis morio L. ssp. morio

Orchis purpurea Hudson
Orchis ustulata L.
Orobanche lucorum A. Braun
Pinus sylvestris L.
Plantago schwarzenbergiana
Schur
Platanthera bifolia (L.) L.C.M.
Richards
Platanthera clorantha
Pleurospermum austriacum (L.)
Hoffm.
Potamogeton trichoides Cham.
et Schlecht.
Primula elatior L. ssp.
leucophylla (Pax.) H. Harrison
ex W.W. Sm. et Fletcher
Pulsatilla grandis Wenderoth
Rhynchospora alba (L.) Vahl.
Ribes spicatum Robson
Rorippa islandica (Oeder)
Borbs
-
R
-
Rosa micrantha Sm.

-
R
R
R
Salix aurita L.
Salix daphnoides Vill.
Salvinia natans (L.) All
Scirpus radicans Schkuhr
Sempervivum zeleborii Schott
Serratula radiata (Welk) Bieb.
Seseli hippomarathrum Jacq.
-
R
R
R
K
E
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
VR
-
134
V
-
Plantago cornuti Gouan.

-
Pleurospermum austriacum (L.)

Hoffm.
Potamogeton trichoides Cham. et
Schlecht.
Primula elatior L. ssp.
leucophylla (Pax.) H. Harrison ex
W.W. Sm. et Fletcher
Pyrola chlorantha Swartz
Rhynchospora alba (L.) Vahl.
Ribes spicatum Robson
-
R
nt
R
R
R
R
Rorippa prolifera (Heuffel)

Weiche
Rubus glandulosus Bellardi
Rumex aquaticus L.
Rumex longifolius DC. in Lam. et
DC.
Sedum caespitosum (Cav.) DC.
Seseli tortuosum L.
Seseli hippomarathrum Jacq.
Silene italica Retz. ssp. italica
Sisymbrium altissimum L.
R
R
R
R
VR
K
V
R
R
R
VR
R
K
R
R
Sorbus aria (L.) Crantz.

Spirea crenata L.
Stellaria palustris Retz.
Symphytum tauricum Willd.
Taxus baccata L.
Thymus serpyllum L.
Traunsteinera globosa (L.)
Reichenb.
Trisetum macrotrichum Hackel
Trollius europaeus L. ssp.
europaeus
Typha shuttlewortii Koch et
Sonder
Vicia peregrina L.
Vicia tenuifolia Roth.
Viola jooi Janka
Wolffia arrhiza (L.) Horkel ex
Wimmer
E
E
E
E
R
nt
V
-
Sisymbrium irio L.
Sisymbrium loeselii L.
Sisymbrium officinale (L.) Scop.
Sisymbrium strictissimum L.
Taxus baccata L.
Thymus comosus Heuffel
Thymus serpyllum L.
-
V
R
-
Trollius europaeus L. ssp.

europaeus
-
R
R
-
Vicia peregrina L.
Viola jooi Janka
-
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
135

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
So 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 (4.2C). 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 So 1949
Ord. Arrhenatheretalia R. Tx. 1931
Al. Cynosurion R. Tx. 1947
As. Festuco rubrae-Agrostietum capillaris Horvat 1951
136
As. Agrostideto Festucetum pratensis So 1949

Corollogy: Piatra Fntnele
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 underlayer, 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 Fntnele
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%.
137
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) i 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.
2.
BELDIE AL., 1977 - Flora Romniei Determinator ilustrat al plantelor vasculare, vol. I-II, Edit. Acad.
R.S.R., Bucureti
CHIFU T., 1995 - Contribuii la sintaxonomia vegetaiei pajitilor din clasele Molinio Arrhenatheretea Tx.
37 i Agrostietea stoloniferae Oberd. in Oberd. et al. 67 de pe teritoriul Moldovei. Bul. Grd. Bot. Iai, 5:
125-132
138
3.
CHIFU T., MNZU C., ZAMFIRESCU O., 2006 - Flora i vegetaia Moldovei, vol. II, Ed. Univ. Al. I.
Cuza Iai
4.
CHIRI V., 2003 - Depresiunea Dornelor: studiu fizico geografic. Edit. Univ., Suceava
5.
CIOCRLAN V., 2000 - Flora ilustrat a Romniei Pteridophyta et Spermatophyta, Edit. Ceres,
Bucureti
6.
ELLENBERG H., 1974 - Indicator values of vascular plants in Central Europe, Scripta Geobotanica, vol. IX,
Verlag Erich Goltze K.G., Gttingen: 1-97
7.
SANDA V., 2002 - Vademecum ceno-structural privind covorul vegetal din Romnia, Edit. Vergiliu,
Bucureti
8.
SANDA V., POPESCU A., ARCU M., 1999 - Revizia critic a comunitilor de plante din Romnia, Edit.
Tilia Press International1, Constana
9.
SANDA V., POPESCU A., BARABA N., 1997 - Cenotaxonomia i caracterizarea gruprilor vegetale din
Romnia. St. i Com. Muz. t. Nat. Bacu, Biol. veget., 14: 2-365
10. SANDA V., POPESCU A., STANCU D. I., 2001 - Structura cenotic i caracterizarea ecologic a
fitocenozelor din Romnia, Edit. Conphis, Bucureti
Table I - As. Agrostideto Festucetum pratensis So 1949

Relev number
1
Altitude (m)
1130
Exposition
E
Slope ()
5
90
Surface of the relev (m)
100
Number of species
43
Caract. as.
3
Alopecurion pratensis
Festuca pratensis ssp. pratensis
1
Phleum pratense
+
Molinion caeruleae et Molinietalia caeruleae
Briza media
+
Gymnadenia conopsea ssp. conopsea
+
Lychnis flos-cuculi
Arrhenatherion
Campanula patula
Centaurea phrygia
1
Cynosurion
Cynosurus cristatus
+
Leontodon autumnalis ssp.
+
autumnalis
Trifolium repens ssp. repens
Arrhenatheretalia
Achillea millefolium ssp.
+
millefolium
Carlina acaulis ssp. acaulis
+
Carum carvi
+
Crepis biennis
+
Dactylis glomerata
Heracleum sphondylium ssp.
+
sphondylium
Knautia arvensis ssp. arvensis
-
2
1125
V
15
95
100
36
3
1125
NE
2
100
100
28
4
1100
E
20
90
100
34
5
1120
NV
10
80
100
24
6
1120
V
10
85
100
27
2
+
3
+
2
+
2
-
1
-
V
IV
+
+
+
-
+
+
+
-
IV
II
II
+
+
+
-
+
-
+
+
IV
IV
1
+
+
+
+
-
+
-
+
-
V
III
II
+
+
+
-
+
+
-
+
+
+
+
+
-
1
+
+
+
V
IV
II
III
III
II
139
Leucanthemum vulgare ssp. vulgare

Tragopogon pratensis ssp. orientalis
Thymus pulegioides
Veronica chamaedrys ssp.
chamaedrys
Anthoxanthum odoratum
Campanula glomerata ssp. glomerata
Euphrasia officinalis ssp. pratensis
Lotus corniculatus
Plantago lanceolata ssp. lanceolata
Polygala vulgaris
Ranunculus acris ssp. acris
Rhinanthus angustifolius ssp.
angustifolius
Rumex acetosa
Trifolium pratense
Vicia cracca
Potentillo Nardion
Arnica montana
Cruciata glabra
Scorzonera rosea
Juncetea trifidi
Campanula serrata
Dianthus deltoides
Hypochaeris uniflora
Nardus stricta
Potentilla erecta
Festuco Brometea
Plantago media
Trifolium alpestre
Variae syntaxa
Alchemilla vulgaris
Galeopsis speciosa
Hypericum maculatum ssp.
maculatum
Stellaria media
Tanacetum corymbosum
Viola tricolor
+
+
+
-
1
+
+
+
+
+
+
+
1
+
-
+
+
-
+
+
+
V
IV
V
III
+
+
+
+
+
+
+
1
1
+
+
+
+
+
+
1
+
+
+
+
+
+
1
1
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
V
II
IV
V
V
IV
III
IV
V
+
+
-
+
+
+
-
+
+
+
+
+
III
IV
III
+
+
+
+
-
+
+
-
II
III
II
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
1
+
+
II
II
V
V
V
+
+
+
+
+
-
+
-
+
+
+
-
+
-
III
V
II
1
+
+
+
-
+
-
+
+
+
+
+
+
-
V
II
III
+
+
+
+
-
+
-
II
II
II
Place and date of the releves: Piatra Fntnele

1 - 27.07.2006; 2, 3, 4 - 20.08.2006; 5, 6 - 1.09.2006
140
Table II - As. Festuco rubrae Agrostietum capillaris Horvat 1951

Relev number
Altitude (m)
Exposition
Slope ()
Surface of the relev (m)
Number of species
Caract. as.
Agrostis capillaris
Festuca rubra
Cynosurion
Bellis perennis
Cynosurus cristatus
Leontodon autumnalis ssp.
autumnalis
Phleum pratense
Plantago major ssp. major
Prunella vulgaris
Trifolium repens ssp. repens
Veronica serpyllifolia ssp.
serpyllifolia
Arrhenatherion
Campanula patula
Centaurea phrygia
1
1130
V
5
95
100
30
2
1140
E
5
95
100
32
3
1140
S
5
95
100
26
4
1100
NV
1
80
100
24
5
1100
NV
3
90
100
37
6
1125
NE
20
90
100
29
7
1150
E
10
90
100
26
8
1148
V
10
95
100
36
9
1145
S
2
100
100
24
10
1150
V
2
100
100
26
11
1150
E
10
90
100
29
1
3
2
3
3
2
4
2
+
4
1
4
3
2
3
2
2
3
1
4
1
4
V
V
+
+
+
+
+
+
+
+
+
-
+
-
+
+
-
+
+
+
-
1
-
II
V
II
+
-
+
+
+
+
+
+
-
+
+
+
+
-
+
+
-
+
+
+
+
+
+
-
+
1
-
1
+
-
+
+
+
+
-
III
II
IV
IV
II
+
+
+
+
+
+
+
+
+
+
-
+
-
+
+
+
+
-
+
-
+
+
-
IV
III
III
141
Phyteumo Trisetion
Hypericum maculatum ssp.
maculatum
Luzula luzuloides ssp. luzuloides
Arrhenatheretalia
Achillea millefolium ssp.
millefolium
Briza media
Campanula glomerata ssp.
glomerata
Carlina acaulis ssp. caulescens
Carum carvi
Leucanthemum vulgare ssp. vulgare
Knautia arvensis
Thymus pulegioides
Tragopogon pratensis ssp. orientalis
Molinietalia
Gymnadenia conopsea ssp.
conopsea
Lychnis flos cuculi
Alchemilla vulgaris
Anthoxanthum odoratum
Centaurea jacea
Euphrasia officinalis ssp. pratensis
Lotus corniculatus
Plantago lanceolata ssp. lanceolata
III
II
IV
+
+
+
-
+
-
+
-
+
-
+
-
+
-
+
-
+
-
V
I
+
+
+
-
+
+
+
+
-
+
+
+
+
+
+
-
+
+
1
-
+
+
+
+
-
+
+
+
+
+
+
-
1
+
+
+
+
+
-
+
+
+
+
+
IV
II
V
II
V
II
II
1
+
+
+
+
+
+
+
+
-
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
1
+
+
+
+
1
+
+
+
+
+
1
+
-
+
+
+
+
-
+
+
+
+
-
V
V
III
I
III
IV
IV
142
Polygala vulgaris
Ranunculus acris ssp. acris
Rhinanthus minor
Rumex acetosa
Stellaria graminea
Trifolium pratense
Festuco Brometea
Anthyllis vulneraria ssp. vulneraria
Echium vulgare
Euphrasia stricta ssp. stricta
Galium verum
Hieracium pilosella
Ranunculus polyanthemos ssp.
polyanthemoides
Trifolium alpestre
Juncetea trifidi
Campanula serrata
Nardus stricta
Potentilla ternata
Scorzonera rosea
Calluno Ulicetea
Antennaria dioica
Arnica montana
Dianthus deltoides
Genista tinctoria ssp. tinctoria
Gentianella austriaca
Potentilla erecta
+
1
+
+
+
1
-
+
+
1
-
+
+
+
+
+
+
+
+
+
+
+
+
-
+
+
+
-
+
+
+
+
+
+
-
+
+
1
III
II
V
II
I
III
+
+
+
+
+
+
-
+
1
-
+
+
+
+
+
+
+
+
-
+
+
+
+
-
III
I
I
II
III
II
+
-
+
-
+
-
+
-
+
+
+
-
+
-
+
-
IV
II
+
+
+
+
+
-
+
-
+
+
-
+
-
+
+
-
+
+
+
-
III
I
III
II
+
+
+
+
-
+
-
+
+
+
-
+
+
+
1
+
+
+
+
1
+
+
+
+
+
+
+
I
II
II
II
II
IV
143
Variae syntaxa
Cirsium vulgare
Cruciata glabra
Pteridium aquilinum
Rumex acetosella ssp. acetosella
Tanacetum corymbosum ssp.
corymbosum
+
+
+
-
+
+
-
+
+
-
+
-
+
+
+
+
-
+
-
+
-
+
-
Place and date of the releves:

1 - Piatra Fntnele, 27.07.2006; 2 - 11 - Piatra Fntnele, 21, 22.08.2006
H - 74,19%
T - 6,45%
T - 11,76%
Ht - 9,68%
H - 70,59%
Ht - 9,8%
Ch - 4,84%
Ch - 3,92%
G - 3,92%
G - 4,84%
Fig. 1. The bioforms spectrum: a) as. Agrostideto-Festucetum pratensis So 1949;

b) as. Festuco rubrae-Agrostietum capillaris Horvat 1951
144
II
II
II
II
I
Circ. - 18,35%
Pont. - 1,47%
Centr. eur. 9,91%
Medit. -0,37%
Atl. - 0,92%
Eur. - 12,11%
Dacic - 0,18%
Euras. 44,77%
% 80
70
60
50
40
30
20
10
0
L
T
Ct
U
R
N
7
b
9 10 11 x
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 So 1949
% 70
Euras. - 50%
Cosm - 8,06%
Eur. - 30,65%
End. - 1,61%
Alp. - 3,23%
Circ. - 3,23%
Carp. balc. 1,61%
Pont. - 1,61%
60
50
40
Ct
30
20
10
0
1
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

ENVIRONMENTAL EDUCATION: EDUCATION FOR TRANSITION TO

SUSTAINABLE DEVELOPMENT
I. M. CIUMAU , NAELA COSTIC
Abstract: It is general accepted that the principles of sustainable development cant 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, Iai, 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, Iai, Romania. and Technische
Universitt Mnchen, Arcistrasse 21, 83000, Mnchen, Deutschland, Wissenschaftszentrum Weihenstephen fr
Ernhrung, Landnutzung und Umwelt, Biowissenschaftlische Grundlagen, Alte Akademie 8, 85354, Freising,
Deutschland, Lehrstuhl fr kologische Chemie und Umweltanalytik Weiehnstephaner Steig 23, 85350, FreisingWeihenstephan, Deutschland.
146
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-ityourself 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
147
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.
148
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
149
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 schoolbased 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,
150
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
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7, 2:79-84
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
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
Chan, K., 2000 - Use of environmental teaching kits in Hong Kong. The Environmentalist, 20:113-121
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
Dahlgren, M.A., berg, 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
Dempsey, R., Gresele, C., Bgeholtz, 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.
Disinger, J.F., 1997 - Environmental education research news. The Environmentalist, 17:153-156
Donahue, T.P., Lewis, L.B., Price, L.F., Schmidt, D.C., 1998 - Bringing science to life through communitybased watershed education. Journal of Science Education and Technology, 7, 1:15-23
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
Fien, J., Poh, I. T.-C., Yencken, D., Sykes, H., Treagust, D., 2002 - Youth environmental attitudes in
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Gough, G.A., Robottom, I., 1993 - Towards a socially critical environmental education: water quality studies
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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
Lucas, A.M., 1980 - Science and environmental education: pious hopes, self praise and disciplinary
chauvinism. Studies in Science Education, 7: 1-26
Lucie, S., 1999 - Environmental Education between modernity and postmodernity: searching for an
integrating educational framework. Canadian Journal of Environmental Education, 4: 9-35
Manring, S. & Moore, S.B., 2006 - Creating and managing a virtual inter-organizational learning network for
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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
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22. Reid, A., Petocz, P., 2006 - University lecturers' understanding of sustainability. Higher Education , 51: 105123
23. Rickinson, M., 2001 - Learners and learning in environmental education: a critical review of the evidence.
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26. Shepardson, D.P., Harbor, J., 2004 - ENVISION: the effectiveness of a dual-level professional development
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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: 307322
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
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34. Wormstead, S.J., Becker, M.L., Congalton, R.G., 2002 - Tools for successful student-teacher-scientist
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152
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 organisms vital functions, as well as to form the readers 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
153
REVIEW
TNASE C., ESAN TATIANA EUGENIA, 2006, Concepte actuale n taxonomia
ciupercilor, Editura Universitii Alexandru Ioan Cuza, Iai: 510 pp. (ISBN 973-703144-X; 978-973-703-144-0)
Rod al unei minuioase munci, Concepte actuale n taxonomia ciupercilor este cel
mai complex i mai complet manual universitar tratnd att clasificarea ciupercilor ct i
morfologia, citologia, ultrastructura, biologia, ecologia i reproducerea lor, cu informaii
utile i asupra utilizrii multora dintre ele.
Aceast carte se adreseaz n primul rnd specialitilor, dar i studenilor de la
facultile de Biologie (cu secii de Biologie, Ecologie i Protecia Mediului, Biochimie,
Biotehnologie), Silvicultur, Agricultur, Horticultur, Farmacie, Medicin veterinar i
uman, Industrii alimentare, precum i celor interesai de cunoaterea i conservarea
diversitii ciupercilor, de protecia patrimoniului documentar, de carte, a monumentelor
istorice, a diverselor tipuri de materiale .a.
Autorii cadre didactice din dou prestigioase universiti ale rii, Iai i
Bucureti au proiectat aceast lucrare sub forma unui tratat pe baza sintezei informaiilor
teoretice selectate din literatura de specialitate, a rezultatelor personale i a experienei
acumulate n domeniul micologiei. Aceast experien, este concretizat n tratarea
minuioas a unor aspecte originale care vizeaz att identificarea acestor organisme, ct i
analiza critic a unor aspecte actuale referitoare la biologia, ecologia, corologia, biochimia
i taxonomia ciupercilor.
Tratatul este structurat n 5 capitole, care reunesc att aspecte teoretice ct i
practice din domeniul micologiei. Ciupercile sunt prezentate ca un grup de organisme
polifiletice reunite n 11 ncrengturi, care aparin la 3 regnuri: Chromista, Fungi i
Protozoa. n aceste categorii taxonomice sunt descrise 1170 specii de ciuperci grupate n
115 familii.
Lucrarea cuprinde 510 pagini, n care sunt inserate 366 de figuri, 120 fotografii
color originale i 30 de tabele. Tratatul a fost realizat de cei doi autori, pe baza celor 580 de
lucrri de specialitate consultate (marea majoritate aprute dup 1990), a rezultatelor
personale i a experienei acumulate de-a lungul multor ani de activitate n domeniu.
Prin analiza critic a cercetrilor din domeniul micologiei generale i aplicate,
inserate n baze de date internaionale sau n reviste i volume ale unor manifestri
tiinifice internaionale, autorii evalueaz reperele i realizrile obinute pn n prezent,
precum i direciile de perspectiv.
n acest context, evideniez experiena i rezultatele autorilor, racordate la
realizrile internaionale, foarte multe comunicate i publicate n reviste de specialitate din
ar i din strintate.
Prof. dr. C. DRGULESCU
154
INSTRUCTIONS TO AUTHORS
The Journal Analele tiinifice ale Universitii Al. I. Cuza din Iai (serie
nou), Seciunea II a. Biologie vegetal, includes original articles of cytology, morphoanatomy, physiology, taxonomy, phytosociology, mycology, phytopathology, along with
book reviews and anniversary announcements.
All the papers must be submitted to our redaction address (Dr. Naela COSTIC,
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 Munii Bucegi. Edit. Acad. Rom., Bucureti
2.
for articles:
DAUB E. M., 1981 - Cercosporin, a photosensitizing toxin from Cercospora species.
Phytopathology, 72: 370 374
155
3.
REDZI S., TUKA M., PAJEVI 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. RUGIN R., TOMA C., IVNESCU L., 2007 Morphological and histo-anatomical
aspects at some dicotyledonate seedlings related to the vascular transition. An. t.
Univ. Al. I. Cuza Iai, 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 tiinifice ale Universitii Al. I. Cuza din Iai, Sec. II a. Biologie vegetal.
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
156

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ANALELE TIINIFICE

UNIVERSITII Alexandru Ioan Cuza

TOMUL LIV, FASCICULA 1

Editura Universitii Alexandru Ioan Cuza Iai

Editura Universitii Alexandru Ioan Cuza din Iai

Dr. Constantin TOMA Universitatea Alexandru Ioan Cuza din Iai

Comisia de refereni tiinifici:

IRINA BERCIU, C. TOMA Histo-anatomical aspects reffering to the

ADRIANA PETRU VANCEA, C. F. BLIDAR, ANCA BACIU African

T. CHIFU, C. MNZU, OANA ZAMFIRESCU Contribution to the study of

Analele tiinifice ale Universitii Al. I. Cuza Iai

THE LEAF STRUCTURE OF SOME NEPENTHES DANSER SPECIES

Botanical Gardens of Iasi, Dumbrava Roie Street, no. 7-9, Romania

Materials and methods

IRINA STNESCU, C.TOMA

IRINA STNESCU, C.TOMA

IRINA STNESCU, C.TOMA

IRINA STNESCU, C.TOMA

IRINA STNESCU, C.TOMA

Analele tiinifice ale Universitii Al. I. Cuza Iai

HISTO-ANATOMICAL ASPECTS REFFERING TO THE VEGETATIVE

Explanation of the plates:

IRINA BERCIU, C. TOMA

IRINA BERCIU, C. TOMA

Analele tiinifice ale Universitii Al. I. Cuza Iai

CONTRIBUTIONS TO THE HISTO-ANATOMICAL STUDY OF THE

Cross-sections - the upper leaf, the middle third

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

Analele tiinifice ale Universitii Al. I. Cuza Iai

MORPHO-ANATOMICAL AND PHYTOCHEMICAL RESEARCHES

hemlock", while menziesii is used in recognition of Archibald Menzies (1754-1842), a

Results and discussions

30. Annual ring 2003 thickness (10x)

The needle and young shoots phytochemistry in P. menziesii:

oil and -pinene (3.88-9.23%), -pinene (12.6-15.22%), -terpinene (21.05-32.87%),

xylem II (I II III: annual rings)

resin ducts from stem

Analele tiinifice ale Universitii Al. I. Cuza Iai

THE ECOPHYSIOLOGICAL REACTION OF SOME VARIETIES OF APPLE

Table I. The strength of Erwinia amylovora attack in 2004

* after Cimpoe, 2001 and Grdinaru, 2002

Results and discussions

Dry substance (%)

Photosynthesis (mg CO2/g/h)

Assimilatory Pigments (mg/g fresh

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

Analele tiinifice ale Universitii Al. I. Cuza Iai

PHYSIOLOGICAL EFFECTS INDUCED BY PURINIC SUBSTANCES AT

The report of chlorophyll a/b:

Analele tiinifice ale Universitii Al. I. Cuza Iai

THE INFLUENCE OF THE Mn2+ IONS EFFECTS ON THE WHEAT (TRITICUM

tefan cel Mare University, Universitii 13 Street, Suceava, Romania

Fig. 1: Germination analysis

Fig. 2: Hypocotyl length analysis

Conclusions and discussions

Analele tiinifice ale Universitii Al. I. Cuza Iai

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

Department of Biology, Faculty of Chemistry-Biology-Geography, West University of Timisoara, Romania

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

Results and discussions

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

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

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

Analele tiinifice ale Universitii Al. I. Cuza Iai