Analele Stiintifice Biologie 3 2008

ANALELE TIINIFICE
ALE
UNIVERSITII Alexandru Ioan Cuza

DIN IAI
(SERIE NOU)
SECIUNEA II
a. BIOLOGIE VEGETAL
TOMUL LIV, FASCICULA 2
2008
Editura Universitii Alexandru Ioan Cuza Iai
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
univ. dr. Constantin TOMA Universitatea Alexandru Ioan Cuza din Iai
univ. dr. Toader CHIFU Universitatea Alexandru Ioan Cuza din Iai
univ. dr. Mihai MITITIUC Universitatea Alexandru Ioan Cuza din Iai
univ. dr. Maria Magdalena ZAMFIRACHE - Universitatea Alexandru Ioan Cuza din
Iai
Profesor univ. dr. Ctlin TNASE Universitatea Alexandru Ioan Cuza din Iai
Confereniar univ. dr. Lcrmioara IVNESCU Universitatea Alexandru Ioan Cuza din Iai
Comisia de refereni tiinifici:

Academician Valeriu COTEA Academia Romn, Filiala Iai
Profesor univ. dr. Constantin TOMA Universitatea Alexandru Ioan Cuza din Iai, membru
corespondent al Academiei Romne, Filiala Iai
Profesor univ. dr. Leontin tefan PTERFI Universitatea Babe-Bolyai din Cluj-Napoca,
membru corespondent al Academiei Romne, Filiala Cluj-Napoca
Profesor univ. dr. Jean Pierre AUQUIRE Universitatea Catolic din Louvain-la-Neuve,
Belgia
Dr. Mary GREGORY Jodrell Laboratory, Royal Botanic Gardens, Kew, Anglia
Profesor univ. dr. Ioana POPESCU Drury University, Springfield, Missouri, SUA
Profesor univ. dr. Andrei MARIN Universitatea din Bucureti
Profesor univ. dr. Ioan BURZO Universitatea Agronomic i de Medicin Veterinar din
Bucureti
Profesor univ. dr. Toader CHIFU Universitatea Alexandru Ioan Cuza din Iai
Profesor univ. dr. Mihai MITITIUC Universitatea Alexandru Ioan Cuza din Iai
Profesor univ. dr. Ursula STNESCU Universitatea de Medicin i Farmacie Gr. T. Popa din Iai
Profesor univ. dr. Ctlin TNASE Universitatea Alexandru Ioan Cuza din Iai
Redactor responsabil: Profesor univ. dr. Constantin TOMA,
membru corespondent al Academiei Romne
Secretar de redacie: Preparator dr. Ramona Crina GALE
Tehnoredactare computerizat: Preparator dr. Ramona Crina GALE
CONTENTS
M. ANDREI, ROXANA MARIA PARASCHIVOIU- Anatomical researches on
the overground vegetative organs of Saxifraga mutata L. subsp. demissa (Schott
& Kotschy) D.A. Webb and Saxifraga paniculata Miller
RAMONA GALE, C. TOMA, LCRMIOARA IVNESCUMorphological and histo-anatomical aspects regarding the floral morphogenesis
in Euphorbia cyparissias L. (Euphorbiaceae Juss.)
16
GENIANA MIHAELA IULIA PREDAN, IRINA GOSTIN
Microsporogenesis and the male gametophyte at Ephedra distachya L.
25
ANCA HEMCINSCHI, RAMONA GALE, C. TOMA

Vegetative anatomy of two Galium L. species (Rubiaceae)
31
MARIA MAGDALENA ZAMFIRACHE, C. TOMA, MARIA DUCA,

SIMONA DUNCA, ZENOVIA OLTEANU, M. TEFAN, RAMONA GALE,
CLAUDIA PDURARIU -A comparative study regarding the morphology and
anatomy of the vegetative apparatus in two Ocimum basilicum L. breeds
39
IRINA STNESCU, GABRIELA VASILE - A comparative numeric analysis of
the secretory trichomes belonging to the foliar limb in some Drosera species
49
CLARA APROTOSOAIE A., URSULA STANESCU, ANCA MIRON,
VIOLETA FLORIA, OANA CIOANCA, MONICA HANCIANU Morphological researches regarding the influence of Topsin M treatments on
Mentha longifolia (L.) Huds. species
56
LUMINIA HUANU-BASHTAWI, C. TOMA - A histo-anatomical study of
the modifications induced by thiophanate methyl on the Calendula officinalis L.
strain
64
NICOLETA IANOVICI - The analysis of daily concentrations of airborne
pollen in the west and southwest of Romania
74
SILVICA PDUREANU - Variability of some pollen morphological traits of
certain taxa in the bordering area of the Ceahlu National Park
80
I. BURZO, V. CIOCRLAN, ELENA DELIAN, AURELIA DOBRESCU,

LILIANA BDULESCU - Research as regard to essential oil composition of
some Artemisia L. species
87
ALEXANDRINA MURARIU,
GIANINA BERECHET,
ANIOARA
STRATU, CAMELIA IFRIM- The ecophysiological peculiarities of some
species of the Acer genus from the Botanical Garden of Iai
93
MARIA
ANIOARA
STRATU,
ALEXADRINA
MURARIU,,
MAGDALENA ZAMFIRACHE, ZENOVIA OLTEANU, LCRMIOARA
OPRIC, C. TNASE, V. C. CHINAN,, C. BRSAN - Physiological and
biochemical aspects in the lignicolous species Gloeophyllum odoratum (Wulfen)
Imazeki and Fomitopsis pinicola (Sw.)P. Karst. (Fungi, Basidiomycota)
collected from Climani National Park (the Oriental Carpathians)
98
SMARANDA VNTU - Somatic embryogenesis in Rubus caesius L.
suspension cultures
105
ROXANA-IULIANA TEODOR, AL. YEPHREMOV Genes of Arabidopsis
thaliana involved in wax metabolism
110
M. A. PORUMB, M. COSTIC Characteristics of planktonic algoflore from
Dorobanti, Aroneanu, Ciric I, II and III lakes (Iasi county)
115
V. SURUGIU - On the occurrence of Zostera noltii Hornemann at the Romanian

coast of the Black Sea
122
OANA ZAMFIRESCU, C. MNZU, T. CHIFU - Contributions to the study of128
the ruderal vegetation from the Ceahlu mountain
IRINA IRIMIA - Contributions to the vegetation study from the Vaslui river
basin (II)
136
Analele tiinifice ale Universitii Al. I. Cuza Iai

Tomul LIV, fasc. 2, s.II a. Biologie vegetal, 2008
ANATOMICAL RESEARCHES ON THE OVERGROUND VEGETATIVE ORGANS

OF SAXIFRAGA MUTATA L. SUBSP. DEMISSA (SCHOTT & KOTSCHY) D.A. WEBB
AND SAXIFRAGA PANICULATA MILLER
M. ANDREI*, ROXANA MARIA PARASCHIVOIU*
Abstract: This paper presents the structure of the flowering stem, the rosette and cauline leaves of Saxifraga
mutata L. subsp. demissa (Schott & Kotschy) D.A. Webb and S. paniculata Miller. The vegetal material was
collected from Bucegi Natural Park. The structural features of the aerial vegetative organs are correlated with
the environmental conditions of these plants. The flowering stem of S. mutata subsp. demissa has
sclerenchymatous sheaths around the isolated vascular bundle as well as around groups of two or three vascular
bundles. The flowering stem of S. paniculata presents some supplementary collateral vascular bundles among
the initial bundles as a result of the enlarging of the stem. We observed isolated phloem groups among the
collateral vascular bundle in the flowering stem of both plants. There is a multilayered sclerenchymatous
pericycle with a mechanical role in the plants in the flowering stem of both analysed taxa. The presence of the
tracheid hydathodes in the rosette and cauline leaves of S. mutata subsp. demissa and S. paniculata is an
adaptation to the high temperature on the limestone rocks in the subalpine and alpine areas. The existence of
the endodermis (with Caspary strips in S. mutata subsp. demissa) and the monolayered parenchyma pericycle
around the vascular bundle in the leaf structure makes the foliar vascular bundle similar to a monobundled
central cylinder. The flowering stem and the cauline leaf have multiseriate glandular hairs and a few
multiseriate and uniseriate non-glandular hairs. The rosette leaves are hairless. The leaves are amphistomatic,
with anomocytic stomata in S. mutata subsp. demisa and tetracytic in S. paniculata.
Key words: sclerenchymatous pericycle, tracheid hydathode, guttation, epithem, multiseriate glandular hairs,
endodermis with Caspary strips in the leaf vein, amphistomatic, collateral vascular bundle.
Introduction
Both studied taxa of the Saxifraga genus are included in the Section Ligulatae Haworth,
but in two distinct subsections: Aizoonia (Tausch) Schott. (includes S. paniculata Miller) and
Mutatae (Engler & Irmscher) Gornall (with S. mutata L. subsp. demissa) [18]. Saxifraga mutata
L. subsp. demissa (Schott & Kotschy) D.A. Webb is a rare and vulnerable plant, endemic to
Romania, already included in the Red List of vascular plants [13]. S. paniculata Miller is a
common plant, widespread in Romania.
Both taxa are perennial, but S. mutata subsp. demissa is monocarpic, usually solitaire or
found in small groups, while S. paniculata is a policarpic, cespitose plant. S. paniculata preferes
sunny limestone rocks while S .mutata subsp. demissa preferes shady places, inside rocks
crevices. The leaves of the two analised taxa are placed alternately on the flowering stem as
well as in a basal rosette.
*
Faculty of Biology, Bucharest University, Romania
Material and methods

The plants were collected in the anthesis from Bucegi Mounts (1530m on 12 of July
2005 for S. mutata subsp. demissa and 1900m on 29 of June 2006 for S. paniculata). The
vegetal material was fixed, preserved, sectioned and coloured according to the usual plant histoanatomical research technique. The cross sections and the tangential/longitudinal ones of the
aerial vegetative organs were analyzed and photographed with a DOCUVAL microscope.
Results and discussions
I. Saxifraga mutata L. subsp. demissa (Schott & Kotschy) D.A. Webb
1. The flowering stem structure. The outline of the flowering stem cross section is
circular (Pl.I:Fig.1). The epidermis, covered by a relatively thick cuticle, is composed of one
layer of isodiametrical cells having thickened walls. Some of the epidermis cells are
transformed in stomata and some in hairs. The stomata have a small air space beneath them and
are located slightly above the rest of the epidermis. The hairs are multiseriate glandular and
multiseriate and uniseriate non-glandular type (Pl.I: Fig.2; Fig.3; Fig.4).
The cortex has 8 to 10 layers of parenchymatous cells with a few intercellular spaces.
The innermost layer of the cortex is the starch sheath having different sized cells with cellulose
walls (Pl.I: Fig.1). The central cylinder presents a multilayered (7-8 layers) sclerenchymatous
pericycle, with a mechanical role. The pericycle cells are small, without intercellular spaces and
have very thick, lignified walls (Pl.I: Fig.1; Fig.4). The pericycle surrounds numerous vascular
bundles (14-16) set in a ring (eustele) (Pl.I: Fig.1; Fig.4). The vascular bundles are collateral.
There are pith rays between the vascular bundles with large lignified cells. The mechanical cells
surround each vascular bundle or groups of two or three bundles forming sclerenchymatous
sheaths (Pl.I: Fig.1; Fig.4). We also observed isolated groups of phloem between the vascular
bundles (Pl.I: Fig.4). The pith has parenchymatous cells (Pl.I: Fig.1).
2. The leaves structure. The structural analysis of the leaves pointed out the following
features:
Both basal rosette leaves and the cauline ones have a bifacial dorsiventral structure with
the two epidermis (upper and lower) and a mesophyll differentiated in palisade tissue, located
under the upper epidermis, and spongy tissue, on the lower half of the lamina. The palisade
tissue has 3-4 layers of hetero-diametrical cells with numerous chloroplasts.
The rosette leaves epidermis is hairless while the cauline leaves epidermis has
multiseriate glandular hairs with a multicellular spherical head. The rosettes leaves as well as
the cauline ones are amphistomatic. The epidermis of the two types of leaves have many
anomocytic type stomata, observed in the tangential (superficial) sections (Pl.II: Fig.1). The
limits between the subsidiary cells can be seen.
The leaves don't have an obvious petiole. The tapered base of the leaves have a
homogenous mesophyll. In the mesophyll veins there are collateral vascular bundles.
Sometimes there are one or two small vascular bundles close to the large vascular bundle of the
midvein (Pl.II: Fig.2). The vascular bundles are surrounded by a multilayered collenchymatous
sheath. A specific feature of the leaf vascular bundle is the presence, under the collenchymatous
sheath, of an endodermis whose cells have Caspary strips on their radial walls. This endodermis
surrounds a monolayered parenchymatous pericycle and thus the mesophyll veins have a
similar structure to a monobundle vascular cylinder.
On both sides of the leaves, near the cartilaginous margin, many hydathodes set in a row
can be observed. These tracheid hydathodes have an epithem with numerous heterodiametric
parenchymatous cells without chloroplasts and with thin walls (Pl.II: Fig.3; Fig.4). This
parenchyma is supplied with water from a group of tracheids which are connected to the
vascular leaf bundles. The epithem opens on the leaf surface with one large aquifer stoma
similar to the other epidermis stomata but without closing movements. This stoma remains
permanently opened. The water eliminated from the hydathodes in the guttation process
represents the adaptation found by these plants in order to create around them a wet
atmosphere, very useful against the high temperature of the limestone rocks in the subalpine
area.
II. Saxifraga paniculata Miller
1. The flowering stem structure. The outline of the flowering stem cross section is
almost circular (Pl.III: Fig.1). The epidermis is composed of one layer of isodiametric cells and
covered by cuticle. In the epidermis there are few stomata (Pl.III: Fig.5) with a small air space
beneath them, located slightly above the other epidermis cells, similar to those of the other
analysed plant. The flowering stem epidermis has some glandular hairs usually with a
multiseriate stalk and a terminal multicellular ovoid gland (Pl.III: Fig.2). It has also very few
uniseriate non-glandular hairs (Pl.III: Fig.6). The separating walls of adjacent cells of the hair
stalk have simple punctuations and may be simple or divaricated (Pl.III: Fig.4).
The cortex has 7 to 9 layers of isodiametric parenchymatous cells with different sized
intercellular spaces. The innermost layer of the cortex is the starch sheath (Pl.III: Fig.1).
The central cylinder has a multilayered (6-7 layers) sclerenchymatous pericycle, with
small cells, without intercellular spaces (Pl.III: Fig.1; Fig. 3). The central cylinder is a eustele
having many vascular bundles set in a ring (Pl.III: Fig.1, Fig.3). The vascular bundles are
collateral. Their size varies because of the presence of supplementary vascular bundles which
are formed at the same time with the widening of the stem. We indicate the presence of some
phloem bundles separated from the collateral bundles, but derived from the latter, which
amplifies the possibility of conducting the photosynthetic products. This observation leads us to
the same conclusion as K. Esau (1965) that conducting tissues are formed whenever they are
needed.
The mechanical cells from around the vascular bundles make up an external cap which
extends also on the sides of the phloem (Pl.III: Fig.3). There are pith rays with large cells (the
external ones present lignified walls) set in 4 or 5 rows between the vascular bundles. The pith
has parenchymatous cells larger than those of the cortex (Pl.III: Fig.1).
2. The leaves structure. The cross sections of the basal rosette leaves and the cauline
ones indicate a bifacial dorsiventral structure. The leaf consists of two epidermis (upper and
lower) and of the mesophyll, differentiated in palisade tissue (with 4-5 layers of heterodiametrical cells) and spongy tissue. The leaves of the S. paniculata Miller are almost sessile,
the lamina has a tapered base which has a petiole-like structure, with an homogenous
mesophyll. The mesophyll differentiation begins in the distal half of the leaf.
Both rosettes and cauline leaves have an amphistomatic lamina, with many stomata of
tetracytic type (Pl.IV: Fig.1). The rosette leaves are hairless while the cauline leaves have
multiseriate glandular hairs with a terminal, ovoid, multicellular gland. The epidermis of the
rosette leaf margins is ciliated at the base. We observed some punctuations in the epidermis and
mesophyll cell walls on the cross sections of the leaf. Thus, the mesophyll can be interpreted as
an aquifer tissue.
The vascular bundles of the mesophyll veins are collateral (Pl.IV: Fig.2). The mesophyll
veins have a similar structure to a monobundle vascular cylinder, the vascular bundle being
surrounded by a monolayered parenchymatous pericycle, an endodermis and two or three layers
of colenchymatous cells. Some of the secondary veins stop on the sides of the leaf at the base of
a passive hydathode (Pl.IV: Fig.3). This tracheid hydathode has an epithem formed by
numerous elongated cells, with thin walls (Pl.IV: Fig.4). In a cross section through the tip of a
cauline leaf we observed a terminal hydathode with numerous small cells and without
intercellular spaces (Pl.IV: Fig.5). The hydathode opens with one large aquifer stoma, on the
upper surface of the leaf, near the margins, at the base of the marginal teeth, in a pit (Pl.IV:
Fig.6). The excretion of water and calcium carbonate through the hydathode is an adaptation of
these plants to their environmental conditions. The presence, number and localisation of
hydathodes represent very important taxonomical criteria in the Saxifraga genus.
Conclusions
The structure analysis of the aerial vegetative organs of Saxifraga mutata L. subsp.
demissa (Schott & Kotschy) D.A. Webb and S. paniculata Miller underlined many similarities
between the two taxa, as following:
1. The presence in the flowering stem of a sclerenchymatous multilayered pericycle, with
mechanical role, can be correlated to the position of this plant on the rocks, ensuring the plant
support.
2. The endodermis and the parenchymatous pericycle present around the leaf vascular
bundle makes the leaf vein similar to a monobundle central cylinder.
3. The tracheid hydathodes existent in the rosette leaves and in the cauline leaves
represent an adaptation of this plant to the environmental condition. The intense guttation is
induced by the high temperature on the limestone substrate during the summer. The water
evaporates on the leaves surface and creates a cool atmosphere around the plant.
4. The large number of vascular bundles in the flowering stem can be correlated to the
tracheid guttation. We also observed isolated phloem groups which amplify the conducting of
the photosynthetic products.
5. The central cylinder is a eustele with collateral vascular bundle.
6. The cauline leaves and the flowering stem are covered with multiseriate glandular
hairs and uniseriate non-glandular hairs.
7. The leaves are amphistomatic.
We also observed several distinct features in the anatomy of the two analysed taxa:
1. In the flowering stem of S. mutata subsp. demissa we observed a few multiseriate nonglandular hairs and in S. paniculata we indicate uniseriate glandular hairs, with divaricated
walls between adjacent cells of the stalk.
2. There are sclerenchymatous sheath around isolated vascular bundles as well as around
groups of two or three vascular bundles in the flowering stem of the S. mutata subsp. demissa.
3. The pith rays of the flowering stem are lignified in S. mutata subsp. demissa and
parenchymatous except for the external ones which are sclerified in S. paniculata.
4. We observed some supplementary collateral vascular bundles among the initial
bundles in the flowering stem of S. paniculata, as a result of the enlarging of the stem.
5. The stomata of the leaves of S. mutata subsp. demissa are anomocytic but those of S.
paniculata are tetracytic.
6. In S. paniculata the hydathodes are found at the base of each marginal tooth while in
the other studied plant they are set in a row, near the cartilaginous margin. In a cross section of
the cauline leaf tip from S. paniculata we observed a hydathode whose epithem is made up of
many cells without intercellular spaces.
7. We observed Caspary strip in the cell wall of the endodermis of the leaves of S.
mutata subsp. demissa.
REFERENCES
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ANDREI M., 1978 - Anatomia plantelor, Edit. Did. .Pedag., Bucureti

ANDREI M., PARASCHIVOIU M. R., 2008 - Protecia i conservarea speciilor genului Saxifraga din Romnia.
In Natura. Seria III, Nr. 2, Bucureti
ANDREI M., PARASCHIVOIU M. R., 2003 - Microtehnic Botanic, Edit. Niculescu, Bucureti
BLAND B., 2000 - Silver Saxifrages, A guide to Encrusted Saxifrages for Gardeners and Botanists, The Alpine
Garden Society, The Friary Press, Dorchester, Great Britain
BOLDOR O., TRIFU M., RAIANU O., 1981 - Fiziologia plantelor, Edit.Did. Pedag., Bucureti
CIOCRLAN V., 2000 - Flora ilustrat a Romniei. Pteridophyta et Spermatophyta, Edit. Ceres, Bucureti
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CUTLER D. F., BOTHA C.E.J., STEVENSON D.W., 2008 - Plant Anatomy. An applied approach, Blackwell
Publishing Ltd., USA
CUTLER D. F., GREGORY M., 1998 - Anatomy of the Dicotiledons, second edition, Vol. IV. Saxifragales,
Clarendon Press, Oxford
ESAU K., 1965 - Plant Anatomy, second edition, John Wiley & Sons, Inc., USA
FAHN A., 1982 - Plant Anatomy, Ed. III, Pergamon Press, Oxford, England
GRINESCU I., 1985 - Botanica, Ed. II, Edit. tiinific i Enciclopedic, Bucureti
HARDING W. 1992 - Saxifrages, A gardeners Guide to the Genus, The Alpine Garden Society, The Friary
Press, Dorchester, Great Britain
OLTEAN M., NEGREAN G., POPESCU A., ROMAN N., DIHORU, G., SANDA V., MIHILESCU S., 1994 Lista roie a plantelor superioare din Romnia. Studii, Sinteze, Documentaii de Ecologie,.1, Academia Romn,
Institutul de Biologie, Bucureti
RVRU M., 1956 - Fam. Saxifragaceae, In Flora Republicii Populare Romne, Vol. IV, p. 85-128, Edit.
Acad. R.P.R., Bucureti
TOMA C., 1995 - Anatomia plantelor I. Histologia i 1977. Anatomia plantelor II. Structura organelor vegetative
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ERBNESCU-JITARIU, Gabriela, TOMA, C., 1980 - Morfologia i anatomia plantelor, E.D.P., Bucureti
WEBB D. A., 1964 - Saxifragaceae, In TUTIN T. G., HEYWOOD V. H., BURGES, N. A., VALENTINE D. H.,
WALTERS S. M., WEBB D. A. Flora Europaea, Vol. I. Licopodiaceae to Plantaceae, University Press,
Cambridge
WEBB D. A., GORNALL R. J., 1989 - Saxifrages of Europe, Cristopher Helm Ltd., Great Britain.
The explanation of figures:

Plate I. S. mutata subsp. demissa: Fig.1. Cross section of the flowering stem showing the epidermis, cortex and the
eustele with sclerenchymatous pericycle (Oc.12,5x; Ob. 25); Fig.2. A multiseriate glandular hair in a cross section
through the flowering stem; Fig.3. One multiseriate non-glandular hair in a cross section through the flowering stem
(Oc.12,5x; Ob. 40); Fig.4. Detail of the flowering stem structure in a cross section showing the collateral vascular
bundles surrounded by sclerenchymatous sheath; one isolated phloem group can be seen (Oc.12,5x; Ob.40); Fig.5.
Tangential section of the flowering stem with a multiseriate glandular hair and a uniseriate non-glandular hair
(Oc.12,5x; Ob. 40).
Plate II. S. mutata subsp. demissa: Fig.1. The lower epidermis of the rosette leaf in an apical view. The simple
punctuations and the stomata shape can be observed (Oc.12,5x; Ob.40); Fig.2. Cross sections through the basal rosette
leaves indicate the midvein structure having a collateral vascular bundle surrounded by a pericycle, an endodermis with
Caspary strips and collenchymatic rings. A large vascular bundle and a smaller one can be seen in the midvein of the
leaf (Oc.12,5x; Ob. 40); Fig.3. Cross section through the basal leaf rosette indicates the tracheid hydathode (Oc.12,5x;
Ob.25); Fig.4. Detail of the tracheid hydathodes with a large aquifer stoma and the epithem (Oc.12,5x; Ob.40).
Plate III. S. paniculata: Fig.1. Cross-section of the flowering stem (Oc.12,5x; Ob.25); Fig.2. Multiseriate glandular
hair in the flowering stem epidermis (Oc.12,5x; Ob.25); Fig.3. Detail of the collateral vascular bundle along with an
independent phloem bundle in the flowering stem cross-section (Oc.12,5x; Ob.40); Fig.4. Uniseriate with divaricated
walls glandular hair in the flowering stem epidermis (Oc.12,5x; Ob.40); Fig.5. Stoma in the flowering stem epidermis
(Oc.12,5x; Ob. 40); Fig.6. One uniseriate non-glandular hair in the flowering stem epidermis (Oc.12,5x; Ob.40).
Plate IV. S. paniculata: Fig.1. Tetracytic stomata of the upper epidermis, in a tangential section through the rosette leaf
(Oc.10x; Ob.40); Fig.2. Detail of the midvein of the rosette leaf (Oc.12,5x; Ob.40); Fig.3. Cross section of the rosette
leaf indicates the connection between the secondary vein and the tracheid hydathode (Oc.12,5x; Ob. 10); Fig.4. Cross
section through the rosette leaf showing a tracheid hydathode which opens on the upper surface of the leaf, in a pit,
towards the margin (Oc.12,5x; Ob.40); Fig.5. Detail of the parenchyma epithem surrounded by a tannin cells sheath in a
cross section through the tip of the cauline leaf; the epithem cells of this terminal hydathode don't have intercellular
spaces (Oc.12,5x; Ob.25); Fig.6. Tangential section of the cauline leaf margin; it can be seen the opening of a
hydathode, with an aquifer stoma, at the base of marginal tooth (Oc.12,5x; Ob.10).
10
M. ANDREI, ROXANA PARASCHIVOIU
PLATE I
4
5
11
PLATE II
12
PLATE III
2
1
13
PLATE IV
1
2
14

MORPHOLOGICAL AND HISTO-ANATOMICAL ASPECTS REGARDING THE

FLORAL MORPHOGENESIS IN EUPHORBIA CYPARISSIAS L. (EUPHORBIACEAE
JUSS.)
RAMONA GALE*, C. TOMA*, LCRMIOARA IVNESCU*
Abstract: Euphorbia cyparissias L is an herbaceous species, wide-spread in the flora of Romania, often
founded in plant cultures and roadsides. This report describes the floral morphogenesis in the cypress spurge.
The vegetative activity of the shoot apex early ends by forming the inflorescence bracts, which have a different
morphology and structure than the nomophylls. Each branch of the dichasial inflorescence ends with a single
cyathium. The order of appearance of the cyathium components is as follows: monostaminate male flowers,
involucre, nectary glands and female flower.
Key words: Euphorbia cyparissias, shoot apex, morfogenesis, dichasium, cyathium
Introduction
Despite the diversity of Euphorbia species, their inflorescence has an homogeneous
organization, whose rendering during the time have launched to contradictory hypothesis.
Tournefort (1700), Linnaeus (1753), Payer (1857) and Baillon (1858) conclude that the true
status is that of a single hermaphroditic flower [4]. Le Maout (1842) adduces a new
interpretation of this curios flower, named cyathium by Warming (1912), the notion being a
long time discussed [7]. According to the actual conception the cyathium represents a
contracted inflorescence, usually constructed by 5 uniparous, scorpioide cymes of
monostaminate male flowers, which encircle a central nude female flower, all being surrounded
by gamophyllous and nectary involucre [2].
Haber [4] analyzes the vascular origin of the cyathium in some Euphorbia species and
adduces an argument to uphold the conception according to which the cyathium is a high
specialized inflorescence. Other researchers [1], [6], [8], [9] investigate the several aspects of
the embryogenesis and the structure of cyathium nectary glands in some representative
Euphorbia species.
The lack of information regarding the floral morphogenesis in Euphorbia species in the
Romanian literature encouraged us to carry this work. The present paper analyzes the
successive stages of the inflorescence development in Euphorbia cyparissias L.
Al. I. Cuza University, Faculty of Biology, Carol I Bd., no. 20A, Iasi, 700506, Romania, ramona.gales@uaic.ro
15
To observe the morphological features of the vegetative shoot apex during its
transformation into the reproductive one, the Euphorbia cyparissias L. plants were analyzed in
vivo at the end of the vegetation period (April), in successive stages of floral morphogenesis.
The material used for the histo-anatomical analysis was represented by the shoot apices
and inflorescences of Euphorbia cyparissias L., which were fixed in FEEA mixture and
preserved in 70% ethylic alcohol. The tissular and cellular modifications of the shoot
reproductive apex and the development of the inflorescence were analyzed on serial crosssections, which were performed using the standard paraffin-embedded protocol applied in plant
histo-anatomical researches. Fixed samples were dehydrated by a passage through
ethanol/water solutions and then embedded in paraffin at 65C for 24 hours. The embedded
material was cut into 13 m thick sections with a rotator microtome. The dried serial sections
were deparaffinized, rehydrated in serial dilutions of ethanol (100%, 90%, and 70%), coloured
with metilen-blue and ruthenium-red, and finally mounted in Canada balsam. All permanent
slides were analyzed in light microscopy, using a Novex (Holland) microscope; the microphotographs were made at the same microscope with a Sanyo digital camera and the sketches
were drawn on a Romanian MC1 microscope with Projektionszeichenspie gel.
Morphological changes of the shoot apex related to the transition from the
vegetative to the reproductive phase. The plastochronic functioning of the shoot apex of
Euphorbia cyparissias L. is very short (approximately one month). During the vegetative phase,
the shoot apex forms numerous leaves with winding disposition.
Towards the middle of April, the tip of the foliated stem becomes convex as a result of a
high number of bracts loosely clustered around the shoot apex, such as the leaves of a scaly
bulb. The bracts form the outside of this bulb, which have approximately the same shape as
the nomophylls, but are smaller than these, will form the involucre of the composed cymose
inflorescence. The inner bracts, which differ from the outer ones by their smaller size and
rhomboidal shape, will form the involucres of the dichasial inflorescences.
The vegetative activity of the shoot apex ends by forming the inflorescence bracts; from
this moment the reproductive apex will form the flower primordium, so that the stem axis will
terminate with a cyathium. The axillary buds will give birth to dichasial branches which may
ramify once or twice, each branch ending with a cyathium.
Structural changes of the shoot apex related to the transition from the vegetative to
the reproductive phase. The initial ring of the shoot apex is consumed by forming the
inflorescence bracts. During this stage, the shoot apex becomes dished as a result of numerous
mitotic divisions which take place in the apical-axial zone.
16
The structure of the stem tip is exclusively given by the bases of the bracts, being
represented by a cellulosic-parenchymatous tissue in which laticifers could be observed
alongside the procambium cordons.
The bract consists of homogenous mesophyll of meatic type, formed by round
parenchymatous cellulosic cells, in which cordons of procambium and laticifers pervade.
Each bud is formed in the axil of a bract, as a result of the numerous divisions of the
dedifferentiated cells of the upper epidermis and of some hypodermic layers from the bract
structure. At the beginning of the floral initiation, the shoot apex enlarges as a result of intense
divisions of the apical-axial meristem, being formed by a wide zone of meristematic cells (with
central nucleus and voluminous nucleolus) which covers a central zone with vacuolizated cells
(with central nucleus and numerous small vacuoles). Immediately below the shoot apex, the
cells of the central-axial zone are disorganized, resulting large aeriferous cavities.
The ontogenesis of the dichasium (Pl. I). In Euphorbia species, the compound
inflorescence is due to the dichasial branching. Each primary axis situated in the axil of an
involucral bract represents, in its first stages, a very simple cymose inflorescence, consisting of
three flowers- a central and two lateral ones with which the secondary axes end [4].
During the reproductive phase, the apex will give rise to a single cyathium. Subsequently
from each axillary bud a primary axis of a dichasium will be formed. In its first stages, the
primary axis presents a circular contour in cross-section; from its outer to its inner part, the
following histo-anatomical zones could be distinguished: 1. a single-layered epidermis
consisting of isodiametrical parenchymatous-cellulosic cells; 2. cortical parenchyma of meatic
type formed by 7-8 layers of cellulosic, thin-walled cells; 3. vascular bundles embedded in a
parenchymatous matrix and exhibiting meristematic tissue (procambium) between a few xylem
vessels and phloem elements; 4. cellulosic-parenchymatous pith, which presents numerous big
aeriferous cavities.
During the dichasium ontogenesis, the vascular system of the primary axis undergoes
several transformations as follows: new vascular elements appear in the external zone of the
cortex (being destined to vascularize the involucre); two meristematic-vascular rings
surrounding a disorganized pith are formed in the lateral parts; the central vascular bundles are
disposed in two opposed arches, the pith being included in their concavities.
In an advanced stage of dichasium development, two axilary buds will be formed on the
primary axis, which will end with a ciathium. Each axilary bud will give rise to a single
ciathium.
The ontogenesis of the cyathium (Pl. II- IV). The cyathium appears in the tip of a
branch such as a bud, having at its base two bracts and being covered by the involucre of the
dichasial inflorescence. Laticifers of the branch pervade in the primordium of the cyathium.
On the flanks of the reproductive apex, meristematic protuberances appear which will
successively form the primordia of stamens. In the first stages of male flower development (Pl.
II), the stamen primordium differentiates a stalked basal region, which will give rise to the
17
pedicel and filament and a trapezoidal upper region, which becomes the anther. The filament
extends concomitantly with the individualization of the two locules of the anther. In the next
stages of the stamen development, the median longitudinal septum and then the two lateral ones
are formed, which will separate each locule into two polinic sacs. In the same time, in the
median part of the anther, the connective begins to form.
It should be mentioned that the forming of the two polinic sacs is not concomitant in the
two locules. The wall of the polinic sacs are not yet completely developed when the
sporogenous tissue is forming.
Concomitantly with the development of the monostaminate male flowers, the involucre
of the cyathium starts to form. Thus, before the forming of the female flower, the cyathium
presents at its external part an involucre of polygonal contour in cross-section and a central axis
surrounded by stamens (in different stages of development).
The apex of the central axis gives rise to the first carpel which closes itself simultaneous
with the forming of a single ovule, which initially has a vertical position. In the next stage, the
stile of the first carpel is formed concomitantly with the appearing of the second carpel whose
evolution is similar with the first one. In the same way the third carpel is formed. The three
separately closed carpels grow together by their edges to form a 3-carpellar, syncarpous, 3locullar ovary with central-marginal placentation (Pl. III).
The central axis of the cyathium grows in length to form the pedicel of the female
flower. Thus, the incompletely developed female flower turns out from the involucre. Each
ovule becomes anatropous, consisting of a funicle, two integuments and nucellus, in which the
embryo sac is not yet formed.
Concomitantly with the development of female flower, primordia of nectary glands
appear in the upper level of the involucre. The development of the four nectary glands is not
simultaneous. As a result of the intense divisions of meristematic cells, the primordium of gland
grows in half-moon shape; the cells of the external layer radialy elongates and will form a
single-layered cellulosic parenchymatous epidermis; the first two layers situated just beneath
the epidermis will form a glandular tissue, and the other ones will form a parenchymatous
tissue, in which some vascular elements of the involucre pervade (Pl. IV).
Conclusions
1. The vegetative activity of the Euphorbia cyparissias shoot apex is early ended by the
forming of the inflorescence bracts, which exhibit different morphology and structure from that
of nomophylls
2. In the axil of the bracts, floriferous buds are formed, which will give rise to the
dichasial inflorescences.
18
3. During the dichasium ontogeny, the conducting system of the primary axis undergoes
several transformations, in order to vascularize the two axilary buds; each of the latest ones
which will give birth to a single ciathium just like the primary axis.
4. The cyathium appears in form of a bud at the base of two bracts.
5. During the ontogeny of the cyathium, the first formed elements are the monostaminate
male flowers, which appear successively on the flanks of the reproductive apex.
6. The involucre is formed from the central axis of the cyathium concomitantly with the
development of the male flowers.
7. In the upper part of the involucre, four nectary glands are formed concomitantly with
the development of the female flower, which is formed from the central axis of the cyathium.
8. The ovary is 3-carpellar syncarpous, 3-locullar with central-marginal placentation; the
carpels, each of it with a single ovule, are formed successively.
9. Initially, the ovules have a vertical position, subsequently they become anatropous.
10. The incompletely developed female flower turns out from the involucre as a result of
the elongation of its pedicel.
REFERENCES
1.
2.
3.
4.
5.
6.
7.
8.
9.
CARMICHAEL J. S., SELBO S. M., 1999 - Ovule, embryo sac, embryo and endosperm development in leafy
spurge (Euphorbia esula), Can. J. Bot., 77, 4: 599-610
EMBERGER L., 1960 - Les vgtaux vasculaires. t. II, Trait de Botanique systmatique (de M. Chadefaud et
L.Emberger), Libraires de LAcadmie de Medicine , Paris
GORI P., 1987- The fine structure of the developing Euphorbia dulcis endosperm, Ann. Bot., 60: 563-569
HABER M. J., 1925 - The anatomy and the morphology of the flower of Euphorbia, Ann. Bot., 39:656-707
KLEIMAN C. 2001 - La Reproduction des angiosperms, Ed. Belin, Paris
PAPP N., 2004 - Nectar and nectary studies son seven Euphorbia species, Acta Bot. Hung., 46, 1-2: 225-234
PARROT G., 1947 - Quelques remarques sur linflorescence dEuphorbia peplus L., Bull. Soc. Bot. France, 94, 9:
424-427
RAJA RAJESWARI RAO K., PRAKASA P. S., 1975 - Embryo development in Euphorbia peplus L., Current
Science, 44, 1: 57-59
WENIGER W., 1917 Development of embryo sac and embryo in Euphorbia preslii and E. splendens, Bot. Gaz.,
63, 4: 266-281
The explantion of figures:

Plate I. A, B, C, D, E, F, G- Serial cross-sections through the dichasial inflorescence of Euphorbia cyparissias
(successive stages of the dichasium ontogeny) (original diagrams)
Abreviations: br.= bract; pr. cyt.= primordium of cyathium; pt. = pith; mt. ts.= meristematic tissue; vs. bd.=vascular
bundle
Plate II. Fig. 1-5. The ontogenesis of a male flower of the Euphorbia cyparissias cyathium: longitudinal sections
through the cyathium in successive stages of the flower morphogenesis (original microphotographs) (scara= 50m).
Plate III. Fig. 1-5. The ontogenesis of female flower of the Euphorbia cyparissias cyathium: longitudinal (2, 3, 5) and
transverse (1, 4) sections through the cyathium in successive stages of the flower morphogenesis (original
microphotographs) (scara= 50m).
Plate IV. Fig. 1-5. The ontogenesis of the nectary glands of the Euphorbia cyparissias cyathium: longitudinal sections
through the cyathium in successive stages of the flower morphogenesis (original microphotographs) (scara= 50m).
19
RAMONA GALE et al.

pt.
PLATE I
vs. bd.
pr. cyt.
br.
A
mt. ts.
E
D
20
RAMONA GALE et al.
PLATE II
locules
male flower primordium
1
2
polinic sacs
filament
385
articulation
pedicel
mature anther
21
RAMONA GALE et al.
PLATE III
carpel
ovule
2
ovule
ovule
anatropous
ovule
3
5
22
RAMONA GALE et al.
PLATE IV
nectary gland primordium
1
2
glandular tissue
Epidermis
parenchimatous-cellulosic tissue
23

MICROSPOROGENESIS AND THE MALE GAMETOPHYTE AT EPHEDRA

DISTACHYA L.
GENIANA MIHAELA IULIA PREDAN*, IRINA GOSTIN**
Abstract: This study describes the microsporogenesis and the male gametophyte in Ephedra distachya L., only
one species of the Ephedraceae family growing spontaneous in Romania. Ephedra distachya L. is a dioecious
plant. Light microscopy (lm) is used to examine the early developmental stages of the male cones and
combined light (lm) and scanning electron microscopy (sem) were used to analyze the mature pollen grains.
The pollen cones are clustered at nodes either sessile or stalked. They are oval-oblong, each is composed of 2-8
decussate pairs of membranous bracts, proximal bracts empty; each distal bract subtending a male flower
composed of 2 basally fused, orbicular or obovate scales (false perianth); sessile anthers on staminal column.
Ephedra distachya pollen is particularly abundant. Ephedra distachya has the polyplicate pollen type
characterized by a pointed-oval shape and 6 longitudinal ridges and 6 valleys, due to alternation of thicker and
thinner exine regions, inaperturate. In polar view, ridges are low and triangular. On the SEM analysis the pollen
wall of Ephedra distachya is simple, so the SEM closely resembles the LM observation.
Key words: Ephedra distachya L., male cones, pollen development
Introduction
Genus Ephedra L. contains 35-45 species, most of them populating the desert or arid
regions. In Romania a single species of Ephedraceae family occurs - Ephedra distachya L. Its
status is rare [12].
Genus Ephedra L. has been much studied from the morphological and anatomical point
of view, information was summarized and included in synthesis works [8, 10, 14, 15, 16]. The
histology of the flowers and strobili of the Ephedra distachya L. plants was examined by Van
Tieghem (1869), Thoday and Berridge (1912) [10], later by Favre-Duchartre M. [5]. Baranec T.
Rehorek and V. studied the reproductive cycle of the Ephedra distachya L. plants, spontaneous
in Slovakia [2] and P. Mehra N. analysed dimensions of the male nuclei of the representatives
in the Ephedra genus [10].
Allison S. D. et al. have studied all the important stages of the Ephedra pollen
development using both photonic and the scanning and transmission electronic microscope [3].
Gamal El-Ghazaly et al. were concerned about pollen grain polarity, the aperture situation and
the pollen tube at 5 species of Ephedra including E. distachya L. [6].
The microsporogenesis and the description of the pollen and the male gametophyte
*
University of Bucharest, Faculty of Biology, Department de Botany and Microbiology, Aleea Portocalelor, nr. 1-3,
060101, Bucureti, Romania; ggentiana@yahoo.com
**
Al. I. Cuza University, Faculty of Biology, Bd. Carol I, no. 20A, 700506, Iai, Romania; irinagostin@yahoo.com
24
development in Ephedra distachya L. provides interesting characterization in this species.

Our study was carried out on samples of Ephedra distachya L. from different places in
Romania: Agigea Marine Dune Reserve (Constanta County), Culmea Pricopanului (Tulcea
County), the Botanical Garden of Iai (cultivated plants) and from Bulgaria (Balchik) [7, 8].
Ephedra distachya L. is a dioecious plant. Male cones were collected during the three spring
months (March, April and May), in successive phaenological phases, from the cones initiation
up to the pollen shedding.
The biological material (male cones) was fixed in 70% alcohol or FAA (formalin 40%, 5
cc: alcohol 70%, 90 cc: glacial acetic acid 5 cc). After being fixed, the material has been
processed in accordance with the paraffin embeding protocol [3], sectioned (transverselly and /
or longitudinally) with a Minot type microtome at 12-15 m thick, colored with Ehrlichs
haematoxylin and embedded in Canada balm. To highlight microspores and young pollen grains
microscopical preparations squash type have been made, that have been colored with acetic
carmine.
All obtained sections were analyzed using light microscope (LM) Docuval type and
photographed with a digital photo camera slr Canon EOS 400D. For the analysis of the pollen
grains at electronic scanning microscope (SEM) a Vega - TESCAN LSH microscope was used
in The Faculty of Biology of The Al I. Cuza University from Iai [12].
Male cones were considered from the morphological point of view with a Belphotonics
stereomicroscope and photographed with the same digital camera.
The description of the pollen grains has been made in accordance with the terminology
adopted by Faegri [4].
The male cones, either sessile or stalked, are found clustered at nodes (fig. 1). The male
cones are green, oval-oblongue, each of them being composed of 2-8 descussate pairs of
membranous bracts, like cups (Fig. 2, 3). The lower pair bracts is sterile, each distal bract
subtending a male flower composed of 2 basally fused, orbicular or obovate scales (primitive
periant) (Fig. 4, 7), an axis (microsporangiophore or anterophore) terminal bearing a number of
anthers, each anther with two micro sporangia (pollen sacs) (Fig. 4). A group of archespores
subepidermal cells in the mature cones is observed in March (fig. 5).
In April, following meiosis I, macrospore dyads have resulted. After meiosis II numerous
tetrads of microspores appear, the wall formation taking place after each stage of meiosis. Soon,
the wall surrounding the 4 microspore cells dissolve and the microspores in the pollen sac are
25
released. The microsporangia wall is composed of three known layers: epidermis, the median
layer and the tapetum layer. The median layer is composed of a single row of parenchimatic
cells and, as the pollen sac grows, its cells are flattened so that they ultimately disappear. The
tapetum has signs of degeneration after the reductional division (Fig. 6) and no longer in the
stage of mature pollen (Fig. 8). At mature pollen sac epidermis persists. Its cells have a regular
appearance and their walls are thickened (Fig. 9). Only in a small portion of the pollen sac
upper part a few parenchimatic cells with thin walls remain. By their breaking a hole will form
that will be issued pollen (poricide opening).
In May, the pollen is divided and forms a small prothalian cell and a large, central one
(Fig. 10). The last divide and formed the second prothalian cell and initial anteridial cell. The
last divide and give a vegetative larger cell, and a generative cell. The generative cell will form
the stalk cell and spermatogene cell, the latter giving rise to gametes. Regarding pollen sacs, the
pollen has 5 cells [8]. The mature pollen grains are yellow, fusiforme and obvious poliplicate:
they have 6 longitudinal ridges (plicae) (sometimes slightly corrugated) and 6 valleys, formed
due to the alternation of thicker and thinner exine regions (Fig. 9, 10). On a polar view, the
ridges are short and triangular. Pollen grains have 53 m long axis and 20 m short axis. At the
electronic scanning microscope (SEM), the pollen surface is no obvious regulate and no
aperture was noticed eather, so the pollen is inaperturate (Fig. 11, 12).
Conclusions
In March, a group of archespore cells appears in the pollen sacs, subepidermaly.
In April the anther wall is composed of an epidermis and a scrap tapetum and the
microspores are free.
In May the anther wall is only represented by the epidermis and numerous grains of
fusiforme and yellow pollen are found inside the pollen sac, in our observation (in 17 May 2007
on Culmea Pricopanului) the pollen grains are uninucleate.
According to the SEM observations, the pollen grains of Ephedra distachya L. have a
simple structure, the SEM observations being very similar to those made on an optical
microscope.
REFERENCES
1.
2.
3.
4.
ANDREI M., RDULESCU D., 1972 - Caiet pentru tehnica preparrii i conservrii materialului biologic.
Tipogr. Univ. din Bucureti
BARANEC, T., V. REHOREK, et al., 1994 - Generative reproduction of ephedra (Ephedra distachya L.) in
Slovakia. Biologia Bratislava, 49 (1): 65-67
DOORES ALLISON S., OSBORN J. M., GAMAL EL-GHAZALY., 2007 - Pollen Ontogeny in Ephedra
americana (Gnetales). International Journal of Plant Sciences 168 (7): 985-997
FAEGRI K., IVERSEN J., 1966 - Textbook of pollen analysis. 2nd Ed. Munksgaard, Copenhagen
26
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
FAVRE-DUCHARTRE M., 1959 - Contribution ltude de la reproduction sexue chez Ephedra distachya. C.
R. Acad. Sci. Paris. 249: 1551-1553
GAMAL EL-GHAZALY, ROWLEY J., HESSE M., 1998 - Polarity, aperture condition and germination in pollen
grains of Ephedra (Gnetales). Plant Systematics and Evolution, 213 (3-4): 217-231
GRINESCU G. P., 1952 - Ephedra L. In: T. SVULESCU (red. princip.). Flora Romniei. 1. Bucureti: Edit.
Academiei Romne
JOHRY B. M., BISWAS C., 1997 - The Gymnosperms. Narosa Publishing House, New Delhi
KRSSMANN G., 1985 - Manual of cultivated conifers. B.T. Batsford Ltd. London
LEHMANN-BAERTS M., 1967 - La morphologie du sporophyte dans le genre Ephedra. La Cellule (Belg.), 67
(5): 7-56
MEHRA P. N., 1950 - Inequality in size of the male nuclei in the genus Ephedra. Ann. Bot., July 1950; 14: 331339
OLTEAN M., NEGREAN G., POPESCU A., ROMAN N., DIHORU G. SANDA V., MIHILESCU S., 1994 Lista roie a plantelor superioare din Romnia. Studii, sinteze, documentaii de ecologie, 1
PLOAIE G. P., PETRE ZOE. 1979 - Introducere in microscopia electronica cu aplicatii in biologia celulara si
moleculara. Edit. Acad. Romane, Bucuresti
SCHNARF K., 1933 - Embryologie der Gymnospermen. In: Linsbauer K. Handbuch der Pflanzenanatomie, II
Abt., 2 Teil, Bd. 2. Verlag Gebruder Borntraeger, Berlin
SINGH H., 1978. - Embryology of gymnosperms. Berlin-Stuttgart
SMARANDACHE D., 2005 - Embriologia plantelor. I. Embriologia arhegoniatelor. Edit. Univ. din Bucureti,
Bucureti.
STEEVES M. W., BARGHOORN E. S., 1959 - The pollen of Ephedra. J. Arnold Arboretum, 40: 221-255

Plate I
Fig. 1 Ephedra distachya L. - Branch with male cones (1 May 2008; Orig)
Fig. 2 Ephedra distachya L. Male cone (20 May 2007) (Oc. 10x; Amplific. 2; Orig.)
Fig. 3. Ephedra distachya L. - Longitudinal section of a male cone - fragment (13 April 2008) (Oc 12.5x; ob. 3.2x;
Amplific. 8; Orig.)
Fig. 4. Ephedra distachya L. - Male flower (20 May 2007) (Oc. 10x; Amplific. 3; Orig.)
Fig. 5. Ephedra distachya L. - Longitudinal section through the anther that outlines the archaesporal cells (17 March
2007) (Oc 12.5x.; ob. 25x; Amplific. 160; Orig.)
Fig. 6. Ephedra distachya L. - Cross-section through the male flower (13 April 2008) (Oc. 12.5x; ob. 10x; Amplific.
6.3; Orig.)
Fig. 7. Ephedra distachya L. - Longitudinal section through the male flower (13 April 2008) (Oc. 12.5x; ob. 10x;
Amplific. 6.3; Orig.)
Plate II
Fig. 8 Ephedra distachya L. - Longitudinal section through the male flower (29 May 2001) (Oc. 12.5x; ob. 3.2x;
Amplific.16; Orig.)
Fig. 9. Ephedra distachya L. - Longitudinal section through the anther - anteral wall and young pollen grains (29 May
2001) (Oc. 12.5x; ob. 40x; Ampific. 16; Orig.)
Fig. 10. Ephedra distachya L. - Unicellular pollen grain (17 May 2007) (Oc. 12.5x; ob. 40x; Amplific.16; Orig.)
Fig. 11. SEM micrograph of Ephedra distachya L. poliplicate pollen (21 May 2008; Orig.)
Fig 12. SEM micrograph of Ephedra distachya L. pollen (21 May 2008; Orig.)
27
GENIANA PREDAN, IRINA GOSTIN
PLATE I
28
GENIANA PREDAN, IRINA GOSTIN
PLATE II
11
10
12
29

VEGETATIVE ANATOMY OF TWO GALIUM L. SPECIES (RUBIACEAE)

ANCA HEMCINSCHI*, RAMONA GALE**, C. TOMA**
Abstract. The structure of two medicinal Galium species (G. album Mill. and G. verum L.) was studied to
determine the range of variation in certain histo-anatomical characters of the vegetative organs. The study
revealed several interesting anatomical data that have not previously been reported in these species, e.g. the
structure of the rhizome, the presence of the trichomes on the adaxial face of G. verum foliar limb.
Key words: anatomy, vegetative organs, Galium
Introduction
Following-up our histo-anatomical investigations on Rubiaceae genre and species [22],
the present study compares the anatomical structure of two perennial species belonging to
Galium genus: Galium album Mill. (syn. G. mollugo L. ssp. erectum (Huds./Briq.) and G.
verum L. The two species are considered medicinal plants [13], [19] and are distinguished from
each other by several morpho-anatomical characters, e.g. the colour of the flowers, the
presence, the frequency and the length of the trichomes.
Among the 37 Galium species from the Romanian flora, G. album and G. verum are
common in the entire country, from the durmast to the subalpine belt [4], [12].
The existing literature on the structure of the representative members of Rubiaceae
family is quite abundant in results from studies dedicated exclusively to them [14], [15], [17],
[18], [21] or from the synthesis treatises on anatomy of Dicotyledons [9] or Angiosperms, in
generally [10]. Some authors investigate the medicinal Galium species [1], [2], [6], [8], others
analyze several aspects regarding the foliar venation [5], the infraspecific variability [7], the
stomata ontogeny [11], the stipules structure [16], the number of stomata in different ecological
conditions [20], [3].
The research material is represented by two species of Galium genus: G. album Mill. and
G. verum L. from the Romanian flora. The material was fixed and preserved in 70% ethylic
alcohol. Cross-sections of the rhizome, aerial stem and leaf were performed using a manual
microtome, coloured with iodine-green and ruthenium-red and embedded in glicero-gelatine.
The superficial sections through the foliar limb were coloured with iodine-green. The obtained
*
Pharmacy AncaFarm, Botosani, Romania

Al. I. Cuza University, Faculty of Biology, Carol I Bd., no. 20A, 700506, Iasi, Romania
**
30
permanent slides were analyzed on a Novex (Holland) microscope and photographed at the
same microscope with a digital photo camera.
The aerial stem
The contour of the transverse section through the aerial stem is quadratic rhomboid,
with rounded ribs, less prominent towards the basis of the organ.
In G. verum, the epidermis presents unicellular trichomes, with very thick walls, whose
frequency decreases towards the upper and the lower level of the stem.
In the four ribs there are cordons of tangential collenchyma and the cortex ends with a
Casparyan endodermis on the entire stem length. In G. verum, between the stem ribs, the outer
cortical layer is collenchymatous.
The stellum presents secondary structure, the conducting tissues being of annual type.
The xylem ring is thicker than the phloem one, comprising numerous libriform fibres with very
thick and intensive lignified walls.
The rhizome
The outline of the transverse section through the rhizome is circular.
In both analyzed species, the rhizome presents secondary structure, resulted only from
the cambium activity (in G. album) or from the activity of both lateral meristems (in G. verum).
In the latest case, the phellogen is differentiated from the Casparyan endodermis; the epidermis
and the cortex are exfoliated.
In G. album, the cortex is relatively thin (5-6 layers); this histological feature is unusual
for a subterranean stem. In the thickness of the cortical parenchyma, crystalliferous cells with
crystal sand and raphides are often present.
The stele is very thick and comprises: 1) a thinner (in G. album) or a thicker (in G.
verum) phloem ring, some of the collenchymatous parenchyma cells containing crystal sand and
2) a single very thick (in G. album) or three (in G. verum) xylem rings, whose thickness
increases from the pith to the secondary phloem.
The secondary xylem consists of much libriform, in which the vessels are irregularly
disposed. Only in G. verum, there are islands of xylem cellulosic parenchyma, some cells
containing crystal sand.
The pith is thin, parenchymatous-cellulosic of meatic type, with very big cells in G.
verum, many of them containing crystal or raphides (in G. album).
The leaf
The sessile leaves are hipostomatic, with stomata of paracytic type.
The epidermis (in front view) presents cells of irregular outline, their lateral walls being
moderately (in G. verum) or mighty (in G. album) wavy. In cross-section, the cells are
isodiametric, being much bigger in the upper epidermis.
31
At epidermis level there are unicellular, short trichomes with very thick wall, more
numerous on the abaxial face. Our observations, in concordance with the anterior published
papers [22], quash the data from Flora of Romania [12], according to which the adaxial face of
G. verum leaves is glabrous.
The foliar limb has bifacial heterofacial structure, the mesophyll being differentiated in
one (in G. album) or two (in G. verum) layers of palisade tissue and 4-5 layers of lacunous
tissue; in the thickness of the latest, cells with crystal sand may be observed. In both species
studied, the outer wall of the epidemic cells is thicker than the others and covered by a thin
cuticle. On the margins of the foliar limb, all the walls of the epidermic cells are thick; a
collenchymatous hypodermis is present.
The median and lateral vascular bundles are surrounded by parenchymatous theca with
isodiametric cells containing slightly chloroplasts.
Conclusions
The structure layout is similar in both analyzed Galium species.
The structural differences between the two Galium species are quantitative, reffering to
the frequency, dimensions and localization of the trichomes, the frecquency of the
crystalliferous cells, the lignification degree of the pith, the number and thickness of the
secondary xylem rings, the sclerification degree of the libriform fibres, the number of the
palisade cells layers, the amplitude of the undulations of the lateral walls of foliar epidemic
cells.
REFERENCES
1.
2.
3.
4.
5.
6.
7.
8.
9.
BORYSOV M. I., 1965- tude de la structure anatomique de Galium ruthenicum Willd., Farm. Zh. Ukrajin. R. S.
R., 39, 5: 59-63
BUTTLER K. P., BRESINSKY A., 1966 Beitrag zur Zitologie von Galium ser. silvatica. Ber. bayer. bot.
Gesellsch Erforsch. heim. Flora, 39: 25-28
CHERMEZON, H., 1910- Recherches anatomiques sur les plantes littorales, Ann. des Sci. nat., Bot., sr. 9.,12:
117-313
CIOCRLAN V., 2000- Flora ilustrat a Romniei. Pteridophyta et Spermatophyta. Ed. Ceres., Bucureti
DARWIN S. P., 1980- Leaf venation and the classification of certain Rubiaceae. ICSEB-II 2ed Internat. Congr.
Syst. and Ecol. Biol., Vancouver (Canada), Jully 17-24, 1980, Abstracts, 176
FISCHER F., 1937- Beitrge zur Pharmakognosie der Plataginalen and Rubialen. Anatomie des Laubblattes.
Thsis. Basel
HENDRICH R., 1977- Bemerkungen zur Variabilitt von Cruciata glabra (syn. Galium vernum). Preslia, 49:
193-201
KOHLMNZER S., 1964- Recherches botaniques et chimiques sur lespce collective Galium molugo L. en
considrant les caryotypes croissant en Pologne. II. Recherches anatomiques. Dissert. Pharm., Pologne, 16, 3:381392
METCALFE C. R., CHALK L., 1972- Anatomy of the Dicotyledons. 2. Clarendon Press, Oxford
32
10. NAPP-ZINN, KL., 1973, 1974 - Anatomie des Blattes. II. Angiospermen, In Handbuch der Pflanzenanatomie,
Bd.VIII, A 1-2, Gebrder Borntraeger, Berlin, Stuttgart
11. PANT D. D., BHARATI M., 1965- Ontogeny of stomata in some Rubiaceae. Phytomorphology (Indic), 15, 3:
300-310
12. PAUC A., NYRDY E. I., 1961- Rubiaceae. In Flora R. P. Romne, 8: 524-589, Ed. Acad. Romne,
Bucureti
13. PERROT E., PARIS R., 1971- Les plantes mdicinales. 1., dit. Presses Universitaires des France, Paris
14. ROBBRECHT E. (ed.), 1994 - Advances in Rubiaceae macrosystematics. Opera Botanica Belgica, Bruxelles, vol.
6
15. ROBBRECHT E., PUFF C., SMET E. (eds.), 1996- Second international Rubiaceae Conference. Proceedings
16. RUTISHAUSER R., 1984- Blattquirle, stipeln und Kollateren bei den Rubieae (Rubiaceae) in Vergleich mit
andern Angiospermen. Beitr. z. Biol. Pflanz., 59, 3:375-424
17. SAINT-JUST S., 1904- Recherches anatomiques sur lappareil vgtatif arien des Rubiace. Thse, Paris
18. SOLEREDER H., 1893- Ein Beitrag zur anatomischen Charakteristik und zur Systematik der Rubiaceen. Bull.
Herb. Boissier, 1: 167-199
19. STNESCU U. i colab., 2004 Plante medicinale de la A la Z, monografie ale produselor de interes terapeutic,
1, Ed. Gh. T. Popa, UMF Iai
20. TOKARZ H. T., SULMA T., BUJEWICZ M., 1969- The number of leaf stomata in Asperula odorata L. plants
derived from ecologically different forest communities and from garden cultivation. Acta biol. med. Soc. Sci.
Gedau, 14: 443-466
21. TOLLE H., 1913- Beitrge zur vergleichenden Anatomie der Rubiaceen. Thesis, Gttingen
22. TOMA C., GOSTIN I., 2000- Anatomical structure of some Rubiaceae species. An. t. Univ. Al. I. Cuza, Iai,
ser. II a (Biol. veget.) 46: 1-10

Plate I. Galium album L. Fig. 1, 2. Cross-sections through the rhizome (x200). Fig. 3 (x40), 4(x200). Cross-section
through the upper third of the aerial stem. Fig. 5, 6. Cross-sections through the middle third of the aerial stem (x200).
Plate II. Galium album L. Fig. 1. Cross-section through the lower of the aerial stem. (x200). Fig. 2, 3. Cross-sections
through the foliar limb (x200). Fig. 4, 5. Superficial sections through the folair limb (x200): 4. lower epidermis; 5.
upper epidermis.
Plate III. Galium verum L. Fig. 1, 2. Cross-sections through the rhizome (x100). Fig. 3 (x40), 4(x200). Cross-section
through the upper third of the aerial stem. Fig. 5. Cross-sections through the middle third of the aerial stem (x200). Fig.
6. Cross-section through the lower of the aerial stem (x200).
Plana IV. Galium verum L. Fig. 1(x40), 2-4 (x200). Cross-sections through the foliar limb Fig. 5. Superficial sections
through the folair limb: lower epidermis (x200).
33
ANCA HEMCINSKI et al.
PLATE I
34
PLATE II
1
2
35
PLATE III
36
PLATE IV
2
3
37

A COMPARATIVE STUDY REGARDING THE MORPHOLOGY AND ANATOMY

OF THE VEGETATIVE APPARATUS IN TWO OCIMUM BASILICUM L. BREEDS
MARIA MAGDALENA ZAMFIRACHE*, C. TOMA*, MARIA DUCA**, SIMONA
DUNCA*, ZENOVIA OLTEANU*, M. TEFAN*, RAMONA GALE*, CLAUDIA
PDURARIU*
Abstract. This study compares the structure and morphology of the vegetative subterranean and aerial organs
in two Ocimum basilicum L. breeds cultivated in Turkey. The object of this study is to emphasize the
infraspecific variation of the plant morphology and anatomy of these two breeds, underlining the importance
of the secretory hairs that produce the volatile oils and concede this species the virtue of medicinal herb and
aromatic plant.
Key words: Ocimum basilicum L., morphology, anatomy, vegetative organs, secretory hairs
Introduction
Ocimum basilicum L. of the Family Lamiaceae is a herb grown as a perennial in warm,
tropical climates. Basil is originally native to Iran, India and other tropical regions of Asia and
it is cultivated in greenhouses or in the field in many other regions too.
Ocimum basilicum L. is an annual species with a vegetataive apparatus composed of a
well ramified fibrous root, a strongly ramified, 60 cm long, four edged erect stem and many
pointy ovate-lanceolate opposite leaves with atenuate serrate edges [1]. The flowers are quite
big, white in colour and arranged in a terminal spike. The four stamens and the pistil are not
pushed under the upper lip of the corolla, but lay over the inferior.
Materials and methods
The material of our investigations is represented by two Ocimum basilicum L. breeds
cultivated in Turkey (to mark out the two species, they were noted with two numbers: 1
nonflowering specimen and 2 flowering specimen). The utilised methods are those currently
used for vegatal anatomy investigations. Cross-sections through the vegetative organs using a
botanical razor and a manual microtome have been executed. These sections have later been
jewelised and tinted using iodine-green and ruthenium-red. The superficial lamina sections
have been tinted using iodine-green. The sections were later analysed and photographed using
a photonic microscope (NOVEX, Holland).
*
Al. I. Cuza University, Faculty of Biology, Carol I Bd., no. 20A, Iasi, 700506, Romania
University of State, Faculty of Biology and Pedology, Chisinau, Moldavia
**
38

The morphology of the vegetative apparatus
Both basil breeds studied present a variable number (5 to 7 for the first breed and 8 to 9
for the second one) of strongly ramified aerial stems of various thickness (those of the first
breed are thicker) that start out at the base of the root. There are leaves on the branches and the
main stems, more numerous on the second breed plants and larger ones on the first breed
plants (Pl. I: Fig. 1).
The structure of the vegetative apparatus
The structure of the subterrranean vegetative organs
The histo-anatomical analysis of the subterranean vegetative apparatus of the two basil
breeds revealed the fact that the main axis which is in the scientific literature considered to be
the main root, has a series of tipical subterranean stem (rhizom) structural features:
1. Lignified pith in the center of the organ, composed of polygonal cells
without meatuses between them and 4 primary ligneous bundles surounded by few ligneous
parenchyme cells also with lignified walls, between them (Pl. I: Fig. 3c);
2. The secondary xylem which is entirely lignified forms two annual clearly
visible rings (Pl. I: Fig. 3d):
- the libriform fibers are prevalent in the first ring (the inner ring). These fibers have
thick and slightly lignified walls. The radial range disposed, small diameter, scarce vessels are
surrounded by few lignified ligneous parenchymatous cells.
- the second ring (the external ring) is thicker, composed of: numerous variable
diameter vessels (bigger than the ones in the first ring); these radial range disposed vessels are
surrounded by few lignified ligneous parenchyme cells, and libriform fibers; these fibers have
strongly lignified thiner walls (compared to those of the first ring).
The felogene formation does not rely on the pericycle (which is the case of many
dicotyledonous plants) but on the differentiation of different layers of the cortex, producing
many layers of often stratified cork, composed of bigger cells than those of the pheloderme and
those of the cortical parenchyme that is still unexfoliated or still persisting between the
periderma successively formed (Pl. I: Fig. 3b). A genuine ritidoma that exfoliates along the
organ is thus formed (Pl. I: Fig. 3a).
Therefore, considering this organ as being a rhizom, the roots that form on it are
endogenous formed adventive roots (Pl. I: Fig. 3e). The structure of these roots is secondary,
as a single result of the cambium activity. The diarche type stellum has a primary structure (Pl.
I: Fig. 3f).
The structure of the aerial vegetative apparatus
The stem of both basil breeds has a primary structure only in the upper third part and a
secondary structure, as a result of the cambium activity, in the other two.
39
The outline of the cross section (Pl. II: Fig. 4, 5) differs in each third of the stem (upper,
middle and lower) and in the two breeds as well. In the upper third part, the outline of the cross
section is rectangular-quadrangular shaped, with 4 generally attenuate costas, some of them
more evident than others, with deeper and narrower valecules berween them in the second
breed. The valecules progresivly grow wider and they become less deep with the thickening of
the organ, so that in a cross-section they show an allmost circular outline.
The epidermis protects the entire surface of the stem (Pl. III: Fig. 6, 7) and it is
composed of sligtly tangentially elongated isodiametrical cells (in the lower third) that have
pericline walls, thicker than the others, a thin cuticle covering the external one. There are very
few stomata present.
There are two types of hairs: 1. uniseriate pluricellular trichomes that are present on the
entire outer surface of the stem; 2. secretory hairs composed of a unicellular or bicellular
hinge, a unicellular pedicle and an unicellular or bicellular gland (Pl. V: Fig. 10a, b, c). The
number of secretory and tectorial hairs per suface unit decreases from the top to the base of the
stem. There are more hairs (tectotial and secretory also) on the first breed stems than on the
second breed stems (Pl. II: Fig. 4, 5).
The parenchymatous-cellulosic cortex, meatus type in the upper third, is thin and
slightly colenchymatic in a hypodermic position (in the first basil breed especially near the
costas) and does not have a special type of endoderma on the exterior. The cortex cells tend to
become tangentially elongated and the air spaces between them become larger as the stem
grows thicker. The anticline division walls are visible inside the cortex inner cell layers in the
lower third of the first basil breed stem.
The primary structure stellum (Pl. III: Fig. 6, 7) follows the general outline of the cross
section and has four large collaterally open type bundles in the four costas and one very small
bundle composed only of phloemic elements between them. The large bundles have radial
ranges of ligneous vessels separated by uniseriate or pluriseriate areas of parenchymatouscellulosic cells and the liberian tissue is composed of pierced tubes and annexe cells.
Belts of sclerenchyma fibers can be observed at the end of the large phloem vascular
bundles in the second basil breed (Pl. III: Fig. 7a) that have in this developing state less
thickened but still cellulosic walls. The fiber walls get progressively thicker and lignified from
the base up to the top of the stem (Pl. III: Fig. 6, 7).
The thickening of the stem is based on the cambium activity that produces a thin
phloem ring on the exterior and a thicker xylem one on the interior. The phelogen becomes
differentiated based on an inner cortical layer at the base of the second breed stem, producing a
single layer of cork (composed of very large cells that have thin walls and little cork) and 1-2
noncolenchymatous pheloderma layers.
The cambium activity is initialy unequal in the circumference of the organ, producing
more secondary ellements (phloem and xylem) in the large bundles (near the costas); thus the
secondary vascular tissue rings are sinuous during this developing stage (Pl. II: Fig. 4b, Fig.
40
5b). The cambium produces subsequently many vascular ellements between the costas so that
both rings (secondary phloem and secondary xylem) become circular (Pl. II: Fig.4c, Fig. 5c).
The secondary xylem ring is almost entirely lignified at the base of the stem (composed
of vessels, libriform fibers, lignified ligneous parenchymatous cells, horizontaly lignified
parenchymatous cells) in the second breed or it has a thick tangential cellulose ligneous
parenchymatous belt only on one side of the organ circumference so that the pith is not situated
in the center anymore but on the side, in the first basil breed.
The pith is a meatus type cellulose-parenchymatous thick pith (Pl. II: Fig. 4a, b; Fig. 5a,
b); the composing cell walls are lignified in the base of the stem (Pl. II: Fig. 4c, Fig. 5c).
The amphystomatic type lamina presents very small dyacitic stomata situated on top of
the epidermis. Its structure is heterofacial bifacial and the mesophyll is differentiated as a very
elongated cell unistratified palisadic tissue and a pluristratified lacunous tissue (4 to 5 layers).
The middle nervure (Pl. IV: Fig. 8a, Fig. 9a, b) is visibly prominent on the inferior side
of the lamina and has a single vascular bundle inside the noncloroplastic noncolenchymatic
fundamental parenchime, which is larger in the first basil breed.
The relatively short uniseriated pluricellular trichomes can be found only on the lamina
middle nervure (Pl. IV: Fig. 9b). There are two types of secretory hairs: a) located in a very
small depression of the upper epidermis, having a bicellular gland (Pl. IV: Fig. 9c; Pl. V: Fig.
10d); b) located in a very large excavation of the lower epidrmis, having a four celled gland
(Pl. IV: Fig. 8b; PL. V: Fig. 10e).
Conclusions
Investigating the vegetative apparatus of the two Ocimum basilicum L. breeds, an
infraspecific variation regarding some morphological and anatomical features has been
observed.
The two basil breeds studied are distinguished by the following morphological features:
1) the size and density of the aerial stem leaves; 2) the developing stage of the subterranean
vegetative apparatus.
The presence of some subterranean stem (rhizom) specific features among the main
subterranean axis indicates that the two basil breeds may be perennial, the species being
considered annual according to the scientific literature.
The structure of the vegetative apparatus in the two basil breeds differes according to
the following features: a. regarding the stem: 1) the outline of the croos-section in the upper
third; 2) the number and density of the trichomes on organ surface unity; 3) the lignification
stage of the secondary structure stellum; 4) the absence or the presence of the protecting
secondary tissues; b. regarding the lamina the developing stage of the vascular tissue in the
middle nervure.
41
The number of secretory hairs on organ surface unit and the number of cells that
compose their gland concede this species the virtue of aromatic and medicinal plant. Our
research has shown that the number of secretory hairs on the vegetative apparatus in the first
basil breed is greater compared to the second one. Both breeds show more numerous secretory
hairs on the lamina and on the tip of the stems, most of the four celled gland hairs being
situated on the lamina.
REFERENCES
1.
2.
3.
4.
5.
CIOCRLAN V., 2000 - Floara ilustrat a Romniei, ed. a II-a. Edit. Ceres, Bucureti
GUULEAC M., 1961 Ocimum L. n Flora R. P. R., Edit. Acad. Rom., Bucureti, 8: 390-393
RUGIN R., TOMA C., 1995 Histo-anatomical researches of some medicinal plants V. Labiatae. An. t.
Univ. Al. I. Cuza Iai, s. a II-a (Biol. veget), 41: 17-22
TOMA C., RUGIN R., 1998 Anatomia plantelor medicinale. Atlas. Edit. Acad. Rom., Bucureti
TOMA-GOSTIN I., TOMA C., IVNESCU L., 2002 Histo-anatomical aspects of Ocimum basilicum L., an
important medicinal plant. 2-nd Conference on Medicinal and aromatic Plants of Southeast european Countries.
Chalkidiki-Greece (Book of abstracts): 140
Explanation of figures:
Fig. 1. The morphology of the aerial vegetative apparatus in Ocimum basilicum L.: first breed (on the left);
second breed (on the right) (original macrophotographs). Fig. 2. The morphology of the subterranean vegetative
apparatus in Ocimum basilicum L.: first breed (on the left); second breed (on the right) (original
macrophotographs). Fig. 3. The structure of the subterranean vegetative apparatus in Ocimum basilicum L.
second breed: a-e. rhizom cross sections: a. general view (unjewelised and untinted section) (x40); b. ritidoma detail
(x200); c. pith and secondary xylem detail(x200); d. secondary xylem detail the border between the two annual rings
(x200); e. the formation of an adventive root (x100). f. adventive root cross section (general view) (unjewelised and
untinted) (original microphotographs). Fig. 4. The structure of the aerial stem on different levels in Ocimum
basilicum L. first breed: aerial stem upper third (a), middle third (b) and lower third (c) cross sections (x40) (original
microphotographs). Fig. 5. The structure of the aerial stem on different levels in Ocimum basilicum L. second
breed: aerial stem upper third (a), middle third (b) and lower third (c) cross sections (x40) (original microphotographs).
Fig. 6. The structure of the aerial stem on different levels in Ocimum basilicum L. first breed (details): aerial stem
upper third (a), middle third (b) and lower third (c) cross sections (x200) (original microphotographs). Fig. 7. The
structure of the aerial stem on different levels in Ocimum basilicum L. second breed (details): aerial stem upper
third (a), middle third (b) and lower third (c) cross sections (x200) (original microphotographs). Fig. 8. The structure
of the lamina in the Ocimum basilicum L. first breed: - middle nervure cross sections a)- unjewelised and untinted
(x100) and mesophillun (b, c ) (x200) (original microphotographs). A secretory trichome and its four celled gland
located inside a lower epidermis excavation (b) and a stomata in the superior epidermis (c) may be observed. Fig. 9.
The structure of the lamina in Ocimum basilicum L. second breed : - middle nervure cross sections a)- unjewelised
and untinted (x100) and mesophillun (b) (x200) (original microphotographs). A tectorial trichome on the adaxial side of
the middle nervure (b) and a secretory trichome and its bicellular gland located inside a small excavation in the superior
epidermis (c) may be observed. Fig. 10. Secretory hairs in an aerial stem cross section in Ocimum basilicum L. (a,
b, c) and in a lamina superficial section (d, e) a. unicellular gland secretory trichome; b. bicellular gland secretory
hairs; c. bicellular gland secretory trichome; the secretory product eliminated between the wall and the cuticle can also
be observed; d. bicellular gland secretory trichome; e. four celled gland secretory trichome (x800) (original
microphotographs).
42
MARIA MAGDALENA ZAMFIRACHE et al.
PLATE I
3. a
3. b
3. f
3. e
43
3. c
3. d
4. a
5. a
PLATE II
4. b
4. c
5. b
5. c
44
6. a
7. a
PLATE III
6. b
6. c
7.b
7.c
45
PLATE IV
8. a
8. b
8. c
9. a
9. b
9. c
46
10. a
PLATE V
10. b
10. c
10. d
10. e
47

A COMPARATIVE NUMERIC ANALYSIS OF THE SECRETORY TRICHOMES

BELONGING TO THE FOLIAR LIMB IN SOME DROSERA SPECIES
IRINA STNESCU*, GABRIELA VASILE**
Abstract. The paper is focused on a comparative numeric analysis of the secretory pedicelate (tentacular)
trichomes of the upper epidermis of the leaf in 16 Drosera species and of the sessile secretory trichomes
belonging to both upper and lower epidermis in the same species. The bigger is the number of the
secretory trichomes, in both epidermises, the more efficient is the plant in capturing and retaining different
organisms. This way, the carnivorous plants can completely benefit by the nutritive compounds resulted
from the preys.
Key words: Drosera, secretory trichomes, comparative analysis
Introduction
Drosera species are characterized by the presence of the secretory pedicelate
(tentacular) and sessile trichomes in the foliar limb. Their structure has been discussed in
many anterior papers [2, 3 and 4]. We proposed to present a comparative numeric analysis,
underlining the importance of the secretory trichomes in the normal manifestation of their
carnivorous character.
We counted the secretory pedicelate trichomes of the upper epidermis and the sessile
trichomes of both upper and lower epidermis belonging to mature leaves (from the basis of
the rosette) in all 16 Drosera species (D. aliciae Hamlet, D. binata Labill, D. brevifolia
Pursh, D. burkeana Planch, D. capensis L.- with three forms: D. capensis Alba L., D.
capensis Narrow Leaf L. and D. capensis Rubra L., D. capillaris Poir, D. cuneifolia
Thunb, D. dielsiana Exell et Laundon, D. intermedia Hayne, D. lovella T. N. Bailey, D.
montana St. Hill and D. spatulata Labill, all belonging to the collection of Alexandru
Borza Botanical Garden, Cluj-Napoca, and Drosera rotundifolia L. from the Natural
Reservation Grdinia Meadow, Suceava district). We have separately noted the number of
the trichomes of 20 microscopic areas, randomly chosen (belonging to the same leaf, to
different leaves of the same individual or to various individuals).
We noted: A=microscopic area, D=diameter, r=radius.
If D = 860 m, r = 430 m, A = x r2 (m)2 , A = 0.580586 mm2
First we noted the number of trichomes depending on their type, location (upper
epidermis or/and lower epidermis) and species and then we calculated their average for the
analyzed area (A = 0.580586 mm2). The next step was to calculate the number of trichomes
corresponding to A= 1 mm2, as follows: Averrage/0.580586
*
Al. I. Cuza University of Iasi, Botanical Garden Anastasie Fatu, Romania, irinastanescu2005@yahoo.com
Al. I. Cuza University of Iasi, Faculty of Biology, Biochemistry and Molecular Biology Laboratory , Romania
**
48

In order to completely analyze the efficiency of Drosera leaves regarding the
capturing of various preys, we considered of great importance the number of the secretory
trichomes of the foliar limb. All investigated Drosera species present pedicelate
(tentacular) secretory trichomes in the upper epidermis and sessile secretory trichomes in
both upper and lower epidermis. A tentacle consists of a various length stalk, longitudinally
traversed by fine tracheids ending in large groups in the glands head. The outer part of the
tentacles heads is composed of 2-3 layers of glandular radial elongated cells (especially
those from the external layer) surrounded by a thin cuticle. The tentacles attract the preys
by secreting sticky sweet mucilage, which contains enzymes too, in order to decompose the
preys body; then, the tentacles, helped by the sessile trichomes, absorb the nutritive
compounds.
The sessile secretory trichomes consist of two parallel rows of two cells each, on top
of the epidermal cells, and two cells between those of the epidermis. These trichomes
absorb the substances secreted by the tentacles, substances which seep on the leaf surface.
Comparing the number of the sessile secretory trichomes of the lower epidermis
(Fig. 1), we sow that D. capensis Alba and D. intermedia: 31.09 and 31.52, respectively.
D. aliciae registrated the lowest value (9.13); D. montana and D. spathulata presented other
similar values: 13.18 and 13.09, respectively.
Comparing the number of the pedicelate (tentacular) secretory trichomes of the
upper epidermis (Fig. 2), we evidenced that the D. binata presented the highest number of
tentacles (14.04), while D. montana registrated the lowest number (3.27). D. capillaris and
D. cuneifolia showed similar values: 8.01 and 7.75, respectively.
An attentive analysis of the Fig. 3 gives us information upon the highest number of
sessile secretory trichomes of the upper epidermis, registered in D. intermedia (25.06), and
the lowest number, in D. binata (6.20). D. capensis and D. capensisNarrow Leaf present
similar values: 21.01 and 20.23, respectively, in one category, in other category being D.
brevifolia and D. burkeana: 13.43 and 13.26, respectively.
In order to check the possible differences or similarities occurring, numerically,
between both types of secretory trichomes belonging to the upper epidermis in all
investigated species, the Anova test - the bifactorial model (without interaction) with an
equal number of observations in the cell, has been applied [1 and 5].
The test alows us calculate the square sums - based on the variability sources, the
factor value and its critical value. Starting with the experimental results obtained, the null
(H0) and the alternative (H1) hypothesis of the test have been formulated.
F = factor; Crit. F = Critical factor
Observation: F < Crit. F - H0 is accepted and H1 is refused;
F > Crit. F - H0 is refused and H1 is accepted.
The hypothesis of Anova test - the bifactorial model (without interaction):
1. H01: Da = Db = Dbr = Dbk = Dc = DcA = DcNL = DcR = Dcp = Dcn = Dd =
Di = Dl = Dm = Dr = Ds
49
H11: Da Db Dbr Dbk Dc DcA DcNL DcR Dcp Dcn Dd

Di Dl Dm Dr Ds, = trichomes averrage, Da = D. aliciae, Db = D. binata, Dbr =
D. brevifolia, Dbk = D. burkeana, Dc = D. capensis, DcA = D. capensis Alba, DcNL =
D. capensis Narrow Leaf, DcR = D. capensis Rubra, Dcp = D. capillaris, Dcn = D.
cuneifolia, Dd = D. dielsiana, Di = D. intermedia, Dl = D. lovella, Dm = D. montana, Dr =
D. rotundifolia, Ds = D. spathulata
2. H02: tt = st
H12: tt st, where tt = tentacular trichomes, st = sessile trichomes
The results of our study have been presented in Table I and Figure 4.
A comparison between the values of the two (calculated and statistical) factors led to
the following conclusions: 1. the calculated value of the factor on columns (0.804) is lower
than its critical value (2.403), so the alternative hypothesis (H11) is refused and the null
hypothesis (H01) is accepted (considerable numeric differences were not registered between
the secretory trichomes in the investigated species); 2. the calculated value of the factor on
rows (33.729) is significantly higher than its critical value (4.543), so the null hypothesis is
refused (H02) and the alternative hypothesis is accepted, that is why we can conclude:
notable numeric differences between both types of secretory trichomes belonging to the
upper epidermis exist.
In order to check the possible differences or similarities occurring between the number
of sessile secretory trichomes belonging to both epidermis of the leaf, the Anova test - the
bifactorial model, with an equal number of observations in the cell, has been applied [5].
The hypotheses are:
1. H01: H01: Da = Db = Dbr = Dbk = Dc = DcA = DcNL = DcR = Dcp = Dcn = Dd
= Di = Dl = Dm = Dr = Ds
H11: Da Db Dbr Dbk Dc DcA DcNL DcR Dcp Dcn Dd
Di Dl Dm Dr Ds , where = averrage of trichomes, Da = D. aliciae, Db = D.
binata, Dbr = D. brevifolia, Dbk = D. burkeana, Dc = D. capensis, DcA = D. capensis
Alba, DcNL = D. capensis Narrow Leaf, DcR = D. capensis Rubra, Dcp = D.
capillaris, Dcn = D. cuneifolia, Dd = D. dielsiana, Di = D. intermedia, Dl = D. lovella, Dm
= D. montana, Dr = D. rotundifolia, Ds = D. spathulata
2. H02: upp.ep. = lr.ep.
H12: upp.ep. lr.ep, where upp.ep. = upper epidermis, lr.ep. = lower epidermis
The results of our study have been presented in Table II.
Comparing the values of both (calculated and statistical) factors, we can affirm: 1.
the calculated value of the factor on columns (6.330) is higher than the critical one
(0.00047), so the alternative hypothesis (H11) is accepted, or, in other words, significantly
differences were registered between the 16 Drosera species, regarding the number of the
sessile secretory trichomes in both epidermis of the limb; 2. the calculated value of the
factor on rows (14.609) is higher than the critical one (0.0016), which alows us to refuse the
null hypothesis (H02) and accept the alternative one; so, we can admit that there are
considerable differences between the two epidermis of the limb, regarding the number of
sessile secretory trichomes.
50
The graphical representation of the number of sessile secretory trichomes belonging

to both (upper and lower) epidermis in the investigated species may be observed in Fig. 5.
Conclusions
The small number of sessile secretory trichomes of the upper epidermis in D. binata
is balanced by the great number of pedicelate (tentacular) trichomes; the tentacles secrete
mucilage rich in sugars and protheolitic enzymes and, then, absorb the compounds resulted
from the degradation of the pray, helped by the sessile trichomes.
In all the other investigated species, the number of sessile trichomes is higher than
that of the tentacles, due to their size differences (a great number of sessile trichomes is
required to absorb the substances accumulated in the upper epidermis and maintain it clean,
proper for respiration).
The lower epidermis presents a higher number of sessile trichomes than the upper
epidermis; in the upper epidermis, the absorbtion of the nutritive compounds is realized by
both categories of trichomes, while in the lower epidermis there are only the sessile
trichomes to absorb the droplets of substances. The greater the number of trichomes, the
better can the plants beneficiate the substances obtained by the preys body and more, the
better can the plants grow and develop.
REFERENCES
1.
2.
3.
4.
5.
FOWLER J., COCHEN L., JARVIS P., 2000 - Practical statistics for field biology, Second Edition, Ed. by
John Wiley & Sons, Ltd., England, 186 207
METCALFE C. R., CHALK L., 1972 - Droseraceae (1: 581-585). In Anatomy of the Dicotyledons,
Clarendon Press, Oxford
STNESCU IRINA, TOMA C., 2008 - Secretory structures of the carnivorous plants belonging to the
Droseraceae family. Proceedings of the 1st International Conference: Environment - Natural Science - food
industry in European Context, Baia Mare, 1: 323-326
TARNAVSCHI I. T., 1957 - Adaptrile morfologice ale plantelor carnivore. Natura, 4: 76-92
VARVARA M., ZAMFIRESCU T., NEACU P., 2001 - Lucrri practice de ecologie, Ed. Univ.
Alexandru Ioan Cuza Iai
51
.c
.c
ap
en
.c
ap
si
ap
en
s
en
si
s
si
"A
s
"N
lb
a
ar
D
ro
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w
ap
Le
en
af
si
s
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ub
r
a
D
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ap
il l
ar
D
is
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un
ei
fo
l
ia
D
.d
ie
ls
ia
D
na
.i
nt
er
m
ed
ia
D
.l
ov
el
D
la
.m
on
ta
D
n
.r
a
ot
un
di
fo
D
li a
.s
pa
th
ul
at
a
na
ifo
l ia
ur
ke
a
re
v
.b
.b
D
.c
.c
ur
ke
a
ifo
l ia
in
at
.c
na
ap
en
ap
si
en
ap
s
si
en
s
si
"A
s
l
"N
ba
ar
ro
D
.c
w
ap
Le
en
af
si
s
"R
ub
ra
D
.c
ap
il l
ar
D
is
.c
un
ei
fo
li a
D
.d
ie
ls
i
a
D
na
.i
nt
er
m
ed
ia
D
.l
ov
el
la
D
.m
on
t
an
D
.r
a
ot
un
di
fo
D
l
ia
.s
pa
th
ul
at
a
D
.b
.b
li c
ia
e
re
v
D
.b
.a
no. of sessile trichomes in the lower epidermis
9.13
in
at
li c
ia
e
.b
.a
10
no. of tentacular trichomes
35
30
31.09
25
5.08
5.77
31.52
26.52
29.28
24.37
27.90
20
19.72
15.42
15.59
3.88
5.77
52
19.29
15
10.51
11.97
11.80
7.06
8.01
3.36
13.18
7.75
4.48
5.77
13.09
Fig.1. Graphical representation of the number of sessile secretory trichomes of

the lower epidermis (in A=1 mm2) in the 16 Drosera species
16
14
14.04
12
10
6.55
7.49
5.51
3.27
4.65
Fig.2. Graphical representation of the number of pedicelate (tentacular) secretory

trichomes of the upper epidermis (in A=1 mm2) in the 16 Drosera species
D.
al
ic
i
D. ae
b
in
D.
br at a
e
D. vifo
bu lia
rk
ea
D.
D
D
ca . ca . ca na
pe
pe
pe
ns
n
n
is s is s is
"A
D. "N
lb
a
ca
r
a
pe row
ns
Le
is
a
"R f
ub
D.
r
a
ca
D. pill
cu ari
ne s
D. ifo
l ia
d
D. iels
ia
in
n
te
rm a
ed
D.
ia
lo
ve
D.
l
l
m
on a
D.
ta
ro
na
tu
nd
D.
if
sp oli
a
at
hu
la
ta
no. of trichomes
D
.c
.c
ifo
l ia
in
at
ur
ke
a
re
v
.b
li c
ia
e
no. of sessile trichomes in the upper epidermis
13.43
.c
na
ap
en
ap
si
ap
en
s
en
si
s
si
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s
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lb
a
ar
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ro
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w
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Le
en
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s
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a
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ar
D
is
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un
ei
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lia
D
.d
ie
ls
i
an
D
.i
a
nt
er
m
ed
ia
D
.l
ov
el
la
D
.m
on
ta
D
na
.r
ot
un
di
fo
D
li a
.s
pa
th
ul
at
a
D
.b
.b
.a
15
30
25
25.06
20
21.01
19.46
20.32
13.69
15.76
9.99
11.37
53
16.45
10
tentacular trichomes
17.14
13.26
10.68
8.96
13.00
5
6.20
Fig.3. Graphical representation of the number of sessile secretory trichomes

of the upper epidermis (in A=1 mm2) in the 16 Drosera species
30
25
20
15
10
sessile trichomes
Fig.4. Comparative representation of the number of both pedicelate (tentacular)

and sessile secretory trichomes of the upper epidermis in the 16 Drosera species
at
a
vi
fo
bu
l
rk ia
D.
D.
D. e a
n
ca
ca
c
a
pe ap
pe
ns en
ns
s
is
is
is
D. "N
"A
lb
c a ar r
pe o w a
ns
is Lea
"R f
D . ub
c a ra
D. pill
cu ari
s
n
D. e ifo
li
d
D . i e ls a
ia
in
n
te
rm a
ed
D.
ia
l
D. ov e
l
D . mo la
ro nta
na
tu
D. n di
s p f ol
a t ia
hu
la
ta
br
e
D.
D.
al
ic
i
D.
D.
bi
n
ae
no. of trichomes
35
30
25
20
15
10
5
0
upper epidermis
lower epidermis
Fig.5. Comparative representation of the number of sessile secretory trichomes

belonging to both epidermis
Table I. Calculated and critical values of the factors
Source of variability
SS
g. l.
Calculated F
SS
Critical F
Rows (type of trichome) 589.446

1
589.446
33.729
4.543
Columns (species)
210.910
15
14.060
0.804
2.403
Total
1062.493 16
SS = sum of squares, g. l. = degree of freedom (g. l. = n-1, where n = values),
SS = average sum of squares

Table II. Calculated and critical values of the factors
Source of variability
SS
g. l.
Calculated F
SS
Critical F
Rows (type of epidermis) 173.911

1
173.911
14.609
0.0016
Columns (species)
1130.431 15
75.362
6.330
0.00047
Total
1482.901 16
SS = sum of squares, g. l. = degree of freedom (g. l. = n-1, where n = values),
SS = average sum of squares
54

MORPHOLOGICAL RESEARCHES REGARDING THE INFLUENCE OF

TOPSIN M TREATMENTS ON MENTHA LONGIFOLIA (L.) HUDS. SPECIES
CLARA APROTOSOAIE A.*, URSULA STNESCU*, ANCA MIRON*, VIOLETA
FLORIA**, OANA CIOANCA*, MONICA HANCIANU*
Abstract. Topsin M is a common systemic fungicide used as protective/curative substance for alimentary
and medicinal plants. That is why it is relevant to evaluate the influence of topsin M upon the
morphological features of Mentha longifolia, a volatile oil producing medicinal plant and parental
species for the hybrid Mentha piperita. We noticed statistical significant variations (oneway Anova) of
the dimensional features of the leaves from topsin M treated plants comparing to the control.
Key words: Mentha longifolia, topsin M, morphological features
Introduction
The abusive and disordered employment of pesticides has the potential to harm
human health and the environment and pose the ecosystems to ecological risk.
Because of possible health effects, widespread use and insufficient data, pesticide
monitoring in plants is necessary. Some of the pesticide monitoring aspects refere to the
investigation of the pestide influence on the morphological features of medicinal plants.
Data from literature report that pesticides may interfere with morphological features
of plants, as following:
smaller and fewer leaves, smaller axial organs (carbamate and
amido-type erbicides) [5];
depigmentations along the nervures, necrosis at edge of the
leaves (urea and sulphonylurea derivates; alchyl N-phenylcarbamates and
alchyl-N-phenylthiocarbamates) [1, 2, 3];
variation of the lenght of the lamina of the leaf and the lenght of
the whole plant (fungicides) [4].
Topsin M is a common systemic fungicide used as a protective/curative substance
for alimentary and medicinal plants. That is why it is relevant to evaluate the influence of
Topsin M upon the morphological features of Mentha longifolia, a volatile oil producing
medicinal plant and parental species for the hybrid Mentha piperita.
*
Faculty of Pharmacy, University of Medicine and Pharmacy Gr.T.Popa, Iasi, Romania
**Botanical Gardens Anastasie Fatu, Iasi, Romania
55
Plant materials were brought from the experimental lots in Anastasie Fatu
Botanical Garden, Iasi. In this experimental area, parallel cultures of Mentha longifolia (L.)
Huds have been made in the period 2001-2003.
Thus, every year there have been two experimental fields: a field which had no
pesticide treatment (control area) and a field which had been treated with pesticide.
The antifungal treatment was achieved in vegetative phase by spraying a wettable
powder of Topsin M 70 PU (TM) (Oltchim Rm. Valcea-Romania) as 0,1% and 0,4%
aqueous solutions.
Investigated samples of Mentha longifolia are presented in table I:
Table I: Samples of Mentha longifolia
Nr.
1.
2.
3.
4.
5.
6.
7.
Sample
Control 2001
Treatment TM 0,1% 2001
Control 2002
Control 2003
Codification
M.l. M 2001
M.l. TM 0,1% 2001
M.l. M 2002
M.l. TM 0,4% 2002
M.l. M 2003
M.l. TM 0,1% 2003
M.l. TM 0,4% 2003
The morphological study has been achieved through the analysis of the parameters
concerning aspect, colour and dimensions of the leaves (officinal product) from both treated
and untreated plants, as following:
- the shape;
- the edges, base and top;
- covered hairs for the both faces of leaves;
- the length and breadth (cm), depending on the leaf insertion: top, middle or the
base of the stem. For each type of leave, for each variant of treatment and for each
control, there have been made 10 measurements of the studied parameter.
The dimensional parameters data were statistically analysed using the Oneway
Anova method and the J.M.P. Programme 5.0.1.2. (SAS Institute, Cary N.C., S.U.A).
Results and discussion
Mentha longifolia leaves from both untreated (Fig. 6) and TM 0,1% (fig. 7) and TM
0,4% (fig. 8) treated plants have the some morphological features of the leave.
The lanceolate or oblong leaves were sessile and arranged opposite on the stem.
The tip of the limb is sharp and the base is narrowed. The edges of the leaves are
convexe and almost parallel and have triangular teeth, 1-4 mm distanced orientated
forward. The nervation is pennate and proeminent on the lower surface. Leaves have
covered hairs placed especially along the nervures on the lower surface. The leaves are dark
green on the upper surface and lighter on the lower surface. The powdered leaves have an
aromatic, characteristic smell.
56
The dimensional parameters of both untreated and treated vegetal material are
shown in Table II, as minimal and maximal registered for each type of leave.
Table II. The dimensional parameters (cm) of the lamina from Mentha longifolia
Top leaves
Middle leaves
Basal leaves
Sample
lenght
M.l. M
2001
M.l.
TM 0,1%
2001
M.l. M
2002
M.l.
TM 0,4%
2002
M.l. M
2003
M.l.
TM 0,1%
2003
M.l.
TM 0,4%
2003
breadth
lenght
breadth
lenght
breadth
2.0 2.5
0.9 1.7
3.0 4.2
1.2 1.6
2.8 4.1
1.0 1.7
2.4 2.9
1.4 - 1.9
3.2 4.2
1.3 1.8
3.0 4.4
1.2 1.9
2.3 2.9
1.1 1.6
3.5 4.3
1.2 1.8
3.0 4.3
1.3 1.6
1.9 2.9
0.8 1.4
3.6 4.2
1.1 1.6
3.0 4.1
1.2 1.6
1.7 2.3
0.7 1.2
3.5 4.3
1.4 1.6
3.0 5.0
1.3 1.9
2.3 3.3
1.2 1.9
3.8 4.4
1.7 2.0
2.9 3.8
1.3 1.8
2.4 4
1 - 1.9
3.9 4.8
1.2 2.1
2.7 3.6
1.3 1.9
The dimensional parameters data were statistically analysed using the Oneway
Anova method and the J.M.P. Programme 5.0.1.2. (SAS Institute, Cary N.C., S.U.A) [6].
Statistical data (x -mean; SD standard deviation; p*- indicator foe evaluation of
statistical significance) were presented in Tables III and IV:
Table III. Biometrical data of Mentha longifolia leaves the length of the lamina (cm)
Top leaves
Middle leaves
Basal leaves
Sample
M.l. M
2001
M.l.TM
0,1% 2001
M.l. M
2002
M.l.TM
0,4% 2002
M.l. M
2003
x
2.23000
SD
0.176698
2.57000
0.188856
2.61000
0.228279
2.49000
0.317805
2.10000
0.240370
p*
0.0006
0.3450
x
3.71000
SD
0.369534
3.78000
0.339280
3.82000
0.342540
3.92000
0.225093
4.02000
0.274064
0.18409
57
p*
0.6643
x
3.22000
3.32000
0.4504
0.16502
SD
0.456557
p*
0.6619
0.545283
0.472464
3.39000
3.43000
0.535516
4.39000
0.645411
0.8614
0.47821
M.l. TM
0,1% 2003
2.77000
0.394546
M.l. TM
0,4% 2003
2.94000
0.602218
0.35409
4.14000
0.206559
4.46000
0.283627
0.15498
3.39000
0.369534
3.17000
0.333500
0.69821
*It is considered that the results are statistically significant if p* is lower than 0.05, in case
of 2001 and 2002 samples, or if p* has a positive value for 2003 samples.
Table IV. Biometrical data of Mentha longifolia leaves the breadth of the lamina (cm)
Top leaves
Middle leaves
Basal leaves
Sample
M.l. M
2001
M.l.TM
0,1% 2001
M.l. M
2002
M.l.TM
0,4% 2002
M.l. M
2003
M.l. TM
0,1% 2003
M.l. TM
0,4% 2003
x
1.24000
SD
0.250333
1.63000
0.176698
1.36000
0.171270
1.15000
0.177951
1.01000
0.172884
p*
x
1.41000
SD
0.137032
1.57000
0.182878
1.48000
0.161933
1.40000
0.200000
1.52000
0.063246
1.84000
0.107497
1.72000
0.234758
p*
0.0400
x
1.30000
SD
0.262467
1.52000
0.278089
1.42000
0.113529
1.47000
0.125167
1.69000
0.196921
1.66000
0.142984
1.61000
0.246982
0.0855
0.0008
0.0150
0.3386
0.14982
0.18511
1.44000
0.195505
1.34000
0.279682
0.08511
p*
0.02982
0.3618
0.19197
0.14197
The statistical data reveal that the following dimensional variations compared to the
corresponding controls are significant:
- increase in lenght and breadth of the top lamina at the 0,1% treated plants;
- increase in breadth of the middle lamina at 0,1% treated plants as well as an
increase of lenght and breadth of the same leave at 0,4% treated plants;
- decrease in lenght of basal lamina at TM 0,1% and TM 0,4% treated plants;
- increase in breadth of the basal leaves at TM 0,1% treated plants.
Conclusions
Antifungal treatment wih Topsin M did not affect morphological features of Mentha
longifolia leaves, except dimensional features.
We noticed statistical significant variations (Oneway Anova) of the dimensional
features of the leaves from Topsin M treated plants comparing to the control, as following:
- lenght and breadth of top lamina increase at 0,1%TM treated plants (fig. 1, 2);
- lenght and breadth of middle lamina increase at 0,4% TM treated plants (fig. 3, 4);
- only the breadth of middle and basal lamina increase at TM 0,1% treated plants;
- the lenght of the basal lamina decreases at TM0,1% treated plants (fig. 5).
58
REFERENCES
1.
2.
3.
4.
5.
6.
GOODWIN S., AHMAD N., 1998 - Relationship between azinphosmethyl usage and residues on grapes and
in wine in Australia. Pesticide Science, 53 (1): 96-100
GUANG-GUO Y., WILLIAMS B., 1999 - Herbicides residues in grapes and wine. J. Environ. Sci. Health B,
34 (3): 397 411
ERSEN F., KRALOVA K., MACHO V., 2000 - New findings about the inhibitory action of
phenylcarbamates and phenylthiocarbamates on photosynthetic apparatus. Pesticide, Biochemistry and
Physiology, 68 (2): 113-118
NITA M,1997. Teza de doctorat, Iasi
SLONOVSCHI V., NITA M., NECHITA A., 2001 - Prezent i viitor n combaterea buruienilor, Edit. Ion
Ionescu de la Brad, Iasi
WAYNE D., 1999 - Biostatistics: a foundation for analysis in the health sciences - 7th ed.
*It is considered that the results are statistically significant if Oneway Anova diagrames of
the samples have a reduced level of superposition
4.5
L varf 2003
4
3.5
3
2.5
2
1.5
Martor
tratam 0.1%
tratam 0.4%
Proba
Samples
TM 0,1%
All Pairs
Tukey-Kramer
0.05
With Control
Dunnett's
0.05
TM0,4%
Fig.1. Oneway Anova diagrame for the lenght of the lamina of the Mentha longifolia top
leaves
59
D varf 2003
1.75
1.5
1.25
1
0.75
0.5
tratam 0.1%
Martor
tratam 0.4%
Proba
Samples
TM 0,1%
All Pairs
Tukey-Kramer
0.05
With Control
Dunnett's
0.05
TM 0,4%
Fig. 2 Oneway Anova diagrame for the breadth of the lamina of the Mentha longifolia
top leaves
L tulp 2003
4.5
3.5
Martor
tratam 0.1%
tratam 0.4%
Proba
Samples
TM 0,1%
All Pairs
Tukey-Kramer
0.05
With Control
Dunnett's
0.05
TM 0,4%
Fig. 3 Oneway Anova diagrame for the lenght of the lamian of the Mentha longifolia
middle leaves
60
2.2
tulp 2003
2
1.8
1.6
1.4
1.2
1
tratam 0.1%
Martor
tratam 0.4%
Proba
Samples
TM 0,1%
All Pairs
Tukey-Kramer
0.05
With Control
Dunnett's
0.05
TM 0,4%
Fig.4.Oneway Anova diagrame for the breadth of the lamian of the Mentha longifolia top
leaves
5.5
Lbazal 2003
5
4.5
4
3.5
3
2.5
Martor
tratam 0.1%
tratam 0.4%
Proba
Samples
TM 0,1%
All Pairs
Tukey-Kramer
0.05
With Control
Dunnett's
0.05
TM 0,4%
Fig.5. Oneway Anova diagrame for the lenght of the lamina of the Mentha longifolia
basal leaves
61
Fig. 6: Control
Fig. 7: Treatment TM 0,1%
Fig. 8: Treatment TM 0,4%
62

A HISTO-ANATOMICAL STUDY OF THE MODIFICATIONS INDUCED BY

THIOPHANATE METHYL ON THE CALENDULA OFFICINALIS L. STRAIN
LUMINIA HUANU-BASHTAWI, C. TOMA
Abstract. The effect of the Topsin M fungicide (thiophanate methyl) on the anatomical structure of the
Calendula officinalis L. strain is evidenced, comparatively with an untreated sample. The histological
modifications caused by this treatment, applied in two different concentrations (of 0.1% and 0.5%,
respectively) have the same stimulation effects as those observed in similar experiments. The crosssections, made at three different levels, were performed through the main branches occurring, in the
moment of fungicide application, in different stages of development - once known that the morpho-histoanatomical type manifestations of pesticides depend on the extent of tissular differentiation as well as on
the development stage of the treated organ.
Key words: Calendula, Topsin M, cytokinin hormone-type action, strain, histo-anatomical modifications.
Introduction
In recent years, considering the secondary effects of pesticides, drastic measures for
reducing their application have been taken, the existing methods for controling different
diseases and pests involving elimination of highly-toxic fungicides, a lower concentration
of those still present on the market, and introduction of some biologically-active principles
[10]. In spite of the success recorded with certain biopreparations, in most cases, total
substitution of fungicides with biological agents did not work [2], integration of chemical
control with the biological one representing the most promising strategy to combat plant
pathogens, with minimum effects on the natural environmental equilibrium [4].
In everyday practice, thiophanate methyl, which is a benzimidazolic fungicide, may
be applied in combination with certain biological agents, such as Pseudomonas fluorescens
or Trichoderma harzianum [5, 12], in which selected mutant strains are resistant to the
action of carbendazime (MBC) the main metabolite of thiophanate methyl and the active
substance in the plant [9]. In their experiments made on Nicotiana tabacum, Garcia et al.
[7] demonstrated that the toxicity of carbendazime is coordinated by the dose-response
relation: at a 50% lower concentration (1.3mM) than the recommended value (2.6mM), the
dry mass, the concentration of carotenoids and some mineral elements (N, K) show a
positive resnet, comparatively with the reference, while a higher concentration (5.2mM) is
phytotoxic. Besides the advantages referring to biomass production, and especially to a
reduced fungicide amount (which is actually a condition of the new regulations on the
integrated system of plant protection), application of a lower dose induces an increase in the
concentration of polyphenolic compounds, comparatively with the reference, and,
especially, higher than that of the treatment applied in the recommended concentration; at a
Al. I. Cuza University of Iai, Faculty of Biology, Bd. Carol I, no. 20A., Iai, Romania
bashtawibio@yahoo.com
63
higher fungicide dose, inhibition of their synthesis may be observed, in spite of the lowest
concentration of polyphenolic compounds recorded in the reference sample [6].
Considering the importance of the phenolics in the host-pathogen interaction, their
ability of granting resistance to pathogens either directly or indirectly, by mediating the
plants Systemic Acquired Resistance as well as reducing the amounts of applied
fungicide is especially interesting for reducing the polluting effects of such substances upon
the environment [13]. Such beneficial effect - of the cytokynin hormone-type action of
the benzimidazolic fungicides - has not been always observed in either practical or
experimental studies, even if they had been applied in recommended doses [1]; more than
that, the utilization of carbendazime, benomyl or thiophanate methyl fungicides was
compromised by the manifestation of some secondary phytotoxic effects [3, 11].
On the basis of such observations, as well as of the idea that most of the
investigations devoted to thiophanate methyl involve mainly study of the metabolism,
toxicity or estimation of the maximum residual potential from the vegetal material, and
considering the influence that fungicides might exert on the morpho-anatomy of the treated
plants [8], the present study analyzes the histo-anatomical modifications possibly induced
in the Calendula officinalis strain by the thiophanate methyl and/or its metabolite (MBC),
comparatively with a non-treated sample, with a well-known structure [14].
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% and
TM70 0.4%), 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. The light micrographs were
performed on a Novex (Holland) microscope, using a Canon 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 in the upper third of the main branches
The reference contour of the cross-section, intensely ribbed (12-13 ribs and an
equal number of valecules), numerous secretory hairs. The bark, cholenchymatized in the
ribs and parenchymatous-assimilatory in the rest, includes 5-6 layers of cells with slightly
thickened walls. Conducting fascicles: 12-13 large or intermediary and 7, respectively,
small ones, each evidencing a primary structure and a girdle of periphloemic fibers with
thin, cellulosic walls (Fig. 1, 2, 3). The medullary rays are parenchymatic-cellulosic, only in
64
front of a few of them occurring small islands (2-3), with very few phloem elements. The
pith is thick, parenchymatous-cellulosic.
TM 0.1% treatment the outline of the cross-section (larger than in the reference)
shows less ribs (9-10), of various size, while the cholencymatic cells have thicker walls
(Fig. 1). The epidermal cells, with thicker external walls, are covered by a thicker cuticle.
On the margin of the bark, tangentially-elongated aeriferous cavities may be observed.
Unlike the reference, more numerous and larger conducting fascicles are present, as
follows: 18 large or intermediary and 8 small ones, all underlining a primary structure (Fig.
1, 2); more than that, more numerous, very small conducting fascicles (6-7), possessing
exclusively liberian elements (Fig. 2, 4), are observed. The cambium is more active, being
formed of more layers (3-4). Many of the xylem vessels appear in process of formation
(Fig. 3). The bark evidences 2-3 small, circular conducting fascicles, each with a thin girdle
of sclerenchymatic fibers at the periphery of the liber (Fig. 4). Some fascicles are collateral
type, structurally similar to those from the central cylinder, others (observed in the main
strain) evidencing liberian elements and even incipien ligneous vessels on the internal side
of the conducting fascicle, as well, actually representing a form of transition towards the
circular hadrocentric vascular bundles, surrounded either partially or totally by liber, at
the periphery. Such abnormal fascicles, resulted from the stimulation activity induced by
thiophanate methyl, have been noticed in different positions at this level of the strain: in the
bark, in the vicinity or in the thickness of the girdles of periphloemic fibers and even inside
the liber from the conducting fascicles of the central cylinder (Fig. 5).
TM 0.4% treatment diameter of the cross-section larger than in the other 2
samples, more prominent ribs (7), alternating with smaller ones (6-7), while the
cholenchyma evidences cells with less thick walls, comparatively with the TM 0.1%
treatment. The same aeriferous cavities as in the TM 0.1% sample appear, yet the secretory
hairs are extremely numerous comparatively with the first treatment, and, especially, with
the reference. The central ring contains 18 large or intermediary and 10 small conducting
fascicles, each with primary structure and with a girdle of periphloemic fibers with thin,
cellulosic walls (Fig. 1, 2); on the lateral side of the conducting fascicles, more liberian
islands (16-17) may be observed (Fig. 2, 4). The cambium is as thick (3-4 layers) and as
active as in the TM 0.1% treatment, numerous xylem vessels being still in the process of
formation; however, the diameter of the vessels is smaller, although the number of vessels
is higher than in the reference (Fig. 3). The bark evidences only one, small conducting
fascicle, of collateral type, similar to those observed in the TM 0.1% treatment.
Cross-sections through the median third of the main branches
The reference the ribs contain a very low amount of cholenchyma, the cell
forming it having slightly thickened walls. Small aeriferous cavities some of them
tangentially-elongated are presented at the periphery of the bark (Fig. 6). The central
cylinder includes 17 large or intermediary conducting fascicles, of open collateral type,
have only a primary structure; the ligneous vessels, separated by cells of cellulosic ligneous
parenchyma, have thickened, yet weakly-lignified walls, a few of them being still under
edification (Fig. 6, 7). The fibers of the periphloemic girdles show slightly cellulosic walls,
while the medullary rays are parenchymatic-cellulosic. The very small conducting fascicles
(8) evidence only liberian elements (Fig. 9).
65
TM 0.1% treatment diameter of the cross-sections is much larger than in the

reference, while the secretory hairs are more frequent. The aeriferous cavities occurring at
the periphery of the bark are very large and considerably elongated in tangential direction
(Fig. 6). The girdle of conducting tissues includes 19 large or intermediary and 5 small
fascicles, all evidencing only a primary structure; during this treatment, more numerous
small fascicles (5) and liberian islands (12) are formed (Fig. 6, 7, 9). Lignification of the
ligneous vessels is weaker, while the fibers of the periphloemic girdles have thin, cellulosic
walls. The cambium, especially the intrafascicular one, is extremely thick (6-7 layers); only
here and there, formation of the interfascicular cambium begins. Cambial activity is much
more intense than in the untreated sample and, consequently, more numerous vessels hardly
appear in a process of edifications (Fig. 8, 9).
TM 0.4% treatment comparatively with the other two samples taken into study,
the walls of the epidermal cells are thicker, being covered by a thicker cuticle. In the ribs,
mention should be made of the presence of typical angular cholenchyma, the cells having
thicker walls than both in the TM 0.1% treated and in the untreated samples. More
numerous conducting fascicles, all with a primary structure, are formed: 22 large or
intermediary and 5-6 small ones, but on the lateral side of the conducting fascicles more
liberian islands (22) may be observed (Fig. 6, 7, 9). The highly active cambium, both intraand interfascicular, includes 6-7 cell layers, which form an almost continuous ring (Fig. 8,
9), while the periphloemic fibers have cellulosic walls, too. In the bark, 1 or 2 neighbouring
conducting fascicles, surrounded by meristematic tissue, are present (Fig. 7).
Cross-sections in the inferior third of the main branches
The reference the ribs are more attenuated while, in the peripheral area, in front of
the more prominent ribs, largely tangentially, elongated aeriferous cavities may be
observed. The bark includes 4-5 layers of cells and the central cylinder has 19 big or
intermediary, 6-8 small conducting fascicles and 8-10 liberian islands (Fig. 10, 15); the first
two categories, of open collateral type, show, each, a thick girdle of sclerenchymatic fibers
with moderately thick, yet intensely lignified walls, at the periphery of the liber. The
vessels of the primary xylem are separated by cells of cellulosic parenchyma, while those of
secondary xylem, by a lignified parenchyma; the libriform elements have slightly
thickened, yet intensely lignified walls (Fig. 10). In some fascicles, the procambium is quite
sufficiently thick (6-8 layers), to become soon an intrafascicular cambium, which is
continuing with the interfascicular cambium, resulted from the dedifferentiation of the
medullary rays (Fig. 12, 15). All conducting fascicles are separated by large medullary rays,
parenchymatically-lignified at xylem level and parenchymatically-cellulosic at liberian
level (Fig. 15). The pith is thick, parenchymatous-cellulosic, the cells showing very thin
walls.
TM 0.1% treatment although the diameter of the cross-sections is larger than in
the reference sample, the small ribs are equally attenuated, with thinner cholenchyma
girdles and cells with less thick walls. The secretory hairs are more frequent and, at the
periphery of the bark, smaller aeriferous cavities may be observed, the cortical parenchyma
being thicker (7-9 layers of cells) (Fig. 10). The treatment stimulates cellular division at
cortical level and even in the medullary rays (only up to the level of the liber), almost all
cells from the bark evidencing more division, both anticline and pericline walls (2, 3 or
66
even 4) (Fig. 14). The large or intermediary conducting fascicles (19-20) are very close to
one another, evidencing much larger sizes, while the phloemic girdles show fibers with
thicker and more intensely lignified walls; more than that, small conducting fascicles (13)
and islands of liberian elements (20) occur among these conducting fascicles (Fig. 10, 11,
15). The secondary xylem shows more libriform, the perimedular parenchyma being
moderately sclerified and lignified. The meristematic tissue forms a continuous ring (8-10
layers), the tracheogenesis process developing still (Fig. 12). The conducting fascicles
xylem, along with the moderately sclerified and lignified medullar rays, form a much
thicker ring than in the reference (Fig. 10). It should be also mentioned that in the bark
there are some atypical conducting fascicles present (2-3) some of which, relatively large or
very small, of open collateral type, surrounded by a plurilayered meristematic tissue, others
being concentric hadrocentric (cortical), each with a thin girdle of sclerenchymatic fibers at
the periphery of the liber, or leptocentric vascular bundles (on the inner side of the ring of
conducting fascicles), surrounded either exclusively by vessels or by vessels and libriforme
fibers (Fig. 16, 17).
TM 0.4% treatment the diameter of the cross-sections exceeds by far that of the
previous samples, yet the cuticle is thinner, while the secretory hairs are more numerous
(Fig. 10). In the much attenuated ribs, less cholenchyma is present, yet the hypodermal
layer is tangentially cholenchymatized along the whole circumference of the strain (Fig.
13). The bark contains approximately 8-10 cell layers, no aeriferous cavities being present
at its periphery, so that the tissues are much more compact. Division of the cortical cells is
intensely stimulated, more numerous division walls being present in the same cell,
especially in those from the periphery of the bark (Fig. 15). The conducting fascicles are
very large or intermediary (20-21) and small (17-19), the last of them having no girdles of
periphloemic fibers; on the lateral side of the conducting fascicles more liberian islands
(20) are occurring. Sclerification and lignification of the libriform elements and of the
ligneous vessels are moderate, while those of the periphloemic fibers are more intense. The
medullary rays are parenchymatically lignified, the cells having slightly thickened walls.
The meristematic tissue forms a much thicker ring (of 10-12 cell layers); more than that, the
cambium is still to be divided, its cell being highly active, which explains why the process
of tracheogenesis is still under development (Fig. 12, 15). The bark evidences the same
peculiar type of fascicle, with the xylem surrounded by plurilayered meristematic tissue.
Conclusions
The most important histo-anatomical reactions from the part of the strains treated
with thiophanate methyl involve a more intense cambial activity and the occurrence of
some liberian-xylemic nodules, in various places of the strains thickness: from outside the
bark (at the bottom of the cholenchymatized ribs) up to the central cylinder (inside the
liber). The intrafascicular cambium is very active, being formed of several cell layers, while
the interfascicular one tends to form, when treated, a continuous, much thicker ring.
The size of the conducting fascicles is much larger than in the reference, their
number exceeding those of the reference; generally, more small conducting fascicles and
more liberian islands result after the treatment. The process of tracheogenesis is still under
67
development, which explains the presence of more immature vessels with thin, celullosic
walls. The treatment with thiophanate methyl stimulates cellular division at cortical level
and, consequently even in the medullary rays (yet only up to the level of the liber) more
division walls (2-3, sometimes 4), usually anticlynic or periclynic, may be observed in the
cells of the bark. As a result of the stimulating action, the much larger-sized conducting
fascicles form, together with the medullary rays, a much thicker ring than that of the
reference, while the secretory hairs are more numerous than in the untreated samples.
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HUTANU-BASHTAWI L., TOMA C., 2008 Contributionsto the histo-anatomical study of the Calendula
officinalis L. leaves treated with thiophanate methyl (Topsin M). An. t. Univ. "Al. I. Cuza" Iai, s. II a ,
fasc.1 (Biol. veget.), 54: 22-32
JAYARAJ J., RADHAKRISHNAN N.V., 2003 - Development of UV-induced Carbendazim-resistant
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KOZERA W., KLEIN M., 1980 - The influence of the fungicides Benlate and Topsin M on the mitotic
process in root meristems of onion setts (Allium cepa L.). Acta Agrar. Silvest., Ser. Agrar., 19: 117-132
MALATHI P., VISWANATAN R., PADMANABAN P., MOHANRAJ D., 2002 - Compatibility of
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MOLINA A., HUNT M.D., RYALS J.A., 1998 - Impaired fungicide activity in plants blocked in disease
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RUGINA R., TOMA C., 1989 - Recherches histo-anatomiques sur quelques plantes mdicinales de la
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68
Ca. of. M
Ca. of. TM 0.1% Ca. of. TM 0.4%
Fig. 1. Cross-sections main branch, upper level (Oc.10x Ob.2)
Ca. of. M Ca. of. TM 0.1% Ca. of. TM 0.4%

Fig. 2. Cross-sections conducting fascicle, upper level (Oc.10x Ob.20)
Ca. of. M
Ca. of. TM 0.1% Ca. of. TM 0.4%
Fig. 3. Cross-sections intrafascicular cambium, upper level (Oc.10x Ob.40x3)
Ca. of. TM 0.1%

Ca. of. TM 0.4%
Fig. 4. Cross-sections upper level, cortical, colateral conducting fascicles
Fig. 5. Cross-sections strain, upper level, TM 0.1 % treatment, concentric conducting

fascicles, present in the bark, in the periliberian fibers and in the liber
69
Ca. of. M
Ca. of. TM 0.1% Ca. of. TM 0.4%
Fig. 6. Cross-sections main branch, middle level (Oc.10x Ob.2)
Ca. of. M Ca. of. TM 0.1% Ca. of. TM 0.4%

Ca. of. TM 0.4%
Fig. 7. Cross-sections middle level normal (a) and cortical (b) conducting fascicle
Ca. of. M
Ca. of. TM 0.1%
Ca. of. TM 0.4%
Fig. 8. Cross-sections conducting fascicle, cambium, middle level (Oc.10x Ob.40)
Ca. of. M
Ca. of. TM 0.1%
Ca. of. TM 0.4%
Fig. 9. Cross-sections interfascicular cambium, middle level (Oc.10x Ob.20)
Ca. of. M
Ca. of. TM 0.1% Ca. of. TM 0.4%
Fig. 10. Cross-sections main branch, basal level (Oc.10x Ob.2)
70
Ca. of. M
Ca. of. TM 0.1% Ca. of. TM 0.4%
Fig. 11. Cross-sections conducting fascicle, basal level (Oc.10x Ob.10)
Ca. of. M
Ca. of. TM 0.1%
Ca. of. TM 0.4%
Fig. 12. Cross-sections conducting fascicle, cambium, basal level (Oc.10x Ob.40)
b
Ca. of. M
Ca. of. TM0.1% Ca. of. TM0.4%
Ca. of. TM0.4%
Fig. 13. Cross-sections angular (a) and tangential (b) cholenchyma, basal level
Ca. of. M
Ca. of. TM 0.1%
Ca. of. TM 0.4%
Fig. 14. Cross-sections bark, basal level (Oc.10x Ob.40)
Ca. of. M
Ca. of. TM 0.1%
71
Ca. of. TM 0.4%
Fig. 15. Cross-sections interfascicular cambium, basal level (Oc.10x Ob.20)
Fig. 16. Cross-sections basal level, TM 0.1% treatment - conducting fascicles of open
collateral type, surrounded by a plurilayered meristematic tissue, occurring in the bark
Fig. 17. Cross-sections - basal level, TM 0.1% treatment - concentric, hadrocentric

conducting fascicles, with ligneous vessels inside, situated in the bark (a), in the vicinity (b)
or in the thickness (c) of the periliberian fibers; concentric, leptocentric conducting
fascicles, surrounded by large xylem vessels and by libriforme elements, present on the
inner part of the conducting fascicles (d) and collateral conducting fascicles, occurring in
front of the medullary rays at the level of the liber (e)
72

THE ANALYSIS OF DAILY CONCENTRATIONS OF AIRBORNE POLLEN IN

THE WEST AND SOUTHWEST OF ROMANIA
NICOLETA IANOVICI*
Abstract. This analysis is meant to determine the way the pollen grains are disseminated in the
atmosphere in Western and South-Western Romania in the year 2004, by using the volumetric method of
collecting, identifying and quantifying data. The monitoring period started on the 16th of February and
ended on the 10th of October 2004 (238 days). 23 pollen types were identified. The early spring spring
period was dominated by the pollen coming from anemophilic trees. In the late summer autumn period
the airplankton was dominated by the airpollen coming from herbaceous plants. For the investigated area
the maximal quantity of airpollen occurred in April. The daily concentration of airpollen for each pollinic
type is expressed in number of pollen grains on m of air (PG/ m). Other parameters taken into account
were: the total daily concentration of airborne pollen, the total monthly variation, and the total annual
variation. The highest daily concentrations were those of Ambrosia (August and September), Artemisia
(August), and Poaceae (May, June, and July). The main airborne polluter was the pollen of Ambrosia
artemisiifolia.
Key words: airpolynic spectrum, pollen types, anemophilic taxa
Introduction
The changes that have occurred in the floristic composition immediately influence
the airplankton composition [24, 17]. The studies of the airpolynic spectrum are of interest
to biologists, but they mainly have an impact on physicians and allergic patients, since
chronological correlations may be established between airpollen concentrations and certain
symptoms of asthmatic and pollinosis patients. To better manage such diseases, pollinic
calendars have been devised in many countries [1, 2, 3, 4, 5, 6, 7, 9, 10, 11, 15, 19, 23, 25,
26, 16, 27, 28, 29]. In Romania, such yearly calendars have been devised for the West and
the Southwest of the country [12, 13, 14]. The aim of this analysis is to describe the
dynamics of the airpollen in Timioara in the year 2004, by taking into consideration the
data obtained using the volumetric method of collecting, identifying and quantifying.
Materials and methods
The monitoring aeropalynologic station (the only one in Romania) belonging to the
Biology Department of The University of Timioara, uses a VPPS 2000 Lanzoni volumetric
pollen-trap. The trap allows the evaluation of the airborne dynamics of the polen in the city
and its surroundings; the results are important for the plains in the west and southwest of
Romania. The apparatus is placed on the roof of the west University building,
approximately 20 meters from the ground, far from industrial areas and the barriers which
might prevent the circulation of air currents. The apparatus applies the principle initially
*
Department of Biology, Faculty of Chemistry-Biology-Geography, West University of Timisoara, Romania
73
presented by Hirst (1952) and the identification is made with a photonic microscope. The
siliconed bands inside the volumetric trap were changed and analyzed weekly [8, 18, 20,
22] using fuxine for coloring. The identification of the pollen was performed on
morphological bases at x400. The daily concentration of pollen is expressed in the number
of pollen grains on m of air (pg/ m). Our bulletins were provided weekly on the following
websites: http://www.pollinfo.ini.hu, http://www.nspolen.com,http://www.polleninfo.org.
In 2004 the monitoring lasted for 238 days, starting on the 16th of February and
ending on the 10th of October. 23 types of airpollen were identified (Tab. I). The plants that
produce allergenic pollen are woody plants, grasses and herbs. The herbs belong to the
Hamamelidae subclass (Urticaceae family), Asteridae (Asteraceae, Plantaginaceae),
Caryophylidae (Chenopodiaceae, Amaranthaceae, Polygonaceae). Most of the anemophilic
woody plants that have an allergenic potential belong to the Hamamelidae subclass
(Platanaceae, Juglandaceae, Moraceae, Urticaceae, Fagaceae, Betulaceae i Corylaceae)
and to the Dileniidae (Salicaceae, Tiliaceae), Rosidae (Aceraceae) and Asteridae
(Oleaceae) subclasses. The Pinophyta (Pinaceae, Taxaceae/ Cupressaceae) are also
anemophilic.
The pollinic range of the month of February included five types of pollen coming
from the following taxa: Corylus (27.75%), Alnus (27.36%), Ulmus (15.91%), Acer
(4.81%) and Taxaceae/Cupressaceae (24.05%). The day with the highest concentration was
the 20th of February (40 PG/m3).
In March we noticed the presence of the following five pollen types in the
airplankton: Populus, Salix, Fraxinus, Betula and Carpinus. The pollen of the
Taxaceae/Cupressaceae represented 26.47% of the total monthly concentration. The day
with the highest concentration was the 30th of March (224 PG/m3); the airpollen of Populus
represented 34% of this concentration.
April 2004 was the month displaying the highest quantity (4017 PG/m3) correlated
with the largest number of taxa which were in the flowering phenophase (16). The highest
concentrations were established for the airpollen of Betula (22.5%), Populus (16.23%),
Salix (15.23%), Fraxinus (10.08%), Carpinus (8.36%). The day when the concentration
was the highest was the 1st of April (318 PG/m3), that can be explained by the presence of
the 9 pollen-producing taxa (Fig.1).
In May, in spite of the presence of 15 pollinic types, the total monthly concentration
of pollen was much lower than the one in the previous month (1489 PG/m3). The pollinic
range was dominated by the pollen of Poaceae (46.4%). As to woody plants, the highest
concentration was that of Pinaceae (13.7%). The highest daily concentration was noted on
the 30th of May (132 PG/m3), the airpollen of Poaceae representing 43%.
In June, eight pollinic types were identified: Pinaceae (4.78%), Poaceae (32.43%),
Urtica (39.33%), Plantago (7.86%), Rumex (6.55%), Tilia (6.84%), Chenopodiaceae/
Amaranthaceae (1.48%) and Ambrosia (0.22%). The pollinic range was dominated by the
airpollen coming from herbaceous plants. The total monthly concentration was higher than
74
the one noted in the previous month (1746 PG/m3). The day with the highest concentration
was the 23rd of June (88 PG/m3), the Urtica type airpollen representing 50% of the total.
Tabel I. Monthly pattern of airborne pollen (%), Timisoara, Romnia, 2004 (airpolynic spectrum)
II
III
IV
VI
VII
VIII
IX
ULMUS
41,75%
58,25%
CORYLUS
26,22%
72,73%
1,05%
ALNUS
25%
71,62%
3,04%
0,34%
TAXACEAE/CUPRESSACEAE
7,61%
55,74%
34,66%
1,99%
ACER
5,51%
16,95%
77,54%
POPULUS
28,04%
71,96%
SALIX
35,80%
64,20%
FRAXINUS
18,07%
79,57%
2,36%
BETULA
8,20%
89,30%
2,50%
CARPINUS
8,60%
90,60%
0,80%
QUERCUS
73,30%
26,70%
PLATANUS
79,50%
20,50%
JUGLANS
87,30%
12,70%
MORUS
79,90%
20,10%
PINACEAE
2,40%
69,20%
28,40%
URTICA
0,04%
5,86%
27,30%
22,30%
37,50%
7%
POACEAE
1,80%
31,70%
25,70%
26,50%
10,70%
3,40%
0,20%
PLANTAGO
28%
28,40%
27%
14,70%
1,90%
RUMEX
27,10%
33%
37%
2,90%
TILIA
17,70%
81,60%
0,70%
CHENOPODIACEAE/AMARANTHACEAE
5%
15,50%
36%
42,50%
1%
AMBROSIA
0,10%
4,80%
33%
61%
1,10%
ARTEMISIA
8,60%
54,40%
36,40%
0,60%
The Poaceae (33.21%) and the Urtica (31.9%) also dominated the pollinic range of
July. The other anemophilic taxa in the flowering phenophase were: Plantago, Rumex,
Chenopodiaceae/Amaranthaceae, Ambrosia and Artemisia. The highest daily concentration
was noted on the 11th of July (119 PG/m3).
In August the second highest quantity of the year was determined (3240 PG/m3).
Only the herbaceous plants were in the flowering phenophase; the pollen of Ambrosia
(31.9%), Urtica (29.2%) and Artemisia (22.6%) dominated the airplankton. The highest
daily concentration was noted on the 31st of August (185 PG/m3), the airpollen of the
Ambrosia type representing 61% of it (Fig.2).
In September, five pollinic types were identified, the highest concentrations
belonging to Ambrosia and Artemisia. On the 8th of September the total concentration
reached 287 PG/m3, the airpollen produced by Ambrosia representing 77%.
75
During the two monitored weeks in October, four pollinic types were identified, with
low concentrations (Ambrosia, Artemisia, Chenopodiaceae/ Amaranthaceae, Poaceae).
Fig. 1. Concentrations of airpollen (PG/m3) measured in April 2004
Fig. 2. Concentrations of airpollen (PG/m3) measured in August 2004
If we consider 30 pg/m3 as a threshold value for the onset of the pollinosis symptoms
for the pollen of all anemophilic plants with an allergenic potential, 11 out of the 23
identified pollen types did not reach this value at any time: Corylus, Alnus, Ulmus, Acer,
Quercus, Pinus, Platanus, Plantago, Rumex, Tilia, Chenopodiaceae/Amaranthaceae. Even
if Urtica produces much pollen, the involvement of this pollen type in the
monosensitization to allergens wasn not proved. For the west and south west of Romania a
real danger is represented by the allergenic airpollen produced by Ambrosia (in August and
September) Artemisia (in August) and Poaceae (in May, June, and July). The number of
days when the daily concentration for these taxa exceeded 30 pg/m3 was of 29 for
Ambrosia, 21 for Artemisia, and 17 for Poaceae. The high daily concentrations are
76
accompanied by the high number of days these pollinic types were present in the
airplankton: 174 days for Poaceae, 111 days for Ambrosia, and 99 days for Artemisia. In
the SW of Romania the most important taxa with an allergenic potential are: Ambrosia,
Artemisia, Poaceae and Betula.
Conclusions
The early spring spring period was dominated by the pollen coming from
anemophilic trees: Acer, Alnus, Betula, Carpinus, Juglans, Morus, Pinaceae, Platanus,
Corylus, Fraxinus, Populus, Salix, Quercus, Taxaceae/Cupressaceae, Tilia, Ulmus (38.6%
of the total annual concentration). During the late summer autumn period the airplankton
was dominated by the airpollen coming from herbaceous plants: Ambrosia, Artemisia,
Chenopodiaceae/ Amaranthaceae, Plantago, Rumex, Urtica (48.7% of the total annual
concentration). The Poaceae produced 12.7% of the total annual concentration. In the
investigated area the daily concentrations reached two highs: in april and in august. The
highest daily concentrations were those of Ambrosia (in August and September) Artemisia
(in august) and Poaceae (in May, June, and July). The pollen season of the plants in the
continental climate ends in october. The main airborne polluter was the pollen of Ambrosia
Artemisiifolia which represented 18.11% of the annual pollinic range.
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Turkey. Israel J Plant Sci, 48: 67-70
BICAKCI A., TATLIDIL S., SAPAN N., MALYER H., CANITEZ Y., 2003 - Airborne pollen grains in
Bursa, Turkey, 19992000. Ann Agric Environ Med, 10: 3136
CARAMIELLO R., POLINI V., SINISCALCO C., MERCALLI L., 1990 - A pollen calendar from Turin
(19811988) with reference to geography and climate. Grana 29: 239249
D'AMATO G., SPIEKSMA F.T.M., 1990 - Allergenic pollen in Europe. Grana, 30, 67-70
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FAEGRI K., IVERSEN J., 1992 - Textbook of Pollen Analysis, Ed. John Wiley and Sons
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pollen sampling in Toledo, Central Spain, Aerobiologia, 22: 5566
GUVENSEN A., OZTURK M., 2003 - Airborne pollen calendar of Izmir - Turkey. Ann Agric Environ Med,
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West University of Timioara, ser. Biology, 5-6: 197-206
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some abundant pollen types, monitored at Vienna, Leiden and Brussels. Grana 30: 309312
16. KAPLAN A., 2004 - Predominant aeroallergen pollen grains in the atmosphere of Ankara, Turkey, Allergy:
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17. LATORRE F., BIANCHI M. M., 1997 - Relacin entre aeropolen y vegetation arbrea en Mar del Plata
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78

VARIABILITY OF SOME POLLEN MORPHOLOGICAL TRAITS OF CERTAIN

TAXONS IN THE BORDERING AREA OF THE CEAHLU NATIONAL PARK
SILVICA PDUREANU*
Abstract: This study presents the variability of four pollen morphological traits from three taxons:
Ranunculus polyanthemos L., Veronica chamaedrys L. and Plantago media L., sampled from four
stationaries found in the bordering area of the Ceahlu National Park. Among the four stationaries, three
of them were affected by polluting noxa agents (dust powder from the cement factory of Taca and
escapement gases), and one was used as control. The four pollen morphological traits taken into account
were: shape of the pollen grain, exine ornamentations, size of the pollen grain, number of germinative
pores/pollen grain. For a real evaluation of these traits, determinations were carried out on 1000
grains/taxon. Being qualitative traits, the shape of pollen grain and exine ornamentations were estimated
by description and microphotographs. As quantitative traits, the size of pollen grain diameter and the
number of germinative pores/pollen grain were determined by biostatistic methods. Measuring the
diameter of the pollen grains was a safe way to separate the diploid against self-tetraploid or mixoploid
plants. On the other hand, there was a tight correlation between the number of germinative pores/pollen
grain and the plant polyploidy degree. The shape of the pollen grain, next to the exine ornamentations did
not show variability from one individual to another of the same taxon, no matter if the stationary was or
not polluted; this proved the genetic strengthen of these two qualitative traits. The biostatistic indices of
the two quantitative traits shaped the limits of pollen variability in the three taxons, which depended not
only on genotype but also on environment.
Key words: pollen grain, exine ornamentation, germinative pores/pollen grain
Introduction
The pollen morphology has a strong taxonomic and phylogenetic significance in the
world of plants. The knowledge of the evolution of pollen morphology brings an important
contribution to plant phylogenetic interpretation. The constant or changing environment
conditions where plants develop are determinant for the appearance of new ecotypes, which
are reflected by pollen morphology [2].
The palinological studies have shown that the pollen morphology has expressed the
polyploidy degree of a certain taxon, which determined the variability of morphopolinic
traits. A polyploidy degree higher than the diploid one is responsible for the formation of
greater pollen grains, with differently placed germinative pores and a diminished fertility
caused by meiosis disturbance [3, 4, 5].
This scientific paper focuses on the limits of the variability of pollen morphology,
characterized by many traits, at certain taxons belonging to different botanic families. We
had in view the influence of polluting noxa agents from the environment on the variation of
morphopalinological traits.
*
University of Agricultural Sciences and Veterinary Medicine Ion Ionescu de la Brad Iasi, Romania
79
The biological material consists in three taxons: Ranunculus polyanthemos L.,

Veronica chamaedrys L. and Plantago media L. They were sampled from four different
stationaries found in the bordering area of the Ceahlu National Park, Neam County. Three
of these stationaries (1, 2 and 4) are affected by polluting noxa agents, while the stationary
3 is used as a control, not being exposed to pollution. Therefore, stationaries 1 and 2 are
polluted with dust powder from the cement factory of Taca, stationary 4 is polluted with
escapement gases from Taca, while the unpolluted stationary 3 is represented by Potoci
Village.
Pollen was sampled from these taxons in the anthesis stage. It was used to
investigate four traits: shape of the pollen grain, exine ornamentations, size of the pollen
grain and number of germinative pores/pollen grain. To determine the shape of the pollen
grain and exine ornamentations, we have used the Tesla electron-scan microscope and
microphotographs have been taken [6].
For the determination of the pollen grain size, micromeasurements have been carried
out on 1000 pollen grains/taxon/stationary. According to the shape of the pollen grains, we
have measured a diameter for the round shaped and two diameters (longitudinal and
equatorial ones) for those ovoid or elliptic shaped. These data helped us to determine the
arithmetical mean and the height of variation. To analyse the variability of the pollen grain
size, the biostatistical indices have been used: standard deviation (S), variation coefficient
(S%) and mean error of the arithmetical mean (S x ) [1].
For establishing the number of germinative pores/pollen grain, determinations on
1000 pollen grains/taxon/stationary have been executed. The method consisted in
introducing the pollen in a mixture of concentrated sulphuric acid (one part) and acetic acid
(two parts) and 3% methylene blue [7].
Results and discussion
The shape of the pollen grains and the exine ornamentations
Ranunculus polyanthemos L. has spherical pollen grains, with a scabrat type exine
ornamentation (fig. 1).
Veronica chamaedrys L. has elliptic pollen grains, with rugulated exine
ornamentation (fig. 2).
In Plantago media L., the pollen grains are approximately spherical and the exine
has a claviform ornamentation (fig. 3) [8].
The pollen sampled from the three taxons was yellow-coloured, no matter what
stationary it has been collected from. In these taxons belonging to distinct families, the
shape of pollen grains and the exine ornamentations were different.
Size of the pollen grains
Measuring the diameter of the pollen grains is a safe way to separate diploid from
self-tetraploid or mixoploid plants. This was due to the fact that doubling the number of
chromosome sets was also correlated to the increase in pollen grain size and to the rate of
non fertile pollen grains.
80
In Ranunculus polyanthemos L., the pollen having a spherical shape, it could be

characterized from the size point of view by measuring one diameter. On the average, it has
30.36 m in the control. In individuals from the stationary 4, the pollen grain has a size
similar to the one of the control ones, while in individuals from the stationary 1, the pollen
grain was smaller. The variability of this trait was low in the control, average for the
stationary 4 and high in the stationary 1 (Tab. I, Fig. 4).
In Veronica chamaedrys L., the pollen has the greatest diameter comprised between
37 and 40 m, with a low variability in the control and average in stationary 4 (tab. II).
The equatorial diameter has a low variability (Tab. III, Fig. 5). The ratio between the
two diameters is between 2.468 and 2.519 m (fig.6).
20
15
10
30
20
10
5
0
control
st. 1
st.4
high diam.
equat. diam.
Fig. 5 Size mean value of pollen grains in Veronica chamaedrys

L.
Fig. 4 Diameter mean value of pollen grain in Ranunculus

polyanthemos L.
22,34
2,52
2,5
micrometers
micrometers
st. 4
40
micrometers
micrometers
control
50
35
30
25
2,48
2,46
22,32
22,3
22,28
22,26
2,44
control
22,24
st. 4
control
Fig. 6 Rate high diameter/equatorial diameter of pollen grain in

Veronica chamaedrys L.
st. 4
Fig. 7 Diameter mean value of pollen grain in PLantago media L.
In Plantago media L., the pollen grain has a diameter of aproximately 22 m, with a
mean variability (tab. IV, fig. 7).
The number of germinative pores/pollen grain
It is well known that there is a tight correlation between the number of the
germinative pores/pollen grain and the degree of plant polyploidy. The number of
germinative pores/pollen grain is also an indicator of the fertility degree of male
gametophyte. That is the reason why we have studied this trait on the six taxons sampled
from different stationeries found in the bordering area of the National Park of Ceahlu.
The characteristic number of germinative pores/pollen grain in Angiospermae, class
Magnoliatae was three. According to different factors, defectively structured pollen grains
(with a lower pore number) or pollen grains with more pores than normal in the species also
appeared, at different rates. These pores are in fact germinative apertures, crevices,
allowing the release of the polynic tube.
In Ranunculus polyanthemos L., the pollen has generally, three pores/grain. It is
surprising that there are 10-15% grains with 4 pores, showing the beginning of polyploidy
(tab. V).
In Veronica chamaedrys the pollen has generally normal values of the number of
pores/pollen grain, namely three. At very low rates (1-2%), pollen has four germinative
pores; therefore, there was a slight tendency of polyploidy in this species (Tab. VI). There
81
were no significant differences between stationaries concerning the number of pores/pollen

grain.
In case of the Plantago media pollen, we have found that most of the pollen grains
had nine germinative pores (Tab. VII) [8]. This situation was also reported in both
stationaries. The increased number of pores/pollen grain showed an advanced polyploidy
process at this taxon.
Table I. The variability of pollen grains diameter in Ranunculus polyanthemos L.
S
S%
No.
Mean Minimum Maximum Variation
(m)
height
value
value
stationary value
Sx
(m)
(m)
(m)
(m)
(m)
Control
30.360
24.150
37.950
13.800
2.377 7.829
0.238
(3)
1
23.137
24.150
41.400
17.250
8.896 38.450
0.889
4
30.136
17.250
39.675
22.425
4.450 14.766
0.445
Table II. The variability of high diameter pollen grains in Veronica chamaedrys L.
S
S%
No.
(m)
height
value
value
stationary value
Sx
(m)
(m)
(m)
(m)
(m)
Control
40.124
31.050
48.300
17.250
2.759 6.876
0.687
(3)
4
37.812
20.700
46.575
25.875
4.041 10.687
0.404
Table III. The variability of equatorial diameter of pollen grains in Veronica chamaedrys L.
S
S%
No.
(m)
height
value
value
stationary value
Sx
(m)
(m)
(m)
(m)
(m)
Control
16.255
6.600
20.400
13.800
2.007 5.387
0.200
(3)
4
15.011
7.250
23.988
16.738
3.500 9.997
0.350
Table IV. The variability of the pollen grains diameter in Plantago media L.
S
S%
No.
(m)
height
value
value
stationary value
Sx
(m)
(m)
(m)
(m)
(m)
Control
22.270
10.350
31.050
20.700
2.742 12.312
0.274
(3)
4
22.322
13.800
25.875
12.075
2.437 10.920
0.243
82
Table V. Nnumber of germinative pores/pollen grain in Ranunculus polyanthemos L.

No.
Mean
% pollen grains with germinative pores
stationary
value
0
1
2
3
4
Control
(3)
1
4
(X )
3.03
83
10
3.01
3.01
0
0
2
1
5
12
83
72
10
15
Table VI. Number of germinative pores/pollen grain in Veronica chamaedrys L.

No.
Mean
stationary
value
0
1
2
3
Control
(3)
4
(X )
2.96
94
2.88
10
86
Table VII. Number of germinative pores/pollen grain in Plantago media L.

No.
Mean
stationary
value
0
1
2
3 4 5
6
7
8
9
10
Control
(3)
4
11
(X )
9.15
78
12
9.07
78
Conclusions
1. The shape of the pollen grain, next to exine ornamentations, does not show
variability from one individual to another of the same taxon, no matter if the stationary was
or not polluted, demonstrating the good genetic strength of these two morphological traits.
2. The size of pollen grains is a species trait. This has a certain variation height
according to eco-physiological conditions of stationaries.
3. If the pollen grains have an elliptic shape, the greater diameter has a higher
variability compared to the equatorial one.
4. In most studied cases, the diameter variability of pollen grains belonging to the
control is more balanced as compared to the one of the individuals from stationaries
affected by polluting noxa. This could be explained by a greater genetic stability of that
trait in areas which are not affected by environment pollution.
5. At Ranunculus polyanthemos L. and Veronica chamaedrys L., the characteristic
number of germinative pores/pollen grains is three, but we have also found pollen grains
with four germinative pores, at different rates, both at the control and at the individuals
83
from pollution affected-stationaries. This proved the presence at these two taxons of
mixoploidy, which developed with different intensity according to the stationary.
6. In Plantago media L., most of pollen grains have nine pores. Next to nine pore
pollen, at this taxon there are also pollen grains with seven, eight, ten and eleven
germinative pores. The increased number of germinative pores shows an advanced
polyploidy in that taxon. The value differences concerning the rate of pollen grains with a
certain number of pores, according to stationary, are insignificant.
7. In Ranunculus polyanthemos L. and Veronica chamaedrys L., we have found
pollen rates with a number of germinative pores lower than three. In the control, these rates
are lower as compared to the other stationaries.
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TARNAVSCHI I.T., SERBANESCU-JITARIU G., MITROIU-RADULESCU N., RADULESCU D., 1990 Monografia polenului florei din Romnia, III: 69-70
Explanation of figures:
Fig. 1 Pollen grains of Ranunculus polyanthemos L. (2300X) (stationary no. 1)
Fig. 2 Pollen grains of Veronica chamaedrys L.(1740X) (stationary no. 4)
Fig. 3 Pollen grains of Plantago media (2300X) (stationary no. 3)
84
SILVICA PDUREANU
PLATE
Fig. 1
Fig. 2
Fig. 3
85

RESEARCHES REGARDING THE ESSENTIAL OIL COMPOSITION OF SOME

ARTEMISIA L. SPECIES
I. BURZO*, V. CIOCRLAN*, ELENA DELIAN*, AURELIA DOBRESCU*, LILIANA
BDULESCU*
Abstract. The chemical composition of the essential oils of Artemisia sp. L. growing in Romania has been
studied. Analysis of the essential oil extracted by hydrodistillation was performed by GC-MS and
emphasised the presence of some major similar compounds in Artemisia austriaca Jack, A. lerchiana
Weber ex. Stechm and A. santonica L.: eucaliptol, camphor, cis-verbenone and borneol or iso-borneol. The
essential oil extracted from another species had a specifically composition. Thus, A. annua L. essential oil
contained 60,20% artemisia ketone; A. dracunculus L. contained 42,34% sabinene and 24,92% methyl
eugenol; A. vulgaris L. contained 41,46 germacrene D and 11,19% -caryophyllene; A. dzevanovskyi
Leonova in Wulf. 57,13% eucaliptol and 20,37% borneol; A. absinthium L. contained 41,65 % -pinene
and 12,77 % myrtenil acetate; A. abrotanum L. contained 12,27 % eucaliptol and 9,23 % borneol.
Key words: Artemisia sp., chemical composition, essential oils.
Introduction
Artemisia genus comprises wild and cultivated species, used in the alimentary and
chemical industry. In the literature only one paper that presents a comparatively study
regarding the essential oil composition extracted from ten Artemisia sp. in Serbia was
found. Knanina et al. [2] noticed a variation of the essential oil depending on specie, the age
of the plant and the ecological conditions.
Lawrence [3] had stated that the essential oil extracted from A. absinthium L. has
two major compounds: cis-sabinil acetate (40,00%) and -thujene (35,00%). Artemisia
annua L. contains 35,70% artemisia ketone and 31,50% 1,8-cineole [4]. In the oil of A.
dracunculus L. the major compound is methyl carveol [6], and in A.vulgaris L. sabinene
and mircene were identified [5].
A. dracunculus L. plants or their extracts are used for food aromatisation, those of A.
absithium L. for the preperation of some drinks, while A. abrotatum L., A. annua L. and A.
vulgaris extracts are toxic.
The aim of the present study, which is carried out for the first time in these plants
grown in Romania, is to make a comparative analysis regarding the essential oils
composition of nine Artemisia sp.
The research has been performed on the following Artemisia species:
A. annua L. - wormwood, from Botanical Garden of U.S.A.M.V. Bucharest.
A. abrotanum L. southernwood, from Botanical Garden of U.S.A.M.V. Bucharest.
*
University of Agronomical Sciences and Veterinary Medicine Bucharest, Romania
86
A. absinthium L. absinthium, Iasi area.

A. austriaca Jack. Alah-Bair, Dobrogea area.
A. dracunculus L. tarragon, from Botanical Garden of U.S.A.M.V. Bucharest.
A. lerchiana Weber ex. Stechm Capul Doloman area, Tulcea
A. santonica L. tartarian southerwood, Muntele de sare area, Buzu.
A. vulgaris L. mugwort, Iai area and a new species in the Romanian flora [1]:
A. dzevanovskyi Leonova in Wulf Capul Doloman area, Tulcea.
The volatile compounds were extracted by hydrodistillation with a Singer-Nickerson
apparatus. The separation and identification of components has been carried out using an
Agilent gas chromatograph, equipped with quadruple mass spectrometer detector. A
capillary column DB-5 (25 m length x 0,25 mm i.d. and 0.25 m film thickness) and
helium as carrier gas were used. The initial oven temperature was 600C, then rising to
2800C at a rate of 40C/min. The NIST spectra bank was used for to identify the volatile
compounds, which were verified with the Kovats indices.
The obtained results are presented in figures 1-9.
The performed analysis permitted us to identify in the obtained mentioned extracts
17 compounds in A. vulgaris sp. and 36 compounds in A. abrotanum, respectively.
From the presented analytical data, it can observed that in Artemisia austriaca, A.
lerchiana and A. santonica there is a similarity regarding the major compounds: eucaliptol,
camphor, cis-verbenone and borneol, or isoborneol, but these are present in different
proportions. Thus, A. lerchiana contains 57.13% eucaliptol, while in A. santonica and A.
austriaca oils camphor, cis-verbenone and eucaliptol were identified; eucaliptol, camphor
and isoborneol, respectively, in aproximately similar amounts (15.8-21.54%). The main
difference concerning these species essential oil composition consists in the fact that the
essential oil extracted from A. austriaca contains a higher amount of higher molecular
weight substances and less volatile: -eudesmol (6,97 %), germacrene D (3,40 %) and relemene (2,64 %).
A relatively similar essential oil composition, almost as in the previous mentioned
species it was noticed at A. dzevanovskyi, the oil included a higher eucaliptol content
(38.42%) and borneol (20.37%) and lower amount of camphen (6.15%) and camphor
(5.44%), respectively.
The others Artemisia analysed species had a specific essential oil composition, that
can alow their identification. So, the A. absinthium oil contained a higher proportion of pinene (41.65 %) and myrtenyl acetate (12.77 %), a small amount of thujonen also being
determined (4.60 %).
The A. annua oil contained appreciable amounts of artemisia ketone (60.20%), next
to a small amount of eucaliptol (13.77%) and -pinene (9.19 %).
Artemisia vulgaris oil was characterised by higher amounts of substances with a
higher molecularly weight and a lower volatility degree, as for instance germacrene D
(41.46%) and -caryophyllene (11.94 %). These substances may assure the characteristic
odour persistence for a longer period of time.
87
The major components of Artemisia dracunculus L. essential oil were sabinene

(42.34%) and methyl eugenol (24.92%). This composition is similar to the one that is
characteristic for the Russian A. dracunculus type.
The essential oil obtained from A. abrotanum contained a higher amount of
eucaliptol (17.59%), but was different compared to the A. lerchiana, A. austriaca, A.
santonica i A. dzevanovskyi oils, because camphor was not identified. Next to eucaliptol,
-terpinene (13.55 %), borneol (13.24 %) and p-cimene (11.30 %) were determined in a
lower amount.
Conclusions
1. Artemisia austriaca, A. lerchiana and A. santonica species are characterised by
similarities regarding the major essential oil compounds: eucaliptol, camphor, cisverbenone and borneol or isoborneol.
2. The essential A. abrotanum oil contained a higher amount of eualiptol, but varied
compared to the Artemisia austriaca, A. lerchiana, A. santonica i A. dzevanovskyi oils,
with the lack of camphor.
3. It has been shown that the A. dzevanovskyi oil contains higher amounts of
eucaliptol (38.42%) and borneol (20.37%).
4. The other Artemisia species analysed had a specific essential oil composition,
which can permit their identification.
5. A. annua essential oil contained a higher amount of artemisia ketone, those of A.
absinthium -pinene, those of A. vulgaris - germacrene D, and those of A. dracunculus sabinene.
REFERENCES
1.
2.
3.
4.
5.
CIOCRLAN V., 2002 - Noi specii n flora Romniei. Buletinul Grdinii Botanice Iai, 11: 97 98.
LAWRENCE B.M., 1992 - Chemical Composition of some Warmwood oils produced in North America.
Perf. Flavor, 17: 42.
LIBBEY L. M., STURTZ G., 1989 - Unusual Essential Oils Grown in Oregon II. Artemisia annua L.,
Journal of Essential Oil Research, 1: 201-202.
MICHAELIS K. et.al., 1982 - Das aetherische Oel aus Blueten von Artemisia vulgaris L. Z. Naturforsch. 37
C: 152-158.
TUCKER A. O., MACIARELLO M. J., 1987 - Plant Identification (oil from fresh leaves). In Proceedings of
the First National Herb Growing and Marketing Conference, Edit. J.E. Simon and L. Grant, Purdue Univ.
Press, West Lafayette: 126-172
88
89
90
91

THE ECOPHYSIOLOGICAL PECULIARITIES OF SOME SPECIES OF THE

ACER GENUS FROM THE BOTANICAL GARDEN OF IAI
ALEXANDRINA MURARIU *, GIANINA BERECHET*, ANIOARA STRATU*,
CAMELIA IFRIM**
Abstract. The paper presents the results of the biochemical and physiological studies carried out in 2005,
on 8 species of the Acer genus, cultivated in the Botanical Garden of Iai in order to emphasize the
ecophysiological peculiarities of the foliar limb, at the pedoclimatic conditions specific to the area. Two
native species (Acer campestre i Acer monspessulanum) and six exotic species (Acer ginnala, A.
negundo, A. negundo var. auratum, A. negundo var. variegatum, A.opalus, A. saccharinum) at which the
print of the ecological factors is recognized in the main physiological manifestations: photosynthesis,
respiration, perspiration, mineral nutrition, was studied. The analysis of the physiological behaviour of the
exotic species, as compared to the native ones emphasized the fact that most of them have adapted to the
conditions of the temperate climate in our country.
Key words: physiological processes, wooden species, adaptation.
Introduction
The Acer genus comprises about 115 species of trees, and more seldom shrubs, with
falling leaves, spread in Europe, Northern Africa, Asia and North America.
The Botanical Garden of Iai is structured on 12 sections, and the themes worked out
for each section have a didactic, scientific purpose, the preservation of the genetic
background of the plants but also a recreative - cultural and hygienical-sanitary purpose
[16].
The species we have analyzed belong to the trees collections of the Flora of the
Globe section: some of them native (Acer campestre i Acer monspessulanum) and others
exotic (Acer ginnala, A. negundo, A. negundo var. auratum, A. negundo var. variegatum,
A.opalus, A. saccharinum), acclimatized and naturalized, of decorative interest.
The concerns of the researchers in our country on these species focused especially on
the fields: morphology [1], [7], [11], [9], [2], [15], histo-anatomy [14], phenology [5],
phytocenology [8], [4], [10], phytopathology [6], phytochemistry [3], [13]. The
physiologycal references concerning Acer genus are a few; they were caried out by
Papadopol S. (1959) and Sprchez Z. (1962).
Starting from these general considerations, the paper presents the results of the
biochemical (water content, assimilating pigments and total mineral elements) and
physiological studies (the intensity of photosynthesis, respiration and perspiration) on 8
species of the Acer genus, cultivated in the Botanical Garden of Iai.
*
Al. I. Cuza University, Faculty of Biology, Carol I Bd., No 20 A, 700506, Iai, Romania
Anastasie Ftu Botanical Gardens, Dumbrava Roie Street, No 7-9, Iai, Romania
**
92
The vegetal material, represented by leaves, was gathered between 8 and 10, 30 a.m,
in the aestival period of 2005, a year characterized from the climatic point of view as
showing a hydric deficit and a thermal excess. We analysed the following physiological
indicators: the water and dry matter (gravimetrical method), the assimilating pigments
(tirban and Frecu spectro-photometrical method), the total mineral elements (ash) (dry
calcination at 4500 C method), the intensity of photosynthesis and respiration (IvanovKosovici method), the intensity of perspiration (Huber - Ivanov method).
Analyzing the ecophysiological behaviour of the exotic species as compared to the
native ones, the data presented in Table I emphasize the fact that these species adapted to
the conditions of the temperate climate in our country, but the action of the high
temperatures and of the relative low humidity (10-20%) in the atmosphere during the day,
produces specific biochemical and physiological changes.
Thus the water losses increase through perspiration leading to a disequilibrium
between the absorption and the perspiration (310 - 357 mg H2O / dm2 / hour: A. negundo
var. auratum, A.opalus and A. ginnala). These results are ecologically explained through
the process of adaptation to stress which is never completed, and the fact that these species
do not impede the perspiration means that they use a more efficient way, namely satisfying
the thermal needs through an increased respiration (1,55 - 2,26 mg CO2 / g / hour). At these
species the ecophysiological reaction of adaptation to stress is not made through the
intensification of the assimilation but through the intensification of the oxido-reduction
reactions, through an increased energetic consumption to the prejudice of the reduction of
the biomass and the dry matter, of which the synthesized organic matters are only 0,640,79% (fig.1).
The species which are resistant to drought (A. negundo var. variegatum, A.
saccharinum ) avoid losing too much water through perspiration (20-30 mg / hour) due to
the specific morpho-anatomical and physiological peculiarities, which constitute
xeromorphism characters.
Regarding the gross photosynthesis, the native species (A. campestre and A.
monspessulanum) which live in their ecological optimum have the biggest intensity of the
net photosynthesis (1,55- 1,88 mg CO2 / g / hour) and the lowest intensity of the respiration
(0,38- 0,47 mg CO2 / g / hour), showing a productive metabolism (14-16% accumulated dry
matter). In the exotic species (A. negundo var. auratum, A. negundo var. variegatum,
A.opalus) the net photosynthesis is maintained at low quotas (0,69- 0,98 mg CO2 / g / hour)
due to the increased respiration (1,54 - 2,45 mg CO2 / g / hour). Although the relation
between the assimilating pigment content and the rapport between them has not found yet
its mathematical expression, it doesnt mean that it doesnt exist but only that it is not
operative due to the complexity of the ecological inter-relations of the factors on the
photosynthesis.
There is undoubtly the fact that at the exotic species in which the intensity of the net
photosynthesis is low, the assimilating pigment content is also diminished (1,63 -2,04 mg /
g fresh substance). We can notice the biosynthesis and the lower accumulation of the
93
chlorophyll b and of the carotenoidic pigments, which influences the net photosynthesis
decrease, with repercussions on the biomass production.
Through the percentage values of the amount of total mineral elements, absorbed and
accumulated in the cells, most of the species belong to the mesotrophic category (8,04 9,17 %).
Conclusions
Based on the results obtained we can estimate that the print of the ecological factors
specific to 2005 causes differentiated influences in the exotic species, as compared to the
native ones.
The species well acclimatized (A. negundo, A. ginnala) are favoured through the
bigger water content, the total assimilating pigments, the increased intensity of the net
photosynthesis and the performances of those which do not integrate in a harmonious
assimilation of the microclimate conditions decrease (A. negundo var. variegatum, A.
negundo var. auratum, A.opalus, A. saccharinum).
REFERENCES
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
BELDIE AL., 1953 - Plantele lemnoase din Romnia. Manual de determinare. Edit. Agro-silvic de Stat,
Bucureti : 342-356.
CIOCRLAN V.,1990 Flora ilustrat a Romniei. Determinarea i descrierea speciilor spontane i
cultivate. vol.I. Edit.Ceres, Bucureti: 416-418.
CODREANU A. MARILENA-VIORICA, ISTUDOR D. VIORICA, DINU V. MIHAELA, DOCIU N.
NICULINA, 2006 - Physico-chemical studies on Acer negundo L. seed oil. 4 th Conference on Medicinal
and Aromatic Plants of South - East European Countries. 28-31mai 2006, Iai: 374 -378.
DONI N., PURCELEAN T., 1975 - Pdurile de leau din R.S.R. i gospodrirea lor. Edit. Ceres
Bucureti.
LUPU I., 1971- Observaii privind comportarea n primii 3 ani a unor specii lemnoase indigene, transplantate
la Grdina Botanic Iai . An. t. Univ. Al. I. Cuza Iai, sec. II. XVII, fasc. 1: 162 - 168.
MITITIUC M., IACOB. V., 1997 - Ciuperci parazite pe arborii i arbutii din pdurile noastre. Ed. Univ.
Al. I. Cuza Iai.
NEGULESCU E. G., SVULESCU AL., 1957 - Dendrologie. Edit. Agro-silvic, Bucureti: 415-432.
PACOVSKI S., 1967- Succesiunea speciilor forestiere. Edit. Agro-silvic, Bucureti: 71-73.
POPA A., GHIU M., OGRUAN I., 1959 - Floricultur i dendrologie. Edit. Agro-silvic de Stat,
Bucureti: 324-325.
SRBU I., OPREA A., TNASE C., 1997- Vegetaia Pdurii parc Grboavele (jud. Galai). Bul. Grd. Bot.
Iai, 6, 2: 311-332.
SVULESCU T., 1958 - Flora R.P.R, VI. Edit. Academiei R. P. R., Bucureti: 228-248.
TTRANU D.I., 1960 - Arbori i arbuti forestieri i ornamentale cultivai n R.P.R. Edit. Agro-silvic,
Bucureti:166-175.
TCACENCO V. LUMINIA, TMA N. VIORICA, POMPONIU A. DANIELA, BERTEANU C.
ELENA, BOTEZATU I. AURICA, 2006 - Studies for identification of some biological active compounds
from indigenous plants with therapeutical value. 4 th Conference on Medicinal and Aromatic Plants of
South - East European Countries. 28-31mai 2006, Iai: 78.
TOMA C., IVNESCU L., 1998 Cercetri privind unele modificri histo-anatomice induse de poluanii
atmosferici asupra aparatului foliar de la specii lemnoase aparinnd familiilor Aceraceae i Oleaceae.
Buletinul Grdinii Botanice Iai, 7: 51-58.
ZANOSCHI V., SRBU I., TONIUC A., 2004 Flora lemnoas spontan i cultivat din Romnia. III.
Edit. Univ. Al. I. Cuza Iai: 138-174.
*** - 1993- Grdina Botanic ( ghid ). Ed. Univ. Al. I. L. Cuza . Iai.
94
Table I. The ecophysiological characteristics of some species of the Acer genus from the Botanical Garden of Iai
Species
Wa
ter
(%)
Acer campestre L.
85.85
67
101
Dry
Assimilating pigments (mg /g.
Gross photosynthesis (mg CO2 / g / h)
mat
fresh substances)
ter
Net
Respiration
Gross
Chl
Ch
Car
Total
photosynthesis (g
oro
lor
ote pigments photosynthesis
%)
phy
op
noi
ll a
hyl
d
l b
2.092 0.760 0.725 3.577
1.55
0.38
1.93
14.15
Acer monspessulanum L.
A. negundo L.
A.negundo var.
auratum (Spaeth)
A.negundo var.
variegatum (Booth)
A.opalus( Mill.)
Acer ginnala Maxim
A. saccharinum L.
83.58
91.02
91.32
13
23
33
45
125
357
1.970
2.207
0.977
1.290 0.705
1.574 0.765
0.429 0.314
3.965
4.546
1.720
1.58
0.94
0.98
0.47
1.54
1.90
2.05
2.48
2.88
16.42
8.98
8.68
8.53
8.19
8.04
7.89
0.97
0.64
90.18
20
192
0.917
0.428 0.286
1.631
0.69
2.45
3.14
9.82
9.17
0.65
85.63
90.03
84.42
54
87
30
310
322
81
1.095 0.518 0.395

2.317 1.190 0.785
0. 818 0.619 0.604
2.008
4.292
2.041
0.82
1.31
1.25
1.55
2.26
0.99
2.37
3.57
2.24
14.37
9.97
15.58
5.55
4.50
8.30
8.82
5.47
7.28
Perspiration
mg
/h
mg
/
dm2
/h
95
Ash
(g
%)
Orga
nic
subst
ance
(g
%)
8.55
5.60
18
16
14
12
10
8
6
4
2
0
er
Ac
tre
es
mp
a
c
A.
la
s su
pe
s
on
m
nu
do
um
tu m
un
rat
ga
eg
au
rie
n
.
a
r
a
er
r. v
ov
Ac
va
nd
o
u
d
g
n
ne
gu
er
ne
r
Ac
e
Ac
dry matter content (g%)
o
er
Ac
total mineral elements content (g%)
lus
pa
A
g
cer
ala
inn
er
Ac
m
nu
a ri
h
c
c
sa
organic substance content (g%)
Fig. 1. The dry matter, total mineral elements and organic substance content in leaves of studied species
96

PHYSIOLOGICAL AND BIOCHEMICAL ASPECTS IN THE

LIGNICOLOUS SPECIES GLOEOPHYLLUM ODORATUM (WULFEN) IMAZEKI
AND FOMITOPSIS PINICOLA (SW.) P.KARST. (FUNGI, BASIDIOMYCOTA)
COLLECTED FROM CLIMANI NATIONAL PARK (THE ORIENTAL
CARPATHIANS)
ANIOARA STRATU*, ALEXADRINA MURARIU*, MARIA - MAGDALENA
ZAMFIRACHE*, ZENOVIA OLTEANU*, LCRMIOARA OPRIC*, C. TNASE*, V.
C. CHINAN*, C. BRSAN**
Abstract. The paper presents the results of the determinations of some physiological and biochemical
parameters (water content, dry matter, total mineral elements, organic substances and soluble proteins and
also the respiration intensity) in the lignicolous species Gloeophyllum odoratum (Wulfen) Imazeki and
Fomitopsis pinicola (Sw.)P.Karst. (Fungi, Basidiomycota). The results obtained emphasize, for both
species investigated, specific variations of the monitored parameters, the values determined presenting
comparable amplitudes for each parameter.
Key words: Gloeophyllum odoratum (Wulfen) Imazeki, Fomitopsis pinicola,(Sw.) P.Karst, physiological
and biochemical parameters.
Introduction
The lignicolous fungi species, because of their particular manner of nutrition,
produce extracellular enzymes which mainly decompose the cellulose and the lignin basic
components of the cellular walls in the woody species but also different substances
resulted from the anthropic activity (hydrocarbons, petrol products, pesticides, residues
resulted from sylviculture etc.) being reduced to non-toxic products [6].
The Gloeophyllum odoratum (Wulfen) Imazeki species and Fomitopsis pinicola
(Sw.) P.Karst species belong to the Poliporacee family [5], they produce the brown rot and
are considered primary decomposers of cellulose, - poly-carbohydrate which, the same as
lignin, is generally resistant to the microbial decomposition.
The paper presents the results of the determinations of some physiological and
biochemical parameters from a more ample study, which focuses on the potential of
mycoremediation of some pollutants from the deposits resulted from mining.
The physiological and biochemical researches were carried out on two
macromycetes species which were assayed from Climani National Park, and namely from
the Pinus mugo and Pinus cembra Reservation (N 470625; E 251427,3; Alt. 1600 m),
which were constituted in control samples.
Al. I. Cuza University, Faculty of Biology, I Carol I Bd., No 20 A, 700506, Iai, Romania
Anastasie Ftu Botanical Gardens, Dumbrava Roie Street, No 7-9, Iai, Romania
**
97
At the end of June (the first assay) only one species was collected, from the same
substratum (coniferous wood), and namely Gloeophyllum odoratum. The fungus was
assayed from three different areas of Climani National Park (area I - Go I; area II - Go II;
area III- GoIII.). In the last decade of July (the II assay) the sample analyzed was
represented by the Fomitopsis pinicola (Fp - II) species.
The climatic conditions specific to the analyzed period (the presence of drought) had
an unfavorable influence on the growth and development of fungi.
The physiological and biochemical parameters studied are: the water and dry matter
content (gravimetrical method) the respiration intensity (Warburg method), the content of
total mineral elements (dry calcination at 4500 C method), the organic substance content
(calculated through the difference between the dry matter content and ash), and also the
soluble proteins content (Bradford method). The quantitative determination of soluble
proteins was achieved on the species of selected macromycetes, as indicator of the protein
metabolism and nucleic acids.
The determinations of parameters mentioned above (with the exception of the
content of soluble proteins) were carried out gradually (whole sporocarp) and differentiated
on component parts of the sporocarp (trama and hymeneal region)
The results obtained regarding the physiological and biochemical parameters are
presented in figure 1-7.
The water content. The presence of water is an indispensable condition for the
achievement of the metabolism processes. Although water is one of the final products of the
breathing process, the process is only triggered at a certain degree of tissue hydration, and
its intensity increases as the water content increases. This is explained through the fact that
the enzymatic complex implied in the oxydo-reduction processes characteristic to breath
can only function in the aqueous medium.
The water content globally determined present moderate values comprised between
33.95 g % (Fomitopsis pinicola) and 50,3 g% (Gloeophyllum odoratum) . Differentially
determined on the areas of the sporocarp, the water content presents values comprised
between 25.49 g % - 58.18 g % (for the trama) and 36.32 g % - 69.47 g % (for the
hymeneal region) (Figure 1-2).
This physiological indicator has a strong connection with: the atmospheric humidity,
the consistency of fructification body, the sensitivity of sporocarp tissues and the mycelium
related to the presence of water in the atmosphere or substratum.
In the analyzed lignicolous species, the water content reflects the coniferous wood
characteristics which is more porous and absorbs more easily the water from the medium.
The dry substance content represents a basic indicator which characterizes the level
of organic and mineral constituents. The species analyzed are characterized through a high
content of dry matter 49.7 % (Gloeophyllum odoratum) - 66,05 % (Fomitopsis pinicola)
and organic substances. The trama is noticed through a higher content of dry matter,
compared to the hymeneal region. (Figure 1, 2).
98
The content of total mineral elements. According to the specialty literature data [1],
the sporocarp body of fungi present a high content of mineral elements (phosphorus,
potassium, calcium, magnesium, sulfur, sodium, iron, zinc) which differ according to
species, age of the fungus, diameter of the pileus, the component parties of the sporocarp,
substratum. In the two analyzed species, the content of total mineral elements globally
determined register low values (0,95 g % - 3,03g %) (fig. 3,4).
This denotes differences as regards the nature of the substratum and the capacity of
exploiting the mineral elements from the substratum that they have at the disposal.
The organic substances in the macromycetes are represented by proteins,
polyglucides, lipids, amino acids, vitamins, phenol compounds, substances which give the
aroma of fungi [1]. The content of organic matter globally determined presents high values
in the two analyzed species (fig. 3, 4). This fact emphasizes an intense metabolic substance,
specific to the saprophyte mode of nutrition and present a special ecological importance
since the contribution of organic matter contributes to the soil formation, to the
improvement of its physical-chemical characteristics.
The content of soluble protein of the biological sample (Gleophyllum odoratum)
assayed from three different areas of Climani National Park, but from the same sublayerconiferous wood, is relatively constant, being comprised between 70,060 mg% and
71,487mg% (Fig. 5). This is fully justified by the fact that it is the same species, assayed
from the same region, without pollution, the only difference being the substratum.
The concentration of soluble proteins in the samples of Fomitopsis pinicola was
lower compared with the Gleophyllum odoratum species (100,132 mg% compared to 70-71
mg%) (fig. 5).
This extremely small difference could however be explained through the species specificity
of the protein metabolism which could be more intense in Fomitopsis pinicola compared to
Gleophyllum odoratum, the samples being assayed from the same area without pollution.
The biochemical studies carried out have lead us to the conclusion that the metabolic
activity - in the present case the protein metabolism presents a specific manifestation being
influenced by a series of external and internal factors.
The respiration intensity is an indicator of the metabolic activity and indirectly, an
indicator of the climatic stress state. The respiration intensity globally determined has low,
but similar values (0.0178- 0,0176 mm3 oxygen/g s.pr./hour).The determinations carried out
by us in the mycorrhiza species have emphasizes higher values of respiration intensity
compares with these lignicolous species (unpublished data). These differences could be
determined by the degree of development of the sporocarp, the cork consistency and its
chemical composition, its hydration degree, the abiotic factors.
The hymeneal region presents an intense respiration in both species, fact determined
by the presence of reproductive structure, located at this level (Fig. 6-7).
The data presented from the specialty literature (Li Xiong, 2000) for the fresh edible fungi
indicate the fact that under normal conditions, their respiration intensity is high, compared
with the one of vegetable species (tomatoes or salad).
The biological samples of Gleophyllum odoratum assayed from the II area presents,
compared with those assayed from the I, III areas, slightly lower values of the analyzed
parameters (with the exception of the respiration intensity and soluble proteins).
99
The analyses differentially carried out on regions of the sporocarp emphasize the fact
that, compared with the trama, the hymeneal region, which comprises the reproductive
formations structures with intense metabolic activity - is characterized through a higher
hydration degree, a more intense respiration, a high content of organic substance and low
content of mineral elements.
Conclusions
The analyzed species are characterized through low respiration intensity, water
content with moderate values, high content of organic substances and soluble proteins, a
low content of total mineral elements, fact which reveals a metabolism specific to the
lignicolous fungi.
The results obtained emphasize, for both species investigated, specific variants of the
monitored parameters, the values determined presenting comparable amplitudes for each
parameter. The particular manifestations of the investigated indicators are mostly
determined, according to us, by the abiotic conditions of the areas where the biologic
material subject to analyses was assayed.
REFERENCES
1.
2.
3.
4.
5.
6.
BERNAS EMILIA, JVORSKA GRAZYNA, LISIEWSKA ZOFIA., 2006 - Edible mushrooms as a source of
valuable nutritive constituents. Acta scientiarum Polonorum tehnologia alimentaria 5 (1): 5-20
BOLDOR O., TRIFU N., RAIANU O., 1981 - Fiziologia plantelor (lucrri practice). Edit. Didactic i
pedagogic, Bucureti.
BRADFORD, M. M., 1976 - A rapid and sensitive method for the quantification of microgram quantities of
protein utilising the principle of protein-dye binding, Anal. Biochem., 72:248-254
LI XIONG., 2000 - Extend Shelf Life of Mushroom by Using Micro-perforated Film (a research proposal)
Department of Food Science, Pennsylvania State University.
5. SLGEANU GH., SLGEANU A., 1985 - Determinator pentru recunoaterea ciupercilor
comestibile, necomestibile i otrvitoare din Romnia. Edit. Ceres
SING A., WARD O.P. (eds), 2004 - Biodegradation and Bioremediation. Springer- Verlag Berlin
Heidelberg: 19 -57.
Acknowledgements
The researches were supported through funds from the project Biotech No. 128: The Ecological
reconstruction through the processes of mycoremediation of soils degraded by mining activities, financed by the
Ministry of Education, Research and Youth of Romania.
100
dry matter
water
100
80
g%
60
40
area I
area II
trama
hymeneal
globally
trama
hymeneal
globally
trama
globally
hymeneal
20
area III
Fig. 1 The content of water and dry matter in Gloeophyllum odoratum
dry matter
w ater
70
60
50
40
g%
30
20
10
0
globally
trama
hymeneal region
Fig. 2 The content of water and dry matter in Fomitopsis pinicola
101
area I
area II
g%
organic substance
total mineral elements
trama
hymeneal
region
globally
trama
hymeneal
region
globally
trama
hymeneal
region
1,6
1,4
1,2
1
0,8
0,6
0,4
0,2
0
globally
g%
80
70
60
50
40
30
20
10
0
area III
70
60
50
40
30
20
10
0
1,5
organic substance
g%
g%
Fig. 3 The content of total mineral elements and organic substances in Gloeophyllum
odoratum
total mineral
elements
0,5
0
globally
trama
hymeneal
region
Fig. 4 The content of total mineral elements and organic substances in Fomitopsis pinicola
soluble proteins
samples analyzed
Fp(II.)
Go(I.3)
Go(I.2)
Go(I.1)
0
20
40
60
80
100
120
mg %
Fig. 5 The content of soluble proteins in Gleophyllum odoratum and Fomitopsis pinicola
102
respiration intensity (mm3 O2 / g fresh material /hour)

0,045
0,04
0,035
0,03
0,025
0,02
area I
area II
trama
hymeneal
region
globally
trama
hymeneal
region
globally
hymeneal
region
globally
0,005
0
trama
0,015
0,01
area III
Fig. 6 The intensity of respiration in Gleophyllum odoratum
respiration intensity ( mm3 O2/ g fresh material / hour)
Fig. 7 Respiration intensity at Fomitopsis pinicola

0,3
0,25
0,2
0,15
0,1
0,05
0
globally
trama
hymeneal region
Fig. 7 The intensity of respiration in Fomitopsis pinicola
103

SOMATIC EMBRYOGENESIS IN RUBUS CAESIUS L. SUSPENSION CULTURES

SMARANDA VNTU*
Abstract: In vitro cultures technologies include many applications: plants improvement,
micropropagation, , unconventional alternatives to exploit secondary metabolites. The regenerative
potential of Rubus caesis L. was evaluated for the establishment of a propagation protocol. Not only the
capacity of differentiation was tested, but also the regenerative potential. The initiation of in vitro cultures
at Rubus sanctus Schreb was achieved from axillary buds, cultivated on different variants of MurashigeSkoog medium. The callus cultures obtained by axillary buds dedifferentiation were multiplicated on agar
medium and liquid medium. The MS medium supplemented with 1mg/l benzylaminopurine and 0,1 mg/l
IAA stimulated somatic embryogenesis at Rubus caesius L. suspension cultures ,whereas the MS medium
supplemented with 1mg/ kinetine generated an intensive proliferative reaction and callus development.
The regeneration of whole plants was obtained in three steps: callus initiation, suspension cultures
initiation, somatic embryogenesis induction.
Key words: somatic embryogenesis, Rubus caesius, suspension cultures
Introduction
In this paper the procedure for the regeneration of Rubus caesius L. plants based on
indirect somatic embryogenesis is presented.The studies are focused on the application of in
vitro methods on this species, well known as a medicinal plant. The fruits are a good source
of natural antioxidants [1], [5], [6].
The initiation of in vitro cultures of Rubus caesius L. was achieved from axillary
buds. The axillary buds were sterilized with ethanol 70 % and then sodium hypochlorite 0,5
% 10-15 minutes. After rinsing with sterile distilled water, the explants were transferred to
MS medium. 4 variants of MS medium were tested. (Table 1 and 2).
The studies of the in vitro behaviour of this species is based on the use of auxins and
cytokinins, in different combinations and concentrations [2], [3], [4]. The variants 1 and 3
(Table 2) stimulated callus induction and callus multiplication, whereas the variants 2 and 4
stimulated somatic embryogenesis and variants 5, without growth regulators induced
embryos development and maturation. The axillary buds explants were cultivated on the
callus induction media at 24 C in complete darkness. After 30 days the callus produced
was used to establish cell suspensions on the same basal MS medium. Suspension cultures
were established by transferring the section of calli 1 cm3 each, from the exponential
growth phase into 250 ml Erlenmeyer flasks containing 100 ml liquid medium. The flasks
were rotated on rotatory shaker at 100 rpm. The cell suspensions were periodically
subcultivated by filtration with a metallic sieve. The next step was the induction of somatic
*
Al. I. Cuza University, Faculty of Biology, 20 A Carol I Bd, Iasi, 700506, Romania
104
embryogenesis. The suspension cultures were transferred on regeneration media and

maintained at 24 C under 16 hours light cycle.
Table 1. Chemical composition of MS medium
COMPOSITION
Concentration (mg/l)
1650
NH 4 NO 3
1900
KNO 3
MACROELEMENTS
440
CaCl 2 2H 2 O
370
MgSO 4 7 H 2 O
170
KH 2 PO 4
6,2
H 3 BO 4
22,3
MnSO 4 4 H 2 O
8,6
MICROELEMENTS
ZnSO 4 7 H 2 O
0,25
Na 2 MoO 4 2H 2 O
0,025
CuSO 4 5 H 2 O
CoCl 2 6H 2 O
VITAMINES
0,025
KI
0,83
Nicotinic acid
0,5
Pyridoxine HCl
0,5
Thiamine
0,1
Mezoinozitol
100
SUCROSE
30 g/l
pH
5,8
105
Table 2. Variants of MS medium

VARIANTS
GROWTH REGULATORS
BAP
IAA
NAA
1 mg/l
1 mg/l
1 mg/l
0,1 mg/l
1 mg/l
1 mg/l
0,1 mg/l
1 mg/l

The main aim of the investigation was to select different cell lines from suspension
cultures for embryogenetic capacity.
The axillary buds of Rubus caesius L. were the explants for callus initiation. Five
different media are used for plant propagation via somatic embryogenesis. These media
differ by the type and level of plant growth regulators.
The begin of callus proliferation was achieved on two variants of MS medium:
variants 1 with a same concentration of benzylaminopurine and indolilacetic acid and
variants 3 with the same concentration of kinetine and naphtalenacetic acid.
The first cell proliferation were observed on the cantact surface of the explant with
nutritive medium (Photo 1, 2). The primary callus cultures were obtained after 4 weeks
from the in vitro cultivation, when the entire explant dedifferentiated (Photo 3).
The callus cultures obtained on agar medium were multiplicated on liquid medium
(Photo 4). The submerse cultures were periodically transferred on fresh medium.
The suspension cultures were cultivated on rotatory shaker at 100 rpm and
maintained in the dark. After 2 weeks, they were transferred on somatic embryogenesis
induction medium (Photo 4, 5).
The main factors for somatic embryogenesis induction were: the selection of cells,
the use of growth regulators in some concentrations and combinations, the photoperiode.
The embryogenic clusters were selected after each stage of subcultivation on
induction media.
Early stages of somatic embryogenesis were observed after a week from the
initiation of the submerse cultures (Photo 6).
The specific steps of the embryo development that were encountered were: globular,
cardo, torpedo and cotyledonary stages. Embryogenesis was induced under various ratios of
auxin to cytokinin.
Despite the selection, the suspension cultures display not only a morphological
heterogenity, but also differences in cell reaction: some cells multiplied and formed
clusters, without regenerative potential, whereas some cells redifferentiated and developed
somatic embryos.
106
The conditions that favor initial somatic embryogenesis may inhibit further
development of the embryos. That is why the variants 2 and 4 are used for initiation of
somatic embryogenesis and variant 5 is utilized to allow somatic embryos development.
The use of MS medium supplemented with excess of benzylaminopurine in
combination with indolilacetic acid and kinetine in combination with naphtalenacetic acid
stimulated the somatic embryo development. The differentiated embryos could be
converted into plants by transferring to the MS medium for maturation, without growth
regulators.
Conclusions
Callus induction was stimulated on MS medium with an equal concentration of a
cytokinine and auxine.
Cell suspensions of Rubus caesius L. were obtained from callus cultures derived
from axillary buds.
The development and maturation of the somatic embryos occurred after 4 weeks in
suspension cultures.
The use a combination of a cytokinine in excess and an auxine led to a increase in
the frequency of embryos differentiation in suspension cultures.
Changing the growth regulators balance in the MS medium has major effects on in
vitro dedifferentiation and redifferentiation at Rubus caesius L.
REFERENCES
1.
2.
3.
4.
5.
6.
BENVENUTI, S., PELLATI, F., MELEGARI, M., BERTELLI, D., 2004-Polyphenols, anthocyans, ascorbic
acid and radical scavenging activity of Rubus, Ribes and Aronia, Journal of Food Science 69(3):164-169.
BUSBY, A. , HIMELRICK, D.G., 1999- Propagation of blackberries (Rubus ssp.) by stem cuttings using
various IBA formulations, Acta Hort, 505: 327-332
FIOLA, J.A., SCHWARTZ, H., 1986-Somatic embryos organogenesis and proliferation in vitro from Rubus
embryos, Acta Hort., 183: 91-98
FIOLA, J.A., HASSAN, M.A., SWARTZ, H.J., BORS, R.H. , MCNICHOLS, R., 1990- Effect of
thidiazuron light fluence rates and kanamycin on in vitro shoot organogenesis from excised Rubus
cotyledons and leaves, Plant Cell, Tissue and Organ Culture, 20: 223-228
MOYER, R.A., HUMMER, K.E., FINN, C.E., FREI, B., WROLSTAD, R. E., 2002- Anthocyanins
phenolics and antioxidant capacity in diverse small fruit: Vaccinium, Rubus and Ribes, Journal of
Agricultural and Food Chemistry, 50: 519-525
NIKITINA, U.S., KUZMINA, L., MELENTEV, A.I., SHENDEL, G.V., 2007- Antibacterial activity of
polyphenolic compounds isolated from plants of Geraniaceae and Rosaceae families, Applied biochemistry
and microbiology, 43(6): 629-634
107
SMARANDA VNTU
PLATE
Photo 1. Callus development
Photo 2. Cell proliferation
Photo 3. Primary callus cultures
Photo 4. Suspension cultures
Photo 5. Cell cluster
Photo 6. Embryogenic cells cultures
108

GENES OF ARABIDOPSIS THALIANA INVOLVED IN WAX METABOLISM

ROXANA-IULIANA TEODOR*, AL. YEPHREMOV*
Abstract: The aerial surfaces of land plants are covered with cuticle that acts as a barrier providing
protection against water loss, pathogen invasion and other environmental aggressions. Besides physical
and chemical barriers, such as a waxy cuticle, plant defense mechanisms also involve a coordinated
activation of cellular responses to limit damage.
The eceriferum (cer) mutants of Arabidopsis define multiple genes required for various steps in cuticular
wax biosynthesis, transport and regulation of lipid-related pathways. Although the basic biochemistry of
wax production has been elucidated, very little is known about its regulation and its contribution to the
natural immunity. This review presents recently cloned wax biosynthetic genes and discusses regulatory
aspects of wax biosynthesis.
Key words: wax, Arabidopsis mutants
Role of cuticle for plants

A distinctive characteristic of all epidermal cell types is the presence of cuticle
covering their outer surface as a continuous lipophilic layer, which forms a barrier over the
aerial organs of land plants during their primary stages of development [7; 14].
Plant cuticle is formed mainly of cutin, a polyester of hydroxy and hydroxy-epoxy
fatty acid derivatives generated from cellular fatty acids [11], embedded in and covered
with a complex mixture of highly hydrophobic soluble materials (aliphatic compounds
comprised mainly of C24-C34 alkanes, alcohols and ketones) called cuticular waxes, as
well as other minor compounds, of an extremely diverse nature [30]. As they are physically
very closely associated, it is difficult to distinguish between the relative contribution of the
cutin matrix and that of cuticular waxes to the physical properties and the biological roles
of the cuticle.
However, in biochemical experiments, the cutin and cuticular waxes are usually
considered and analyzed separately, due to the soluble properties of the waxes versus the
cutin polymer, which remains insoluble. Such a structure gives the cuticle a set of highly
protective features and these were studied initially as to limiting nonstomatal water loss and
gaseous exchanges, controlling the absorption of lipophilic compounds, and providing
mechanical strength and viscoelastic properties [2; 25].
Additionally, the cuticle also functions in normal plant developmental processes,
including the prevention of postgenital organ fusion and pollenpistil interactions [17; 28],
as well as protecting the plant from biotic and non-biotic environmental stress factors [27].
Pruitt et al. [24] and Sieber et al. [28] have suggested that cuticle permeability also
influences cell-to-cell communication by enhancing or attenuating the passage of signal
molecules. For example, such signals could be required for organ adhesion, when they
Molecular Plant Genetics, Max Planck Institute for Plant Breeding Research, Carl-von-Linne Weg 10, Cologne,
Germany
109
would be moving across the cuticle, or for mediating signaling between trichomes and
stomata, when moving within the developing epidermis [18; 13].
Arabidopsis mutants offer information on the regulation of cuticle development
Mutants with deficient or altered wax coatings have been identified due to their
nonglaucous or glossy phenotype. Thus, there are no reports of mutants that lack wax
completely, indicating the vital function that waxes play for the normal development of
plants.
On the other hand, changes in a minor wax component are less likely to lead to a
clearly discernable phenotype and therefor have not been reported. Similarly, plants that
overproduce waxes are not easily detected by visual screenings.
In Arabidopsis, as well as in other species, the mutants that have been identified to
be defective in wax and/or cutin formation facilitated the identification of enzymes
associated with the cutin and wax pathways. Some of the enzymes catalyzing various steps
in the wax pathway have been characterized or their function has been proposed based on
the phenotype of the corresponding mutants.
The identification of eceriferum (cer) mutant lines described mainly by Koornneef
et al. [12] and by McNevin et al. [20] has led to the isolation and characterization of various
genes associated with cuticular wax metabolism in Arabidopsis. CER1 intended to encode
an aldehyde decarbonylase [1]. Several genes playing a role in the fatty acid elongation
pathway that generates very long chain fatty acid (VLCFA) wax precursors have also been
characterized. They include the FATTY ACID ELONGATION1 homologs (FAE1) [9],
FIDDLEHEAD (FDH) [38; 24], 3-KETOACYL-CoA SYNTHASE [33], CUT1/CER6, and
CER60 [21; 5]. CER6 has been suggested to be the key condensing enzyme for wax
biosynthesis in Arabidopsis, due to its expression throughout all stages of stem and leaf
development, as well as in the inflorescence [8]. CER2 encodes a CoA-dependent
acyltransferase, a component of the fatty acid elongase complex, apparently located in the
nucleus [37; 14]. The cer2 mutant shows reduced levels of the decarbonylation pathway
products and it accumulates C26 and C28 acyl groups, primary alcohols, and wax esters but
the precise function of the gene is still unknown. Furthermore, its nuclear localization is
very intriguing for a protein of the fatty acid elongase complex. Many of the cer mutants
remain still to be characterized and the isolation of their corresponding genes might bring
valuable information on the mechanisms of wax metabolism.
Several reports have also provided insights into the biosynthesis of cutin monomers
in plants. Chen et al. [3] reported the isolation of the WAX2 gene and showed that the
protein it encodes for has 32% similarity to CER1 and contains certain regions with
homology to sterol desaturases and short-chain dehydrogenases/reductases. It was
suggested therefore that WAX2 plays a metabolic role in both wax and cutin synthesis, thus
pointing to a link between wax and cutin metabolism. ADHESION OF CALYX
EDGES/HOTHEAD (ACE/HTH) is proposed to be an oxidase catalyzing the formation of
dioic acids from -hydroxy acyl-CoAs [13; 16]. The Arabidopsis LACERATE (LCR) gene
[36] encodes a cytochrome P450; enzyme activity assays using the recombinant LCR
protein showed that it could efficiently catalyze the formation of -hydroxy fatty acids
(ranging from C12 to C18:1). Expression of LCR gene is predominant in inflorescence and
110
siliques, as well as in roots and young seedling tissue and it is the first cytochrome P450 hydroxylase for which a mutant has been isolated. Results of microarray analysis conducted
in our group (Yephremov et al., unpublished data) on three independent cuticular mutants
has revealed that a palmytoil protein thyoesterase (PPT) is almost ten fold up-regulated in
three mutants, as compared to wild type. In humans, PPT is a lysosomal long-chain fatty
acyl hydrolase that removes fatty acyl groups from modified cysteine residues in proteins,
and the defective enzyme causes infantile neuronal ceroid lipofuscinosis, a recessive
hereditary neuro-degenerative disorder [34]. In plants, acyl-acyl carrier protein (ACP)
thioesterases play an essential role in chain termination during de novo fatty acid synthesis
and in the channeling of carbon flux between the lipid biosynthesis pathways [10].
Epidermal differentiation and implicitly cuticle formation is essential for the general
development of the whole plant, starting from the very early embryo stage. This fact is
supported by the characterization of the abnormal leaf shape 1 (ale1) mutant of
Arabidopsis, which shows impaired cuticle formation, adhesion of endosperm and embryo,
as well as fusion of cotyledons and leaves. The corresponding ALE1 gene encodes a
member of the subtilisin-like serine protease family and it is preferentially expressed during
seed development, showing a weak transcript expression in young embryo and a strong one
within the endosperm cells closely surrounding the developing embryo [32]. Three
aminoacid residues (aspartic acid, histidine and serine) are consistently conserved in the
catalytic regions of subtilisin-like serin proteases.
In animals, such proteases activate precursors of hormones, growth factors, or
receptors involved in the control of various developmental processes, including embryonic
patterning and proper epidermal differentiation. Although many members of this family of
proteases were reported in plants [29; 26], little is known, with few exceptions, about their
precise role. In addition to the developmental factors controling the synthesis of cuticular
lipids, environmental signals such as light intensity, photoperiod [19; 35], humidity [31],
chilling [22; 23] and seasonal variation [6; 4] have also been shown to ifluence wax
biosynthesis. In 1984, Sutter [31] described the dramatic response of wax production to
environmental cues, during tissue culture, observing that when the relative humidity is
high, wax production is low. When tissue-culture-grown plants are transfered to an
environment with less humidity (growth chabinet or greenhouse), production of wax is
stimulated and within a rather short period of time of a few days only, the plant synthesizes
a complete protective layer of wax.
Conclusion
The fact that cuticular wax is ubiquitously present is testimony to its essential role in
the adaptation of plants to the aerial environment, with all its implications. On the other
hand, the fact that environmental cues have an influence upon wax composition and
quantity is evidence that wax production is an actively regulated process.
An active regulatory netword is indicated also by the high diversity of proteins that
have been shown, through the Arabidopsis mutants, to be involved in the process of wax
biosynthesis. Although the biosynthesis of plant cuticular components has been studied for
over four decades, we still know little about the factors regulating the partitioning of fatty
111
acid precursors and the synthesis of waxes with the synthesis of cutin and with other
cuticular compounds. However, the cloning of wax biosynthetic genes and the further
characterization of the respective proteins promises to bring valuable insights into the very
deep regulatory mechanisms of wax and cuticle development.
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113

CHARACTERISTICS OF THE PLANKTONIC ALGOFLORE FROM

DOROBANTI, ARONEANU, CIRIC I, II AND III LAKES (IASI COUNTY)
M. A. PORUMB* , M. COSTIC**
Abstract:After the setting of Ciric river by building some barrages, the lakes Dorobanti, Aroneanu, Ciric
I, II and III were formed, their total surface being of about 123 ha. When the lakes were formed, processes
of colmating and accumulating used waters started, these waters were coming from the side localities and
from the Ciric recreation area. Water quality from these aquatic mediums was evaluated on these
conditions, knowing the diversity of phytoplankton algae, the phytoplankton evolution and the specific
structure of algoflore. Researches concerning phytoplankton of the Ciric river area (May, 2006) show the
diatoms dominance (BACILLARIOPHYTA) at Dorobanti mijloc and Parau Ciric Pod stations, of the
CHLOROPHYTA group at Ciric II, Ciric I and Aroneanu stations and the big heaviness of the
CYANOPHYTA group at Dorobanti Dig station (in this moment, this station is attested by the algologic
indicators as being very polluted). During the first cold period of the year 2007, biodiversity is reduced
(because of the pollution), and the genus analysis shows the dominance of the green algae
(CHLOROPHYTA) at the first 4 lakes: at Dorobanti station the biggest numerical heaviness is presented
by Monoraphidium tortile (W.et G.S.West) Kom.-Leg. 21.18% of the total algae; at Aroneanu station
Monoraphidium contortum (Thur.) Kom.-Leg - 25.81% and Monoraphidium tortile (W.et G.S.West)
Kom.-Leg. 18.54% are dominant; at Ciric I, Monoraphidium contortum (Thur.) Kom.-Leg 48.88%, at
Ciric II, Monoraphidium contortum (Thur.) Kom.-Leg 55.66%, and at the last lake, Ciric III, the biggest
numerical heaviness was observed by Diatoma elongatum (Lyngb.) Agardh. (Bacillariophyta) 47.70%
of the total planktonic algae determined at that station. Numerical densities of phytoplankton from the 5
studied lakes varied between the values of 1,105 3,830 exemplars algae/ml in March, 2007, and 15,603
73,759 exemplars algae/ml in November, the same year. These values show an important pollution
degree of the investigated ecosystems. As it follows, researches are necessary for a period of several years
in order to point out the implied processes and phenomena complexity and evolution in those aquatic
ecosystems that are affected by anthropic pollution.
Key words: Ciric lacustrine complex, phytoplankton, algal taxons, water quality.
Introduction
Ciric recreation complex is made of a series of artificial barrage lakes, in falls, that
are alimented by Parau Ciric and by rains, and it has a recreation function: Dorobanti (70.00
ha), Aroneanu (23.00 ha), Ciric I, II, III with a total surface of 30.00 ha and 2.70 m in
maximum depth. Ciric Complex, which is situated on Valea Ciricului between Dealul Ciric
in the East and Dealul Sorogari in the West, is at 3.5 km up the confluence with Bahlui
River. From a physical-geographical point of view, it is placed at the contact between two
big subunits of Podisul Moldovenesc (the Moldavian Plateau): Campia Moldovei
(Moldavian Plain) and Podisul Central Moldovenesc (Central Moldavian Plateau). This
lacustrine Complex is 1.5 km long and its tail is near Aroneanu village.
*
**
The Station of Biological Researches Petre Jitariu Piatra Neam, Romania

Faculty of Biology, Al. I. Cuza University of Iasi, Romania
114
The trophic level of the Ciric lakes surface water is of 3rd category in quality
(according to STAS 4706/88), with a more accentuated degradation degree during the hot
period of the year when high temperatures determine a massive development of green-blue
algae and of euglenophycea, and finally, a more accentuated pollution that has also
implications in the underground waters in the area. The main causes of the water quality
depreciation come from the diffused pollution and from direct evacuations of used waters
that come from the economic agents in the area, the tourist activity and private proprieties,
these facts being amplified by the lack of canalization in the area.
Phytoplankton indicates the water quality of an aquatic ecosystem by its qualitative
and quantitative composition. Season conditions interfere with existent pollution, the result
being a phytoplankton that reflects the whole existent conditions.
Phytoplankton samples were collected each semester, within 2006 2007, from the
stations: Parau Ciric (1); Dorobanti (2); Aroneanu (3); Ciric I (4); Ciric II (5) and Ciric III
(6). Quantitative results of the samples that were studied at microscope by the above
mentioned proceedings, are appreciated by calculi, and a formula that includes the surface
of the microscope lamella, the volume of the drop under lamella (0.03 ml a drop that
comes from a graded and calibrated dropping glass of 1 ml), the ocular field diameter, the
number of analyzed microscopic fields and the quantity of sedimented sample or/and
centrifuged.
Algal biomass is determined by the establishment of cellular volumes (in microns)
of the counted algae and the conversion of these volumes in grams/m3, starting from
Dussart raport, 1966 (Limnologie. Letude des Eaux Continentales, Gauthier Villars,
Paris): 1,000,000 microns3 = 0,000001 grams.
For the algal biomass calculus, the lists of cellular volumes from the literature are
used for each genus, lists that are completed with original lists.
Microscope observations were made using the phase contrast a technique by which
fine details can be identified, these details being difficult to see by common proceedings.
In the documentary research regarding the results of planktonic algae there were
mainly used the series of Polish determinators Flora Slodkowodna Polski, of authors:
SIEMINSKA [13], STARMACH [14, 15, 16, 17, 18]. There were also consulted the works
of HINDAK [3,4,5], KOMAREK and AGNOSTIDIS [7], JOHN and collab.[6], that were
completed by other determinators and with the latest revisions of some genera and races of
different systematic groups of algae. There were also used some works from the ecological
literature of specialty [2, 8, 10, 11, 12]. Work methods were applied in a critical manner,
according to the necessities of the theme.
115

Researches regarding phytoplankton of the Ciric river area that were made in 20062007 show significant differences between the studied aquatic ecosystems, from an
algological point of view.
Qualitative data show the following qualitative structure of planktonic algal
communities in the 6 stations of assay:
CYANOPHYTA- Anabaena flos-aque Breb. ex Born et Flah (4), Anabaena solitaria Kleb.
(2,4), Anabaena spiroides Kleb (2,4,5), Anabaena sp. (4), Aphanizomenon flos- aque Ralfs
ex Born et Flah (4), Anabaena sp. (4), Gloeocapsa punctata Nag. (4,5), Gloeocapsa turgida
(Kutz.) Hollerb. (2,4,6), Gomontiella marthae Claus (2), Merismopedia tenuissima Lemm.
(3,4,5), Microcystis incerta (Lemm.)Lemm. (2,5), Microcystis pulverea(Wood) Mig. In
Lemm. (5), Oscillatoria limnetica Lemm. (2), Oscillatoria planctonica Wolosz. (3),
Oscillatoria granulata Gordner (1), Pseudanabaena sp. (2), Romeria leopoliensis (Rocib.)
Koczw. (4), Romeria gracilis Koczw. (2), Spirulina laxissima G.S. West (2,4,5), Spirulina
meneghiniana Zanard. (4), Spirulina raphidioides Geitl (2,5), Spirulina sp. (4,5),
Synechococcus elongatus (Nag.) Nag. (2,4);
CHRYSOPHYTA- Chrysococcus sp. (2,3), Hymenomonas roseola Stein (2), Mallomonas
sp. (4,5), Ochromonas sp. (2,4,5,6);
XANTHOPHYTA- Tribonema monochloron Pascher et Geitler (1,2)
BACILLARIOPHYTA- Achnanthes minutissima Kutz. (1, 2, 3, 5, 6), Achnanthes sp. (1, 2,
6), Amphora ovalis Kutz. (1, 2, 5, 6), Caloneis sp. (2), Coconeis pediculus Ehr. (2, 3, 4),
Coconeis placentula Ehr. (2), Cyclotella compta (Ehr.) Kutz. (2,5), Cyclotella
meneghiniana Kutz. (5), Cyclotella sp. (2.), Cymbella amphicephala Naeg. (1), Cymbella
ventricosa Kutz. (5), Diatoma anceps (Ehr.) Kirchn. (5), Diatoma elongatum
(Lyngb.)Agardh (6), Diatoma elongatum var. actinastroides Krieger (6), Diatoma
elongatum var. tenue (Agardh) Van Heurck. (3,6), Gomphonema constrictum Ehr. (6),
Gomphonema olivaceum (Hornem.)Breb. (1,2,3,4,6), Melosira granulata (Ehr.) Ralfs (2),
Navicula cryptocephala Kutz. (1,2,6), Navicula placentula (Ehr.) Grun. (1), Navicula
radiosa Kutz. (1,2), Navicula rhynchocephala Kutz. (2), Navicula viridula Kutz. (1,2),
Navicula sp. (1,2,5), Nitzschia acicularis W.Smith (1,4,5), Nitzschia capitellata Hust. (4),
Nitzschia closterium (Ehr) W.Sm. (5), Nitzschia circumsuta (Bailey) Grunov (1), Nitzschia
linearis (Ag.) W.Sm. (1,2), Nitzschia palea (Kutz.) W. Smith (1,2,3,4,5), Nitzschia
sigmoidea (Ehr.) W.Smith (1), Nitzschia tryblionella Hantz. (Kutz.) Grun. (1), Nitzschia sp.
(3,4,5,6), Rhoicosphaenia curvata (Kutz.)Grun. (2,4,5,6), Rhizosolenia longiseta Zach. (1),
Rhizosolenia eriensis H.L.Smith (5), Rhopalodia gibba (Ehr.) O.Muller (4), Surirella
angustata Kutz. (1), Surirella ovata Kutz. (1), Surirella sp.(6), Synedra acus Kutz. (3),
Synedra nana Meist.(4), Synedra tenera W.Sm. (2), Synedra ulna (Nitz.) Ehr. (3,5),
Synedra vaucheriae Kutz. (3), Synedra sp. (3,5);
PYRROPHYTA- Chroomonas acuta Utermohl (2), Chroomonas nordstedtii Hansa. (1,2),
Peridinium cinctum Ehr. (5);
CHLOROPHYTA- Chlorella vulgaris Beijer. (1,2,3,4,5), Chlorogonium tetragamum
Bohlin(4), Chlamydomonas sp. (3,5), Cladophora sp. (4), Closteriopsis acicularis
(G.M.Smith) Belcher et Swale (2), Closterium acutum Breb. (1), Coelastrum sphaericum
116
Naeg. (2,5), Coelastrum sp. (2), Crucigenia rectangularis (Nag.) Gay (1), Crucigenia
tetrapedia (Kirchn.) W.et G.S.West (1,2,5), Dictyosphaerium pulchellum Word (5),
Didymocystis fina Komarek (5), Golenkinia radiata Chodat (5), Hyaloraphidium contortum
Pasch. et Korsch. (4), Keratococcus bicaudatus (A.Br.) Boye.-Pet. (5), Kirchneriella
aperta Teiling (2), Kirchneriella contorta (Schm.) Bohl (2, 3, 4), Kirchneriella irregularis
(Smith) Korsch. (6), Kirchneriella obesa (W.West) Schm. (1,4), Kirchneriella subcapitata
Korsh. (1, 2), Koliella longiseta (Wisl.)Hind. (3, 4), Koliella planctonica Hind. (2, 3, 4, 5,
6), Koliella spiculiformis (Wisch.)Hind. (1, 3, 4, 5, 6), Koliella sp.(3,4), Lagerheimia
genevensis (Chodat) Chodat (2, 3), Monoraphidium arcuatum (Korsch.) Hind. (2, 3, 4, 5),
Monoraphidium contortum (Thur.) Kom.-Leg. (2, 3, 4, 5, 6), Monoraphidium griffithii
(Berkeley) Kom.-Leg. (3, 5, 6), Monoraphidium komarkovae Nygaard (4, 5),
Monoraphidium minutum (Nag.) Kom.-Leg. (2, 3, 4, 5, 6), Monoraphidium pusillum
(Printz) Kom.-Leg.(2), Monoraphidium tortile (W.et G.S.West) Kom.-Leg. (2, 3, 4, 5, 6),
Monoraphidium sp. (2, 5), Nephrochlamys agardhianum Nag.(2), Nephrochlamys sp. (3),
Oocystis lacustris Chodat (1, 2, 5), Oocystis marsonii Lemm. (2), Oocystis sp. (2),
Pediastrum tetras (Ehr.) Ralfs (1), Scenedesmus acutus Meyen (5), Scenedesmus
acuminatus (Langerh.) Chodat. (2,3), Scenedesmus bicaudatus Deduss. (4, 5), Scenedesmus
dispar Brebis. (2, 4, 5), Scenedesmus ecornis (Ehr.ex Ralfs) Chodat (5), Scenedesmus
linearis Kom. (1, 2, 5, 6), Scenedesmus opoliensis Rich. (2, 5), Scenedesmus quadricauda
(Turp.) Brebis. 1, 3, 4, 5, 6), Scenedesmus sp. (3, 5), Schroederia nitzschioides (G.S.West)
Korsch. (3, 4, 5), Schroederia spiralis (Printz) Kors. (1, 5), Schroederia sp. (1), Siderocelis
ornata (Fott) Fott (2), Staurastrum sp. (4), Stichococcus bacillaris Nag. (1, 2), Tetraedron
caudatum (Corda) Hansg. (1), Tetraedron minimum (A. Braun) Hansg. (3), Tetraedron
trigonum (Naeg.)Hansg. (1, 2, 4), Tetrastrum glabrum (Roll.) Ahlst.et Tiff. (3), Ulothrix sp.
(4);
EUGLENOPHYTA- Euglena acus Ehr (4, 5), Euglena clavata Skuja (2, 3), Euglena
gasterosteus Skuja (2, 3), Euglena limnophila (2, 3), Euglena matvienkoi Popova (3),
Euglena polymorpha Dang. (4), Euglena proxima Dang. (2, 3), Euglena spathirhyncha
Skuja (3), Euglena texta (Duj.) Hubn. (2, 3), Euglena tripteris (Duj.) Klebs. (3), Euglena
sp. (2, 3, 4), Lepocinclis acuta Prescott (3), Lepocinclis ovum (Ehr.) Lemm. (2, 3),
Lepocinclis sp. (2, 5), Peranema sp. (4), Phacus pleuronectes (Ehr.) Duj. (4), Phacus
pyrum (Ehr.) Stein (4), Phacus sp. (2), Trachelomonas verrucosa Stokes (2, 3),
Trachelomonas volvocina Ehr. (2,5), Trachelomonas sp. (3);
The diversity degree of planktonic algoflore is, generally, determined by the water
quality existent conditions.
Quantitative data (Table I) show the diatoms dominance (BACILLARIOPHYTA) in
Dorobanti mijloc and Parau Ciric Pod stations, of the CHLOROPHYTA group in Ciric
II, Ciric I and Aroneanu stations and the big heaviness of the CYANOPHYTA group in
Dorobanti Dig station (in this moment, this station is attested by the algologic indicators
as being very polluted).
117
Table I. The phytoplankton of the Ciric lacustrine complex (nr. exempl../ml)

Nr
Station/Phylum
Parau
Dorobant-
Dorobant-
Aroneanu-
Ciric-
mijloc
dig
dig
255 674
709
Ciric I
Ciric
II
Pod
1
Cyanophyta
Chrysophyta
Bacillariophyta
51 773
Chlorophyta
355
Euglenophyta
Total algae
709
2 482
142
8 510
3 901
5 319
1 064
6 028
3 546
638
4 610
5106
355
52 837
8 510
260 639
12 056
10 993
5 886
During the samples assay in May, general season conditions and the moment
atmospheric situation of the barrages area have determined the dispersal in the
phytoplankton mass of some elements from the algal periphyton. In July, 2006 (Table II)
some important differences are found in the total number of algae and the distribution of
planktonic algae groups, according to the assay station.
Table II. The phytoplankton of the Ciric lacustrine complex - July 2006
(nr.exempl../ml)
Nr
Station/Phylum
Parau
Dorobant-
Dorobant-
Aroneanu-
Ciric I-
Ciric
Ciric-
mijloc
dig
dig
dig
Ii-dig
Pod
1
Cyanophyta
319
71
532
3 191
6 028
7 092
Chrysophyta
142
284
319
709
709
709
Xanthophyta
425
284
Bacillariophyta
425
248
446
1 064
3 546
6 383
Pyrrophyta
71
71
709
Chlorophyta
2411
3050
355
7 801
3 901
Euglenophyta
Total algae
3 793
71
922
8 156
4 610
2 482
4 008
2 269
14 184
22 694
20 567
Massive algal development is stated in the following stations: Aroneanu dig, Ciric
I dig and Ciric II dig, this fact proving the presence of big charges of biogenous
substances at these stations, the existence of a very big pollution, respectively. In stations:
118
Aroneanu dig; Ciric I dig; Ciric II dig is also stated an important development of bluegreen algae (CYANOPHYTA), which indicates the presence in these stations of an excess
of nutrients of organic and inorganic nature, which help these algae to develop and they are
represented here by some genera that proliferate in very intense pollution conditions.
EUGLENOPHYTA group is a very important indicator of water quality due to its
mixotroph nutrition manner. These algae very much develop in water if it is polluted with
toxic substances, especially organic ones, in big quantities. So, important numerical
development of euglenoides at station 4 - Aroneanu dig, but also at Ciric I dig; Ciric II
dig show the fact that important pollution sources are overflowed and collected here. In
the studied lakes, in phytoplankton, in the first cold period of the year 2007 (Table III), 53
taxons were identified that belong to 6 phylums; they are distributed as it follows: 19
taxons in Dorobanti lake; 23 in Aroneanu lake, 14 in Ciric I lake, 21 in Ciric II lake and 15
in Ciric III lake.
Table III The phytoplankton of the Ciric lacustrine complex - March 2006
(nr.exempl../ml)
Nr.
Station/Phylum
Dorobant
Cyanophyta
106
Chrysophyta
Bacillariophyta
468
Pyrrophyta
42
Chlorophyta
489
Euglenophyta
Total algae
Aroneanu
Ciric I
Ciric II
299
170
Ciric III
106
42
319
85
2 149
3 404
4 808
425
3 830
5 275
2 765
255
2 340
42
64
1 105
2 638
This determined number of taxons reflects a reduced biodiversity that is mainly

owed to pollution. Analysis of dominant genera from the 1st of March, 2007 shows the
following: at Dorobanti station Monoraphidium tortile (W.et G.S.West) Kom.-Leg
(CHLOROPHYTA) is dominant, it represents 21.18% of the total algae; at Aroneanu
station Monoraphidium contortum (Thur.) Kom.-Leg - 25.81% and Monoraphidium tortile
(W.et G.S.West) Kom.-Leg 18.54% are dominant; at Ciric I, Monoraphidium contortum
(Thur.) Kom.-Leg represented 48.88%, at Ciric II Monoraphidium contortum (Thur.)
Kom.-Leg was registered with values of 55.66%, and at the last lake, Ciric III, Diatoma
elongatum (Lyngb.)Agardh (BACILLARIOPHYTA) was dominant with 47.70% of the
total algae determined at that station. Except for Diatoma elongatum (Lyngb.) Agardh.
algae, which is -oligosaprobe, the other 2 dominant genera mentioned above are mesosaprobe, and this fact situates the water quality in that saprobity degree. Algological
analysis results show a significant pollution degree in the 5 studied hollows, at Ciric II and
Ciric I lakes, especially.
119
- In the 5 studied lakes in November, 2007 (Table IV), 51 taxons were identified in
phytoplankton.
Table IV The phytoplankton of the Ciric lacustrine complex November 2007
(nr.exempl./ml)
Nr.
Station/Phylum
Dorobant
Aroneanu
Cyanophyta
26 596
2 218
Chrysophyta
Bacillariophyta
6 383
Chlorophyta
40 425
Euglenophyta
355
Total algae
73 759
Ciric I
Ciric II
Ciric III
18 440
25 532
50 709
2 482
355
2 482
1 064
9 929
709
10 993
21 986
12 411
11 347
15 603
51 064
73 404
30 496
This number of taxons reflects a quite reduced algal biodiversity, which is mainly
owed to pollution.
Conclusions
Although Dorobanti Lake presents the biggest algal numerical diversity, the
dominance of green algae (CHLOROPHYTA) in detriment of CYANOPHYTA group
shows, comparatively, the fact that it is not the most polluted lake of the studied
ecosystems. Ciric II Lake is more polluted compared to Dorobanti Lake as, in the
conditions of a total algal density of close values, the high number of green-blue algae
(CYANOPHYTA) of Ciric II lake shows an important organic charge of water and a
significant pollution, respectively. Lakes Ciric I, II and III have also an accentuated
pollution level which is demonstrated not by the high numerical algal densities only, but
especially by the presence of some algae of the CYANOPHYTA group with a big cellular
volume and a high biomass/individual (e.g.: Aphanizomenon flos-aquae Ralfs ex. Born et
Flah;
Numerical densities of phytoplankton in the 5 studied lakes have oscillated between
the values of 1,105 3,830 exemplars algae/ml in March, 2007, and 15,603 73,759
exemplars algae/ml in November, the same year. These values show an important pollution
degree of the investigated ecosystems. In comparison, in this sense, we mention that 5 lakes
from the course of Bistrita river have phytoplankton values of 383 9,382 exemplars
algae/ml [10], Cuiejdel lake had 1,060 10,116 exemplars algae/ml within 2000 2004
[11], and 9 aquatic ecosystems of reduced productivity in the Danube Delta had values
between 684 2,271 exemplars algae/ml [9] limits;
Although results concerning the pollution spectrum in the area are significant, there
are necessary some researches for a period of various years in order to highlight the
complexity and evolution of implied processes and phenomena in the respective
120
ecosystems and in order to elaborate and apply these ecosystems reconstruction that are
affected by the anthropic pollution so badly;
The water quality of the 5 lakes which is shown by the dominant indicative genus of
algae is of -mesosaprobe type;
The measures that will be taken in order to improve water quality will refer to the
reduction of pollution sources and to the improvement of canalization, water treatment and
general salubrization conditions.
REFERENCES
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
DUSSART B., 1966- Limnologie. Letude des Eaux Continentales, Gauthier Villars, Paris
GERMAIN H., 1981- Flore des diatomees (Diatomophycees)- eaux douces et saumtres du Massif
Armoricain et des contres voisines d,Europe occidentale, Socit Nouvelle des Editions Boube 11, place
Saint-Michel, 75006-Paris
HINDAK Fr.,1978 (ed)-Slodkovodne rissy, Slovenske Pedagogiske nakladeistvo, Bratislava
HINDAK Fr., 1977- Studies on the chlorococcal alge (Chlorophycceae) I, Biologicke Prace, 4, XXIII,
Bratislava.
HINDAK Fr.,1980- Studies on the chlorococcal alge( Chlorophycceae)II, , Biologicke Prace,6, XXVI,
Bratislava.
JHON D.M., WITTON B.A., BROOK A.J. (Editors), 2003 (Second Edition) - The Freshwather Algal Flora
of the British Isle, University Press, Cambridge
KOMAREK J., AGNOSTIDIS K., 1986 Modern approach to the classification System of cyanophytes.2.
Chroococcales. Arch. Hydrobiol. Suppl. 73, 2 (Algological Studies 43), 157-226 Sttugart
PORUMB M. A., 1986- Clasificarea nivelurilor de troficitatae a 18 iazuri din judetele Cluj, Alba si Bistrita
Nasaud pe baza criteriului algologic, Lucr. Simpoz. A III-a Comfrinta Nationala de ecologie Arad, 4-7
iunie (1986), 349-351
PORUMB M. A., 2000 - Cercetri privind algele planctonice din Delta Dunrii n arealul dintre Bra Sulina
i Bra Sf. Gheorghe. Studii si Cercetari, Muzeul de t. Naturale Piatra Neam, 10: 35-46.
PORUMB M. A., 2006- Cercetari privind algoflora planctonica a lacurilor de baraj din cursul mijlociu si
inferior al raului Bistrita, Stud. Si Cercet., Muz. St. Nat. Piatra Neamt, 10: 53-65
PORUMB M.A., 2006- Studiu privind algele din lacul de baraj natural Cuejdel-Neamt, Stud. Si Cercet. Muz.
St. Nat. Piatra Neamt, 10: 65-77
PREMAZZI G., DALMIGLIO A., CARDOSO A.C., CHIAUDANI C., 2003- Lake management in Italy:
The implications of the Watyer Framework Directive, Lakes Reservoirs: Research and Management, 8: 4159
SIEMINSKA J., 1966- Bacillariophyceae, Flora Slodkow. Polsky
STARMACH K., 1966- Cyanophyta. Glaucophyta. Flora Slodkow, Polsky, 2
STARMACH K., 1968- Chrysophyceae. Flora Slodkow. Polsky, 7
STARMACH K., 1972- Chlorophyta III. Zielenice nitkowate. Flora Slodkow. Polsky
STARMACH K., 1974- Cryptophyceae, Dinophyceae, Raphidophyceae. Flora Slodkow. Polsky, 4
STARMACH K., 1980- Chrysophyceae, Flora Slodkow. Polsky
STARMACH K,. 1983- Euglenophyta. Flora Slodkow. Polsky
Acknowledgements
The researches were supported from the funds distributed within the CEEX project no. 634: The
terrestrial and aquatic peri-urban ecosystems from Ciric river basin, from the north of Iai municipality, financed
by the Ministry of Education and Research of Romania.
121

ON THE OCCURRENCE OF ZOSTERA NOLTII HORNEMANN AT THE

ROMANIAN COAST OF THE BLACK SEA
V. SURUGIU*
Abstract. The finding of a small Zostera noltii patch (~20 m2) at the Romanian Black Sea coast is
reported. The systematics as well as the geographical distribution of this species is briefly reviewed. Some
ecological characteristics of this species are also provided. The causes of the declining of Zostera noltii
beds at the Romanian coast of the Black Sea are discussed.
Key words: seagrass beds, Zostera noltii, distribution, Romanian coast, Black Sea
Introduction
The dwarf eelgrass Zostera noltii (Hornem.) Toml. & Posl. belongs to the family
Zosteraceae Dumortier, 1829. This family together with the families Cymodoceae,
Posidoniaceae, and Hydrocharitaceae forms an ecological group of aquatic angiosperms
adapted to live in the marine environment [5]. Representatives of these families are
collectively called seagrasses due to their grass-like appearance.
At the Romanian coast, there is also present another species of the genus Zostera, the
common eelgrass Z. marina L. [6, 12]. In the past both these eelgrass species covered with
a lush growth the bottom of marine lagoons Sinoie, Zmeica and Golovia [14]. Isolated
patches were reported also at Cape Midia [2] and Agigea [3]. At that time one even spoke
about the exploitation of eelgrasses and their use as surrogate for artificial wool, as stuffing
material for pillows and mattresses and for packing up eggs, pieces of furniture and other
fragile objects [1, 11].
In the last 40 years, due to pollution and eutrophication, seagrasses have declined
drastically in abundance, not only at the Romanian coast, but also in the entire Black Sea.
The aim of this paper is to reveal the actual status of Zostera noltii beds at the Romanian
coast of the Black Sea. Although Zostera noltii is distinguishable from Z. marina, in many
cases the authors have cited these plants simply as Zostera, irrespective of whether they
refer to one or another species. In order to prevent the confusion between these two species
of eelgrass an identification key is provided.
The dwarf eelgrass patch was identified at Mangalia (4348'18.0"N;
02835'31.9"E), between 1.3-1.9 m deep, on sandy substrate, in a small embayment formed
by a dyke (Fig. 1). The patch is approximately 7 m long and 5 m wide. Samples of Zostera
*
Al. I. Cuza University of Iai, Faculty of Biology, Bd. Carol I, no. 20A, 700506, Iai, Romania
e-mail: vsurugiu@uaic.ro
122
noltii were taken by snorkelling on 26th May 2005, 30th June 2005, 4th August 2006, and 8th
August 2006. Voucher specimens were herborised and deposited in the Herbarium
collection of the Faculty of Biology, Alexandru Ioan Cuza University of Iasi. The in situ
photographs were taken with a digital ReefMaster DC 310 underwater camera. All the
specimens collected were checked against the species description from the speciality
literature [6, 9].
Fig. 1 Location of the Zostera noltii meadow at Mangalia (4348'18.0"N; 02835'31.9"E).

Systematics
Zostera (Zosterella) noltii Hornemann, Fl. Dan. 12(35): 3, tab. 2041 (1832)
(Dwarf eelgrass)
Fig. 2, 3
Zostera marina auct. non L.Phucagrostis minor Cavol., Phucagr. Anth. XIV, Pl.
2 (1792) nom. invalid.Zostera trinervis Stokes, in Bot. Mat. Med. 4: 319 (1812)
nom. illeg.Zostera nana Mertens ex Roth, Enum. Pl. Phan. Germ. 1: 8 (1827)
nom. invalid.Zostera minor (Cavol.) Nolte ex Reichenb., Icon. Fl. Germ. 7: 2
(1845).Zostera pumila Le Gall in Congr. Sc. France 16: 96, 144 (1850).
Description.Colour is grass-green. Rhizomes 0.5-2 mm in diameter, with 1-4 roots at each
node. Internodes 4-35 mm long. In cross-section the strengthening fibre bundles occur in
the innermost layer of the outer cortex of the rhizome. Sterile shoots (leaves) alternately
arranged and flattened. The leaf sheath at the base clasps the stem but is not fused into a
tube, 4-5 cm long. Leaf blades 6-22 cm long and 0.5-1.5 mm wide, with approximately (1)3
123
irregularly spaced veins. The tips of leaves initially rounded, but, as the plant matures, they
become notched (emarginate), often asymmetric. Generative shoots lateral, unbranched or
with a few branches near the base, shorter and narrower than the sterile shoots; with 1-6
spathes. Spathal sheath 12-20 mm long and 1.3-2 mm wide. Spadix lanceolate with 4-5
staminate flowers and 4-5 pistillate flowers. Fruit ellipsoid, 1.5-2 mm long; pericarp dark
brown. Seeds smooth, white in colour, 1.5-2.0 mm long, excluding the style. 2n = 12.
Biology.Hermaphrodite, perennial herb. Flowering period extends from June to August
[6]. In the British Isles the plant retains its leaves throughout the winter. Main method of
reproduction is by vegetative growth. However, seedling germination appears to be also
important. In the Black Sea peaks of asexual reproduction occur in spring and autumn,
when regrowth of the rhizome system is the most intense [7].
Habitat.Intertidal, between mean high water neap and mean low water neap. In waters
with reduced salinity Zostera notii grows deeper and may become permanently submerged.
Thus, in the Black Sea the species occurs from 0.6 m down to 8-10 m deep. In
Mediterranean the dwarf eelgrass may grow down to 20 m deep. Inhabit sandy or muddy
sand substate. Euryhaline species, very tolerant to desiccation. The lower salinity limit is
about 15 g l1. The plant is restricted to sheltered sites such as estuaries, salt marshes, bays
and lagoons. At Mangalia it was found in association with Cladophora sericea (Fig. 2).
Geographical distribution.The species is distributed along the Atlantic coasts of Europe
and northern Africa, extending from southern Scandinavia (Norway) to the tropic of Cancer
(Mauritania and Senegal). It also occurs around the British Isles, in the Baltic Sea,
Mediterranean Sea, Adriatic Sea, Black Sea, Sea of Azov, Caspian Sea and Aral Sea [4].
It the Black Sea dwarf eelgrass was recorded in the Sevastopol Bay, along the Caucasian
coast (gulf of Anapa, gulf of Novorosiisk, gulf of Gelendjik), gulf of Karkinit, Tendrovsky,
and Burgas Bay [8].
At the Romanian coast the presence of Zostera noltii meadows was previously reported at
Mamaia, Agigea, Cape Midia, Mangalia, lake Razim, lake Golovia and Sinoie lagoon [2,
3, 7, 14].
Key to the Zostera species from the Black Sea
1. Rhizome thin, with 1-4 roots at each node. Leaves 6-22 cm long and 0.5-1.5 mm wide,
with one principal vein; leaf-sheaths open with open margins overlapping; leaf tip
emarginate. Generative shoot lateral. Retinacula present. Seeds smooth .........................
................................................................................................. Zostera (Zosterella) noltii
- Rhizome thick, with numerous roots at each node. Leaves 50-150 cm long and 3.07.0(9.0) mm wide, with 3 to 5(9) principal veins; leaf-sheaths closed, tubular, rupturing
with age; leaf tip obtuse to slightly mucronate. Generative shoot terminal. Retinacula
absent. Seeds with 16-25 longitudinal ridges ........................... Zostera (Zostera) marina
In the Black Sea marine phanerogames are represented by 6 species: Zostera
marina, Z. noltii, Potamogeton pectinatus, Ruppia maritima, R. spiralis, and Zannichellia
major [8]. However, only the two species belonging to the genus Zostera, commonly
124
known as eelgrasses, are considered to be fully confined to the marine environment. The
remaining species forms the so-called eurysaline group, an ecological group of flowering
aquatic plants tolerant of considerable changes in salinity from full-strength seawater to
freshwater [5].
The eelgrass biocoenosis is an important element of shallow-water coastal benthic
environments [9]. Seagrass beds are among the most productive marine communities. The
standing biomass of dwarf eelgrass in the Black Sea ranges from 126 g m2 (Gulf of
Karkinit) to 380 g m2 (Gulf of Anapa) [12]. The production of Zostera noltii is estimated
to 15 g fresh weight/kg/day.
As key primary producers, the eelgrasses represent an important source of food for many
organisms, especially in the form of detritus. Although, Zostera noltii is grazed directly by
waterfowl and by some fish species [6]. The dense, matted root system of eelgrasses
stabilise the soft sediments, and thus reduce coastal erosion. Also, seagrass beds increase
habitat diversity, providing shelter for a wide variety of marine organisms [2, 9].
Zostera noltii occurs on sedimentary substrata, in areas sheltered from water
motion (currents and waves). Because Romanian coast is exposed to north-south
alongshore current and to strong winter storms, it offers very few suitable conditions for
eelgrass grow. Extensive carpets of eelgrasses were present only in the quiet brackish water
lagoons. However, sparse Zostera shrubs were reported in front of Agigea at 0.8-1 m depth
[3, 7]. Denser mats were found at Cape Midia [7]. Recently Teac et al. [13] indicated on
the presence of small areas (approx. 3030 cm) populated by small eelgrass Zostera sp.
(most probably Z. noltii) at Mamaia Casino and Mangalia, on a sandy-silty texture
substrate, in the proximity of the protective wave-breaking dam.
The decline of seagrasses in relatively open areas of the Romanian coast, as well as in the
entire Black Sea, is due principally to the eutrophication. The nutrient enrichment of water
increased the phytoplankton density, thus decreasing the transparency and diminishing the
amount of light that can reach the bottom. The light penetration is also reduced by the
siltation of near-shore sediments due to littoral works and to the construction of protective
dams. Another cause for the decline of eelgrasses is the collapse of fishing in the Black Sea
which reduced grazing on the epiphytes that live on the grass blades. The overgrowth of
epiphytes thus prevents or reduces light intensity at the surface of the grass blade. Thus, the
blades of the dwarf eelgrass observed at Mangalia were densely covered by various
microscopic algae.
The freezing of the sea may also have dramatic deleterious effects due to the ice scour [9].
However, the severe frost of the sea occurred during the 2005-2006 winter had no
detectable impact upon the size and density of the meadow from Mangalia.
As a result of a gradual reduction of salinity in all Romanian littoral lakes from
polyhaline to ahaline over the last 40 years, vast fields of eelgrasses have disappeared
completely.
Because at the Romanian littoral Zostera noltii occurs very sparsely and is
threatened to extinction it must be put under protection.
.
125
Fig. 2 Zostera noltii meadow at Mangalia.
REFERENCES
1. BACALBAA-DOBROVICI N., 1951 - Posibilitatea valorificrii ierbii de mare (Zostera) n R.P.R. Bul. Inst.
Cercet. Pisc., Bucureti, 10(1): 25-32.
2. BCESCU M., MLLER G.I., GOMOIU M.-T., 1971 - Ecologie marin. Cercetri de ecologie bental n
Marea Neagr. Analiza cantitativ, calitativ i comparat a faunei bentale pontice. Ecologie marin, IV, Ed.
Academiei R.S.R., Bucureti
3. BOTEZ M., CRUU S., BOCEC A., CALINICENCO N., 1937 - Le plan de la Station Zoologique Maritime
Regele Ferdinand dAgigea Constantza (Roumanie): Topographie, Biotopes et Biocnoses littoraux.
AnnalesScientifiques de lUniversit de Jassy, 23(2): 1-4.
4. DEN HARTOG C., 1970 - The Sea Grasses of the World. North-Holland Publishing Company, Amsterdam
5. DEN HARTOG C., KUO J., 2007 - Taxonomy and Biogeography of Seagrasses. In: A.W.D. Larkum, R.J. Orth,
C.M. Duarte (Eds.), Seagrasses: Biology, Ecology and Conservation, XVI, Springer Verlag, Berlin
6. GRINESCU I., NYRDY E.I., PAUC A., PRODAN I., ERBNESCU I., YAHARIADI C., 1966 - Flora
Republicii Socialiste.Romnia., XI,Ed. Academiei R.S.R., Bucureti
7. MIHNEA P., 1965 - Biocenoza faciesului cu Zostera. Lucrare de diplom, Univ. Al. I. Cuza Iai, Facultatea
de Biologie-Geografie
8. MILCHAKOVA N.A., 1999- On the status of seagrass communities in the Black Sea. Aquatic Botany, 65: 2132.
9. PHILLIPS R.C, MEEZ E.G., 1988 - Seagrasses. Smithsonian contributions to the marine sciences, No. 34,
Smithsonian Institution Press, Washington, D.C.
126
10. ROBERTSON A.I., MANN K.H., 1984 - Disturbance by ice and life history adaptations of the seagrass
Zostera marina. Mar. Biol., 80: 131-142.
11. RUDESCU L., 1956 - Cercetri privitoare la exploatarea i prelucrarea ierbii indigene de mare. Rev. Ind.
Lemnului, celulozei i hrtiei, Bucureti, 7: 354-359.
12. SKOLKA H., 1977 - Algues macrophytes et Phanerogames des mers saumtres pontocaspiennes. In: E.A. Pora
& M.C. Bcescu (eds.), Biologie des eaux saumtres de la mer Noire, pp. 59-69. Institut Roumain de
Recherches Marines, Constana.
13. TEAC A., BEGUN T., GOMOIU M.-T., PARASCHIV G.-M., 2006 - The present state of the epibiontic
populations to the biocenosis of stone mussels in the shallow water off the Romanian Black Sea coast. GeoEco-Marina, 12: 53-66.
14. TEODORESCU-LEONTE R., LEONTE V., MATEI D., OILEANU B., 1956 - Observaii asupra
complexului Razelm-Sinoie n perioada 1950-1952. Ann. Inst. Cerc. Pisc., 1: 1-50.
Acknowledgements
I am much indebted to my colleague Dr. C. Mnzu for checking the identification of the species and for
helpful advice
Fig. 3 The habit of Zostera noltii (Hornem.) Toml. & Posl.
127

CONTRIBUTIONS TO THE STUDY OF THE RUDERAL VEGETATION FROM

THE CEAHLU MOUNTAIN
OANA ZAMFIRESCU*, C. MNZU*, T. CHIFU*
Abstract: The study presents four types of ruderal phytocoenoses identified in the Ceahlu Mountain and
classified through the analysis of a appropriate number of relevs in the following vegetation associations
[4] [8]: Potum supinae Brun-Holl 1962 em. Gutte 1969 and Rumicetum alpini Beger 1922, of all.
Rumicion alpini Rbel ex Klika in Klika et Hada 1944, ord. Rumicetalia alpinae Mucina in Karner et
Mucina 1993. cl. Mulgedio-Aconietea Hada in Klika 1948, Sambucetum ebuli Felfldy 1942, all. GalioAlliarion (Oberd. 1957) Lohmeyer et Oberd. in Oberd. et al. 1957, ord. Lamio albi-Chenopodietalia bonihenrici Kopecky 1969, and Telekio-Petasitetum hybridi (Morariu 1967) Resm. et Raiu 1974, all.
Petasition officinalis Sillinger 1933, ord. Convolvuletalia sepium R. Tx. 1950 em. Mucina 1993, cl.
Galio-Urticetea Passarge ex Kopecky 1969. For each association we give the description of the site and
climatic conditions, the characteristic and associate species, and the synthetic table based on the analyses
of the carried out relevs.
Key words: weeds, mountain zone, anthropic impact
Introduction
The existence of weed associations in the national park area is related to the
grazing activity, especially the station of the animals in a certain space, and to the anthropic
impact, which is hard to control because of the numerous touristic paths that cover the
mountain.
Although not complete, our ruderal formation inventory indicates the presence and
the wide distribution of weed communities, and states the need of an action plan for the
management of the area.
The Ceahlu Mountain, whose central part was designated a national park, is
located in the centre of the northern part of the Oriental Carpathians [1] [9], in the western
extremity of Neam County. The boundaries of the mountain are Bistricioara River towards
north, Izvoru Muntelui-Bicaz reservoir towards east, Bicaz River towards south, and the
streams Pntec and Bistra towards south-west [5].
The studied communities lay between 530 m and 1200 m of altitude, which
corresponds to the mixed forest and coniferous forest zones.
We used the classical method for vegetation research, by J. Braun-Blanquet,
completed and adapted to the local conditions [2]. The relevs were sampled various
conditions in regard to the altitude, exposition, slope, and periods of the vegetation season,
University Al. I. Cuza Iai, Faculty of Biology, Carol I, no. 20A, 700506, Iai, Romania.
128
with the aim to achieve the complete picture of the grass layer composition. Consequently,
the relevs ware analysed for the identification of the associations to which they belong.
The survey has been accomplished form May to September. According to the
Phytosociological Code of Nomenclature, the description of the associations resulted from
the analyses of five to ten relevs [2].
We found the following associations:
Class Mulgedio-Aconitetea Hada in Klika 1948
Order Rumicetalia alpini Mucina in Karner et Mucina 1993
Alliance Rumicion alpini Rbel ex Klika in Klika et Hada 1944
1. Ass. Potum supinae Brun-Holl 1962 em. Gutte
1969
2. Ass. Rumicetum alpini Beger 1922
1. Ass. Potum supinae Brun-Holl 1962 em. Gutte 1969 (tab. I)
- the association is spread in the mountain zone, on stripped and compacted
grounds where animals are stationed for a long time, around sheepfolds and households.
- it grows on soils that are rich in nitrates, and on which the original vegetation
still resists.
- there are many species of the class Molinio-Arrhenatheretea that stands for the
fact that the communities of this association form small patches in the meadows of Festuca
rubra and Agrostis tenuis.
- the phytocoenoses are dominated by Poa supina, which forms a dense layer
together with the species Alchemilla vulgaris agg., Veronica persica, Polygonum aviculare,
Stellaria nemorum, Plantago major, P. media, Potentilla anserina, Stellaria media,
Geranium pusillum, Cerastium alpinum, Urtica urens etc.
- this layer is penetrated here and there by the high stems of some species like:
Cirsium vulgare, Festuca rubra, Sisymbrium officinale, Matricaria discoidea, Trifolium
pratense, Campanula abietina, Ranunculus acris ssp. friesianus, Rumex acetosa, Cirsium
arvense, Galeopsis speciosa etc.
2. Ass. Rumicetum alpini Beger 1922 (tab. I)
- it is a nitrophilous mountain association that occurs on grounds stripped for a
long time, on sufficiently humid soils.
- on these grounds, the original vegetation disappeared almost completely.
- these communities grow along some streams, on soils rich in nutrients resulted
from the vegetation matter decomposition or brought from higher regions.
- in the Ceahlu Mountain, the communities with Rumex alpinus are rare and
limited to small areas [6].
- the species richness is small
- Rumex alpinus dominates the communities together with Urtica dioica and
Ranunculus repens [7].
129
- other highly constant species are: Chenopodium bonus-henricus, Stellaria media,

Veratrum album, Poa pratensis, Myosotis scorpiodes etc.
Table I. Associations of the alliance Rumicion alpini Rbel ex Klika in Klika et Hada
1944
Association
Altitude (m.s.m.)
Orientation
Slope (degrees)
Vegetation cover (%)
No. of surveys
Caract. ass.
Poa supina
Rumex alpinus
Rumicion alpini et Rumicetalia alpine
Capsella bursa-pastoris
Chenopodium bonus-henricus
Galeopsis tetrahit
Stellaria media
Urtica dioica
Mulgedio-Aconitetea
Milium efffusum
Rumex obtusifolius
Veratrum album ssp. lobelianum
Hypericum maculatum
Veronica serpyllifolia
Ranunculus repens
Achillea distans
Alchemilla vulgaris agg.
Molinio-Arrhenatheretea
Plantago major
Festuca rubra
Agrostis capillaris
Taraxacum officinale
Lolium perenne
Polygonum aviculare
Prunella vulgaris
Potentilla anserina
Trifolium pratense
Ranunculus acris ssp. friesianus
Rumex acetosa
130
Potum supinae
650-1200
V-E
0-10
80-100
7
Rumicetum alpini
920-1200
V
0-10
80-100
5
V
II
II
V
V
II
I
I
IV
I
III
III
V
I
II
II
I
II
I
II
II
I
I
V
II
I
V
I
II
V
IV
II
II
I
I
I
I
I
I
I
I
I
Association
Lotus corniculatus
Plantago lanceolata
Cynosurus cristatus
Poa pratensis
Trifolium repens
Variae syntaxa
Cirsium vulgare
Stellaria nemorum
Sisymbrium officinale
Veronica persica
Arctium lappa
Veronica chamaedrys
Senecio rupestris
Elsholtzia ciliata
Matricaria discoidea
Urtica urens
Myosotis alpestris
Matricaria recutita
Arctium minus
Plantago media
Cerastium alpinum
Campanula abietina
Geranium pusillum
Malva pusilla
Cirsium arvense
Polygonum hydropiper
Lappula squarosa
Galeopsis speciosa
Myosoton aquaticum
Myosotis scorpioides
Geranium phaeum
Torillis japonica
Stachys sylvatica
Potum supinae
I
I
I
IV
Rumicetum alpini
I
III
II
I
III
II
II
II
II
II
I
I
I
II
I
I
I
I
I
I
I
I
I
I
I
-
II
I
I
I
I
I
I
I
I
II
I
I
I
I
Class Galio-Urticetea Passarge ex Kopecky 1969

Order Lamio albi-Chenopodietalia boni-henrici Kopecky 1969
Alliance Galio-Alliarion (Oberd. 1957) Lohmeyer et Oberd. in
Oberd. et al. 1957
3. Ass. Sambucetum ebuli Felfldy 1942
Order Convolvuletalia sepium R. Tx. 1950 em. Mucina 1993
131
Alliance Petasition officinalis Sillinger 1933

4. Ass. Telekio-Petasitetum hybridi (Morariu 1967)
Resm. et Raiu 1974
3. Ass. Sambucetum ebuli Felfldy 1942 (tab. II)
- occurs in dumping grounds, stream shores near localities, ruderal meadows with soil rich
in organic matter.
- the characteristic species Sambucus ebulus forms dense clusters, which sometimes are
almost pure or it can associate with ecologically different species.
- over 70% of the species are mesophilous whereas the rest are mesohygrophilous,
hygrophilous, xerophilous, and xeromesophilous.
- the frequent species are: Urtica dioica i Galium aparine
4. Ass. Telekio-Petasitetum hybridi (Morariu 1967) Resm. et Raiu 1974 (tab. II)
- occurs on stream shores on alluvial deposits, on shaded or partly shaded areas with highly
humid substrata.
- 25% of the species of the floristic composition are hygrophilous and mesohygrophilous,
higrofile i mezohigrofile, but the great majority of the species are mesofilous.
- the characteristic species, Petasites hybridus, is also dominant and covers the area with its
broad vigorous leaves.
- the subconstant species are: Mentha longifolia, Cirsium oleraceum, Telekia speciosa,
Urtica dioica, Ranunculus repens etc
- we consider that the studied phytocoenoses belong to the subassociation typicum because
of the high cover of the characteristic species, Petasites hybridus.
Table II. Associations of the alliance Galio-Alliarion (Oberd. 1957) Lohmeyer et Oberd. in
Oberd. et al. 1957 and Petasition officinalis Sillinger 1933
Association
Altitude (m.s.m.)
Orientation
Slope (degrees)
Vegetation cover (%)
No. of surveys
Caract. ass.
Sambucus ebulus
Telekia speciosa
Galio-Alliarion et
Lamio-Chenopodietalia
Aegopodium podagraria
Campanula trachelium
Cardamine impatiens
Chelidonium majus
Sambucetum ebuli
550-950
E, N, SE
0-15
80-100
10
Telekio-Petasitetum hybridi
530-890
S, V, N, SE
0-10
80-100
8
V
-
III
I
I
I
-
132
Association
Geum urbanum
Glechoma hederacea
Lamium maculatum
Lapsana communis
Aegopodion podagrariae et
Impatienti-Stachyon
Anthriscus sylvestris
Geranium robertianum
Heracleum sphondylium
Impatiens noli-tangere
Rumex obtusifolius
Petasition officinalis et
Convolvuletalia sepium
Angelica sylvestris
Carduus personatus
Chaeropyllum hirsutum
Cirsium oleraceum
Filipendula ulmaria
Myosoton aquaticum
Petasites hybridus
Poa trivialis
Balloto nigrae-Robinion
Ballota nigra
Torilis japonica
Galio-Urticetea
Carduus crispus
Eupatorium cannabinum
Galium aparine
Salvia glutinosa
Urtica dioica
Geranium phaeum
Artemisietea et
Stellarietea mediae s.l.
Arctium lappa
Artemisia absinthium
Tussilago farfara
Arctium minus
Convolvulus arvensis
Echium vulgare
Onopordon acanthium
Salvia verticillata
Sambucetum ebuli
I
II
I
I
I
I
II
I
I
I
I
-
II
II
I
I
II
III
II
II
IV
I
I
I
I
III
I
IV
I
I
I
I
III
-
I
II
I
II
II
I
I
I
133
Association
Cirsium arvense
Polygonum convolvulus
Bunias orientalis
Anthemis tinctoria
Nepeta cataria
Veratrum album ssp. lobelianum
Equisetum arvense
Daucus carota
Ranunculus repens
Poa pratensis
Medicago lupulina
Rumex crispus
Potentilla anserina
Alchemilla vulgaris agg.
Potentilla reptans
Elymus repens
Mentha longifolia
Bromus commutatus
Achillea millefolium
Equisetum telmateia
Poa chaixii
Lythrum salicaria
Carex hirta
Plantago major
Plantago lanceolata
Trifolio-Geranietea s.l.
Origanum vulgare
Verbascum lichnytis
Inula conyza
Veronica chamaedrys
Solidago virgaurea
Variae syntaxa
Lepidium campestre
Sambucus nigra
Euphorbia cyparissias
Poa nemoralis
Hypericum maculatum
Stachys sylvatica
Galeopsis speciosa
Sambucetum ebuli
I
I
I
I
I
I
I
-
II
II
II
I
I
I
I
I
I
I
I
I
I
I
III
II
II
III
II
II
II
I
I
-
II
I
I
I
I
I
I
I
I
I
I
I
134
Association
Vicia sepium
Hypericum hirsutum
Scrophularia nodosa
Lysimachia vulgaris
Orobanche caryophyllacea
Carex remota
Campanula rapunculoides
Lycopus europaeus
Valeriana tripteris
Euphorbia amygdaloides
Rubus idaeus
Sambucetum ebuli
I
I
I
-
I
II
I
I
I
I
I
I
REFERENCES
1.
2.
3.
4.
5.
6.
7.
8.
9.
BURDUJA C., 1968 - Muntele Ceahlu. Flora i vegetaia. Ocrot. nat., Bucureti, 6: 63 92.
BRAUN-BLANQUET, J., 1964 Pflanzensoziologie, 3, Aufl., Springer, Wien, 865
CHIFU T., MITITELU D., DSCLESCU D., 1987 - Flora i vegetaia judeului Neam. Mem. Sec. t. Acad.
Rom. Seria IV, 10(1): 281 302.
CHIFU, T., MNZU, C., ZAMFIRESCU, O., 2006 - Flora i vegetaia Romniei, vol. 2. Vegetaia. Ed.
Univ. Al. I. Cuza, Iai.
GRINESCU I., 1924 -Consideration gobotaniques sur le mont Ceahlu ( Carpates Orientales). Bul. Soc. St.
Cluj, 2(2): 104 112.
NYARADY E., 1924 -Contribuii la cunoaterea vegetaiei i florei muntelui Ceahlu. Bul. Grd. Bot. i Muz.
Bot. Cluj, 4: 2 3.
RVRU M., 1936 - Nouti din flora Muntelui Ceahlu, Distr. Neam. Bul. Grd. Bot. Muz. Bot. Univ. Cluj,
16 (1 4): 78 85.
SANDA V., POPESCU A., STANCA D., 2001 -Structura cenotic i caracterizarea ecologic a fitocenozelor
din Romnia. Ed. Conphis.
ZANOSCHI V., 1971 - Flora i vegetaia masivului Ceahlu. Tez de doctorat. Cluj Napoca
135

CONTRIBUTIONS TO THE VEGETATION STUDY FROM THE

VASLUI RIVER BASIN (II)
IRINA IRIMIA
Abstract: The paper presents 6 vegetal associations, 2 belonging to the Bidentetea tripartiti R. Tx. et al.
ex von Rochow 1951 class, 3 to the Galio Urticetea Passarge ex Kopeck 1969 class and 1 to the
Epilobietea angustifolii R. Tx. et Preising ex von Rochow 1951 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 association taken into account were classified in the following
phytocoenosystem, taking into account the recent papers on phytosociological classification
[2, 6, 11-16]:
BIDENTETEA TRIPARTITI R. Tx. et al. ex von Rochow 1951
BIDENTETALIA TRIPARTITI Br.-Bl. et R. Tx. ex Klika et Hada 1944
BIDENTION TRIPARTITI Nordhagen 1940 em. R. Tx. in Poli et J. Tx. 1960
1. Ass. Polygono lapathifolii Bidentetum tripartiti Klika 1935
2. Ass. Bidentetum cernui Kobendza 1948
GALIO URTICETEA Passarge ex Kopeck 1969
LAMIO ALBI CHENOPODIETALIA BONI HENRICI Kopeck 1969
GALIO ALLIARION (Oberd. 1957) Lohmeyer et Oberd. in Oberd. et al. 1957
3. Ass. Sambucetum ebuli Felfldy 1942
CONVOLVULETALIA SEPIUM R. Tx. 1950 em. Mucina 1993
SENECION FLUVIATILIS R. Tx. 1950
4. Ass. Urtico Convolvuletum Grs et Mller 1969
5. Ass. Galegetum officinalis Dobrescu et Vialariu 1981
EPILOBIETEA ANGUSTIFOLII R. Tx. et Preising ex von Rochow 1951
ATROPETALIA Vlieger 1937
ATROPION Br.-Bl. et Aichinger 1933
6.
Ass. Eupatorietum cannabini R. Tx. 1937

Alexandru Ioan Cuza University, Faculty of Biology. B-dul Carol I, 20 A, Iai, Romania
136
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 [5].
Ass. Polygono lapathifolii Bidentetum tripartiti Klika 1935
(Syn.: Bidentetum tripartiti Libbert 1932, Bidentetum tripartiti sesu auct.)
Chorology: Dneti, Deleni, Fereti, Munteni de Sus (Leocov M., 1972) Moara
Grecilor (Mititelu D., 1975), Vaslui, Vleni (Mititelu D. and collab., 1996), Vrriei hill
Ecology: It is a nitrophilous association growing in marshes, on the border of the
water and in wet micro-depressions where grows on areas rich in decomposing organic
materials.
The phytocoenological characterization: The characteristic species is Bidens
tripartita which covers 65-80%. We also find other characteristic species of the Bidention
alliance, of the order and class of: Bidens cernua, Polygonum lapathifolium, Lycopus
europaeus etc (tab. I).
After the analysis of the surveys undertaken, the following was noticed:
- the spectrum of bioforms indicates the dominance of hemicryptophytes (50%),
followed by terophytes (38,88%), hydrohelophytes (5,56%) and chamaephytes (5,56%);
- the phytogeographical spectrum shows us the predominance of Eurasian
elements (55,56%), followed by those circumpolar (27,77%), cosmopolite (11,11%) and
European (5,56%);
- the spectrum of ecological indices shows the prevalence of heliophilous species
(38,88%), amphitolerant to temperature (33,34%), mesohydrophilous (33,34%),
amphitolerant to soil reaction (72,22%) and which grow on soils with a high concentration
of mineral nitrogen (38,88%).
Observations: The association was mentioned as part of the studied area, by M.
Leocov (1972) presenting only the floristic composition of the association and by D.
Mititelu (1975, 1996) without having a table with floristic surveys.
Ass. Bidentetum cernui Kobendza 1948
(Syn.: Bidentetum cernui Slavni 1951)
Chorology: Micleti (Mititelu D.,1975), Vaslui
Ecology: The association comprises annual hydrophilous species that appear as
stripes on the border of the water where it grows on areas rich in decomposing organic
materials.
The phytocoenological characterization: The characteristic and dominant
species is Bidens cernua together with Bidens tripartita, Echinochloa crus-galli, Myosoton
aquaticum etc. We may also find characteristic species of the class PhragmitiMagnocaricetea that emphasize the hydrophilic character of the association (tab. II).
137
- the spectrum of bioforms indicates the dominance of hemicryptophytes,
(44,45%), followed by terophytes (33,35%), hemiterophytes (5,55%), hydrohelophytes
(5,55%), phanerophytes (5,55%) and geophytes (5,55%);
elements (50%), followed by those circumpolar (27,77%), cosmopolite (11,11%), Pontic
(5,56%) and European (5,56%);
- the spectrum of ecological indices shows the prevalence of heliophilous species
(35,29%), mesothermal (35,30%), developing on humid wet soils (29,42%), which grow
on soils with a high concentration of mineral nitrogen (41,18%), amphitolerant to soil
reaction (52,95%).
Observations: The association was quoted by D. Mititelu (1975) without floristic
surveys.
Ass. Sambucetum ebuli Felfldy 1942
Chorology: Micleti (Mititelu D.,1975), Codeti, Dneti, Soleti, Tcuta,
Vaslui, Vleni (Mititelu D. and collab., 1996), Chirceti, Coropceni, Emil Racovi
Ecology: This association was encountered under the form of compact clusters, of
variable dimensions, near the households, on the place of abandoned sheepfolds, where the
animals stood, and the substrate is rich in organic substances in decomposition.
We noticed a fast evolution of this associations extension within the territory
taken in the study, in the meadows that are frequently grazed by the sheep, appearing under
the form of groves, and on the border of the roads, thus forming green, high fences.
species is Sambucus ebulus, which forms a layer with coverage of 75-100%. Because of the
strongly developed system of rhizomes, this species plays an important role in
consolidating the eroded lands (tab. III).
The floristic composition of the association is relatively rich in species (51
species), noticing that besides the species characteristic to the Galio-Urticetea class, the
also appear species from the Festuco-Brometea class and Molinio-Arrhenatheretea class,
which come from the neighboring meadows.
- from the spectrum of bioforms we notice the predominance of
hemicryptophytes (58,48%), followed by terophytes (15,09%), hemiterophytes (13,21%),
geophytes (5,67%), phanerophytes (5,67%) and chamaephytes (1,88%);
elements (49,05%), followed by the European (9,43%), cosmopolite (11,32%), circumpolar
(7,54%), continental Eurasian (5,65%), central European (3,78%), adventive (3,78%),
Pontic-Mediterranean (3,78%), Pontic-Balkan (1,89%), Asian (1,89%) and Mediterranean
ones (1,89%);
- from the spectrum of ecological indices we notice that the species which form
the association are heliophile (35,55%), amphitolerant species towards the temperature
138
indece (37,77%), being spread in the central Europe (28,88%), which grow on dry up to
humid soils (4-28,88%, 5-24,45%), amphitolerant towards the reaction of the soils (60%)
and the contents in mineral nitrogen (24,44%).
Observations: The association has been mentioned from the area taken in the
study, but without having presented a table with floristic surveys.
Ass. Urtico Convolvuletum Grs et Mller 1969
(Syn.: Urticetum dioicae Steffen 1931)
Chorology: Codeti, Vleni, Soleti, Vaslui, Dneti, Micleti, Tcuta (Mititelu
D. and collab., 1996)
Ecology: The association was encountered on the places where garbage is thrown,
therefore on the lands rich in decomposing organic substances, usually near the villages,
parks, and even at the border of forests.
species is Urtica dioica, which covers a surface of 90-100%, besides which the species
Convolvulus arvensis also grows. In the floristic composition, few species are present,
accompanying or being characteristic for other classes as well (tab. IV).
- the spectrum of bioforms reveals us the predominance of hemicryptophytes
(63,62%), followed by geophytes (13,64%), terophytes (13,64%) and hemiterophytes
(9,10%);
- the phytogeographical spectrum indicates us the predominance of Eurasian
elements (45,45%), followed by the cosmopolite (22,73%), circumpolar (13,64%),
European (9,10%), Mediterranean (4,54%) and continental Eurasian ones (4,54%);
- from the spectrum of ecological indices we notice that the species which
compose the association are heliophile (42,10%), mesothermal (36,84%), spread in central
Europe (36,84%), which are developed on dry up to moderately humid soils (31,57%), with
a high content of mineral nitrogen (36,85%), amphitolerant towards the soil reaction
(52,64%).
Observations: The association was mentioned from the area, but without
presenting floristic surveys.
Ass. Galegetum officinalis Dobrescu et Vialariu 1981
(Syn.: Senecio biebersteinii-Galega officinalis Borza 1960 n.n.)
Chorology: Vaslui
Ecology: It forms compact phytocoenoses situated at the border of rush-beds or in
some slightly lowland, damp-humid or even with excess of humidity places, on glazed
soils.
The phytocoenological characterization: Besides Galega officinalis, which is the
most important one, there are also the species characteristic to the classes Galio-Urticetea,
Artemisietea i Stellarietea. Among the species with a high constancy we mention
Taraxacum officinale, Tanacetum vulgare, Polygonum dumetorum, Lolium perenne,
Stellaria media etc. (tabel V).
139
- the spectrum of bioforms indicates the predominance of hemicryptophytes

(58,06%), followed by terophytes (22,58%), geophytes (16,13%) and hemiterophytes
(3,23%);
- the phytogeographical spectrum ; reveals the preponderance of Eurasian
elements (51,61%) and cosmopolite (22,58%), followed by the circumpolar (16,12%),
Pontic-Mediterranean (3,23%), Mediterranean (3,23%) and continental Eurasian ones
(3,23%);
- the spectrum of ecological indices indicates species which love the light
(40,74%), which are developed on humid soils (22,22%) and rich in mineral nitrogen
(25,92%). The species are amphitolerant as regards the temperature (48,15%) and soil
reaction (59,25%).
Observations: The association is quoted for the first time from the area taken in
the study.
Ass. Eupatorietum cannabini R. Tx. 1937
Chorology: Dobrov, Poiana, Slobozia (Dobrescu C., 1978), Brnova, Poiana cu
Cetate, Poieni (Mititelu D. and collab., 1995)
Ecology: This association in encountered in different biotypes with humid and
shadowed substrate: in forest cuts, in glades, forest margins and bushes, parks, along some
rivers, confluents of Vaslui river, on soils rich in nutritive elements.
The phytocoenological characterization: Besides the characteristic species
Eupatorium cannabinum there also appear species from the classes Galio-Urticetea,
Artemisietea vulgaris i Molinio-Arrhenatheretea. Among these, we mention Sambucus
ebulus, Geum urbanum, Epilobium hirsutum, Urtica dioica etc. (tab. VI).
- from the spectrum of bioforms, we notice the predominance of
hemicryptophytes (65,21%), followed by terophytes (13,05%), hemiterophytes (17,39%)
and geophytes (4,35%);
- the phytogeographical spectrum reveals the predominance of Eurasian
elements (65,21%), reflecting the temperate character of climate, followed by the
circumpolar (17,39%), cosmopolite (8,70%) and adventive ones (8,70%);
- from the spectrum of ecological indices it results that the species comprising
the association love the light (33,33%), are mesothermal (42,86%), being spread in the
central Europe (66,66%), which are developed on moderately-humid up to humid soils (623,81%, 8-23,81%), with a high content of mineral nitrogen (33,33%), amphitolerant
towards the soil reaction (42,85%).
REFERENCES
1.
2.
3.
BELDIE AL., 1977 - Flora Romniei - Determinator ilustrat al plantelor vasculare. Vol. I-II. Ed. Acad.
R.S.R., Bucureti
CHIFU T., MNZU C., ZAMFIRESCU O., 2006 - Flora & vegetaia Moldovei (Romnia). Vol. II. Ed.
Univ. Al. I. Cuza, Iai: 519-551, 666-687
CIOCRLAN V., 2000 - Flora ilustrat a Romniei - Pteridophyta et Spermatophyta, Ed. Ceres, Bucureti
140
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
DOBRESCU C., 1978 - Completri la cercetrile fitocenologice din Podiul Central Moldovenesc. An. t.
Univ. Al. I. Cuza Iai, s. II a (Biol.), 24: 11-13
ELLENBERG H., 1974 - Indicator values of vascular plants in Central Europe. Scripta Geobotanica, Vol.
IX, Verlag Erich Goltze K.G., Gttingen: 1-97
GEIELBRECHT TAFERNER L., MUCINA L., 1993 - Bidentetea tripartiti In: MUCINA L.,
GRABHERR G., ELLMAUER T. Die pflanzengesellschaften sterreichs, Gustav Fischer Verlag Jena
Stuttgart New York, Bd. I: 90-109
LEOCOV M., 1972 - Contribuii la studiul agro- i geobotanic al buruienilor din bazinul Vasluie. Tez de
doctorat, Instit. Agron. Ion Ionescu de la Brad Iai, Facultatea de Agricultur
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 Mariana, 1996 - Noi contribuii la flora i vegetaia judeului Vaslui. St. i Cerc.
Muz. Piatra-Neam, 8: 193-211
MUCINA L., 1993 - Galio Urticetea In: MUCINA L., GRABHERR G., ELLMAUER T. Die
pflanzengesellschaften sterreichs, Gustav Fischer Verlag Jena Stuttgart New York, Bd. I: 203-251
MUCINA L., 1993 - Epilobietea angustifolii In: MUCINA L., GRABHERR G., ELLMAUER T. Die
pflanzengesellschaften sterreichs, Gustav Fischer Verlag Jena Stuttgart New York, Bd. I: 252-270
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. Ed. Vergiliu,
Bucureti
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
SANDA V., POPESCU A., STANCU D., 2001 - Structura cenotic i caracterizarea ecologic a
fitocenozelor din Romnia. Ed. Conphis, Bucureti
Tab. I. Ass. Polygono lapathifolii Bidentetum tripartiti Klika 1935
Number of survey
Altitude (m.s.m.)
Cover of the vegetation (%)
Surface of survey (m)
Number of species
Associations characteristics
Bidens tripartita
Polygonum lapathifolium
Bidention tripartiti
Bidens cernua
Polygonum mite
Rumex conglomeratus
Bidentetalia et Bidentetea
Lycopus europaeus
Echinochloa crus-galli
Myosoton aquaticum
Mentha longifolia
Ranunculus sceleratus
Phragmito-Magnocaricetea
Ranunculus repens
Alisma plantago-aquatica
Epilobium hirsutum
Juncus inflexus
Agrostis stolonifera
Lysimachia nummularia
Mentha pulegium
1
94
80
10
9
2
330
70
10
9
3
94
80
10
5
4
90
50
10
11
5
220
80
10
8
3
+
4
+
4
-
3
+
4
-
V
III
+
+
+
+
-
1
-
+
-
II
II
I
I
1
-
+
+
+
+
-
+
+
+
+
+
+
+
+
-
III
III
III
III
I
+
+
+
-
1
+
-
+
+
1
-
IV
II
II
1
-
+
-
+
+
+
+
+
-
III
II
I
I
141
Place and date of the surveys:

1,3. Vaslui, 20.08.2003, 12.08.2003; 2. Vrriei hill, 23.08.2003; 4.
Moara Grecilor, 12.08.2003; 5. Fereti, 08.2001
Tab. II. Ass. Bidentetum cernui Kobendza 1948
Number of survey
Altitude (m.s.m.)
Number of species
Bidens cernua
Bidention tripartiti
Bidens tripartita
Bidentetalia et Bidentetea
Echinochloa crus-galli
Myosoton aquaticum
Polygonum lapathifolium
Ranunculus sceleratus
Rorippa austriaca
Phragmiti-Magnocaricetea
Alisma plantago-aquatica
Lythrum salicaria
Typha latifolia
1
94
100
10
11
2
94
80
10
4
3
94
60
10
7
4
94
65
10
6
5
94
70
10
12
+
+
+
-
+
-
+
-
+
+
V
II
+
-
+
-
+
+
+
+
1
+
-
+
+
+
+
-
IV
III
II
II
I
+
+
-
+
-
+
+
II
II
I
1
+
+
+
-
2
-
+
-
IV
I
I
+
+
-
I
I
I
Juncus inflexus
Cichorium intybus
Variae syntaxa
Arctium lappa
Lathyrus tuberosus
Salix alba

1-5. Vaslui, 12.08.2003
Tab. III. Ass. Sambucetum ebuli Felfldy 1942
Number of survey
1
2
3
Altitude (m.s.m.)
190
210
260
Exposition
NE
S
S
Slope ()
1
1-2
1
100
100
75
25
25
25
Number of species
18
14
12
Sambucus ebulus
5
5
4
Galio-Alliarion et Lamio albi-Chenopodietalia
Aristolochia clematitis
+
Veronica chamaedrys
+
Galio-Urticetea
Galium aparine
+
Glechoma hederacea
+
Geum urbanum
+
+
Onopordetalia
Berteroa incana
+
+
1
Tanacetum vulgare
+
Artemisia absinthium
+
Carduus nutans
-
142
4
260
SV
2
100
25
18
5
180
NV
1-2
100
25
23
6
210
7
210
90
20
11
90
20
12
+
-
+
+
III
III
+
+
-
1
1
-
+
+
III
III
III
+
+
-
III
II
I
I
Artemisia vulgaris
Arctium tomentosum
Urtica dioica
Ballota nigra ssp. nigra
Elymus repens
Carduus acanthoides
Conium maculatum
Rumex obtusifolius
Erigeron annuus
Helianthus tuberosus
Leonurus cardiaca ssp. villosus
Carduus crispus
Torilis arvensis
Conyza canadensis
Polygonum aviculare
Cardaria draba
Cirsium arvense
Lathyrus tuberosus
Atriplex tatarica
Sonchus oleraceus
Achillea setacea
Eryngium campestre
Salvia nemorosa
Galium humifusum
Poa angustifolia
Galium verum
Scabiosa ochroleuca
Bromus inermis
Erodium cicutarium
Galium mollugo
Cichorium intybus
Lolium perenne
Centaurea jacea
Plantago lanceolata
Lotus corniculatus
Juncus inflexus
Achillea millefolium
Lysimachia nummularia
Dactylis glomerata
Poa pratensis
Prunus spinosa
Rosa canina
Elaeagnus angustifolia
Artemisietea vulgaris
+
+
1
+
+
+
+
Stellarietea mediae
+
+
+
+
+
+
+
+
+
+
+
Festuco-Brometea
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
Variae syntaxa
+
+
+
+
+
+
-
+
+
+
+
+
+
-
+
1
+
-
+
2
+
+
+
+
III
III
III
III
III
III
II
I
I
I
I
I
+
+
-
II
II
II
II
II
II
I
I
+
+
+
-
III
II
II
II
I
I
I
I
I
I
I
+
+
+
+
+
+
-
+
+
+
+
+
III
III
III
II
II
I
I
I
I
I
+
+
II
I
I

1. Chirceti, 6.08.2003; 2. Coropceni, 27.07.2003; 3,4. Emil Racovi, 6.08.2003; 5.
Codeti, 07.2002; 6,7. Dobrov, 1.07.2004
Tab. IV. Ass. Urtico Convolvuletum Grs et Mller 1969
Number of survey
Altitude (m.s.m.)
Number of species
Urtica dioica
1
150
100
25
9
2
220
100
15
10
3
160
100
20
12
4
150
100
10
10
5
94
100
20
12
143
Petasition officinalis et Convolvuletalia sepium

Galium aparine
+
+
+
Cucubalus baccifer
+
+
+
+
+
Sambucus ebulus
+
+
Galio-Urticetea
+
+
+
Veronica chamaedrys
+
+
+
Glechoma hederacea
+
+
+
+
+
Carduus acanthoides
+
+
Elymus repens
+
+
Elymus hispidus
+
Arctium tomentosum
Leonurus cardiaca ssp. villosus
Stellarietea mediae
Capsella bursa-pastoris
+
+
Lathyrus tuberosus
+
Lepidium ruderale
+
-
+
+
-
IV
III
III
II
+
-
IV
III
II
+
+
+
IV
II
II
I
I
I
+
+
+
III
II
II
Bellis perennis
Lolium perenne
Verbena officinalis
Geum urbanum
Agrimonia eupatoria
+
+
+
+
Variae syntaxa
-
+
-
+
+
+
-
III
III
II
+
-
I
I

1, 2. Codeti, 5.08.2005; 3. Vleni, 5.08.2005; 4. Soleti, 5.08.2005; 5.
Vaslui, 5.08.2005
Tab. V. Ass. Galegetum officinalis Dobrescu et Vialariu 1981
Number of survey
1
2
Altitude (m.s.m.)
94
94
90
100
10
10
Number of species
12
14
Galega officinalis
5
5
Senecion fluviatilis et Convolvuletalia sepium
Polygonum dumetorum
+
+
+
Calystegia sepium
+
Galio-Urticetea
Urtica dioica
+
Ranunculus repens
+
+
+
Tanacetum vulgare
+
Lathyrus tuberosus
+
Elymus repens
+
Artemisia vulgaris
Verbena officinalis
Cannabis sativa ssp. spontanea
1
Equisetum arvense
+
Stellarietea mediae
Stellaria media
+
Linaria vulgaris
-
144
3
94
95
10
13
4
94
70
10
12
5
94
55
10
16
+
-
+
-
III
II
I
+
+
-
+
-
II
II
I
+
+
+
+
+
+
+
-
+
+
+
+
-
+
+
+
+
+
+
+
V
IV
III
III
III
III
II
II
+
-
+
+
III
II
Cirsium arvense
Lolium perenne
Lotus corniculatus
Mentha longifolia
Trifolium repens
Inula britannica
Bidens tripartita
Lycopus europaeus
Xanthium strumarium
Polygonum aviculare
Molinio-Arrhentheretea
+
+
+
+
+
Bidentetea tripartiti
+
+
+
Variae syntaxa
+
+
-
+
+
+
-
+
+
-
+
+
+
+
-
III
III
III
II
II
I
1
-
II
I
I
I
I

1-5. Vaslui, 20.08.2003
Tab. VI. Ass. Eupatorietum cannabini R. Tx. 1937
Number of survey
Altitude (m.s.m.)
Number of species
Eupatorium cannabinum
Atropion et Atropetalia
Sambucus ebulus
Cirsium vulgare
Epilobietea angustifolii
Conyza canadensis
Galio-Urticetea
Urtica dioica
Epilobium hirsutum
Erigeron annuus
Geum urbanum
Myosoton aquaticum
Sambucus ebulus
Lapsana communis
Arctium lappa
Dipsacus fullonum
Ranunculus acris
Lythrum salicaria
Symphytum officinale
Prunella vulgaris
Mentha pulegium
Equisetum telmateia
Stachys sylvatica
Scrophularia nodosa
1
370
70
50
7
2
350
75
50
9
3
393
100
50
9
4
393
100
25
10
5
393
100
25
12
+
+
+
-
+
-
III
II
II
+
+
1
+
-
+
+
-
+
+
-
IV
III
I
+
-
+
+
-
+
-
+
+
-
+
+
+
+
IV
II
I
I
I
I
+
+
+
-
+
+
+
+
-
III
II
II
II
I
I
+
+
-
+
-
+
+
III
II
I
+
+
+
+
Querco-Fagetea
+
-

1. Dobrov, 12.07.2003; 2. Poieni, 08.2002; 3-5. Brnova, 08.200
145
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ANALELE TIINIFICE

UNIVERSITII Alexandru Ioan Cuza

TOMUL LIV, FASCICULA 2

Editura Universitii Alexandru Ioan Cuza Iai

Editura Universitii Alexandru Ioan Cuza din Iai

Comisia de refereni tiinifici:

ANCA HEMCINSCHI, RAMONA GALE, C. TOMA

MARIA MAGDALENA ZAMFIRACHE, C. TOMA, MARIA DUCA,

I. BURZO, V. CIOCRLAN, ELENA DELIAN, AURELIA DOBRESCU,

V. SURUGIU - On the occurrence of Zostera noltii Hornemann at the Romanian

Analele tiinifice ale Universitii Al. I. Cuza Iai

ANATOMICAL RESEARCHES ON THE OVERGROUND VEGETATIVE ORGANS

Faculty of Biology, Bucharest University, Romania

Material and methods

ANDREI M., 1978 - Anatomia plantelor, Edit. Did. .Pedag., Bucureti

The explanation of figures:

M. ANDREI, ROXANA PARASCHIVOIU

M. ANDREI, ROXANA PARASCHIVOIU

M. ANDREI, ROXANA PARASCHIVOIU

M. ANDREI, ROXANA PARASCHIVOIU

Analele tiinifice ale Universitii Al. I. Cuza Iai

MORPHOLOGICAL AND HISTO-ANATOMICAL ASPECTS REGARDING THE

The explantion of figures:

RAMONA GALE et al.

RAMONA GALE et al.

male flower primordium

RAMONA GALE et al.

RAMONA GALE et al.

nectary gland primordium

Analele tiinifice ale Universitii Al. I. Cuza Iai

MICROSPOROGENESIS AND THE MALE GAMETOPHYTE AT EPHEDRA

development in Ephedra distachya L. provides interesting characterization in this species.

The explanation of figures:

GENIANA PREDAN, IRINA GOSTIN

GENIANA PREDAN, IRINA GOSTIN

Analele tiinifice ale Universitii Al. I. Cuza Iai

VEGETATIVE ANATOMY OF TWO GALIUM L. SPECIES (RUBIACEAE)

Pharmacy AncaFarm, Botosani, Romania

The explanation of figures:

ANCA HEMCINSKI et al.

ANCA HEMCINSKI et al.

ANCA HEMCINSKI et al.

ANCA HEMCINSKI et al.

Analele tiinifice ale Universitii Al. I. Cuza Iai

A COMPARATIVE STUDY REGARDING THE MORPHOLOGY AND ANATOMY

Results and discussions

MARIA MAGDALENA ZAMFIRACHE et al.

MARIA MAGDALENA ZAMFIRACHE et al.

MARIA MAGDALENA ZAMFIRACHE et al.

MARIA MAGDALENA ZAMFIRACHE et al.

MARIA MAGDALENA ZAMFIRACHE et al.

Analele tiinifice ale Universitii Al. I. Cuza Iai

A COMPARATIVE NUMERIC ANALYSIS OF THE SECRETORY TRICHOMES

Results and discussions

H11: Da Db Dbr Dbk Dc DcA DcNL DcR Dcp Dcn Dd

The graphical representation of the number of sessile secretory trichomes belonging

no. of sessile trichomes in the lower epidermis

no. of tentacular trichomes

Fig.1. Graphical representation of the number of sessile secretory trichomes of

Fig.2. Graphical representation of the number of pedicelate (tentacular) secretory

no. of sessile trichomes in the upper epidermis

Fig.3. Graphical representation of the number of sessile secretory trichomes

Fig.4. Comparative representation of the number of both pedicelate (tentacular)

Fig.5. Comparative representation of the number of sessile secretory trichomes