Rezumate Volum DePOSTERE 52 (1)

A New Approach of the Academic Research in BiologyBook of abstracts Editors: Octavian Popescu, Mădălin Enache, Medana Zamfir, Sorin Ştefănuţ Text set: Sorin Ştefănuţ Cover: Lucian Apetre Scientific Committee: Prof. Dr. Octavian POPESCU – Director of Institute of Biology Bucharest; Dr. Medana ZAMFIR – Scientific Secretary of Institute of Biology Bucharest; Dr. Nicolae MIRANCEA – Head of Plant and Animal Cytobiology Department; Dr. Sorin ŞTEFĂNUŢ – Head of Ecology, Taxonomy and Nature Conservation Department; Dr. Mădălin ENACHE – Institute of Biology Bucharest. EDITURA ARS DOCENDI – UNIVERSITATEA DIN BUCUREŞTI Editură cu profil academic şi cultural, recunoscută de CONSILIUL NAŢIONAL AL CERCETĂRII ŞTIINŢIFICE DIN ÎNVĂŢĂMÂNTUL SUPERIOR Şos. Panduri 90, sector 5, Bucureşti Tel./Fax: +4 021 410 25 75 www.arsdocendi.ro [email protected] Descrierea CIP a Bibliotecii Naţionale a României 50 YEARS OF ACADEMIC RESEARCH IN BIOLOGY. Conference ( 2010 ; Bucureşti) 50 years of academic research in biology : the anniversary conference of the Institute of Biology : Bucharest, 2010 : book of abstracts / Institute of Biology Bucharest. - Bucureşti : Ars Docendi, 2010 ISBN I. Institutul de Biologie (Bucureşti) 061.62(498 Buc.):57 Copyright © Institute of Biology Bucharest, 2012 Printed in Romania, Institute of Biology Bucharest Institute of Biology Bucharest A New Approach of the Academic Research in Biology Book of abstracts ARS DOCENDI 2012 UNIVERSITY OF BUCHAREST . D. ANDRONIC 30 MISSING-LINK CARBAMOYL PHOSPHATE SYNTHETASE E. ENACHE 34 MICROBIAL DIVERSITY IN THE SUBTERRANEAN ICE DEPOSITS OF SCĂRIŞOARA CAVE A. AND CHALLENGES M. C.IBB INFRASTRUCTURE DEVELOPMENT FOR ENHANCING THE BIODIVERSITY INVESTIGATION CAPACITY. BRAD. A. G. C. TEODOSIU and M. I. PASCU.Z.CONTENTS THE PROJECT . POPA. STANILOVA 25 CLONAL PROPAGATION AND GENETIC IMPROVEMENT OF SELECTED SMALL FRUIT SPECIES THROUGH BIOTECHNOLOGY APPROACHES A. RUSU. IN ACCORDANCE WITH GLOBAL CLIMATE CHANGES . PERERA.M. BAS – FACILITIES. H. B. HILLEBRAND-VOICULESCU. GUY-EVANS. OREN 17 UNRAVELING OF THE COCOA BEAN FERMENTATION PROCESS OPENS NEW PERSPECTIVES FOR CHOCOLATE PRODUCTION LUC DE VUYST 18 SPECIES AND SPECIATION O. T. PERŞOIU. POPA. IŢCUŞ. DIMITROVA. T. D. ARDELEAN and S. GHIŢĂ 38 . ONAC and C.I. A. KIBÉDI-SZABÓ. GUSSEV. TAXONOMY AND NOMENCLATURE OF PROKARYOTES A.R. TSONEVA. IVANOVA. LIBIAKOVÁ and M. LATEČKOVÁ 28 NUCLEOTYPIC EFFECTS IN VITRO CONDITION OF DIFFERENT CABBAGE GENOTYPES INFECTED WITH CAULIFLOWER MOSAIC VIRUS L. BOSSEVA. GAJDOŠOVÁ. CH. STOEVA PLANT BIOTECHNOLOGY DEVELOPMENT IN THE INSTITUTE OF BIODIVERSITY AND ECOSYSTEM RESEARCH. N.DIBIOCLIM 11 INVITED LECTURES AND ORAL PRESENTATIONS WHAT’S IN A NAME? THOUGHTS ABOUT SPECIES CONCEPTS. Y. POPESCU 22 23 PLANT CONSERVATION TOWARD SOCIETY D. ARDELEAN. EVANS.P. E. C PURCAREA 33 HALOARCHAEA FROM SOME ROMANIAN MAN-MADE AND NATURAL HYPERSALINE ENVIRONMENTS G. V. ACHIEVEMENTS. PURCĂREA 37 DYNAMICS OF MICROBIOTA IN MARINE MICROCOSMS (INDOOR): RESULTS AND PERSPECTIVES I. S. GEORGIEV and T. R. VADRUCCI. ISTRATE and G. S. WALTHER and E.THE GROWTH OF Ulocladium chartarum IN MICROGRAVITY: ON RANDOM POSITIONING MACHINE AND ON BOARD OF INTERNATIONAL SPACE STATION GOMOIU. KERESZTES. A. A. CH.B. I. VĂDINEANU 54 BIODIVERSITY DATA TRANSFERRED IN ECOSYSTEM SERVICES STUDIES M. E. C. D.C. D. PURICE 55 PRELIMINARY ASSESSMENT ON THE EFFECT OF URBANIZATION ON ECTOMYCORRHIZAL DIVERSITY D. V. M. ALDEA. COMAN. C. MUNTEAN. CATANA. BANCIU. ANDREI. MIHĂILESCU. CHATZITHEODORIDIS 39 DIVERSITY OF ARCHAEAL COMMUNITY IN THE CHEMOCLINE OF A HYPERSALINE LAKE IN THE TRANSYLVANIAN BASIN (ROMANIA) Ş. GEORGIEV. HONCIUC and D. HELEPCIUC. F. GHEORGHE and I. BANCIU 40 STATINS AS CANDIDATES FOR TUMOR-TARGETED NANOMEDICINES M. V. D. ONETE. F. BOSSEVA. GUSSEV. PREDA and A. NICOLAE.E. ALEXE and H. MITOI. BARICZ. ANGELOV. V. HOLOBIUC. M.C. ION. CRISTIAN. E. MIHAI. Z. F. R. IVANOVA. JERCA . LICARETE. Fraxinus excelsior AND Quercus robur IN FOREST ECOSYSTEMS UNDER THE INFLUENCE OF ANTHROPOGENIC PRESSURE A. BANCIU 42 EX SITU APPROACH TO CONSERVATION OF LILIACEAE IN BULGARIA T. MIRANCEA 49 51 INVESTIGATING THE IMPACT OF CYANOBACTERIAL BLOOMS ON AQUATIC ECOSYSTEMS OF THE DANUBE DELTA P. G. Y. I. G. MIHĂILESCU 57 59 PROJECT SOP ENVIRONMENT: CONSERVATION STATUS MONITORING OF SPECIES AND HABITATS FROM ROMANIA UNDER THE 17 ARTICLE OF THE HABITATS DIRECTIVE 2011-2012 S. VOICHITA.E. I. M. COGĂLNICEANU 46 TELOCYTES – INFRASTRUCTURE AND FUNCTIONAL ROLE N. S. C. CRISTIAN and M. TSONEVA. CRISTEA . ALUPEI. STOEVA 44 PLANT IN VITRO SYSTEMS FOR EX SITU CONSERVATION AND BIOTECHNOLOGICAL APPLICATIONS I. DIMITROVA and T.-A. M. AIFTIMIE-PĂUNESCU. MOGÎLDEA 56 CITES AND NATURE CONSERVATION IN REPUBLIC OF KENYA S. MINODORA. SPAAK and C. SANDU CONTRIBUTIONS TO THE KNOWLEDGE OF LITTER DECOMPOSITION RATES OF PLANT LITTERS OF Acer pseudoplantanus. MOLDOVEANU 73 THE DYNAMICS OF EXTRACELLULAR β . RADU.GLUCOSIDASE AND ALKALINE PHOSPHATASE IN AQUATIC ECOSYSTEMS FROM THE DANUBE DELTA IN 2011-2012 INTERVAL I.D. METHODS – OLD. RÎŞNOVEANU 69 LONG-TERM DYNAMICS (1993-2011) OF THE BENTHIC MACROINVERTEBRATES COMMUNITIES IN THE INNER DANUBE DELTA C. CIOBOIU 83 . MANU 78 METHODS USED IN THE ECOLOGICAL RESEARCH OF SOIL MITES (ARACHNIDA . G. COMAN and G. PĂCEŞILĂ 74 STRUCTURAL-FUNCTIONAL PARTICULARITIES OF THE ECOSYSTEMS OF A PLAIN CATCHMENT AREA (CASE STUDY) O. PURICE and O. MAICAN and D. RÎŞNOVEANU. PURICE 82 THE ECOLOGICAL SUCCESSION IN TERMS OF THE TROPHIC STRUCTURES IN FOREST PLANTATIONS D. CURRENT AND PERSPECTIVES D. A. S. BREZEANU 76 MODERN METHODS USED IN TAXONOMY AND ECOLOGY OF SOIL FAUNA M. CRISTOFOR 71 THE DYNAMICS OF CYANOBACTERIAL BLOOMS IN LAKE ROŞU REFLECTED IN LONG-TERM STUDIES M. TAXONOMY AND NATURE CONSERVATION POSTERS LONG-TERM ECOLOGICAL RESEARCH OF ZOOPLANKTON COMMUNITY IN LAKE ROŞU (DANUBE DELTA) L. PARPALĂ 65 SPATIAL DISTRIBUTION OF THE ROTIFERS DIVERSITY IN SFÂNTU GHEORGHE BRANCH L. IGNAT and S. FIERA.ACARI) – HISTORY AND PERSPECTIVES – V. HONCIUC 79 THE COMMUNITIES STRUCTURE OF INVERTEBRATES FAUNA FROM RAPE AND ALFALFA CROPS C.C. PURICE 80 THE EPIGEIC INVERTEBRATES IN THE ECOLOGICAL STUDIES IN ROMANIA. BÎRSAN. G. RADU. E. FLORESCU 67 LINKING THE STRUCTURE OF THE WEED-BED FAUNA COMMUNITIES AND THE ENVIRONMENTAL VARIABLES IN SFÂNTU GHEORGHE ARM OF THE RIVER DANUBE E.TURBOVEG DATABASE USED IN BUCEGI MASSIF STUDIES C. NICOLAE 61 ECOLOGY. CIOBOIU and G. MAICAN and C. POPA and A. GROSU-TUDOR. M.M. TSONEVA and V. ONETE 91 92 NON-NATIVE VASCULAR AQUATIC MACROPHYTES IN BULGARIA V.S. ZAMFIR. D.SPECIFIC DIVERSITY OF ENTOMOFAUNA (COLLEMBOLA. GEORGIEV and S. COLEOPTERA) FROM SĂRATA .. C. COBZARU 95 THE ESTIMATION OF COMMUNITY INTEREST MAMMALS IN ROMANIA O. M. R. AXINI and FAWAZ AL-AZKI 85 THE GRAYLING (Thyallus thymallus L. ADAMESCU. CRISTEA 87 88 BIRD RINGING CAMP FROM LETEA. JURJ. (BRASSICACEAE) IN PARÂNG MOUNTAINS (ROMANIAN CARPATHIANS) R. M. ION BIODIVERSITY OF THE FUTURE IN MILLENIUM SEED BANK M. ROMANIA M. 1758). M. FIERA 84 BIOGEOGRAPHYCAL DATA ON MOLLUSKS SPECIES IN DOBROGEA. WOUTERS and L. M. IONESCU. COTOVELEA. CAZACU. G. FEDORCA. G.MONTEORU FOREST (BUZĂU COUNTY) S. DE VUYST 104 . A DECLINING SPECIES I. DANUBE DELTA BIOSPHERE IMPORTANT SITE FOR THE STUDY OF BIRDS MIGRATION I. TSONEVA CONSERVATIONALLY IMPORTANT VASCULAR AQUATIC MACROPHYTES IN BULGARIA S. ŞTEFĂNUŢ 89 90 FIRST RECORD OF Draba dorneri HEUFF. A. COBZARU. MIHĂILESCU. GEORGIEV 93 NATURA 2000 SITES IN THE LOWER OLT HYDROGRAPHIC BASIN KEY SPECIES AND HABITATS FOR THE INTEGRATED MANAGEMENT OF THE AREA S. CISMASIU 103 LACTIC ACID BACTERIA FROM FERMENTED VEGETABLES WITH POTENTIAL APPLICATIONS IN FOOD INDUSTRY AND HEALTH S. ION and I. POCORA THREATENED BRYOPHYTES IN ROMANIA S. GRIDAN 97 MICROBIOLOGY POSTERS THE SELECTION OF THE ACIDOPHILIC BACTERIA BELONGING TO THE Acidiphilium GENUS WITH HIGHER ENZYMATIC ACTIVITY IN THE PRESENCE OF HEAVY METAL IONS C. IONESCU. CHIŞAMERA and V. HELEPCIUC and M.R. S. ROMANIA AND THEIR BIOTECHNOLOGICAL POTENTIAL R. PERŞOIU.C.E. PURCĂREA 114 PLANT AND ANIMAL CYTOBIOLOGY. D. N. PRUNESCU 117 LONG-TERM CULTURES IN MINIMAL GROWTH CONDITIONS IN Dianthus trifasciculatus KIT SSP. M. ARDELEAN. HELEPCIUC.P. PASCU. ANDRONIC. D. NEAGU. STANCU 108 HALOPHILIC BACTERIA FROM DIFFERENT HYPERSALINE HABITATS IN SLANIC PRAHOVA. BREZEANU and C.M. HILLEBRAND-VOICULESCU. PERERA. HRISTEA 120 DEFENSE-RELATED ENZYMES GENE EXPRESSION IN Cucumis sativus L. IORDACHE and D. M. TEODOSIU and M. I. CIUCĂ. APOSTOL .E. GRIGOROV 123 ANTIOXIDANT ACTIVITY OF POLYPHENOLS FROM Vitis vinifera CVS. SMEREA and T. CÎRNU . MITOI. BIOTECHNOLOGY POSTERS FIBROLAMELLAR LIVER CARCINOMA LIKE TUMOR IN AN EXPERIMENTALLY IRON LOADED MOUSE P. PASCU. HELEPCIUC 119 GREEN SYNTHESIS OF SILVER NANOPARTICLES USING PLANT CELL CULTURES G. A. MITOI and G. PRUNESCU and C. C.M. POPA. PURCĂREA 113 CELL GROWTH AND DGGE ANALYSIS OF MICROORGANISMS FROM SCĂRIŞOARA CAVE ICE SEDIMENTS C. A.E. isabelle LONG-TERM CALLUS CULTURE UNDER THE INFLUENCE OF SOME ELICITORS TREATMENT R. SARCHIZIAN .E.STUDIES CONCERNING THE MORPHOLOGICAL AND FUNCTIONAL VARIATIONS OF SOME BACTERIAL CELLS UNDER CONTAMINATION STRESS IMPACT M. BRAD and C.I. COGĂLNICEANU. EVANS and C. G. IŢCUŞ. M. MITOI 125 . S.M. VOICHITA. ENACHE 110 METALIC NANOPARTICLES CITOTOXICITY AGAINST PHOTOTROPHIC AND HETEROTROPHIC BACTERIA I. F. GUY-EVANS. H. D. T. MITOI and F.E. CORNEA 122 DIRECT SOMATIC EMBRYOGENESIS AS POSSIBLE WAY OF VIRUS ELIMINATION FROM SEEDLESS GRAPEVINE L. COJOC. CÎRSTEA 106 ORGANIC SOLVENT RESISTANCE MECHANISMS IN Vibrio alginolyticus IBBCT2 M. APOSTOL 112 STRUCTURE-FUNCTION INVESTIGATION OF Methanobrevibacter smithii FULL-LENGTH CARBAMOYL PHOSPHATE SYNTHETASE E. DAMIAN. MIHAI. parviflorus AS EX SITU PRESERVATION TOOL I. I. PLANTLETS INDUCED BY MICROORGANISMS F.E. A. M. HOLOBIUC. ŞTEFĂNESCU and D. V. M. I.E. SPANIEL. C. BREZEANU 131 . BANCIU and F. SOARE 129 ELECTRONOMICROSCOPICAL ANALYSIS OF THE Usnea barbata (L. BANCIU. HELEPCIUC. ALDEA. DOMŻALSKA. VOICU and A. BREZEANU and L.E. S. L. A. MITOI. F. (LICHENOPHYTA) SYMBIOTROPHIC FORMATIONS REGENERATED IN “IN VITRO” CULTURE D.AN EFFICIENT METHOD FOR SOMATIC EMBRYOGENESIS INDUCTION AND PLANT REGENERATION FOR Serratula bulgarica ACHTAROFF ET STOJ.E. AIFTIMIE-PĂUNESCU 127 GENETIC STABILITY ASSESSMENT OF Scilla autumnalis REGENERANTS C. M.E.) MOTT. HELEPCIUC 128 COMPARATIVE STUDIES BETWEEN FERNS GAMETOPHYTE AND SPOROPHYTE BY BIDIMENSIONAL ELECTROPHORESIS AND POLYPLOIDY ANALYSIS F. EX SITU GERMPLASM CONSERVATION A.C. 874.IBB INFRASTRUCTURE DEVELOPMENT FOR ENHANCING THE BIODIVERSITY INVESTIGATION CAPACITY.693.000 lei Target: Development of IBB infrastructure for enhancing the biodiversity investigation capacity. acquisition of RDI software and hardware) already started a direct contribution to the development of RDI studies of the .2. from microorganisms to plants and animals. research center) Period of implementation: 01.2.THE PROJECT .2012 Project budget: 42. in view of the preservation and the stable management of natural resources and biodiversity. and in accordance with the European Research Area.267 lei Ineligible expenditure: 8. The activities developed in this project (acquisition of modern and performing research equipment. IN ACCORDANCE WITH GLOBAL CLIMATE CHANGES DIBIOCLIM General data of the project: Beneficiary: Institute of Biology Bucharest of the Romanian Academy POS-CCE: O.785 lei National Budget: 7.052 lei FEDR: 25. expansion and organization of laboratories and spaces for Research Development and Innovation (RDI). considering the global climate changes.860.2009 – 31.08.1. in accordance with global climate Investment The project contribute to develop research capacities by enhancing the scientific and technical potential. The main interest of all research departments of the Institute of Biology is the study of species biodiversity from all categories of organisms. – Development of existent R-D infrastructure and creation of novel R-D infrastructures (laboratories.428.03. in accordance with European strategies in research fields. of high interest worldwide – the quantitative and qualitative evaluation of species.and time. European research areas are directed to the evaluation and prediction of biodiversity changes and the characterization of ecosystem dynamics. and offer the possibility of new research domains and directions. with important impact on the advancing of both scientific knowledge and applications in different economy areas (environment. Microbiology researches focuses on (1) the evaluation of taxonomical and physiological diversity of microorganisms isolated from environments characterized by normal and extreme conditions of temperature. including terrestrial and marine environments. medicine). Novelty and impact The current project successfully implemented by the Institute of Biology directs to a very prominent and up-to-date scientific area. evaluation of the diversity of species. population and ecosystem diversity. of natural and human related ecosystems and socio-ecological complexes for the environment protection and durable development. and the conservation of their biodiversity. The major Ecological researches in our institute focused on taxonomical studies of invertebrates and plants isolated from terrestrial and aquatic ecosystems. ecosystems and complexes of natural and human related environments. industry. The Institute of Biology has an excellent research tradition. the production and efficiency of these terrestrial and aquatic ecosystems. the monitoring of biomarkers. the analysis of space. marked by prestigious names of national and international recognition.evolution of populations. hydrostatic . the proposed studies concerning plant and invertebrates biodiversity will be directed in accordance with the international tendencies and strategies. and is known for its constant involvement in priority research directions considered worldwide.environment. Considering the global climate changes. The successfully implementation of this project for infrastructure development started to determine a substantial research progress regarding species diversity. 428.Q TRAPTM LC/MS/MS system and software .000. heavy metals and carbohydrates.Scanning Electron Microscopy and software .X-ray Spectrophotometer and software .052 Lei (11.Atomic Absorption Spectrophotometer and software . 90% of this costs representing a direct investment in infrastructure and RDI equipment.Ultracentrifuge and software . as well as microbial biomarkers for polluted environments. pH.Genetic Analyzer ABIPrism 3500 (Applied Biosystems) and software . Major research equipments purchased into frame of DIBIOCLIM project .Atomic Force Microscopy and software .Transmission Electron Microscopy and software . high concentrations of salt. The total budget was 42. starting in March. 2009.pressure. Period of implementation and costs This Project has been implemented during a period of 42 months. and (2) the identification of cellular and molecular structures and metabolites with bio(nano)technology potential.000 €). . INVITED LECTURES AND ORAL PRESENTATIONS . . 91904 Jerusalem. taking into account phenotypic. The Hebrew University of Jerusalem. and Plants). E-mail: aharon. the number of names of species. Fungi.oren@mail. The two codes are not compatible in many aspects. Israel. The nomenclature of one group of prokaryotes is still highly problematic. However. the names of the higher taxa of prokaryotes (phyla. To illustrate all this I intend to present the halophilic Archaea of the family Halobacteriaceae as an example. genera. based on culture-independent studies using molecular markers (in particular 16S rRNA genes) it is clear that most species of Bacteria and Archaea are still awaiting to be isolated and described. kingdoms or domains) are currently not covered by the rules of the Code. TAXONOMY AND NOMENCLATURE OF PROKARYOTES A. The nomenclature of the prokaryotes. there is no consensus on how to define and to delineate such species. OREN Department of Plant and Environmental Sciences. and the operational definition of a species based on DNA-DNA hybridization is also not always satisfactory.huji. families. 17 . Currently the description of new species is based on a polyphasic approach.WHAT’S IN A NAME? THOUGHTS ABOUT SPECIES CONCEPTS. chemotaxonomic and genotypic properties. Thanks to central registration and indexing. A new beginning was made in 1980 with the publication of the "Approved Lists of Bacteral Names". showing how species concepts and nomenclature have changed over the years and how the occurrence of lateral gene transfer events and possibly even the formation of recombinant hybrids complicate the delineation of species.il The number of different species of prokaryotes with names with standing in the nomenclature has recently surpassed ten thousand. The Institute of Life Sciences. While we have exact information about the number of prokaryote species with validly published names described in the literature. Comparison of small-subunit ribosomal RNA sequences is insufficient to delineate species. and these are classified in about two thousand genera and more than three hundred families. as regulated by the rules of the International Code of Nomenclature of Prokaryotes ("The Bacteriological Code"). a group traditionally covered by the International Code of Botanical Nomenclature (now renamed as the International Code of Nomenclature for Algae. and solutions are currently been sought for the problem. No generally accepted species concept exists yet for the prokaryotes. orders and classes with standing in the nomenclature of prokaryotes is known at any moment. These numbers represent only a very small fraction of the true number of different prokaryote taxa that exist in nature. and that is the Cyanobacteria. is extremely well-ordered.ac. Department of Bioengineering Sciences. AAB are necessary for exothermic oxidation of ethanol (produced by the yeasts) and lactic acid (produced by the LAB) to acetic acid under aerobic conditions. B-1050 Brussels. A particular succession of microbial activities carried out by yeasts. etc. proteases. on-farm processes that differ in methods (heaps. caused by air ingress due to pulp degradation. either manually or mechanically. Cocoa bean fermentations are still spontaneous. Diffusion of 18 . uncontrolled. LAB that develop with the yeasts simultaneously are necessary for citric acid conversion. They also contribute to pulp removal through pectinolysis. Yeasts are important for a fast and consistent ethanol production through anaerobic carbohydrate (glucose) fermentation. Pleinlaan 2. which is fermented for approximately four to six days.) and practices (agricultural and operational practices on the farm) and hence generally result in end-products of variable quality. Belgium Cocoa beans originate from the fruit pods of the cocoa tree. next to lactic acid production by carbohydrate (fructose and glucose) fermentation and mannitol formation through fructose reduction under microaerophilic conditions. it is the cocoa pulp surrounding the cocoa beans that undergoes a fermentation process. They have an astringent and bitter flavor and taste and have to be processed before they can be roasted and converted into chocolate. acetic acid. Penetration of ethanol. platforms. and heat into the beans kills the embryos and disintegrates the beans internally.UNRAVELING OF THE COCOA BEAN FERMENTATION PROCESS OPENS NEW PERSPECTIVES FOR CHOCOLATE PRODUCTION LUC DE VUYST Research Group of Industrial Microbiology and Food Biotechnology. for instance the cocoa pod surfaces. Vrije Universiteit Brussel. lactic acid bacteria (LAB). thereby producing lactic acid and acetic acid. and acetic acid bacteria (AAB) guarantees a successful cocoa bean fermentation process. hence initiating enzymatic reactions involving glycosidases. Once the cocoa pods are opened. thereby being removed and hence facilitating drying. boxes. These microorganisms grow fast and selectively because of the nutritious environment and low-pH conditions of the cocoa pulp-bean mass. the cocoa pulp-bean mass is inoculated by microbiota from the direct environment. and polyphenol oxidase. This post-harvest processing includes fermentation and drying. Actually. Theobroma cacao L. This combined fermentation and drying initiates physical and enzymatic processes inside non-germinating cocoa beans to develop color and flavor precursors. a well-maintained and clean fermentation (baskets. independent of the fermentation method applied (heap. Enterobacteriaceae may contribute to pectinolysis. and sensory analysis of chocolates produced thereof. it turned out that always the same species of yeast (Saccharomyces cerevisiae). it has been shown that high-quality fermented dry cocoa beans were obtained when mature. ghanensis. LAB. Weissella fabaria. Whereas fructophilic LAB contribute to the initial fructose fermentation and reduction. Brazil. provided that good agricultural practices are applied under correct circumstances of fermentation and drying. and AAB (Acetobacter pasteurianus) dominated. and Malaysia) by means of a multiphasic approach. citric acid conversion and gluconic acid production early in the fermentation while Glucon(aceto)bacter may be responsible for late gluconic acid production. Acetobacter fabarum. encompassing both culture-dependent and culture-independent microbiological analyses of temporal fermentation samples. Weissella ghanensis. temporal metabolite target analysis of cocoa pulp-bean mass and fermented dry beans. Although different new species were discovered when analyzing spontaneous cocoa bean fermentation processes. LAB (Lactobacillus fermentum). unraveled that a restricted species diversity of yeast. in addition to a removal of the placenta before fermentation. Ecuador. albeit that Hanseniaspora opuntiae. Ivory Coast. For instance. Fructobacillus pseudoficulneus. This species diversity has been confirmed by constructing a 16S rRNA gene clone library as well as by metagenomic analysis of cocoa bean fermentation samples. fresh and healthy cocoa pods and beans were used. etc. boxes. a regular mixing of the cocoa pulp-bean mass during fermentation. cerevisiae 19 . A. such as Lactobacillus cacaonum. and AAB communities was necessary for a successful fermentation. Several of the prevailing microorganisms were tested in appropriate cocoa pulp simulation media to unravel their functional roles as well as in small plastic vessels containing fresh cocoa pulp-bean mass as to evaluate their capacity to dominate the cocoa bean fermentation process. Lactobacillus fabifermentans. Lactobacillus plantarum. Leuconostoc paramesenteroides. or platform) and the cocoa population used (Criollo. senegalensis as well as Enterobacteriaceae were present at the start of the fermentation process.polyphenols and alkaloids out of the beans reduces bitterness and astringency of well-fermented cocoa beans. box. and Acetobacter senegalensis. Forastero.) and drying infrastructure. and A. Nacional). It turned out that the use of cocoa pulp simulation media allowed to simulate spontaneous cocoa bean fermentations and confirmed the functional roles of S. Acetobacter ghanensis. Detailed investigation of the cocoa bean fermentation process in several cocoa-producing countries (Ghana. and a controlled operation and duration of the fermentation and drying processes. Pichia kudriavzevii. and A. the inoculated starter culture species were able to outgrow the natural contamination of the cocoa pulp-bean mass and they prevailed during fermentation. triculture fermentations in a cocoa pulp simulation medium confirmed the interactions between S. fermentum 222 and A. Furthermore. For the production of a standard bulk dark chocolate. and A. pasteurianus 386B and another composed of L. It resulted in chocolates with masked bitterness and/or without unfermented flavour. these data allowed developing a starter culture composed of appropriate strains of S. cerevisiae. and A.(conversion of glucose into ethanol). pasteurianus to improve the cocoa bean fermentation process. Two starter culture mixtures were tested during cocoa bean heap and box fermentations. The addition of the LAB/AAB starter culture to the fermenting cocoa pulp-bean mass accelerated the cocoa bean fermentation process regarding citric acid conversion and lactic acid production through carbohydrate fermentation and resulted in chocolates with standard flavour profiles. whereas cocoa-specific AAB species could be distinguished into preferential lactic acid oxidizers and simultaneous lactic acid and ethanol oxidizers. Lb. Lb. pasteurianus 386B. In all starter culture-added cocoa bean fermentation processes. namely one composed of S. Also. different starter cultures were implemented on several farms in two different cocoa-producing regions (West Africa and Southeast Asia). L. fermentum. cerevisiae. pasteurianus. L. cerevisiae. L. and A. fructose and lactic acid into lactic acid and acetic acid and of fructose into mannitol). and A. the vessel fermentations confirmed the dominance of these species when inoculated into fresh cocoa pulp-bean mass. pasteurianus for reliably improved cocoa bean fermentation processes regarding speed of fermentation (from six to four days). it has been shown that cocoa-specific LAB species could be distinguished into fructose/non-fructose lovers and citric acid/non-citric acid convertors. the fermentation being finished within four days. fermentum (conversion of glucose. cerevisiae H5S5K23. In conclusion. fermentum. fermentum 222. Finally. The application of these starter cultures resulted in fermented dry cocoa beans that gave concomitant milk and dark chocolates with a reliable flavour. Ultimately. This enabled a fast and consistent ethanol production by yeasts. uniform quality of the fermented dry cocoa beans (avoiding variations in fermentation degree and flavour deviations typically seen in the 20 . Taken together. pasteurianus (conversion of ethanol and lactic acid into acetic acid and further oxidation of lactic acid and acetic acid into carbon dioxide and water). fermentum. as yeasts produced not only ethanol but also volatiles that contributed to the final cocoa flavour. the addition of a yeast strain to the LAB/AAB starter culture was necessary. these research results ultimately led to the composition of a mixed-strain starter culture consisting of appropriate strains of S. independent of cocoa-producing region or fermentation method. 21 . and flavourful chocolates produced thereof. This starter culture is ready now as to test it for commercial exploitation in cocoa bean fermentation and chocolate production.case of spontaneous cocoa bean fermentation processes). responsible for the significant amount of current biological diversity. this complex process. The speciation is an evolutionary process through which is made a gradual transformation of one ancestral species into two or more new species. POPESCU Institute of Biology Bucharest. Romanian Academy The fundamental unit of biological diversity is the species. In this context. Furthermore. There are several distinct species concepts. 22 . biological and phylogenetic properties. is currently one of the most important biological challenge. my lecture will discuss certain species concepts and selected contemporary data concerning speciation study. followed by the acquisition of some reproductive isolating mechanisms. A species is designated binomial by gender and specific epithet name. the species is a taxonomical rank represented by a group of individuals closely related to each other in terms of morphological. Essentially.SPECIES AND SPECIATION O. Bulgarian native flora comprises about 4000 taxa of which 12. STOEVA Institute of Biodiversity and Ecosystem research. stakeholders and NGOs. has been a main focus of politicians. nature habitats and environment issues. Flag species of conservation value. which declines due to the globalization of agriculture. The Convention on Biological Diversity and the two recent derivative documents: the updated Global strategy for Plant Conservation (2011-2020) and Nagoya Protocol on Access and Benefit sharing of genetic resources (2010) set out advanced targets and provides modern tools to ensure sound plant conservation policies on international and national level. The conservation of biodiversity. Various examples for developed strategies and actions taken in Bulgaria that perform different levels of 23 . S. putting enormous pressure on natural habitats – is Man. which could capture social involvement. Among the endemic species.8 % are Balkan and Bulgarian endemics. 75% of agricultural crop varieties have disappeared and three quarters of the world’s food come from only 12 plant species. We are facing a global ecological crisis. According to FAO estimates. The importance and urgency of the issues related to biodiversity conservation requires the strengthening of the link between scientific data and traditional knowledge preserved by the local communities and the finding of their present interpretation in the modern reality. but also agrobiodiversity. Y. Various strategies. including wild plant species and crops. The loss of biodiversity does not only affect wild species. GUSSEV. CH. action plans and other tools have been developed to preserve biological resources. economically and/or socially importance are most suitable to attract public interest and engagement.PLANT CONSERVATION TOWARD SOCIETY D. are main target of participatory approach in nature conservation. According the Red List of Bulgarian Vascular Plants (2009) 176 endemics (108 Balkan and 68 Bulgarian) have been assigned an IUCN threat category – CR. T. 165 are legally protected by the national Biological Diversity Act. TSONEVA. IVANOVA. Bulgarian Academy of Sciences. scientists. GEORGIEV and T. Moreover Bulgaria is one of the top five countries in terms of crop wild relatives richness in Europe. BOSSEVA. V. DIMITROVA. Bulgaria is among the countries with richest biodiversity in Europe. Plant species. which involves land and sea ecosystems and where the main actor – who also bears the most responsibility. Bulgaria Biodiversity loss is a major concern of the human society in the last decades. EN or VU. efficiency in the preservation of wild and agro-biodiversity are provided so to represent current status of plant conservation in the country. 24 . The biodiversity-engaged community and especially its scientific part need to reinforce its identity and build up interdisciplinary groups to be able to launch more efficiently long-term and funding-supported measures. The objects of the investigations are rare and threatened plant species from the Bulgarian flora. Bulgaria Plant biotechnology is a new research area in the Institute of Biodiversity and Ecosystem Research with the Bulgarian Academy of Sciences. Its development began eleven years ago with the establishment of the Biotechnological Laboratory of Medicinal Plants in the frame of the 5year project “Alternative Approaches of Bioproduction of Alkaloids and Active Substances from Bulgarian Rare and Threatened Medicinal Plants” (2001-2006) supported by the NATO Science for Peace Programme. analytical for phytochemical analyses. close to the GLP requirements. populations of which need to be strengthened. aiming at rapid plant multiplication by means of different in vitro techniques. consisting of several special sectors corresponding to the interdisciplinary scientific studies: laminar box for in vitro manipulations in sterile ambience. glass-ware cleaning. BAS – FACILITIES. allowing their graduate adaptation to lower humidity. the most perspective for in vitro cultivation are some law-protected medicinal plants with market demand and resource deficit. STANILOVA Institute of Biodiversiy and Ecosystem Research. extraction of bioactive compounds. with modern equipment and furniture. The laboratory is compact. (Amaryllidaceae). ACHIEVEMENTS. which was crucial for the successful completion of the whole procedure of plant propagation. Among them. Besides. Bulgarian Academy of Sciences. the laboratory was extended with an extra room for ex vitro adaptation of the in vitro obtained plants. The first in vitro cultivated species in our laboratory was the medicinal plant Leucojum aestivum L. AND CHALLENGES M. cultures’ room. the biosynthesis of active substances by in vitro cultures of medicinal plants is desirable for many advantages. and auxiliary rooms for media preparation.PLANT BIOTECHNOLOGY DEVELOPMENT IN THE INSTITUTE OF BIODIVERSITY AND ECOSYSTEM RESEARCH. which gathering from the wild populations is forbidden or limited. In vitro propagated medicinal plants are suitable planting material for an establishment of field plantations as an alternative way to ensure plant biomass for production of drugs. Recently. Sofia. microscopy for karyological and histological control. The facilities were designed and organized to serve a new research direction. which is used as raw material for extraction of the alkaloid galanthamine and production of the medicine Nivalin® by the Bulgarian pharmaceutical factory Sopharma 25 . and so forth. 1±1. 2 MSc thesis and 3 PhD thesis have been developed in the Biotech lab in the frame of several projects with national or international funding. Other successfully in vitro multiplied medicinal plants are the critically endangered in Bulgaria species Alchemilla mollis (Buser) Rothm. some critically endangered species according to the IUCN criteria with problematic seed germination as Fritillaria meleagroides Patrin..) and Geum bulgaricum Pancic. The acclimatized plants on the experimental field plot kept the features of the parental plants. The cultures’ features studied during 3 years of subcultivation revealed clone specific dynamics of the biosynthetic activity in spite of the unchanged cultivation conditions and medium composition. (Rosaceae) and three Bulgarian high-mountain endemics from the same genus: A. and some other psammophytes from the white and grey dunes along the Bulgarian Black Sea Coast. We developed successfully both approaches: rapid in vitro micropropagation and in vitro biosynthesis of alkaloids. Until now. similar to the commercial antioxidant butylated hydroxytoluene. due to difficult financial situation. are ready to be transferred to their natural habitats as well as to the Botanical garden of BAS. Scientific network has been established between biotechnologists from several Balkan 26 ..Georgiev. similar to that of the commercially important wild populations. jumrukczalica Pawł and A. s. affected by the anthropogenic pressure. which is not a common practice in our country. last summer and distinguished with high antioxidant activity (IC50 13. & Jav. It is worth to note that the success of the next steps related to the ex vitro adaptation and acclimation of the plants to open air conditions increased significantly after the setting of the phytotron.&T.. Among them are: Pancratium maritimum L. plants of the endemics Centaurea davidovii Urum. IC5012. the most important challenges. Several in vitro clones were selected for their high propagation rate and galanthamine content over 1 mg/g DW. Alchemilla achtarowii Pawł. several endemics from the alpine area as Papaver degenii (Urum. Several hundreds of ex vitro adapted Alchemilla plants are well developing on the field plot and the first 80 A. C. Nowadays. reached flowering stage and formed normal seeds. Seeds of many threatened plant species included in the Red Book of Bulgaria and protected by the Biodiversity Act have been in vitro germinated and multiplied as a first step of ex situ conservation. All of them survived the winter at 1500 m a. l. pseudaxillaris Stef. mollis plants were easily acclimated on Vitosha Mt. A. bundericensis Pawł. are related to the opportunities to keep young people in the Academy and to establish partnerships between the scientific team and the business world.9 μg/mL. Astragalus physocalyx Fisch.AD. Thus.65 μg/ml). achtarowii Pawł. and Astragalus physocalyx Fisch. The biotechnological approach of in vitro plant multiplication in the institute is on the threshold of a new stage.countries. and exchange of experience is in progress. 27 . including Romania. Utilization of genetic engineering could ensure relatively quick acquiring of desired useful agronomic traits without cardinal changes of cultivar genotype. identical with mother genotype. In vitro adventitious organogenesis was studied in different Vaccinium and Rubus species and cultivars. which can be avoid by tissue cultures. Classical breeding is in fruit trees limited by their long reproductive cycle and high heterozygosity what make the breeding process very time-consuming. Tissue cultures represent the effective tools not only for rapid mass propagation of high quality planting material needed for large-scale cultivation. anthocyanin pigments. Micropropagation via direct shoot regeneration from dormant buds is well suited to obtain genetically homogeneous planting material. therefore an increase in effort to establish the large-scale cultivation of productive cultivars is noticed. however the results of these studies showed that development of efficient and widely applicable regeneration system in these species is complicated because the regeneration ability is highly genotype-specific and it is often impossible to regenerate plants from tissue derived from mature woody plants. as well as for genetic engineering. 28 . They are widely cultivated in various parts of the world.CLONAL PROPAGATION AND GENETIC IMPROVEMENT OF SELECTED SMALL FRUIT SPECIES THROUGH BIOTECHNOLOGY APPROACHES A. bioactive organic substances with antibacterial and anticancer effects. mainly due to high sensitivity of their cells and tissues to selection antibiotics commonly used in genetic transformation. Increasing demand for berry fruits on the world market evocates also necessity to create new genotypes with improved attributes. GAJDOŠOVÁ. considerably limited till now. LIBIAKOVÁ and M. but also for germplasm conservation and improvement. and also as an excellent source of antioxidants. lack of initial material. Genetic transformation and following regeneration of transgenic plants is in Vaccinium and Rubus spp. LATEČKOVÁ Institute of Plant Genetics and Biotechnology of the Slovak Academy of Sciences. G. Traditional vegetative propagation by cuttings has some limitations. Slovak Republic The species of Vaccinium and Rubus genus belong among commercially and biologically important small fruits because of their high content of vitamins. such as rapid spreading of diseases. In vitro plant regeneration via adventitious organogenesis is an effective system for the production of plant material. dependence on seasonal growth or low propagation coefficient. influence of age and type of explants on efficiency of genetic transformation and selection procedure were followed. 29 . testing for influence of selection antibiotics commonly used in genetic transformation on regeneration ability of the respective cultivars.The present paper is focused on description of reliable system for micropropagation and adventitious regeneration of Vaccinium and Rubus spp. selected cultivars. In our research the focus was put on testing of factors influencing successful micropropagation and regeneration of adventitious shoots (type of explants. type and concentration of growth regulators). genetic transformation of selected cultivars by Agrobacterium tumefaciens and regeneration of transgenic plants. For optimization of transformation and regeneration conditions for studied cultivars the duration of co-cultivation of plant tissue with Agrobacterium. basic culture medium. influence of plant genotype. Transformation of plants was done by use of different strains of Agrobacterium tumefaciens and the transformation efficiency was evaluated with the aim to select the appropriate strain of Agrobacterium tumefaciens. The development of viral infection influence correlates with selective methylation of different genomic DNA regions that leads to changes in their transcriptional activity (Boyko A. DNA genomic nucleic acid component). Republic of Moldova Impact of stress-derived stimuli. These modifications would start with changes in DNA methylation and chromatin conformation patterns and. it was recognized the contribution of in vitro condition in induction of genetic instability and variability. isometric virions. According the proposed objectives. inducing photosynthesis decreasing. Also. it could lead to a genetic mutation causing fixation of a new treats. Moldavscaia and Iuniscaia. gene expression modification.5% glutaraldehyde on cacodylate buffer. Brassica oleracea var. including viral pathogens. Preservation of genome stability and integrity is the primary task for every organism. The tissue were dehydrated in series of ethanol and propylene oxide and embedded in Epon’s resin. capitata.. Chisinau. Kovalchuk I. The experimental plants were infected with cauliflower mosaic virus (CaMV. Cabbage plants were grown in field following a standard technique. As biological materials. activation of transposing elements. respiration increasing and carbohydrate levels alteration. including plants. Postfixation was done in 1% OsO4. For in vitro culture were used the leaf explants from plants presented positive results to viral particle presence and which developed a systemic infection. It is known that the viral infections dramatically affect plant physiology. 2011). the morphogenetic calli pieces were fixed for transmission electron microscopy.. ANDRONIC Institute of Genetics and Plant Physiology of the ASM. Ultrathin 30 . On basis of studies has been established that viral infection can contribute to chromosome breaks. result in a wide range of modification triggering local and global responses via multiple (frequently intercrossing) transduction pathways. pH 7. The stereologic technique adapted for electron microscopy was used for morphometric analysis. using solution of 2. chromatin condensation and cells proliferation. if the pathogen pressure continues over several generations.NUCLEOTYPIC EFFECTS IN VITRO CONDITION OF DIFFERENT CABBAGE GENOTYPES INFECTED WITH CAULIFLOWER MOSAIC VIRUS L. cultivars Haricovscaia. The objective of this study is the morphometric evaluation of nuclear structures in vitro condition from cabbage calli cells derived from virus infected and non-infected plants. on perception structures in plants. were the cultivated cabbage. in dependence of the host genotype.033±0.84 % 0.105 36.177±0.219±0.025±0.0028** 34.053±0.028±0. of var.235±0. the infection.011* 33.261 Relative volume of nuclear structures Nucleus Membrane chromatin 0. Although the relative value of chromatin distributed in nucleplasm varied significantly in cells derived from virus-infected donors (47% to 22-24% in control).81 % 0.252 Coef. CaMV 0.009 35.044±0.034±0.0009 18.008 33.0007 13.01.90 % 0.268 Coef.0019 47.0007 21.52 % 0.10 % 0.215±0.82 % Haricovscaia Control Coef. we assume that the observed deviations could result due to a position effect variation inducted by viral infection. inducted by the CaMV. nucleo-cytoplasmic rate. of var. 0.85 % 0.09 % 0.334*** Coef.63 % 0.0007 23.sections were stained with uranyl acetate and lead citrate.032±0. **.significant at P ≤ 0. Table 1. CaMV 0.0006 22.013** 37. Moldavscaia Control 0. CaMV 0.25 % Nucleplasmic chromatin 0. 31 . *** . Sections were examined with transmission electron microscope (AM).25 % 0. As morphometric parameters were used: the relative volume of cells.0020 ** 47.018±0. of var. nucleus. conducts to the emphasize in the chromatin distribution.49 % 0.185±0.65 % 0.013* 37.026±0. chromatin (membrane associeted and nucleplasmic).08 % 0. of var. On basis of the obtained data. nucleo-cytoplasmic rate and its coefficient of variation. generated and by the in vitro culture (table).61 % 0. *.001 For all three cabbage cultivars were established the increase of the membrane chromatin relative volume.0002** 33.0019 * 47. the modifications of means value were not been statistically confirmed for all analyzed genotypes.020±0.330*** Coef.05. In study were used only the cells and nuclei in which a not denoted the viral particles. 0.01 % 0.019±0. of var. Morphometric changes of nuclear parameters in vitro condition in noninfected and CaMV infected cabbage cells NucleoGenotype Treatments cytoplasmic rate 0. According the obtained data.168±0.94 % 0. Iuniscaia Control 0.318** Coef.0009 19.0028** 37. of var.048±0.45 % 0. 139: 935–948.. pp.W. 32 . Genetic and Epigenetic Effects of Plant–Pathogen Interactions: An Evolutionary Perspective. Laval V.. Plant Physiol. Yun B.. Kovalchuk I.. . 1. Love A. Milner J.J. Cauliflower Mosaic Virus. Engages Three Distinct DefenseSignaling Pathways and Activates Rapid Systemic Generation of Reactive Oxygen Species. Molecular Plant. Loake G.J.. 2005. a Compatible Pathogen of Arabidopsis. 1–10.. Boyko A.. 2005). It is proposed that cauliflower mosaic virus triggers a rapid systemic response that precedes virus movement and results in a rapid local and systemic induction of reactive oxygen species (Love et al. 2011.. 2.J.which mechanism is not clear. the major human intestinal archaeon. expressed in E. The putative small size CPSase was isolated and characterized in order to identify a putative missing link for this class of enzymes. In addition to a bacterial type full-length (120 kDa) synthetase subunits. USA The genome of Methanobrevibacter smithii. ATP-dependent partial reactions indicate that MS-s that could catalyzed a CPSase specific reaction that occurs in three steps.Z. N. MS-s represents the smallest active CPSase characterized so far that might correspond to the postulated ancestral kinase which evolved into fulllength CPSases. and purified by affinity chromatography. coli. Romania. GUY-EVANS2. Kinetic parameters reveal a higher apparent affinity for ATP and ammonia as compared to other large CPSases.MISSING-LINK CARBAMOYL PHOSPHATE SYNTHETASE E. a major enzyme in the biosynthesis of arginine and pyrimidine nucleotides. and appears to be a methanogen specific ammoniametabolizing enzyme. PERERA2. MI. coli CPSase. Chemical cross-linking and size-exclusion chromatography indicate a dimeric and tetrameric structure.R. Detroit. 2Eastern Michigan University. inspite of a reduced primary sequence conservation. KIBÉDI-SZABÓ1. suggesting a particular role MS-s in ammonia trapping in the cell. C PURCAREA1 1 Institute of Biology Bucharest. Structural analysis and 3-D molecular modeling reveal a similar folding with that of the carbamate-phosphorylating domain of the E. contains several genes coding for carbamoyl phosphate synthetase (CPSase). Department of Biochemistry and Molecular Biology. C. 3 Wayne State University School of Medicine. USA. H. 33 . EVANS3. Department of Microbiology. MI. this methanogen possesses a reduced size (41 kDa) homologous subunit (MS-s). The recombinant MS-s was cloned. Bucharest. D. Ypsilanti. Romanian Academy. Department of Chemistry. POPA1. some exploitation areas were abandoned resulting in various man-made salt lakes with different depths (3. recommend them as a group of wide biotechnological potential (10. we studied some potential biotechnological applications of some haloarchaea strains: . ENACHE Institute of Biology Bucharest. Shepherd Bath. 12). forming a part of the domain Archaea. Balta Albă lake in Buzău county. formed in the Neogene period are located at around 100 km north of Bucharest. Romanian Academy Hypersaline environments (lakes. The salt deposits in Slănic Prahova and Telega (Doftana–Telega). as well as their capacity to produce different compunds (biopolymers. The physiological and biochemical features specific to haloarchaea. solar salterns. These hypersaline environments are populated by halophilic microorganisms. Salt exploitation in these areas started around the year 1685 by using the bell type exploitation technology.the isolation and characterization of S-layer from haloarchaeal cell envelope and preliminary investigations related to the binding of S-layer to some porous silicon substrates. etc. The results showed the predominant presence of Haloferax species in the investigated salt lakes. Hypersaline environments are widely distributed also in Romania either in solid or liquid forms: salt lakes and salt mines located in Prahova county. having natural or man-made origin.0–2. This suggests that members of this genus play an important role in ecology of salt lakes. hypersaline lakes from Ocna Sibiului. carotenoid pigments. mainly haloarchaea.the synthesis of exopolysaccharides. etc) of industrial interest. are a group of extremely halophilic microorganisms. enzymes and halocins. Dead Sea. halorhodopsin.5M NaCl) and are able to grow up to saturation (13). TEODOSIU and M. Haloarchaea. enzymes. . lipids. Bride Cave and Telega (Palada) (1).) are found in nature throughout the world. Red Bath.40 m) and widths known today as Green Bath. . bacteriorhodopsin. Shepherd Bath.HALOARCHAEA FROM SOME ROMANIAN MAN-MADE AND NATURAL HYPERSALINE ENVIRONMENTS G. Red Bath. Also. halocins. Techirghiol lake nearby to Black Sea coast. etc. which require high salt amounts for growth (at least 2. After the 18th century. Green Bath and Telega lakes and in the salt crystals taken from subterranean salt mine Unirea by a polyphasic approach using biochemical and molecular methods. even though the largest number of 34 .the response to some heavy metals and strains capacity to reduce their concentration. The aim of our studies was to evaluate the diversity of haloarchaea present in Bride Cave. salt deposits. members of the family Halobacteriaceae. the maximum EPS yield (0. Zn and Ni by the tested strains (11). some strains showed identical patterns. prahovense and Hfx. Hfx. mannose. noricense. mediterranei (2). and they showed a variety of action spectra.5M (4. Green Bath.1M NaCl. observable in hypersaline lakes from the surface of Slănic salt deposit. The TEM examination of the isolated S-layer showed the existence of the monomolecular crystalline lattice with a highly ordered arrangement in the dense form. sp. The enzyme produced by Hfx. 2. According to data resulted from TLC and FT-IR analysis. The Haloferax members. 8). fructose. TL5 strain has a higher susceptibility for Zn ions compared with Hfx. The investigated haloarchaea strains were susceptible to Zn and Hg but moderately resistant to Cr and Ni. were identified also in subterranean rock salt. 7). Red Bath and Bride Cave. GR2 strain isolated from the Bride Cave lake. being classified as tolerant. pH 6. prahovense was a heteropolysaccharide containing mainly glucose. TL5 strain and the collection strain Hfx.475 g%) was obtained in medium with 3% glucose and 2M NaCl. and some isolates appear to be closely related to Hbt. sp. GR1 was influenced by NaCl concentrations in the growth media and had a maximum activity at 3M NaCl. The synthesis of EPS enhanced the reduction activity of Cr. sp.Halorubrum species was identified in hypersaline environments (6. Halocins were also detected in some strains isolated from Romanian salt lakes. showed higher activity with increasing concentration of MgCl2 in the presence of 2. sp. The activity was lost at NaCl concentrations below 2. uronic acids and sulfate and have a thermal stability until to 2070 C. prahovense. The enzyme of Hfx.0 mM Cr. sp.5 mM Ni and Pb. The exopolysaccharide biosynthesis activity was detected in some Haloferax strains isolated from Telega lake such as. The biochemical characterizations by protein content and chemical treatment have demonstrated the presence of S-layer in the Hfx. but with decreasing numbers (9).4M NaCl. Amylases produced by Haloferax and Halorubrum strains isolated from Techirghiol lake. mediterranei tolerated 5. Zn and Ni ions from media with high salinity.5M NaCl.5M NaCl. The microbiota of subterranean rock salt from Slănic area were characterized by the presence of some Halorubrum and Haloarcula species. Hfx. galactose.5 and 500 C and lost its activity bellow 1. sp. When compared as halocin producer and targets. Hfx. The amylases were produced by some strains isolated from Bride Cave and Techirghiol lakes. Extracellular lipase activity was detected in some strains isolated from lakes Shepherd Bath. TL5. Cr. the polymer of Hfx. Both strains showed the capacity to reduce the concentration of Pb. while in relaxed 35 . In the case of Hfx. GR1 showed maximum activity at 3. supporting the tight clustering of strains in phylogenetic tree reconstructed from 16S rRNA gene sequences (5). but activity decreased with increasing Mg concentrations at 3. . Popescu G. Cojoc R. J. The S-layer proteins attached to all investigated plates of porous silicon. but it seemed that the hydrophobic surface was more favorable. 11: 3–7. Micro and nanoengineering. Dumitru L.M. Popescu G. (eds) Progress in nanoscience and nanotechnologies. Romania. 173–197.. pp. Acad Române.)... GR 2 (JCM 13922): isolation. Wien. NewYork. Ed.. Enache M. 4: 261–267. Adaptation of Microbial Life to Environmental Extremes. Dumitru L. The effect of NaCl concentrations on the growth and lipase activity at Haloferax sp. Enache E. Adaptation of Microbial Life to Environmental Extremes.. 2012. 7. Dumitru L. 6.. 8. Series B. Dumitru L.form after treatment with 4M urea. Inst. Faghi A.. 3. Acad.. Biochem. In: Helga Stan-Lotter & Sergiu Fendrihan (eds. Teodosiu G. Inst.. Symp. Biol.).. Itoh T. 2002. NewYork. 4. 11. 2009. Zarnea G. Halophilic archaea isolated from man-made young (200 years) salt lakes in Slănic.. 11. In: Helga Stan-Lotter & Sergiu Fendrihan (eds. Teodosiu-Popescu G. Dumitru L. Eur. Oren A. nov. 23: 791–795. 2008. 1994.. Enache M. Proc.. S-layer of Haloferax sp.. Eq. Enache M. 12. Teodosiu G. 1–19. Series B. 2010. Springer. Enache M. 36 . Cent. In: Kleps I. on both hydrophilic and hydrophobic surfaces. 1: 27–32. Rodriguez-Valera F.. Kamekura M. Seria Vol. Romania. Popescu G. Industrial and environmental applications of halophilic microorganisms... Novel Research Results and Application. Environ.. Kamekura M.. Springer. Kleps I. 57: 393–397.. Wien. Syst. 6: 233–236.. Proc. GR1. Dumitru L... 1990.. 9. 10. Proc. 31:825–834. Rom. The effect of Na+/Mg2+ ratio on the amylase activity of haloarchaea isolated from Techirghiol lake... characterization and binding to silicon nanostructurated substrates.. Soc. 5.. The treatment of silicon plates with hexamethyldisilazane increased the amount of attached S-layer protein (3). The tolerance of extremely halophilic archaea to heavy metals. Biosorption of some heavy metals from media with high salt concentrations by halophilic Archaea. Proc...C.. Dascalu D. GR 1. Dates from historic salt exploitation at Slanic-Prahova (in Romanian). J. 2004a. 27: 68–71. Rom. Rev Muzeelor. Popescu G. Ignat T.. Evol. Itoh T.. 58: 135–147. Dumitru L. Biol.. Halophilic microorganisms from man-made and natural hypersaline environments: physiology.. 2012. Enache M. Zarnea G.. 2007. Enache M. Int... a low salt environment. Kamekura M. 2. Prahova.. 2007.. 1. 2004b. Teodosiu G. Drăgănescu L. an extremely halophilic archaeon isolated from a Romanian salt lake. pp. Stan-Lotter H. Teodosiu G. pp. Biotechnol. ecology and biotechnological potential. Itoh T.. 2009... Kamekura M. Microbiol. Biotechnological potential of halobacteria. Acad... Physico-chemical boundaries of life. Bucharest. 3: 388-395. 146–152. 13. Enache M.... Biotechnol... Haloferax prahovense sp.. Ion A. Novel Research Results and Application. Halocin HF 1 a bacteriocin produced by Haloferax sp. Dumitru L. Technol.. Biol. BRAD4. B. 165: 1-10. A. Well characterized geo-chronologically (3..P. Teacher C. We are interested in characterizing the microbial diversity of this underground glacier. PURCĂREA2 Emil Racoviţă Institute of Speleology. Romania. 4. Romania.P. Sowers T. IŢCUŞ2. 37 . ONAC4 and C. C. ARDELEAN2. Florescent microscopy was performed on light exposed ice samples to examine the presence of phototrophs. 116: D02111. Pazdur A. 11(2): 640-656.. 2011. Perşoiu A. Onac B. Moreover. Ice genesis and its long-term dynamics in Scărişoara Ice Cave. in order to correlate the composition of the microbial communities (prokaryotes and eukaryotes) trapped in different aged ice layers with the corresponding climate pattern... Here we report the presence of both prokaryotic and eukaryotic microorganisms in different aged ice layers growing within this range of temperatures.. RUSU2. 3. 10° and 15°C. Onac B. E. Bucharest. 2009.-V. 2011. Comparison of the microbial diversity at different depths of the GISP2 Greenland ice core in relationship to deposition climates. Bojar A.MICROBIAL DIVERSITY IN THE SUBTERRANEAN ICE DEPOSITS OF SCĂRIŞOARA CAVE A. Cluj-Napoca. Ice samples of different ages were investigated by cultivation on several media and C-sources (BIOLOG Ecoplates) at 4°. HILLEBRAND-VOICULESCU1... very little is known about subterranean ice biotopes – an isolated and light deprived cold environment. Scărişoara Cave (Romania) hosts the oldest and largest underground perennial ice block in the world (2). 3Ştefan cel Mare University. 4Emil Racoviţă Institute of Speleology. Romanian Academy. Romania 1 While extensive studies have been performed on various exposed ice deposits.P. viable cyanobacteria and phototrophic eukaryotes were found in the light exposed recent ice sediments.... Microbiol. 2Institute of Biology Bucharest. Wynn J. A. PASCU2.. JGR Atmospheres.. PERŞOIU3.. Romania. Evaluating the paleoecological potential of pollen recovered from ice in caves: A case study from Scarisoara Ice Cave.. POPA2.G. Romania. Environ. 4) it represents an important paleoclimatic archive. The Cryosphere Discuss. I. Genomic DNA was extracted from each sample and the corresponding amplified SSU-rRNA fragments were analysed by DGGE. D. The diversity of these communities correlated with the age of the ice and the growth conditions. Stable isotope behavior during cave ice formation by water freezing in Scarisoara ice cave. Perşoiu A. Feurdean A. Review of Paleobotany and Palynology. 1. Perşoiu A. 4: 1909–1929. T.M. Brenchley J. Pazdur A.. 2.. Suceava. Romania. Miteva V. 2010. Holmgren K. Special attention is devoted to the isolation and cultivation of phototrophic microorganisms growing together with heterotrophic microorganisms in gasoline supplemented microcosms. 38 .) both at population level and at single cell level by automated image analysis.I.45µm) sea water)] supplemented in different concentrations with gasoline and inorganic nutrients. ii-dead cells (propidium iodide positive cells). together with new co-workers. concerns: a) the increase in microcosms diversity by the use of dispersants. esterase activity etc.DYNAMICS OF MICROBIOTA IN MARINE MICROCOSMS (INDOOR): RESULTS AND PERSPECTIVES I. b) monitoring of metabolic activity by more diverse methods (resazurine reduction. c) quantification of pollutant consumption in microcosms and d) analysis of microbial diversity by molecular techniques. 2Maritime University Constanţa In this paper we present our results concerning microbial cell density dynamics [i-total cell count (acridine orange. GHIŢĂ2 1 Institute of Biology Bucharest. The relationships between basic and applicative aspects will be also taken into account. iii) cells able to grow and to multiply (Kogure method) over 24 hours] and metabolic activity (CBO5) in indoor (18⁰ C) marine microcosms (non filtered or filtered [(0. Sybr GreenI. The perspectives. Romanian Academy. CTC reduction. DAPI). ARDELEAN1 and S. In Spaceships and Space Stations fungal species can develop submerged mycelium in the depth of different substrates. WALTHER2 and E. performed on Random Positioning Machine (RPM) and of microgravity during Space Experiment performed on board of International Space Station (ISS). In this case. science team of IBB took pictures at FD4 and FD8. Switzerland. each biocontainer was introduced in a pouch of protective foam and NOMEX fabric. on the growth of Ulocladium chartarum colonies. Results suggest that EST is very important as a step of strategy to perform space experiments with fungi but the real effect of microgravity can be studied only on board of ISS. Department of Geological Sciences. grown on culture plates (provided by Institute for Space Science. Magurele) were integrated in three biocontainers (provided by European Space Agency) which were finally sealed. I.THE GROWTH OF Ulocladium chartarum IN MICROGRAVITY: ON RANDOM POSITIONING MACHINE AND ON BOARD OF INTERNATIONAL SPACE STATION GOMOIU1. chartarum (provided by Institute of Biology. Interestingly was that the strongest positive effect of microgravity was put in evidence on 1 day old colonies. The duration of space experiment was 14 days. On RPM. including tools containing small amounts of organic compounds acting as biodeteriogens and risk factor for crew health. 3National Technical University of Athens. The positive effect of microgravity is better expressed on ISS experiment then on EST. Bucharest) having different ages. the Crew took pictures at FD5 (flight day) and FD9. culture plates were inoculated just before handover. Ulocladium chartarum colonies developed as response to microgravity a complex colony morphotype. Spores and hyphae were not spread in microgravity due to the fact that they are attached on the nutrient. On ISS. mycelium and spores viability as well as changes at the cellular level were investigated. Romanian Academy. S. School of Mining and Metallurgical Engineering. To assure a high protection during experiments. 2Space Biology Group. two days longer then EST due to NASA decision. due to the fact that secondary metabolites acted as toxic compounds in completely isolated environment. ETH Zürich. VADRUCCI2. Photo sessions had been also performed by science team of IBB for ground controls. CHATZITHEODORIDIS3 1 Institute of Biology Bucharest. Athens. All colonies developed sporulated aerial and submerged mycelium in both experiments but only submerged mycelium was viable after landing. for all colonies. The study of Ulocladium chartarum colonies after de-docking and landing clearly suggested that the growth took place in microgravity. launch was planned to take place one day after handover. never mentioned in similar conditions. Colonies of U. 39 . Greece The effect of simulated microgravity during Experiment Sequence Test (EST). Microbial communities living in the chemocline. 2 Department of Molecular Biology and Biotechnology. 2010). Cluj-Napoca. MUNTEAN1. Institute of Biological Research. To achieve the proposed objectives we used culture-dependent and culture-independent methods. 2Institute for Interdisciplinary Research in Bio-NanoSciences. 2. dissolved oxygen and conductivity) were determined using a portable water quality meter monitor. 2000 m2 and a depth of 33. ALEXE4 and H. etc. physico-chemical analysis (salinity. 23°47’17’’ E). 2010). Babeş-Bolyai University. Romania. 46°35’09’’ N. thus leading to a density-based stratification in the water body. Romania. As a consequence of their tectonic evolution the salts reached the surface in several lower pressure areas. total organic carbon. G. pH. A. BabeşBolyai University. 3National Institute of Research and Development for Biological Sciences (NIRDBS). ANDREI1. It has a conical contour due its placement in a former medieval bell-shaped salt mine (Alexe. BARICZ1. Saline lakes in Transylvania have an almost circular distribution at the inner periphery of the Carpathian Mountains. and the majority was formed after the collapse of abandoned salt mines (Alexe. Cluj-Napoca.-A.DIVERSITY OF ARCHAEAL COMMUNITY IN THE CHEMOCLINE OF A HYPERSALINE LAKE IN THE TRANSYLVANIAN BASIN (ROMANIA) Ş. The salinity recorded in the water column ranged from 67 g/l at the top to over 300 g/l at 4 m below the surface. V. The strong salinity gradients of these hypersaline lakes prevent the mixing of water layers. Romania 1 The halite deposits of the Transylvanian Basin in Romania were formed as massive evaporites following the sea level drop in the former Paratethys basin during the Middle Miocene.). Samples were collected with a submersible electric pump and stored in sterile 2 L polypropylene bottles. Cluj County. KERESZTES1. This study aims to investigate the diversity of Archaea that possibly inhabit the chemocline of the thalassohaline Ocnei Lake (Turda area. Z. M. in which diverse microorganisms have the possibility to thrive. compared to those from the upper (mixolimnion) and lower (monolimnion) water layers are exposed to a variety of ecological niches on a small spatial scale. chlorophyll extraction and total cell counts 40 . They were used for: biomass fixation on polycarbonate filters. C. This anthropo-hypersaline lake has a surface of approx. Cluj-Napoca. BANCIU1. Romania. Babeş-Bolyai University. Romania.7 m. In situ measurements (water temperature. determination of major ions. COMAN3. 4Faculty of Geography. isolation of archaeal strains on selective media. together with bioinformatics and statistical analysis. Faculty of Biology and Geology. Cluj-Napoca. Rossello-Mora R.. DNA was extracted from the filtered biomass. 2010. The highest number of prokaryotic cells was recorded throughout the chemocline and in the hypolimnion. and used in the PCR amplification of partial 16S rRNA genes. Our clone library revealed phylotypes belonging to Halobacterium. 2010. Acknowledgements: This work was supported by a grant of the Romanian National Authority for Scientific Research. The Halobacterium phylotype is the dominant member in the archaeal clone library (94% coverage) and is the only taxon that could be isolated by culture-based methods. Cluj-Napoca (in Romanian). 2000. Antón J. Amann R. 4th Ed. Microbiol. Handbook of microbiological media. Likens G. Environ.5-3 m) of water column. 1991. Alexe M. Wetzel and Likens... 1. We found that genetic variability of Archaea (at the sampling site) is restricted to members of the Halobacteriaceae. Measurements of chlorophyll-a and total organic carbon indicated that the primary productivity is most probably located in the upper layer (0. project number PN-II-ID-PCE-2011-3-0546. Halorubrum and to the recently described genera Halonotius and Halolamina. Rodriguez-Valera F. 2. 41 .(Atlas.. New York.G. 3. 3rd Ed. 66 (7): 3052-3057. CRC Press... 2000. Limnological Analyses. Washington D... Antón et al. Based on the physico-chemical profiling of the water column in Ocnei Lake and the total cell counts. Appl. A study on the salt lakes of the Transylvanian Basin. A clone library was constructed and the sequences obtained were compared to those stored in GenBank (NCBI) for taxa identification. Atlas R. our results showed a direct relationship between the microbial abundance and salinity. Wetzel R.. 2010. Extremely halophilic bacteria in cristallizer ponds from solar salterns.E. Cluj University Press.C. Springer-Verlag. 2000).... CNCS – UEFISCDI. 4.M. we used the subcutaneous B16. To assess the pleiotropic effects of statins that can be responsible for statin-induced cytotoxicity in vitro. CRISTIAN1 and M. “Babes-Bolyai“ University. despite the increasing data regarding the antitumoral activities of statins.1. ALUPEI1. Our in vitro data have shown that only the lipophilic statin had strong inhibitory actions on proliferation and viability of B16. such as angiogenesis and inflammation. their complete mechanisms of action still remain obscure. Romania 1 Statins as inhibitors of 3-hydroxy-3-methylglutharyl coenzyme A (HMG-CoA) reductase. Faculty of Biology and Geology. with a polydispersity value lower than 0. To study the anticancer potential of statins in vivo. Cluj-Napoca.1 μm. Mean particle size of the liposomes was determined by dynamic light scattering and found to be 0. with the effects induced by a hydrophilic statin. the mice were sacrificed and tumors were isolated. Therefore. LICARETE2. BANCIU1. Romania. However. the rate-limiting enzyme in de novo cholesterol synthesis. E. on antioxidant defense mechanisms of tumor cells. 2Molecular Biology Centre.v. On day 15. Free statins and liposomal statins were administered i. this study aims to investigate the underlying mechanisms of the cytotoxic effects of statins on B16. F. Cluj-Napoca. simvastatin (SIM). statins were encapsulated in long-circulating liposomes. we studied the statin actions on the expression of the hypoxia-inducible factor 1α (HIF1α). Institute for Interdisciplinary Research in Bio-Nano-Sciences. after statin encapsulation in long-circulating liposomes. These liposomal formulations enable accumulation of the statins at sites of malignancy.C.F10 murine melanoma cells in vitro and on established murine melanoma models in mice. Statin treatments started at day 11 after tumor cell inoculation. can exert tumor cytotoxic actions. as well as on tumor cell capacity to support vital processes for tumor development in vivo. We have compared the effects of one lipophilic statin.F10 melanoma cell proliferation and viability at concentrations of each statin ranging from 20-100 µg/ml. These cytotoxic effects were mainly mediated via suppressive actions of SIM on HIF-1α expression and nonenzymatic antioxidant levels. Liposomal statins were prepared by lipid film-hydration method.2.B. as free drugs as well as liposomal statin formulations. followed by multiple extrusion steps.F10 melanoma cells. “BabesBolyai“ University”.STATINS AS CANDIDATES FOR TUMOR-TARGETED NANOMEDICINES M. pravastatin (PRV). as well as 42 . To study the antitumor activity of liposomal statins and free statins.2 Department of Molecular Biology and Biotechnology. at a dose of 5 mg/kg at days 11 and 14 after tumor cell inoculation.F10 murine melanoma model. In vitro cytotoxicity was investigated in regard with B16. This leads to high reduction in isoprenoids levels which in turn determines an inhibition of the synthesis of several intracellular antioxidants. our results offer promise for anticancer therapies with statins in combination with antitumor cytokines (IFN-γ. These limited antitumor effects of PRV might be explained by the hydrophilic nature of this drug and by the lack of a drug transporter in the membrane of B16. with a higher extent for liposomal SIM (85% inhibition of tumor growth) compared to liposomal PRV (60% inhibition of tumor growth). In conclusion. IL-12p70). 43 . PRV did not have any cytotoxic activity on B16. such as inflammatory cells infiltrated in tumor mass.F10 murine melanoma cells.the rate-limiting enzyme in de novo cholesterol synthesis.due to the inhibition of superoxide dismutase activity in cancer cells.F10 melanoma cells. No antitumor actions were noted after statin administration as free forms. SIM induced the reduction of angiogenic/inflammatory capacity of tumor cells that can strengthen and support its cytotoxicity. Our in vivo data suggested that both liposomal statins had strong inhibitory activity on tumor growth.1α expression via quenching the activation of Ras proteins. We noted only a delayed and lower suppressive action of PRV compared to the SIM on the expression of HIF-1α and on production of proangiogenic/ proinflammatory proteins in tumor cells. the antitumor activity of both statins was enabled by liposomal formulations that can allow passive accumulation of drugs at the sites of malignancy. vital for membrane stabilization and suppression of HIF. In addition. The hydrophilic statin. Altogether. The strong cytotoxic capacity of SIM might be related to the inhibition of 3-hydroxy-3-methylglutharyl coenzyme A (HMG-CoA) reductase . These beneficial actions might be exploited in future in vivo experiments that will aim to investigate the antiangiogenic potency of liposomal statins on other cell types than tumor cells. Sofia. Gagea (13). Figure 1. STOEVA Institute of Biodiversity and Ecosystem research. BOSSEVA. GUSSEV. S. Ornithogalun (12) and Colchicum (11). IVANOVA. living and in vitro collections are extensively used for the conservation Liliaceae species. V. most of them economically important with ornamental.EX SITU APPROACH TO CONSERVATION OF LILIACEAE IN BULGARIA T. DIMITROVA and T. Richness of Liliaceae in Bulgarian flora. According to the IUCN criteria. Most abundant genera are Allium (41). Bulgaria Family Liliaceae comprises 140 species of Bulgarian flora. ANGELOV. Y. edible and medicinal value. 1). Many of them are slow-growing and have reproduction impediments that require development of species-specific procedures. Nearly half of Liliaceae species are of conservation importance and 20 are included in Bulgarian Red Data Book (2011). GEORGIEV. 13 are critically endangered. Legally protected by the Bulgarian Biodiversity Act (2002) are 43 species and 17 are under the regulations of the Medicinal Plants Act (2000). Ten of the genera are monotypic (fig. Bulgarian Academy of Sciences. 5 are endangered and 1 is vulnerable. G. D. TSONEVA. Propagation and conservation protocols are developed predominately for breeding programs of economically important 44 . CH. Ex situ storage of germplasm in seed banks. such protocols are missing or insufficient.species. In vivo and in vitro propagation have been applied to selected species from Tulipa. The effectiveness of the applied procedures. cytogenetic and isozyme analysis are presented. However. 45 . Fritillaria and Ruscus and results for seed germination. Seed collections of 22 species are deposited in Millennium Seed Bank and since 2011 a Research seed bank has been launched in IBER-BAS. Case studies for different conservation approaches applied to Liliaceae are presented. scientific value of the obtained data and implications for further conservation measures are discussed. for many other taxa. especially for local endemics. Lillium. preferably in the country of origin”. biosynthesis of useful secondary metabolites by plant cell cultures. D. In this context. MITOI. Cucurbitaceae. R. COGĂLNICEANU Institute of Biology Bucharest. BANCIU. electrophysiology. Achillea schurii. We have established reproductive multiplication protocols (by somatic embryogenesis. an extreme halophile plant. ALDEA. Primulaceae. Our objective was to develop reproducible and efficient protocols for collecting. VOICHITA. Germination experiments on Nitraria schoberi L. although the European Plant Conservation Strategy (GSPC target 8) recommends that “60% of threatened plant species should be introduced in ex situ collections. CATANA. Plumbaginaceae. HELEPCIUC. in vitro stress phenomena. Asteraceace. MIHAI. revealed that the first and the compulsory step for their ex situ conservation have to be the dormancy breaking. Isoenzyme patterns (catalase. Campanulaceae. Gentianaceae. Knowledge about breaking dormancy and optimizing seeds germination is useful in plant conservation. Despite existing in situ conservation activities Romania still does not have such a strategy. M. I. HOLOBIUC. AIFTIMIE-PĂUNESCU. ISTRATE and G. Liliaceae. belonging to Caryophyllaceae. We focused during the last decade on plant cell differentiation and morphogenesis. esterase. F. medium and long term. adventitious shoot regeneration) for several Vascular species listed in the Romanian Plant Red Book. R. Convolvulaceae. A. C. peroxidase. Biotechnological approaches for plant ex situ conservation. axillary shoot proliferation. Analysis of the genetic diversity within native populations is a prerequisite for ex situ conservation being useful both for sampling and prioritizing strategies.microbe interactions. Scrophulareaceae. Romanian Academy The main objective of our team is to use plant in vitro systems for fundamental research and biotechnological applications.E. micropropagation and storage of endangered plant species for short. C. we have started an in vitro collection for endangered plant conservation. Papaveraceae. The continuous decline of biodiversity and the growing problem of climate change had led to an increasing interest for ex situ plant conservation. genetic transformation. Our current research topics include endangered plant ex situ conservation. and apoptosis. and Marsilea quadrifolia. 46 . Convolvulus persicus. alkaline phosphatase) and RAPD analyses have been used to characterize intra-populational variability in some wild populations of Centaura pontica.PLANT IN VITRO SYSTEMS FOR EX SITU CONSERVATION AND BIOTECHNOLOGICAL APPLICATIONS I. CRISTIAN.E. and the study of plant . F. Polypodium vulgare). Athyrium filix-femina. Melissa officinalis. To assess the genetic stability of the in vitro regenerants. Ruscus aculeatus. Bioactive compounds extracted from plant cell cultures are already widely used. We successfully established active in vitro plant collection based on medium term preservation protocols in several species from Caryophillaceae. Synthetic seeds defined as "encapsulated meristems or somatic embryos which functionally mimic seeds able to develop into seedlings under sterile conditions” were also obtained in Dianthus and Gentiana spp. Microbial and fungal cultures or different fungal extracts were applied to activate plant defense systems. For any in vitro conservation program of rare and/or endangered plants a method for medium and long term storage is required. different strategies were used each of them with advantages and limitations. Atrichum undulatum. In vitro manipulation of plant cell cultures for screening of useful biocompounds is a research direction that will be developed in the future in our lab. but are influenced by ontogenic development and physiological status. Fabaceae families. Reboulia hemisphaerica. Marchanthia polymorpha). Athalamia hyalina. liverworts: Bucegia romanica. The identified anatomical differences represented reversible adaptive responses to the in vitro culture conditions. Fragaria x ananassa. Glycine max. But the most spectacular and the most studied is a long-term (20 yrs) stable callus culture of Vitis vinifera which is producing anthocyanin pigments. Plant in vitro systems for secondary metabolites biosynthesis. Asplenium trichomanes. Scilla autumnalis). we succeeded to cultivate in vitro: lichens (Xanthoria parietina. Gentiana punctata.Brassicaceae. and ferns (Marsillea quadrifolia. For the first time in Romania. for several species (Dianthus callizonus. Usnea barbata). RAPD analyses are sensitive in detecting variations among individuals between and within species. Cetraria islandica.dehydration. droplet freezing) have been tested on plant material from our collection. In our lab we have obtained callus cultures of Ecballium elaterium. Isozyme analyses provided a convenient method for detecting genetic changes. Alyssum borzaeanum. The RAPD profiles of Marsilea quadrifolia showed a genetic stability among micropropagated plants. For example. vitrification. the in vitro methodologies were tested previously on common species. Long term preservation methods (encapsulation. Gentianaceae and Scrophulariaceae families. isozyme analyses and reverse 47 . Marsilea quadrifolia. bryophytes (mosses: Bartramia halleriana. we studied in vivo and in vitro variability of histological traits of aerial vegetative organs. We used optic and electron microscopy analyses. As a working principle for the lower plants. In vitro regenerants from nine plant species were characterized by enzyme variability for detecting possible somaclonal variations. Plant-microbe interactions. resveratrol and pycnogenol. Defense-specific changes were observed in treated plants. like cell wall strengthening. 48 .transcription-PCR (RT-PCR) methods to detect cellular and molecular responses. and expression of some defense-related genes. activation of new isoforms. the term telocytes (TCs) was introduced as replacement for the ICLC. Moreover.. Diagramatic representation of telocyte ultrastructural phenotype. Recently. Two long telopodes originate from the cell body where the nucleus is located. 2011) as is also the case of such phenotype in stromal adipous tissue (personal ultrastructural unpublished data). 2011). b. 2011. a rough endoplasmic reticulum and. Gherghiceanu and Popescu.TELOCYTES – INFRASTRUCTURE AND FUNCTIONAL ROLE N. Each telopode is represented by alternations of podomer-podome. Usually. telopodes follow a convoluted aspect (Popescu. Fig. 2012 a. TCs are represented by a special phenotype of interstitial (stromal) cells. inside of the perinuclear cytoplasm. by their telopodes. Sometimes. similar to canonical (gastro-intestinal) interstitial cells of Cajal (ICCs) have been described by Popescu’s group (Popescu et al. dyctiosomes of Golgi apparatus are located. 2005) in Bucharest (Romania). endoplasmic reticulum and caveolae (Ca2+ uptake/release units). the delivered macromolecules from these microvesicles are considered to play a paracrine role by sending signals to 49 .. Romanian Academy A new type of interstitial (stromal) cell termed interstitial Cajal-like cells (ICLCs) because of their morphologic aspect. A synapse is depicted (elipsoidal area) between the telocyte and an adjacent extension of another cell.and/or hetero-cellular junctions. 1. A telocyte (TC) has a cell body where an ovoidal nucleus is located and two or more (maximum five) cell prolongations (Fig. Cell-body prolongations termed telopodes are very long (tens up to hundreds of micrometers) with a moniliform aspect. TCs were identified and described in different normal tissue types (Popescu. Telopodes may perform homo. As a general rule. 2012. Cantarero et al. Rusu et al. 2011. 1).. MIRANCEA Institute of Biology Bucharest. Each telopode start abrouptly from the cell body as slender cell extension with moniliform aspect: thin segments less than 200 nm thickness (podomeres) alternate with dilated segments (podoms). at first glance. TCs release small microvesicles (mean diameter of 180 nm). eventually. podoms accommodate mitochondria. c. J. Nicolescu M. 27(6): 807-816. 2005. mast cells. Cell Tissue Res. we describe for the first time TCs as interstitial cell phenotype inside of the human skin tumor stroma (manuscript in preparation).. Cell. 2012. that some particular aspects are detectable. and vimentin. Histopathol. 1.. Dermengiu D. unfortunately.. 4. C-kit. 15: 26-30.A.... TCs establish homocellular junctions by specialized somatic synapses leading to a 3-D network inside of the tissue stroma....... 193: 403-411. 16(2): 401-405.. Biol. being a matter of debate and..S. In many respects.. 6. Pop F... J. 5...S. Mănoiu V. Hostiuc S. Popescu L... 8. Gherghiceanu M.. Popescu L.I... 2011.S. In spite of the fact that TCs give positive immunohistochemical reaction for CD34. Rusu M.. but mention must be made.. Creţoiu D. Popescu L. Luesna B. Biomed. Due to their heterocellular junctions (heterocellular connexions).. Vâlcu M. Hostiuc S.. Mol. TCs are considered to be involved in intercellular signalling – in pathophysiology. 194(4): 359-367. the ultrastructural cell phenotype identification remains the only possibility to discriminate TCs from other stromal cell mimicing the telocytic phenotype. Junquera E. Rusu M.. especially concerning the number of heterocellular junctions.. Histol.C. Rusu M.C. TCs from the tumor stroma share general ultrastructural characteristics to those described in normal human skin. Ion D.M. Jianu A. so that. 2012b. 348: 265-279. 2012c. Mănoiu V. As stromal cells..M. Med. Mirancea N.. Gherghiceanu M.M. Mirancea N. Anat.. Med.M. Radu E.. Engin. 3. Cell Mol. J. Using TEM investigations.. 9(3): 714-730.C. Ann..C. TCs are located in strategic positions: often they are in close vicinity or in direct contact with endothelial cells. 2011. 2.C. Păduraru D. Lală A. 50 . Didilescu A.. Păduraru D. Cantarero C. Pop F. Paduraru D. Anat. naked nerve edges etc... Mirancea N. Ann. Rusu M.. 2011. J.. The possible functional role of the TCs as tumorstroma cells remains largely unknown. Mănoiu V. so far there is no report about a reliable immunolabeling specific to TCs.neighboring cells.I.. 7. 2012a. further investigations are required.C. 5(2):83-92.. Internat. Cell Mol.. Med. Gurca G. Jianu A.C.M. SPAAK1 and C. shifting the chemical equilibrium in the lake from harmless ammonium to highly toxic ammonia. The nutrient pollution of the Danube River led to an accelerated eutrophication of the shallow lakes in the Danube Delta in the 80’s. mass mortality of fish can also be caused by cyanobacterial toxins. Lyngbya. can have tremendous impact on aquatic ecosystems and wildlife. EAWAG. with drastic consequences for the trophic networks: massive cyanobacterial blooms occurred. whether toxic or not. Monitoring lakes to follow the pollution and recovery process takes decennia and requires large investments. SANDU2 1 Swiss Federal Institute of Aquatic Science and Technology. Dübendorf. In eutrophic water bodies mass mortality of fish can occur as a consequence of oxygen depletion. Oxygen depletion can also occur after raise in pH due to increased photosynthesis. especially in the very dry and warm years occurred in the last decade. Their presence may not only affect drinking water quality and recreational use by reduced transparency.and human health. Nostoc and Planktothrix have been found to be able to produce toxic secondary metabolites that represent a health hazard to human and animal health. and in the recent years. affecting the seasonal succession of plankton and macrophytes communities. the history of pollutants and species compositions 51 . CH. Cyanobacterial blooms. Cylindrospermopsis. Nodularia. the nutrients stored in the sediments and the increasing impact of climate change favored new episodes of cyanobacterial blooms. Romanian Academy The goal of this research project is to emphasize the impact of harmful cyanobacterial blooms on aquatic ecosystems of the Danube Delta and develop methods to predict future patterns of such blooms in order to identify protection strategies to reduce health hazards in the future. Field studies have shown that cyanobacteria negatively influence zooplankton biomass and community composition compared to other microalgae such as chlorophytes and flagellates. which influences the whole food web. Sediments are an excellent archive of biotic remains and biomolecules under cold and anaerobic conditions. Aphanizomenon. but over 40 species from the genera Anabaena. Although apparently between 1990-2000 the aquatic ecosystems seemed to recover slightly. However.INVESTIGATING THE IMPACT OF CYANOBACTERIAL BLOOMS ON AQUATIC ECOSYSTEMS OF THE DANUBE DELTA P. 2Institute of Biology Bucharest. impacting the food web and having possible adverse effects on livestock. bad odour and taste. principally by high microbial oxygen demand during bloom die-offs. Microcystis. the discovery of gene clusters responsible for the biosynthesis of cyanobacterial toxins (microcystins. The goal of this research project is to reconstruct: i) the history of cyanobacterial blooms. nodularin. cylindrospermopsin. Hence. offering a novel window into past aquatic ecosystems and the environmental changes that triggered community dynamics. This knowledge is crucial to predict the risk for harmful cyanobacterial blooms in the context of environmental change and their consequences for lake food webs. in some instances being also possible to derive a toxicity index based on the quantity of toxic genes. The identification of a broad range of cyanobacterial genetic markers allow their use to detect.using DNA markers started to be deduced from lake sediments. based on total lipid extracts from sediment layers and subsequent analysis by gas-chromatography-massspectrometry (GC-MS). quantify and study structural. also. Quantitative real-time PCR (qPCR) has been increasingly applied to monitor potentially toxic cyanobacteria population in diverse aquatic environments. contamination by petroleum-hydrocarbons may also emerge as molecular signature in lipid biomarkers. anatoxin-a and homoanatoxin-a) allows the identification of toxigenic strains in the environment. moreover. saxitoxin (STX). An alternative method to reconstruct past cyanobacterial dynamics in sediments targets lipid biomarkers. These methods can also be applied to ancient DNA in lake sediments. The molecular distribution of lipid biomarkers can also reflect different aspects of the biogeochemical processes linked to lake contamination and cultural eutrophication. the project will focus on 4 major objectives: • development of methods to reconstruct cyanobacteria dynamics from lake sediment cores • relate the cyanobacterial blooms with other biotic and abiotic factors that may influence their development • resurrect Daphnia and cyanobacteria from sediment samples to test for adaptations and eventual parasites prevalence 52 . functional and evolutionary aspects of cyanobacteria and their communities in the field. and iii) the effects of cyanobacteria on locally adapted zooplankton in the Danube Delta. The hydrogenisotopic composition of individual lipids from aquatic organisms can be related to the composition of their growth environment. The detection of patterns associated with N-fixing cyanobacteria allows the reconstruction of historical changes in N-limitation of the aquatic ecosystem. ii) the occurrence of toxic genes from lake sediments. especially in the context of the expected increasing impact of climate change. 53 .• analyse the sediment cores of selected Danube Delta lakes and develop conceptual model to forecast future blooms and consequences based on climate. where eutrophication of lakes and rivers is still of high concern. discharge and pollution scenarios The methods developed during the project and its results will represent valuable tools for water managers in Eastern European countries. Bucharest Unicersity Litter decomposition is a key component of nutrient cycling and of terrestrial productivity. Litter decomposition resulting from trees and herbaceous layer reinforces the organic matter enrichment of soil and releases necessary nutrients for plants’ growth and soil CO2 fluxes. respectively different nutrients fertilization. VĂDINEANU3 Faculty of Biology. Litter decomposition process was investigated in forest ecosystems using of plant litters Acer pseudoplantanus. component of the national Long Term Socio-Ecological Research Network.CONTRIBUTIONS TO THE KNOWLEDGE OF LITTER DECOMPOSITION RATES OF PLANT LITTERS OF Acer pseudoplantanus. PREDA2 and A. The experimental research was conducted in Neajlov catchment. understanding and explaining the mechanisms involved in litter decomposition under the influence of anthropogenic pressure. The aim of this paper is to make contributions in identifying. 54 . E. Determination of litter decomposition rates has a crucial role in identifying factors that influence nutrient availability. Fraxinus excelsior and Quercus robur and the results of decomposition rates depending on anthropogenic pressure exerted. JERCA1 . particularly forest ecosystems and global carbon cycle. Fraxinus excelsior AND Quercus robur IN FOREST ECOSYSTEMS UNDER THE INFLUENCE OF ANTHROPOGENIC PRESSURE A. All studies have been focused on the biodiversity of a region. For maintaining these values people need conservative strategies for minimizing loss of biodiversity. Debates regarding both the form of the relationship between species richness and ecosystem processes and the mechanisms underlying these relationships have been along time. changes in ecosystems’ good and services provided to society have high economic impact. studies of climate change especially showed that changes in biodiversity at all the organizational levels determined changes of the processes and the conditions from the local. changes in organisms’ community composition and vulnerability to invasion. medicinal or genetic resources for human beings are directly reduced by changes in species diversity and abundance. intellectual. trophic interactions influence the abundance of species that control and modify fluxes of materials and energy affecting the ecosystem processes. HONCIUC and D. to regional and world level. going finally to affect ecosystem goods or services and to alter controls on critical ecosystem processes. More than the presence or absence of the species integrated in the ecosystem. fuel. Developing the project RO1567IBB01/2012 “Researches regarding the relationship between biodiversity and functions in some ecosystems from Romanian Carpathians” we try to clarify some aspects of the relationships between biodiversity and ecosystem functions at regional level by transferring biodiversity data obtained in the last decades in the ecosystem services studies. ONETE. structural materials. These processes and conditions of natural ecosystems that support and sustain human life are defined as ecosystem services. MINODORA. leading to reduced control of the ecosystem processes. In our day biodiversity is defined at the organizational level of life as the totality of genetic. Romanian Academy In the former time scientists used the terms „species diversity” and „species richness”. 55 . Also. biodiversity has great impact on human values (cultural. species and ecosystem diversity. NICOLAE. the ecosystem characteristics and processes are determined by the non-additive interaction among species. For instance.BIODIVERSITY DATA TRANSFERRED IN ECOSYSTEM SERVICES STUDIES M. M. aesthetic and spiritual).C. In the last decades. V. For human society. The biodiversity generates ecosystem services. The food. D. PURICE Institute of Biology Bucharest. which have extramatrical mycelium of contact and short distance exploration type. The aim of this study is to investigate the ectomycorrhizal (ECM) diversity from areas affected by anthropogenic disturbance compared to those from less disturbed systems. The research was carried out in the framework of Synthesys Project GB-TAF-564/2012 and RO1567-IBB03/2012 Project. The first data set regarding colonizing root tips revealed that in the stands of varying levels of anthropization the ECM species diversity is lower that in the reference site. Three sites. the highest frequency have the species belonging to genera of Russula. but their function differ in ecosystems with different anthropization levels. were identified. Tuber and Cenococcum.PRELIMINARY ASSESSMENT OF THE EFFECT OF URBANIZATION ON ECTOMYCORRHIZAL DIVERSITY D. with various levels of disturbance. It is known that fungi are key organisms in nutrient cycling and ecosystem stability. that have Quercus as dominant host tree. A reference site of a mature forest was selected for comparison. Quercus associating ectomycorrhizal fungi communities were sampled from each site. Romanian Academy The conversion of natural ecosystems in urbanized ecosystems increase ecological footprint and have strong negative impacts on the ability of natural capital to produce resources and services. MOGÎLDEA Institute of Biology Bucharest. This data are fundamental for understanding the differences between ECM communities’ functions in ecosystems with different degree of anthropization and for identification of the level of anthropogenic disturbance from which the ectomycorrhizae role is highly perturbed. The urban ecosystems exhibit a simplified structure and reduced function compared to natural ecosystems. The preliminary data revealed that in the reference site. 56 . under the United Kingdome control. Roan and Sable Antelope. MIHĂILESCU Institute of Biology Bucharest. it is very important to use the scientific studies 57 . jewels and rhino corn for medicinal purpose. developing the human elephant conflict resolution. increase the conflicts between human population and wild animals. for examples: African Elephant. Leopard. a number of species have endangered status according with IUCN categories of species protection. especially in Kenya. the European countries and the African countries have common purpose for world species conservation. In Africa. increase the local and international community participation. due to this reason. Conclusions: Increasing the research studies about wild animals’ behavior is the proper mechanism for human animals’ conflict resolutions. Developing the applied conservation measures according with ecological criteria’s for establish the rules for living closing to the animals is the main challenge. The outcomes of species and their habitat protection projects are: increase the protected areas and wildlife sanctuary according with national and international regulation. developing the basic infrastructure and operations. species specific programmes such as Monitoring the Illegal Killing of Elephants . especially in the countries from south-east Asia.MIKE) must find external funding. management of species conservation oriented to research. Romanian Academy The Convention on International Trade in Endangered Species of Wild Fauna and Flora – CITES is the specific legal instrument for conservation of species that should be protected in natural habitats for their survival. increase the uncontrolled trade with ivory objects. In the last fourteen years. The frequent reducing population of wild animals is related with illegal hunting. mostly from donor countries and regional organizations such as the European Union. The conservation activities (training. Main anthropic pressures on species and their habitats are: reduction and fragmentation the species habitat due to expansion of human population. In respect of CITES convention. decrease the area of wild natural habitats. Lion. White Rhino. the many Mammals species request special management for their protection. The important nature conservation developing in Kenya was start in 1975 with Environmental Law Protection. increase the conservation education. law enforcement.CITES AND NATURE CONSERVATION IN REPUBLIC OF KENYA S. IFAW and Kenya Wildlife Service. Future important step in conservation: developing the public awareness campaign to inform people both about the non-medicinal importance of rhino corn and about negative effect of developing the manufacturing of ivory objects and jewels. 58 . CEEweb for Biodiversity.financial and organizational support of the European Commission.for analyzing the possible securing critical corridors and dispersal areas of the wild endangered animals. Acknowledgement: Project “Implementation of CITES and EU Wildlife Trade Regulations in CEE region” . a structured brief report on the general implementation of the directive in each Member State.Annex D: Reporting format for the conservation status of a habitat type . . For all of the Member States the reporting format request the general description of the main measures taken (overview at national level. activities taken including legal measures. Main achievements under the Habitats Directive are represented by the reporting period with a special emphasis on the Natura 2000 network.PROJECT SOP ENVIRONMENT: CONSERVATION STATUS MONITORING OF SPECIES AND HABITATS FROM ROMANIA UNDER THE 17 ARTICLE OF THE HABITATS DIRECTIVE 2011-2012 S. estimate based on partial data with some extrapolation and/or modelling.Annex E: Evaluation matrix for the conservation status of a habitat type .it defines the information required for each habitat type covered by Annex I of the directive on biogeographic level. I. or absent data can be reported. providing links to important information sources. the following can be used: complete survey or a statistically robust estimate. The deadline for submitting the 3rd report to the Commission is the 30 June 2013. links to online resources.defines the method and criteria on how to assess the conservation status of each habitat type on biogeographic level. systematic studies. IV and V of the directive on biogeographic level. Romanian Academy The Habitats Directive defines a six-year cycle for reporting. MIHĂILESCU. estimate based on expert opinion with no or minimal sampling. ION. M. 59 .Annex C: Evaluation matrix for the conservation status of a species – defines the method and criteria on how to assess the conservation status of each species on biogeographic level.). . . etc. The Reporting Format for 2007-2012 consists of: . Regarding the criteria represented by the Method used /Population size. being for Romania the first assessment of conservation status based on best available data for each species and each habitat type.Annex A: General reporting format .Annex B: Reporting format for the conservation status of a species it defines the information required for each species covered by Annex II. GHEORGHE and I. CRISTEA Institute of Biology Bucharest. . fish (93). like the distribution and biology. distribution maps for species and habitats of Community importance in the standard format GIS ETRS. amphibians and reptiles (15 137). scrubs/ bogs and mires/ swamps/ rocky formations/ forest (80). assessments for species and habitats of Community interest -species of plants (102). like coastal and marine (9). caves (3). scrubs/ bogs and mires/ swamps/ rocky formations/ forest (44). which aims by 2015. invertebrates (192). mammals (72). regular grid of 10x10 km. metafile database with records made in the field. Monitoring plans are made for all species and habitats of Community interest.22). bats (107) and habitats. 28) and habitats. marine (4 species of mammals and 9 species of fish). the preparation of the National Report according to Article 17 of the Habitats Directive 92/43/EEC. as follow -species of plants (105 species). sets of data in the reporting format. like coastal and marine (10). fish (35). published in 500 copies each. the particularities of each species or habitat. as well as the implementation of the Natura 2000 network nationwide. differentiated for each biogeographical region. amphibians and reptiles (15 and 19 respectively). halophytic/dunes/ grasslands/ freshwater (33). Acknowledgement: „Conservation status monitoring of species and habitats from Romania under the 17 Article of the Habitats Directive” 20112015. halophytic/dunes/ grasslands/ freshwater (79). Summary Report on the state the conservation of species and habitats of Community interest in Romania.The project goal is to protect biodiversity and natural heritage and to improve the natural environment through proper monitoring of the conservation status of species and habitats of Community interest in Romania. the following: database with existing information on the status of species and habitats of Community importance. caves (1). introduced in national monitoring system (SIMSHAB). invertebrates (73). bats (Chiroptera. monitoring guides for species and habitats. marine (21). During 2011-2012. being developed by taking into account the requirements of the European Commission. following the conclusions of the report and the information dissemination activities. mammals (terrestrial. database of publications on species and habitats of Community interest. the project has recorded some progress which will contribute to the final objectives of the project. project financed by SOP Environment 60 . Hennekens and J.TURBOVEG DATABASE USED IN BUCEGI MASSIF STUDIES C. ArcGIS. Since 1994. There is a fixed standard structure of the table. there could extend both the header data (e. ArcView.D. Mulva. There are releveés from Koelerio-Corynephoretea (incl. In the first of these. Sedo. Vegetation Database of the Dry Grasslands from the Transylvanian Basin. PC-ORD.Scleranthetea. but it easily to extend it by defining an extra field. Syntax-5. alpine grasslands of Bucegi Massif are included (RO1567-IBB01/2012). Schamineé) in 2001. NICOLAE Institute of Biology Bucharest.g. Vegetation Database of Dry Grasslands in the Southeast Romania. Excel. Each database can have its own extension. Canoco. Database Beech Forest from Romanian Carpathians and Vegetation Database of Oak Forest of Romania. there are filters available to export the selected relevés to enable further analysis with various programs (e. Kobresio myosuroidis-Seslerietea caerulea) and Festuco-Brometea classes. and export of vegetation data (relevés). selection. TURBOVEG has been accepted as the standard programme for the European Vegetation Survey. Elyno-Seslerietea (Seslerietea albicantis. This is a very useful tool for the processing of phytosociological data. Juice. In this way. MS Access. On the other hand. For the beginning they were included 72 releveés of natural and seminatural grasslands of studied teritorry. GoogleEarth). designed for the storage. in Romania there are known in this moment five vegetation database: Vegetation Database of Dry Grasslands in the Southeast Romania. add altitude or total cover) and the species data. Oak-Hornbeam Forest Database of the Transylvanian Basin. Festucetea vaginatae). Data can be imported manually (separate relevés or tables) and automatically (import of data files). 61 . Romanian Academy The computer software package TURBOVEG (for Microsoft® Windows®) was developed by two Dutch researchers (S.g.Twinspan. According to this. 62 ECOLOGY, TAXONOMY AND NATURE CONSERVATION POSTERS 63 64 2359 wet weight μg/l/24 h. Between 2011 and 1983 (the eutrophy peak) a high similarity (68%) was noticed. All quantitative parameters reached the highest levels (1008 ind/l. In 2011. biomass and productivity recorded significant increases (668 ind/l. abundance. Between 1975-2011. led to structural and functional changes of entire foodweb. 65 . 7231 wet weight μg/l. In the second period. the species richness values were diminished. By applying the simple correlation between planktonic producers and consumers in 2011. resulted in the completion of several distinct stages of trophic status succession. biomass and productivity are lower (194 ind/l. 842 wet weight μg/l/24 h. a strong relationship was found. in 1975 – 2011 interval. respectively).588778. The question is: can this be a new stage of hypertrophy in the shallow lakes of Danube Delta? To detect the differences among years based on specific richness. the hypertrophy period with maximum anthropogenic impact (1981-1990). while the Pearson correlation between phytoplankton and zooplankton abundance was very highly significant (R2=0. 860 wet weight μg/l. The consequence was powerful "water bloom"phenomena due to excessive algal development. This fact confirms the variations of zooplankton parameters in different trophic conditions. These processes. Their increased intake favored the replacement of macrophyte type producers dominance with algal type. the Lake Roşu shows a special case. the evolution of Danube Delta lacustrine ecosystems was characterized by three significant stages: the reference period (before 1980) characterized by meso-eutrophy. Romanian Academy The pressure of natural and anthropogenic factors on shallow lakes in the Danube Delta. 19338 wet weight μg/l. and the recovery period (after 1990) defined by a decline in trophic level. a single factor ANOVA test was applied. respectively). Future research will elucidate the causes. The Bray Curtis analysis was used to assess the similarity degree among the annual values of zooplankton biomass. 104 wet weight μg/l/24 h.05) for all parameters. In the reference period the species richness was much higher than in eutrophic-hypertrophic period. while the values of abundance. The responsible factor for this process is the dynamics of nutrient concentrations. biomass and productivity of zooplankton. The obtained values indicate statistically significant differences (p<0. The abundance. follows the general trend of deltaic shallow lakes. respectively). PARPALĂ Institute of Biology Bucharest.LONG-TERM ECOLOGICAL RESEARCH OF ZOOPLANKTON COMMUNITY IN LAKE ROŞU (DANUBE DELTA) L. the zooplankton dynamics of Lake Roşu. In terms of trophic levels. 550458. The zooplankton biomass is highly significant related to phytoplankton biomass (R2=0. lead us to conclude that the zooplankton dynamics is directly dependent on ecosystem trophic status.000841).P=0. Ecological research conducted between 1975-2011. 66 .001541). P=0. such as decreasing of the species number. natural sectors and canals. major hydrotechnical changes in the original structural configuration occured. The annual K dominance analysis highlights the following species: Brachionus calyciflorus dorcas Gosse.SPATIAL DISTRIBUTION OF THE ROTIFERS DIVERSITY IN SFÂNTU GHEORGHE BRANCH L. Romanian Academy Rivers are dynamic ecosystems. as Rényi diversity profiles. Rényi diversity profiles easily order from high to low the diversity for each area. These issues can be solved by using in addition.g. One sample may contain a higher species richness while the other has a better Shannon Index. Earlier research results at Ceatal Sf Gheorghe (1981-1985. the natural sectors and meanders 67 . being in permanent anthropogenic changes simultaneous with society development. riverbank morphology. 1991-1992) emphasized 105 species of rotifers. water flow. while the autumn 2008 was the most unfavourable season for rotifers development for the entire Sfântu Gheorghe branch. by cutting off six meanders (the bends areas of the river) and consequently shortening the river with around 32 km. During the 80’s. Instead. The seasonally graphic representation of K dominance curves for each area illustrates the diversity dynamics during the research. Synchaeta oblonga Ehrenberg. which provide many goods and services. in Sfântu Gheorghe branch (Danube River). turbidity. FLORESCU Institute of Biology Bucharest. the river systems played a fundamental role. the technical engineering changed the relationship between biocoenoses and hydrogeomorphological conditions by creating new areas (new canals) with different features from the rest of the branch. a diversity ordering method. highly complex. Despite the fact that the diversity indices are widely used in ecological research. Our research conducted between 2008 and 2010 has shown the consequences of these works. Keratella quadrata Gosse. while the recent results point out a decrease to 83 species. erosion processes). density and biomass. they have presented among the highest values of abundance in the entire branch. for all three types of ecosystems: meanders. Thus. This kind of human impact induced severe effects on the environmental conditions (e. these works consisted in straightening out the branch. Due to their importance. giving a general view for the entire survey period. The lowest curve of the diversity profile belongs to the canals representing the areas with smalest diversity of the Sfântu Gheorghe branch. comparing the different sites or samples may not be so accurate. water residence time. The highest diversity was found in the autumn of 2009. In order to meliorate the navigation. we applied oneway ANOVA single factor analysis using the Shannon Wienner. Simpson’s dominance and Evenness values. The results have shown no statistically significant difference between natural sectors and meanders (P>0. making impossible to distinguish the diversity differences between them. For confirmation. 68 .05).curves tend to overlap when α reach the value of 1. LINKING THE STRUCTURE OF THE WEED-BED FAUNA COMMUNITIES AND THE ENVIRONMENTAL VARIABLES IN SFÂNTU GHEORGHE ARM OF THE RIVER DANUBE E. 2004). Multivariate statistical analysis allowed the identification of key control variables explaining the spatial and temporal variability of the studied communities. 2University of Bucharest. RÎŞNOVEANU2 1 Institute of Biology Bucharest. The aim was to shorten the navigation route with about 32 km and increase the fluid and the solid flows (Popa. modifying its morphology with important consequences on the structure and functioning of aquatic communities. Faculty of Biology. The effects of the past hydrotechnical works are still poor documented in the scientific literature. compared with those in the newly built canals. In order to capture and characterize the weed-bed fauna response to the particular conditions of the habitats in different morphological sections of the watercourse. RADU1. COMAN1 and G. including the development of hydrotechnical works along the Sfântu Gheorghe arm of the River Danube. The research project entitled “The impact of hydrotechnical changes on the ecological systems of the Sfântu Gheorghe arm in the wider context of the sustainable development” (2008-2010. Vădineanu. Along the Sfântu Gheorghe arm. particularly during the period 1983-1989. Understanding the communities’ mechanisms of response and identifying the main factors that induce and control the structural differences of the trophodynamic modules of the weed-bed fauna in different habitats along the river arm represented important objectives of the research to which this paper is addressed. › greater morphological complexity of the aquatic or riparian plant associations (as well as of their component taxa) in the reference sections and meanders could have been a better premise for the 69 . A. six meanders were cut and new canals were built. taxonomic richness and diversity of the weed-bed communities are generally higher in the free-flowing sections (reference) and in the cut meanders of the Danube arm. Romanian Academy) has attempted to fill this gap in knowledge. 1996. the communitie’s structure was characterized and related to the variation of physico-chemical parameters. Romanian Academy. Department of Systems Ecology and Sustainability The Danube Delta – an important ecological system belonging to the World Natural Heritage – has been transformed over time by different types of human activities. The results have shown the following: › numerical density. g. the total phosphorus and total nitrogen.the variability of these communities’ structure in space (e. 70 . and the water transparency).› development of the associated weed-bed fauna communities in these areas. the dissolved organic carbon. the water temperature and the concentrations of the dissolved oxygen in the water column) while others are explaining .in certain proportions .g. some of the monitored environmental variables are well explaining the inter-seasonal and interannual variations of the weed-bed fauna communities’ structure (e. the organic matter and the chlorophyll in the water column. Tipulidae). RÎŞNOVEANU2. Compared to other weighty wetlands. constant and dominant components in freshwater systems. Ceratopogonidae. consisting in sediment-dwelling species. Brăila Ecological Research Station Benthic macroinvertebrates. Benthic communities are also a cost-effective and commonly use indicators in long-term ecological research and water quality monitoring programs. The Lower Danube Wetlands System (LDWS) that includes the Inner Danube Delta (IDD) is one of the largest and still well preserved wetlands systems in the world. covering a total area of 8. Eleven taxa were identified (Nematoda. Chiriloaia varied between 777 and 10. transition towards hypertrophy of the aquatic ecosystems due to increasing anthropogenic disturbance. and has affected the functions of the LDWS and the ecosystem services it provides (buffering capacity for nutrients and pollutants. G. Gordiidae. Fundu Mare. Hirudinea. Faculty of Biology./m2. Oligochaeta. Benthic samples were analyzed over the 19 years period (1993-2011) in two shallow lakes (lakes Chiriloaia and Fundu Mare). 71 . Results revealed a simplified structure of the studied communities. are major. Trichoptera. BÎRSAN1. CRISTOFOR3 1 Institute of Biology Bucharest. Amphipoda. 2University of Bucharest. In L. production). Gastropoda./m2. Oligochaetes and chironomids were frequently co-dominants in both. along the higher trophic levels. has resulted in important changes of both water quality and structure of the food webs. sustaining the ecosystem services provision. Nevertheless. Chironomidae. macroinvertebrates’ total densities as well as diversity and similarity indexes were used in order to assess in time and space variations within the macroinvertebrate communities structure. the abundance of 9 taxa recorded ranged between 3. 3 University of Bucharest. representative for the IDD. They are particularly important in highly eutrophic systems. G.LONG-TERM DYNAMICS (1993-2011) OF THE BENTHIC MACROINVERTEBRATES COMMUNITIES IN THE INNER DANUBE DELTA C.750 ind.C. the boast of this area is given by the largest expanse of reed bed and pelican colonies. They are significantly contributing to energy flow at the sediment-water interface as well as in the nutrients cycling.521 – 8. part of regional and global biodiversity.307 km2. The total abundance recorded by the 11 taxa in L. Romanian Academy. E.134 ind. RADU1. Communities’ taxonomic composition. IGNAT3 and S. characterized by high densities of energy and mass flows from sediments to the benthic fauna and consequently. Ephemeroptera. in the last three decades of the 20th century. 72 .70 to 1.63 in L. The low values of Shannon diversity index (0.17 in L.38 to 0.91 in L.67 in L. Chiriloaia and 0. Fundu Mare) and Pielou equitability (0.26 to 0.37 to 0. Chiriloaia and 0.number and biomass. Fundu Mare) suggest an advanced simplification level of benthic communities under the pressure of hypertrophy. ) Wittrock. a characteristic stage of the deltaic ecosystems in the critical period after 1980. The Mc Naughton and Wolf dominance index of phytoplankton biomass between 1982-1985 exceeded the threshold of 0. 73 . The Lake Roşu is representative for the lacustrian-type of ecosystems. the chance of intense proliferation of cyanobacteria group increased.72 wet weigh mg-l) have shown new eutrophication signals. Anabaena scheremetievi Elenkin. the herbivorous zooplankton being forced to feed on detrito-bacterial aggregates to be able to survive. they induce a bottleneck in the carbon and energy flow of the plankton food web. Oscillatoria tenuis Agardh. involves increased concentration of nutrients and also the nitrogen factor limiting role of phytoplankton development. becoming dominant in the ecosystem. The hypertrophy. Romanian Academy Cyanobacteria became increasingly dominant as concentrations of total phosphorus and total nitrogen increased during the eutrophication of Danube Delta ecosystems. including intense cianobacterial blooms episodes. and the ecosystem tended to reach a functional regime.5 in all seasons. especially in the warm seasons. inedible algae. Monthly averages in July-September period exceeded 100-125 times the mentioned limit.THE DYNAMICS OF CYANOBACTERIAL BLOOMS IN LAKE ROŞU REFLECTED IN LONG-TERM STUDIES M. yearly averages of biomass exceeded 10-30 times the "water blooms” threshold.) Kirchner. Microcystis pulverea (Wood) Migula. due to a lower nutrient pressure. The edibility degree of these species is very low. the high values of phytoplankton biomass (78. more stable. being the largest lake (1375 ha) of the fluvio-maritime delta. with a wide range of possible biological impacts including potentially toxic effects and impacts on food web functionality. After 1980. Anabaena hassalii (Kütz. MOLDOVEANU Institute of Biology Bucharest. In general. The temporal dynamics of Cyanobacteria blooms is variable. While in 2001 the diatoms and cyanobacteria biomass decreased. The spectacular values of cyanobacterial abundance and biomass between 19801990 triggered also the dominance of potential toxic species. in 2011. In the new environmental conditions. Taking into account these reasons. the dominant species belong to toxin-forming Cyanobacteria: Microcystis aeruginosa Kützing. Microcystis flos-aquae (Wittr. As large. our work aimed to highlight the dynamics of cyanobacterial blooms in Lake Roşu at a wider temporal scale (1975-2002) and update the existing information by presenting the evolution of Cyanobacteria in the year 2011. while for alkaline phosphatase the average values were 1080 nmol p-nitrophenol L-1h-1 in Roşu Lake and 795 nmol p-nitrophenol L-1h-1 in Mândra pond. This paper presents some characteristics of two enzymatic activities β-glucosidase and alkaline phosphatase .. due their size. while alkaline phosphatase is a nonspecific enzyme that catalyzes the hydrolysis of a large variety of phosphate esters (Janson et al. 1991). while for alkaline phosphatase the maximum activity was reached in summer. playing therefore an important role in the transfer of matter and energy through aquatic ecosystems (Harbott et al. Extracellular enzymatic hydrolysis is the first step in mineralization of most of the polymeric constituents included in the composition of the organic matter (polysaccharides. In order to use the energy stored in this macromolecules.8330 nmol pnitrophenol g-1h-1 for β-glucosidase and 6470 nmol p-nitrophenol g-1h-1 for alkaline phosphatase. cannot be used directly by heterotrophic microorganisms (Chróst. both in 2011 74 . organoesters) by heterotrophic microorganisms.. The resulted monomeric compounds can be directly transferred from the environment to microbial cells. also. Nowadays. The seasonal dynamics of β-glucosidase activity shows maximum activity in autumn. 1988). Both β-glucosidase and phosphatase activities recorded high values in water column and sediment. In sediment the high average values were. the recording of enzymatic activities can be used as a method to assess the intensity of metabolic activity of the microbial communities.. In water column the average values for βglucosidase activity were 855 nmol p-nitrophenol L-1h-1 in Roşu Lake and 630 nmol p-nitrophenol L-1h-1 in Mândra pond. 2000). Romanian Academy In the aquatic ecosystems most part of the organic matter consists of polymeric macromolecules that.THE DYNAMICS OF EXTRACELLULAR β . β-glucosidase is a specific enzyme that act in the final step of cellulose degradation (Nicolescu et al.GLUCOSIDASE AND ALKALINE PHOSPHATASE IN AQUATIC ECOSYSTEMS FROM THE DANUBE DELTA IN 2011-2012 INTERVAL I. PĂCEŞILĂ Institute of Biology Bucharest. proteins. heterotrophic microorganisms produce extracellular hydrolytic enzymes able to catalyze dissociation of macromolecular compounds to their monomeric components outside of the cells. after the fall of vegetation. recorded in Roşu Lake .from water and sediments of two aquatic ecosystems from Danube Delta: Roşu Lake and Mândra pond. 2005). In aquatic ecosystems microbial enzymatic activity depends on a multitude of abiotic and biological factors (temperature. macrophytes development. phytoplankton blooms. pH. etc. 75 . Annual average values of the two studied enzymatic activities were higher in 2012 than in 2011 for both water and sediment samples. the highest values were recorded in autumn for both β-glucosidase and alkaline phosphatase activities. oxygen content. In sediment.and 2012.) that may influence the amount of available substrate as well as the hydrolysis kinetics. Future investigations will be focused on the correlations between the extracellular enzymatic activity and the dynamics of the other environmental parameters. Draft of the Preajba Valley hydrographical basin from Oltenia Plain (Cioboiu 2002). 2Institute of Biology Bucharest. Romanian Academy. 3rd. 2nd. This small surface includes a great variety of ecosystems: hillocks covered by hay fields and pastures. covers a surface of 30 Km2. namely the case of the Preajba Valley system. The composition and distribution of the biocoenotic and population structures of the Preajba Valley system functions in accordance with the 76 . streams. Craiova. due to its structural-functional characteristics.STRUCTURAL-FUNCTIONAL PARTICULARITIES OF THE ECOSYSTEMS OF A PLAIN CATCHMENT AREA (CASE STUDY) O. rivulets. Fig. 1). a vast diversity of geographical. and ecological structures. and small reservoirs. Located within a plain area. Bucharest Generally. the system that belongs to the Jiu hydrographical basin. chemical. BREZEANU2 1 The Oltenia Museum. due to its dimensions. CIOBOIU1 and G. swamps. agricultural fields and a complex hydrographical network – springs. a catchment area includes. hydrological. defining in this way the complexity of the river system within a hydrographical basin. This complex organization induces the structural-functional characteristics and complexity of the ecosystems (Fig. Starting from the definition of a river system. and 4th order. rivers can be of the 1st. Nature Sciences. Ujvary. 1. physical. we bring to your attention a more or less particular case. taking into account the hierarchy of the river systems and the classification established by Strohler (apud. 1959). Thus. 25 3. Chironomida. of individuals/sq. Heteroptera. Ostracoda 1.5 66 100 66 16. The Preajba Valley system is atypical due to its geographical location (plain area) and small surface (30 Km2). Ostracoda. Zoobenthos is made up of 13 large groups of invertebrates. Ephemeroptera. In case of phytoplankton.5 113. Ciliata.25 7. In case of the reservoirs. Testaceae. Gamarida.5 16. Zoobenthos structure (mean values) Taxonomic group Chironomidae Gamaridae Ostracoda Heteroptera Gastropoda Bivalvia Cladocera Copepoda Efemeroptera Plecoptera Isopoda Oligocheta Hirudinea TOTAL No. some comprising a great number of individuals (Chironomida 4.7 5. there have been identified species of Diatomeae. in the area of the rhitron (including the springs and rivulets).5 33 Gastropods represent one of the main groups of the benthos structure (there have been identified 18 species).5 226.particularities of the ecosystem organization.5 160 213./m2.600 ind.7 5.750 ind.5 53 26. Plecoptera 226 ind.753 353. Copepoda.5 16.5 20 6172 Abundance % 38.95 0. the structure of the biocoenosis is made up of representatives of all groups specific to the aquatic ecosystems (pelagic and benthic communities). Cladocera.5 66 16. In the area of potamon (the area that covers the hilly and plain sector of the Prejba Valley hydrographical basin).5 49. m 2.85 100.35 0./m2.0 Frequency % 100 16./m2. the ichtyofauna is represented by 10 species.25 5.45 1.5 1./m2) (Table 1). there have been identified more than 70 alga species and in case of zooplankton. Ephemeroptera 213 ind. Table 1. Rotiphera.35 18.746.45 0.5 186.8 8. Thus. but it gathers all the structuralfunctional characteristic of a large dimension hydrographical system.65 3. 77 . cyprinids being dominant.5 293 26.5 16. 65 species of Ciliata. immuno assays and ELISA (enzyme linked immuno-sorbent assay). Mathcad. Modern ecological methods are based especially on molecular analysis and on statistical ones. SPSS. C12/C13). in the last decades. Infostat.MODERN METHODS USED IN TAXONOMY AND ECOLOGY OF SOIL FAUNA M. cybertaxonomy. 78 . which use the specifity of antibodies to detect an antigen. Molecular analysis could be used in revealing the trophic preferences. the combination of stable isotope and fatty acid analysis with gas-chromatographycombustion-isotope-ratiomonitoring-mass-spectrometry (GC-C-IRM-MS). Molecular applications have had particular success in facilitating the identification of taxonomically difficult species. were the most used for the last ten years. BiodiversityPro. new research methods have been used. All these modern research methods provide a more detailed and complex study on soil fauna. Romanian Academy In order to clarify some systematic problems and to highlight the ecological role of the soil fauna groups. MANU Institute of Biology Bucharest. elucidating phylogenetic relationships. fatty acids (FAs). Traditional taxonomy techniques are now improved with modern methods as: field emission low temperature scanning electron microscopy (FE-LTSEM). Approaches with a polymerase chain reaction (PCR) step to amplify dietary DNA are highly effective. Statistical modeling programmes as: Statistica. digital illustration techniques. Canoco. which is one of the best studied stress response systems in the context of heavy metals. SAS. Other modern molecular methods are used to establish indicators for soil quality such as: terminal-restriction fragment length polymorphism (T-RFLP) analysis and the metallothionein system. in genomic mapping. Systat. FISH (fluorescence in situ hybridization). Analysing the trophic interrelationships in soil food web was made using modern techniques as: stable isotope analysis (N15/N14. in understanding of population structures. immunological techniques. which provides a wide variety of statistical and graphical techniques. making possible a better highlight of their ecological role and of their ecological services. in establishment of genetic diversity. A new used programme is R. HONCIUC Institute of Biology Bucharest. as well as functional indicators such as productivity. reference database. investigated. etc. production and energy budget. along with aspects of a task for the evaluation of the hypothesis as of suprasistems issues and biological systems that integrates and soil fauna. diversity.METHODS USED IN THE ECOLOGICAL RESEARCH OF SOIL MITES (ARACHNIDA . number the vertical distribution. dominance. frequency. soil fauna in different ecosystems from Romania. As is known. and biomass.ACARI) – HISTORY AND PERSPECTIVES – V. phase 2012-Identify ecosystems which will be considered in the study and calibration methodology of work. with reference to the similar research carried out in similar areas in Europe. By using the methods he showed with the highest fidelity the structure of soil fauna in the ecosystems. On the basis of the results obtained in the research conducted so far. Through the use of quantitative methods in research verified made far get quantitative parameters as: numerical abundance. we plan to test them using the methods of modern ecology (mathematical modeling. dispersion. Romanian Academy The work is part of the research project-Research on the relationship between biodiversity and ecosystem function in some of the Romanian Carpathians. 79 . These methods have been used in research achievement until now in all kinds of ecosystems in Romania. and through the use of qualitative methods to obtain structural parameters: coefficient of variation. molecular methods. soil fauna. density. relative abundance. is using the most appropriate methods for quantitative and qualitative assessment of the taxonomic groups of soil fauna of both methods rely on statistical calculation. In this context the work aimed at a presentation of the research methods used in environmental research.) that will elucidate questions concerning ecology and the evolution of biological system and aboveindividual. p at Frumuşani and 48.each with 2. known as a crop pest. dominant were: Dermestidae (8. and in the alfalfa crop.5 ind. to identify both the useful and crop pest arthropods and finally to characterize the invertebrate communities in terms of their numerical abundance.22 ind. S. recedent (Corylophidae and Endomychidae ./u. PURICE Institute of Biology Bucharest. had a status of 80 . most of the families of Coleoptera found in this culture. ind. FIERA. Springtails were sampled with a Macfadyen cylindrical corer. Propylaea quatuordecimpunctata and Coccinella septempunctata (Coleoptera: Coccinellidae). This study aims to establish the structure of arthropod communities (Collembola. MAICAN and D. at Singureni only 711. Gonioctena fornicata (Coleoptera: Chrysomelidae). from alfalfa crop was also present in our samples. The entomofauna of alfalfa and rape crops was collected with an entomological net. Establishing the quantitative and qualitative structure of the epigean fauna was performed using pitfall (Barber) traps filled 2/3 with 4% formaldehyde solution. 19 families of beetles were identified in the studied crops.07%) and Chrysomelidae (5.08%). the other left families of beetles.3. 12 families and 15 families at Singureni). with the relative numerical abundance between 1. identified in the alfalfa crop from Singureni are known to be the most common species of predatory coccinelids that are characteristic to this crop. In the culture of rape we could determine species belonging to Phyllotreta and Chaetocnema genera (Chrysomelidae: Alticinae).) from two localities: Singureni (Giurgiu County) and Frumuşani (Călăraşi County). Coleoptera). focusing on invertebrates’ knowledge from rape (Brassica napus L.p.THE COMMUNITIES STRUCTURE OF INVERTEBRATES FAUNA FROM RAPE AND ALFALFA CROPS C. Scarabaeidae (7.) and alfalfa crops (Medicago sativa L. Curculionidae. Romanian Academy Our investigations have been conducted under a bilateral project between Romania and Republic of Moldova.p.03% were classified as subdominant (Bostrychidae . eudominant were: Carabidae (58. In the rape culture from Frumuşani. As we expected.each with 1. at Singureni). Coccinellidae. In terms of the average density number.10%).59%) and Staphylinidae (10. The largest number of families was encountered in the cultures of rape (at Frumuşani.01%). Byturidae .02%) and subrecedent (Cantharidae./u./u.03%).05%).01% and 3. the highest values were registered in the rape crops (16. while in the alfalfa crop from Singureni we found only 11 families. Dermestidae (18. in terms of their numerical abundance. known as a predatory species. only the family of Carabidae (with a frequency of 83. Staphylinidae and Scarabaeidae (each with 50% as frequency). relatively constant were Staphylinidae and Silphidae (with frequencies of 66.4. The accessory class was represented by 4 families and the accidental class by 5 families. were the families Curculionidae (39. 34 species of carabid beetles (Fam. Hemisotoma thermophyla (Isotomidae). Carabidae) have been identified in the studied crops. We notice the presence of predatory species. Silphidae (15. Here the eudominant families were: Carabidae (32. from the rape culture and in the other one crop the most dominant. Agonum dorsale at Frumuşani and Carabus glabratus at Singureni. these species are also part of the predatory invertebrate epigean fauna. Protaphorura sakatoi (Onychiuridae) and Orchesella flavescens (Entomobryidae) were dominant in the culture of rape from both localities. found together with Pterostichus cylindricus and Callistus lunatus.37%) and the left 7 families were subdominant (Tenebrionidae) and recedent (5 families). 15 species of Collembola were identified.2. we could identify similar features as found in the rape culture from Frumuşani. Carabidae (26.89% as frequency) and Carabidae (77. 81 . constant were Curculionidae (88.5%).58%). from numerical density of invertebrates populations’ point of view and their spectrum of activity represent a major economic importance in reducing the attacks caused by pests naturally. the rest of beetles were accessory or had an incidental status in the integrating biocenosis. were the dipters. Tomoceridae and Sminthuridae). followed by Homoptera and Hymenoptera. both in the structure of dominance and constance classes.89%).56%) and Byturidae (12.55. From the insects collected by sweeping the vegetation with nets. eudominant.37%. Our acquired data.4. eudominant in number density. In the alfalfa crop from Singureni we also identified Cicindela germanica.6 % Curculionidae . belonging to 7 families (Hypogastruridae. Entomobryidae. Beetle fauna was within four classes of constancy.67%) and Byturidae (frequency . Chrysomelidae (16.99%).67%) and the left 11 families were subdominant (Byturidae . Staphylinidae . the most dominant were the beetles.09%).3%) and subrecedent (another 8 beetle families). these two species were eudominant in terms of their density number and frequency in the rape crops in both localities.56%). Regarding the alfalfa crop.33%) was constantly present.dominant (9.accidental groups. the family Silphidae . Onychiuridae. relatively constant were the families of Dermestidae. Isotomidae. In the rape culture from Singureni.78% as frequency). but with an accidental status. Neanuridae. Entomobrya handschini and Lepidocyrtus paradoxus (Entomobryidae) were the most dominant species of the alfalfa crop. METHODS – OLD. using various protocols. In the qualitative approach the methods are more subjected to the sampling error risk (it is about the time and intensity sampling. because the field trips are time limited and because they are quite costy. PURICE Institute of Biology Bucharest. the light traps and feromone traps quite rare used. This approach is used with restrictions in protected areas requiring non-destructive sampling methods.THE EPIGEIC INVERTEBRATES IN THE ECOLOGICAL STUDIES IN ROMANIA. CURRENT AND PERSPECTIVES D. There are two main directions of study for the structure of invertebrates’ populations: qualitative and quantitative approach. In this cathegory of methods. These methods require expensive equipment (respirometers. These molecular analyses are also costy and require extra. The quantitative approach involves fauna sampling. The same reasons are valid for the method of markingrecapture necessary to establish the dispersal power and the fauna potential in recolonisation of vairous disturbed areas. when more than one person is taking the samples). Another necessary approach in the functional ecology studies is the molecular analysis of invertebrates’ material. molecular analysis) are critical elements necessary in the ecological studies in order to draw some applicative conclusions for environmental protection and environmental management. invertebrates are a major part both as structural and functional contribution in the integrating coenoses. to confirm the species identity (in case the classical taxonomical tools are not enough) and to establish the gut content (the food spectrum) of some generalist consumers. trophic structure and energetic budget. secondary production & productivity. growing chambers) and are long-time studies.training and knowledge in this domain. Romanian Academy In the ecological studies. This is why in the ecological studies. during a determied period of time (enough to provide a dataset signifficant statistically). the chosen methods of study are decissive for the accuracy of study’s purpose. These methods cover the coenoses structure and the methods already used are classical ones but cover all the aspects necessay in the ecological studies. 82 . As a conclusion. the classical methods are still ued successfully and the lack of practice in the ecosystem functioning studies (energetic budget. A lack in the ecological studies is represented by the ex-situ methods related to interspecific relationships. Romanian Academy. 83 . in comparison with a much older plantation from the Bucharest surroundings (Codrii Vlăsiei). depending this time on the plantations age. the epigeic invertebrates have generally the main composition. To have more precise and complex conclusions on the succesional evolution in plantations. It is about the predator-prey relationships (Carabidae-Gasteropoda. are far from being signifficant at coenotic level. The groups of invertebrates exclusively fitophagous are dominant numerically and quite constant during the summer season. CIOBOIU2 1 Institute of Biology Bucharest. The preliminary data allow some partial conclusions: the differences in the structure and relationships are hard to be noticed at short time intervals (4-9 years. For now. as the implementation model they attempt to mimic as accurately as possible the patterns of the natural forests. an example is the short study (during the summer season) on a group of plantations of 4-9 years old from the Criş and Mureş basins. But regarding as succession. PURICE1 and O. regardless the plantations age. further studies are necessary in similar sites but with different ages. the evolutionary differences. even noticed. Carabidae-Isopoda) and interspecific competition Carabidae-Araneae. talking about the sites from Criş and Mureş basins).THE ECOLOGICAL SUCCESSION IN TERMS OF THE TROPHIC STRUCTURES IN FOREST PLANTATIONS D. 2Muzeul Olteniei Craiova It is known that plantations. There are also more or less signifficant differences in the interspecific relationships (trophic). At the epigeous level. the invertebrate fauna answers / reflects in a particular way the succession processes at the local scale. the plantations have a go . we can say that at a scale of 10 years.the time at least different in comparison with the ecological structures they imitate. To illustrate this idea. the correlation coefficient (r) shows the interspecific trends at the middle of the lifecycle for the most of the studied invertebrates. First information show that at the supraspecific level. In this last case. exclamationis. 57/2007 on the regime of natural protected areas. FIERA Institute of Biology Bucharest. Cetoniidae. Cassidinae (Cassida viridis. The taxonomic studies on coleopterogical material collected between 2011and 2012 revealed the presence of some taxa belonging to Chrysomelidae. Staphylinidae and Scarabaeidae families. 84 . in the hilly and mountainous regions).MONTEORU FOREST (BUZĂU COUNTY) S.SPECIFIC DIVERSITY OF ENTOMOFAUNA (COLLEMBOLA. it is noted the presence of Lucanus cervus (Coleoptera: Lucanidae) . in the context of research on the taxonomy and chorology of the flora and fauna species from Romania (made in the project no. O. Orchesella pannonica. MAICAN and C. Chrysolina varians – common species on Hypericum sp. the conservation of natural habitats. Romanian Academy The paper presents data on specific diversity of the entomofauna (Collembola. multifasciata) and Isotomidae (Isotomiella minor) families. From the Cerambycidae family (longhorned beetles) the following species were recorded: Xylotrechus antilope. Thaumanura carolii. COLEOPTERA) FROM SĂRATA . Orchesella pannonica (Collembola) is recorded for the second time in the Romanian fauna. Among valuable faunistical elements. RO1567-IBB03/2012). Silphidae.protected species listed in the “Council Directive 92/43/EEC on the Conservation of natural habitats and of wild fauna and flora” and in the “Government Emergency Ordinance no. exclusively phytophagous) were represented by species from the following subfamilies: Chrysomelinae (Gonioctena fornicata. Also. Entomobryidae (Lepidocyrtus paradoxus – dominant species. Anoplodera sexguttata ab. According to IUCN Red List categories and “European Red List of Saproxylic Beetles” (Nieto & Alexander. Gastrophysa polygoni. The Class Collembola was represented by species belonging to Neaunuridae (Neanura minuta. Cryptocephalinae (Cryptocephalus quinquepunctatus – a rare species in the Romanian fauna) etc. Coccinellidae. Cerambycidae. Cassida vibex). Judolia cerambyciformis and Cerambyx scopolii. Buprestidae. Anisorus quercus. The leaf beetles (Chrysomelidae family. 2010). the species Gnorimus nobilis (Cetoniidae) and Cerambyx scopolii (Cerambycidae) are classified as “Least Concern” (LC).). Coleoptera) in the forest of Querco petraeae-Carpinetum from Sărata-Monteoru (Buzău county)..  Deutonura albella etc. wild fauna and flora”. BIOGEOGRAPHYCAL DATA ON MOLLUSKS SPECIES IN DOBROGEA, ROMANIA M. AXINI1 and FAWAZ AL-AZKI2 1 “Monachus” Group of Scientific Research and Ecological Education; 2 Faculty of Sciences, Department of Geology, University from Lattakia, Syria Dobrogea region is located on the northern Balkan Peninsula in southeastern Central Europe (44017`03,77``N, 28021`53,27``E). It occupies an area of approximately 23.142 km2, of which 15.570 km2 are located in Romania (making up 6.52% of the total area of Romania) and 7.572 km2 in Bulgaria. By its characteristics different from the rest of the country (geographical position, presence and influence of Black Sea climate and soil structure, history and evolution of land with consequences for the history and evolution of its biodiversity), Dobrogea is an geographical "entity" characterized by a mixture of flora and fauna elements of Southern, Eurasian, Ponto-Caspian, European origin, etc.; many species are rare or endemic to this geographical region, or at limit of areal. The plenty and the diversity of original ecological and genetic fund which Dobrogea was so generous endowed, it is more expressive by the fact that Romania is only 2.36% of Europe surface. In this geological, geographical and ecological context, the species of mollusks have appeared and developed. Currently, 309 living species occurs in Dobrogea, which inhabit both terrestrial, aquatic, underground and marine environmentals. Of these, many species are endemic or relict. The Danubian and Pontic species of gastropods - Esperiana esperi (FÉRUSSAC 1823) and Esperiana acicularis (FÉRUSSAC 1829), both the Fam. Thiaridae (Order Mesogastropoda); they are Pontian-Southeast European relicts. Identification of these species in continental zones of Dobrogea, as Conacu-Negreşti Valley, demonstrates that these areas are former branches of the Danube River which connects between it and Black Sea, or the presence of millions years ago of ParaThethys Sea, and later, of Sarmatian Sea. This is demonstrated by geology and geography data, as well as by our field observations (the form of the canyons and of the valleys). On limestone slopes with “rocks to day”, on plateaus, hills and canyons in the south-west of Dobrogea is worth mentioning many specimens of the gastropod Helicella obvia dobroudschae (CLESSIN 1886), Fam. Helicidae (Order Stylomatophora), xerophilous species, endemic to Dobrogea. Chondrula tridens (O. F. MÜLLER 1774), Zebrina detrita (syn. Helix detrita) (MÜLLER 1774), and Zebrina varnensis (L. PFEIFFER 1847), all 85 three species of the Fam. Enidae (Order Stylomatophora), are found on the limestone walls with “rocks to day”, in less numerous specimens. The presence in areas without forests of Zebrina detrita (syn. Helix detrita) large snail, and Chondrula tridens (relatively common species in the foliage of oak forests) demonstrates the existence of pubescent oak forests (Quercus pubescent) in these zones of Dobrogea, which subsequently disappeared from various natural and man-made causes (for example, formation of the lake from the Conacu-Negreşti Valley, recent geological time lake, formed by natural damming result of catastrophic fooding). These are xerophilous species belonging to the genera of West Asian origin. Zebrina varnensis is a xerophilous species, highly resistant to drought, found on sunny slopes covered with herbaceous vegetation, in sandy areas. This species is a regional endemism spread of the Black Sea coast of Romania and Bulgaria. Presence of this species in continental zones of Dobrogea demonstrates once again the same thing above mentioned for Esperiana genus. The analysis of the zoogeographical elements of mollusks from Dobrogea again demonstrates that this region is a mixture of Central and South European-Anatolian, European, Dobrogean Endemic, Regional Endemic, European-Turanian, Holarctic, Holopalearctic, North Mediterranean, North Mediterranean-Turanian, Mediterranean-Lusitanian, Pontian, Ponto-Mediterranean, Pontian-Southeast European, Caspian Relict, Preglacial Relict, South European-Turanian, Transpalaearctic, West and Central Eurosiberian, West and Central Palearctic, West-Palearctic, etc. elements. In ecological terms, living mollusks in Dobrogea are: calciphilous, crenobiont, eurybiont, hygrophilous, of freshwater, lithophilous, mesophilous, pelophilous, phytophilous, pomatophilous, rhithrophilous, stenoepibathic, of stagnant water, terrestrial, trogloxene, xerophilous, α-βmesosaprobic, β- mesosaprobic, o-β-mesosaprobic, etc. species. The presence of millions years ago of ParaThethys Sea, and later, of Sarmatian Sea led to the formation of Dobrogea territory, an area predominantly limestone. Fossil marine species are present (for example, Mactra sp.) in the rocks from the lakes, in the canyons and ravines, on limestone walls and on Sarmatian coasts with “limestone rocks to day”. The presence of these species help us to reconstruct of paleo-environment and natural history of extinct and existing species of mollusks in Dobrogea. T 86 THE GRAYLING (Thyallus thymallus L., 1758), A DECLINING SPECIES I. CRISTEA Institutul de Cercetari si Amenajari Silvice The paper presents the results of the research made between 20092011 with the main purpose of carrying out estimate studies on the distribution area of the species in the Romanian rivers and of the habitats populated by the grayling. Although they have registered an increasingly decline in number and a continuous regress of the distribution area, the grayling populations still maintain in some rivers whose hydrological regime has not been modified by anthropic activities. However, recent observations on grayling populations and habitats certify the decline of the populations in many of the mountaineous rivers where this species was traditionally present. The main reason for the reduction in number or even the disappearance of the species is the hydropower works carried out in the upper segments of the rivers by which the adults’ migration route towards the breeding areas is interrupted. The regulation of the river streams and the embankment represent other reasons which cause important changes in the structure of ichtyofauna, reducing the area or access to the habitats favorable to the species and exposing the populations to additional selection pressures. Last, but not least, climate changes, and especially the dryness of the climate have caused the flow reduction in the majority of rivers and the restriction of the optimal habitat for the grayling. The site works undertaken have consisted in the inventory and monitoring of ichthyofauna species in the habitats populated by the grayling in order to estimate the population parameters. Works for the description of the aquatic habitats favorable to the grayling, monitoring of water quality, characterization of the trophic structure of the rivers, identification of the main sources and forms of water pollution have also been carried out. As a result of these activities, the distribution maps of the grayling populations in the Romanian rivers have been drafted and a database related to the habitats populated by this species has been created. 87 From conservation point of view. Cuza” University. 3“Al. 4 stations being placed in reed beds. for nesting and wintering. followed by Acrocephalus scirpaceus (31 birds) and Phylloscopus collybita (27 birds). breeding and population demography of birds.BIRD RINGING CAMP FROM LETEA. I. ringed by the same team in the past years. 2"Grigore Antipa” National Museum of Natural History. Danube Delta Biosphere is on the one of the most important migration route in Europe. Alcedo atthis. POCORA3 1 Institute of Biology Bucharest. behavior. one for Athene noctua and one for Parus major. Ficedula parva. All the birds’ parameters related to physiognomic and physiologic characteristics were taken (measurements. This technique is one of the most effective methods to study the biology. CHIŞAMERA2 and V. Romanian Academy. body fat index. All the birds were controlled for ectoparasites and released. we captured 7 protected species by the Birds Directive (annex I): Acrocephalus melanopogon. We capture and ringed 260 birds of 32 species in 5 days. Any record of a ringed bird. the Eurasiatic pathway. Picus canus. age) and an individual ring with code was place on the foot of every bird. The most abundent species captured was Acrocephalus schoebaenus (37 birds) and Parus caeruleus (32 birds). 88 . weight. Most of the birds were Passeriformes-25 species. COBZARU1. We perform a ringing camp in Letea. will bring a lot of information about its life. having 7 trapping station with 200 m of ornithological nets. DANUBE DELTA BIOSPHERE . bouth sedentay species. We had two recaptures. Luscinia svecica and Lanius collurio. Iaşi Scientific bird ringing is a research method which consists in the individual marking of birds. ecology.IMPORTANT SITE FOR THE STUDY OF BIRDS MIGRATION I. movement. through recapture or on the occasion of its final recovery as a dead bird. sex. offering the best conditions for birds on migration. in autumn 2012. G. of which 4 are hornworts. In the present the Red list of Bryophytes from Romania includes 375 threatened species. Erzberger P. Until now. BUCA] (Erzberger.80 10. S.00 50.72 5.00 Liverworts No. The area distribution of bryophytes is more rapidly changed than of the phanerogams.63 43.75 58.39 2. 113 Endangered and 104 Vulnerable species (Table 1).96 2.03 Mosses No. 2011.66 10.77 100.00 33. and Marsupella apiculata was recently reported from Călimani Mountains. 158 are Critically Endangered.00 100. conf. Höhn & Pócs 2012). leg. in this time.12 11.14 100.36 12.00 0. Number of threatened bryophytes in Romania IUCN category CR EN VU NT LC DD NE TOTAL Threatened (CR+EN+VU) Hornworts No.00 0. Ştefănuţ.53 10.00 25.06 47.27 89 .00 0. Table 1. Răchitiş Peak.THREATENED BRYOPHYTES IN ROMANIA S. ŞTEFĂNUŢ Institute of Biology Bucharest. & I.93 3. & T. Romanian Academy Bryophytes are much more sensitive to air pollution than higher plants and disappear earlier than many phanerogams due to the increasing habitat destruction. of species 158 113 104 79 462 43 21 980 375 % 16.68 7. Bryological researches in Romania have known a real progress in the last 20 years. Höhn M.00 0. Goia (2012).61 8.15 4.55 2. two new species of bryophytes have been published every year in Romania. Bryophytes are good indicators of climate changes. A number of 979 species was published by Ştefănuţ S.00 39. Statistically. 980 species of bryophytes were published from Romania.69 9. of species 124 96 81 73 333 30 21 758 301 % 16.00 38. of species 32 17 23 6 128 12 0 218 72 % 14. The assessing of the threatened bryophytes status in Romania is very important in achieving conservation strategies and designation of important national sites for bryophytes conservation (IBrA – Important Bryophytes Area).50 0.00 25.00 100. of species 2 0 0 0 1 1 0 4 2 % 50. & det..71 Bryophytes No. 2012 [EGR. Pócs. 218 liverworts and 758 mosses species. ION Institute of Biology Bucharest.FIRST RECORD OF Draba dorneri HEUFF. Known only from few localities. Notes on habitat characteristics that provide habitat for D. project no. Romanian Academy Draba dorneri Heuff. It occurs in small crevices on the glacial caldera walls of Mândra Lake. (BRASSICACEAE) IN PARÂNG MOUNTAINS (ROMANIAN CARPATHIANS) R. This work was supported by the national project “Monitoring the conservation status of species and habitats from Romania under Article 17 of the Habitats Directive”. dorneri and some conservation issues are also discussed. on a north-eastern facing cliffs between 2250 and 2300 m elevation. 90 . (Brassicaceae) is an endemic plant restricted to Southern Carpathians (Romania). the discovery of a new population from Parâng Mountains is reported here. 130537 financed by European Fund of Regional Development and implemented by Institute of Biology Bucharest – Romanian Academy with Ministry of Environment and Forest Biodiversity Department of Romania. They target plants and regions most at risk from climate change and the everincreasing impact of human activities. It is focused on global plant life faced with the threat of extinction and plants of most use for the future.000 species of plant are faced with the threat of extinction. The Millennium Seed Bank Project is an international conservation project coordinated by the Royal Botanic Gardens. By 2020. Major partnerships exist on all the continents. West Sussex. its purpose is to provide an "insurance policy" against the extinction of plants in the wild by storing seeds for future use. the letter and the spirit of the 1973 Convention on International Trade in Endangered Species of Wild Fauna and Flora (“CITES”). 60. The Millennium Seed Bank Partnership is the largest ex situ plant conservation project in the world. associated benefit sharing and traditional knowledge. Kew's Millennium Seed Bank Partnership aims to save plants world wide with a focus on plants most at risk and most useful for the future. enabling the countries involved to meet international objectives such as the Global Strategy for Plant Conservation and the Millenium Developmental Goals of the United Nations Environment Programme. ONETE Institute of Biology Bucharest. and relevant national and regional laws and regulations concerning biodiversity including laws relating to access to plant genetic resources. the 1992 Convention on Biological Diversity (“CBD”). Plants provide the air we breathe and clean water and we all rely on plants for food. collections are kept in the country of origin with duplicates being sent to the Millennium Seed Bank Project for storage. they have already successfully saved seeds from over 10% of the world's wild plant species. They also save the seeds of the world's plant life faced with the threat of extinction. Together with their partners in 50 countries worldwide. their aim is to secure the safe storage of seed from 25% of the world’s plants. and those that could be of most use in the future. the Nagoya Protocol on Access to Genetic Resources and the Fair and Equitable Sharing of Benefits Arising from their Utilization to the Convention on Biological Diversity. Launched in the year 2000 and housed in the Wellcome Trust Millennium Building situated in the grounds of Wakehurst Place. the International Treaty on Plant Genetic Resources for Food and Agriculture (“ITPGFRA”). 91 . Kew. Where possible. Romanian Academy Today. The storage facilities consist of large underground frozen vaults preserving the world's largest collection of seeds. The seeds that are saved are conserved outside their native habitat.000 to 100.BIODIVERSITY OF THE FUTURE IN MILLENIUM SEED BANK M. Rumex maritimus. E. C. Lindernia dubia. This publication focuses on the group of non-native vascular aquatic macrophytes in Bulgaria. According to our recent estimates the following non-native vascular aquatic macrophyte species are occurring in Bulgaria: Acorus calamus. Azolla filiculoides. One species of them is freefloating on the water surface. Najas graminea. Elodea canadensis. GEORGIEV and S. Paspalum distichum. three species are submerged plants and the other 12 are emergent plants. Eclipta prostrata. Bulgarian Academy of Sciences. Arundo donax. nuttallii. Mimulus guttatus. The term ‘aquatic macrophytes’ refers to large plants visible to the naked eye and having at least their vegetative parts growing permanently or periodically in aquatic habitats. strigosus. Sagittaria latifolia. Heteranthera rotundifolia. TSONEVA Institute of Biodiversity and Ecosystem research. Cyperus esculentus. 92 .NON-NATIVE VASCULAR AQUATIC MACROPHYTES IN BULGARIA V. As a result of field studies and an analysis of literary sources only two species (Elodea canadensis and E. nuttallii) are estimated as a potentially invasive species in Bulgaria. Portulaca oleracea. Bulgarian Biological Diversity Act (BBDA). Regionally Extinct are 5 species. Endangered and Vulnerable) are 59 species. Presented and analyzed are summarised data about plants listed under European and national policy instruments and documents such as the Habitats Directive. Aquatic macrophytes with protection status in Bulgaria. The group of vascular aquatic macrophytes comprises about 9% of the total diversity of vascular plants in Bulgaria and is one of the most vulnerable parts of it. TSONEVA and V. 93 . Red List of Bulgarian vascular plants. Fig. The term ‘aquatic macrophytes’ refers to large plants visible to the naked eye and having at least their vegetative parts growing permanently or periodically in aquatic habitats. and Red Data Book of the Republic of Bulgaria. 2 shows the distributions of species amongst the different categories in the Red List of Bulgarian vascular plants.CONSERVATIONALLY IMPORTANT VASCULAR AQUATIC MACROPHYTES IN BULGARIA S. 1 illustrates the count of species among the different protection documents. This publication aims to highlight the conservation value of the group of vascular aquatic macrophytes in Bulgaria. Fig. Bern Convention. Bulgarian Academy of Sciences. GEORGIEV Institute of Biodiversity and Ecosystem research. Under the threatened categories (Critically Endangered. Figure 1. The present work discusses also the major threats to the vascular aquatic macrophytes and gives some examples of successfully applied active protection measures. Red List species counts of aquatic macrophytes in Bulgaria 94 . Figure 2. Coracias garrulus. We consider that a global view must be the first step in terms of management. COBZARU Institute of Biology Bucharest. is the last natural landscape. Cionia nigra are nesting in the sites and more than 20. Sterna albifrons. between Izbiceni and the flow in the Danube. in the lower basin of the river Olt were designated a number of areas of biodiversity conservation concern. Protected species like Phalacrocorax pygmaeus.979 ha for SPAs. Now there are 15 water dams between Râmnicu Vâlcea and Izbiceni with a complete regulation of the water course between the two localities.NATURA 2000 SITES IN THE LOWER OLT HYDROGRAPHIC BASIN . beginning in the 1970s and 1980s. Starting with the expansion of Natura 2000 network in Romania. but their populations are threatened by habitat loss. MIHĂILESCU. Burhinus oedicnemus.529 ha and a high fragmentation of habitats. The avifauna from the Olt inferior basin is young and has been transformed together with the transformation of the landscape. Cerambyx cerdo. landscape fragmentation and the loss of dead wood habitats.080 ha for SCIs and 74. with the construction of the water dams and regulations for electricity purposes. changing from a dominant alluvial forest bird fauna to predominant open water and land fauna. The SCIs network in the studied area comprise from small units of 66 ha. the protected areas having a lot of common elements with the same 95 . shelters invertebrates like Lucanus cervus. covering more than 24 300 km2. ION and I. to large areas like ROSCI0354 Platforma Cotmeana with a surface of 12. We identified for the entire area from database and regular field trips the habitats and species which are most critical for conservation. it remains one of the most important site for wildlife and natural habitats. Romanian Academy The hydrographic basin of the river Olt is one of the largest in our country. as the ROSCI0174 Pădurea Studinița which shelters a pure Eastern white oak wood (91AA) with Ponto-Sarmatic deciduous thickets (40C0*). The natural landscape of the river has been shifted dramatically. despite of economical interests for the pit run.KEY SPECIES AND HABITATS FOR THE INTEGRATED MANAGEMENT OF THE AREA S. There are 12 Sites of Community Importance (SCIs designated under Habitat Directive) and 3 Special Protection Areas (SPAs designated under Birds Directive) with a total surface of 55. Most of the woody habitats.000 water birds are wintering at the flowing of the Olt river. Himantopus himantopus. Morimus funereus. M. The mouth of the river. Here more than 80% of the area is represented by community interest wood habitats considered to be in a favorable state. which. Acknowledgement: Project RO1567-IBB04: Identificarea şi cunoaşterea habitatelor naturale şi a speciilor endemice.conservation needs. The second step is the individual approach where the specific conservation measures must be taken to be integrated in the management plans of the sites. 96 . rare şi/sau periclitate din România. A. Ursus arctos 13. The monitoring activity. 7. 12. FEDORCA². 3. allowing analyze at the European level. M. GRIDAN¹´² 1 Department of Forest Sciences. CARNIVORA: 10. 97 . is an obligation defined by the European Directives (Habitats and Birds). Lynx lynx. Monitoring and evaluating the conservation status can be achieved by processing the scientific information and the results will be aggregated and evaluated at different spatial scales. 19. Mustela putorius. 15. Castor fiber. Sicista subtilis. Spermophilus (Citellus) citellus. Mustela eversmanni. COTOVELEA. M. R. Monitoring suppose the development of continue and permanent program in order to provide a series of observations in time. IONESCU. 9. JURJ². 6. 18. The species habitat in Romania will be considered and after a stratified and randomly analysis will result grids (10X10 km)for all species: RODENTIA: 1. Cricetus cricetus. Dryomys nitedula. 20. Lutra lutra. C. 14.THE ESTIMATION OF COMMUNITY INTEREST MAMMALS IN ROMANIA O. IONESCU¹´². ARTILODACTYLA 21. G. Muscardinus avellanarius. Mesocricetus newtoni. Canis aureus. ADAMESCU. Rupicapra rupicapra. so that it can permit the establishment of the conservation status and the trends of species evolution. M. Martes martes. Sicista betulina. CAZACU. 2. In this context. * Canis lupus. Vormela peregusna. 5. 16. 11. as an observation which is repeated on long term is needed in order to track the conservation status of species and their trends on long term. 17. the monitoring activity is defined as the whole observation measures undertaken in order to ensure that the favorable conservation status are maintained in the monitoring period. POPA² and A. Mustela lutreola. Transilvania University of Brasov. and the dates which will be reported must be compatible and comparable. 4. Felis silvestris. 8. Bison bonasus (extincted in wild). 2Forest Research and Management Institute The activity of monitoring the species and the habitats of conservative interest. 22. Microtus tatricus. the reports of the projects in which studies and researches have been conducted (The National Authority of Management POS environment. Direct observations c. The landuse from CORINE LANDCOVER As a starting point in this activity there will be used : a information from the hunting units database regarding the animals which have been hunted in the interval 2007-2012 (The Ministry of Environment and Forests). centralized information at country level concerning the damages created by large carnivore (The Ministry of Environment and Forests. 98 . Photo . ICAS. The Ministry of Environment and Forests) In order to establish the population size/the number of individuals there will be used different methods as: a.). b. c.Figure 1. Environmental Agencies. Capturing and recapturing of some individuals Selected grids will be covered in order to determine the conservation status of species habitat and data with GPS tracking will be collected. Points were individuals or tracks were observed will be marked with GPS point. Identifying the tracks when there is a fresh snow layer b.trapping d. ITRSV. 2. 1996)  Bioregions Species habitat X in  Bioregion Y  Species habitat X in  Bioregion Y  Grid with distribution [10x10km] ……….History Period: 1950‐2012  References  Raported presence  Hunting units  Distribution map of  species X at national level  Actual Potential  Species habitat  (Peterken. 5.  Grids stratified randomly What do we whant with the field dat?  (based on Article 17)  1. 3. Area Population (size)  Species habitat  Perspectives  Evaluation of conservation  status  Parameters/variables  99 . 4. Selected grids (stratified and randomized) for Rupicapra rupicapra Photo 1. Rupicapra rupicapra 100 .Figure 2. MICROBIOLOGY POSTERS 101 102 THE SELECTION OF THE ACIDOPHILIC BACTERIA BELONGING TO THE Acidiphilium GENUS WITH HIGHER ENZYMATIC ACTIVITY IN THE PRESENCE OF HEAVY METAL IONS C.M. CISMASIU Institute of Biology Bucharest, Romanian Academy The uncontrolled discharge of industrial waste waters into the river systems containing metallic ions induce profound changes of water quality, which are translated in disrupting the ecosystems due to perturbation processes in the trophic chain, with inhibiting mineralization, accumulating of heavy metals in elevated concentrations in certain the aquatic organisms and are ultimately to humans. The present of metallic ions in the solution and the need for their removal has opened two important ways to approach of the research, namely: their extraction in order to recover of heavy metal ions and industrial waste water treatment to reduce the concentrations of heavy metal ions to values mentioned in international standards. The study of the interactions between acidophilic heterotrophic bacteria and heavy metal ions can occur through active processes, involving the metabolic sequences of living microorganisms or passive processes, independent of cellular metabolism. The residual biomass of heterotrophic bacteria belong to the Acidiphilium genus proved an affinity for a wide variety of heavy metal ions such as Cr6+, Cr3+, Cu2+, Zn2+, Fe3+ and Ni2+. In these contexts, it became necessary studying starch degradation capacities of the acidophilic heterotrophic bacteria cultures in the presence of heavy metal ions. Growth experiments were carried out at different species type, heavy metal ion type and contact times between cell and these ions. The comparative studies regarding the influence of the metallic ions solution on the growth and the starch degradation activity is developed with a maximum intensity up to 14 days of incubation periods at the same substrate concentration. High percentages degradation of substrate were obtained using populations of heterotrophic bacteria compared with purified strains, which confirms the adaptation of populations to the concentrations of heavy metal ions used. 1. Pandey B., Fulekar M.H., 2012. Bioremediation technology: A new horizon for environmental clean-up, Biology and Medicine, 4(1): 51-59. 2. Wang J., Chen C., 2009. Biosorbents for heavy metals removal and their future, Biotechnology Advances, 27: 195-226. 3. Johnson D.B., Hallberg K.B., 2008. Carbon, iron and sulfur metabolism in acidophilic micro-organisms, Advances in Microbial Physiology, 54: 201-255. 103 Green tomatoes. survival under low pH environment. D. GROSU-TUDOR1. or pears). The LAB isolates (139) were identified through (GTG)5-PCR fingerprinting and 16S rRNA gene sequencing. Colonies were randomly picked up. The most common vegetables used are cucumbers. Pleinlaan 2. to test the newly isolated LAB for the production of bacteriocins and exopolysaccharides. The preservation is based on the property of lactic acid bacteria (LAB) to ferment sugars to lactic acid. Leuconostoc citreum. The aims of our study were: to isolate and identify the LAB involved in these spontaneous fermentations of vegetables. sometimes mixed with fruits (water melon. Also. Leuconostoc mesenteroides. and the antimicrobial activity in the presence and absence of prebiotics. Vrije Universiteit Brussel. catalase-negative isolates were stored at 75°C in liquid MRS medium supplemented with 25% (v/v) of glycerol as cryoprotectant and used in our further experiments. whereas enterobacterial counts were low in most cases. to evaluate the probiotic characteristics of the new isolates. Belgium Fermented vegetables represent a frequently used food in Romania.S. apples.LACTIC ACID BACTERIA FROM FERMENTED VEGETABLES WITH POTENTIAL APPLICATIONS IN FOOD INDUSTRY AND HEALTH S. as represented by their MRS counts. and bell peppers. LAB were prevalent in all end-samples. Brine and vegetable samples were collected from 20 fermentations carried out at household level in the region of Vălenii de Munte (Chiojdu) and Bucharest. The most frequently isolated LAB were Lactobacillus plantarum and Lactobacillus brevis. M. salt tolerance. The strains were further screened for the ability to produce exopolysaccharides and for their antagonistic activity against a set of nine 104 . indicating good fermentation quality. purified and tested for catalase production and Gram-staining. Faculty of Sciences and Bioengineering Sciences. ZAMFIR1. B-1050 Brussels. Samples were analysed for acidification (pH measurements) and plated on VRBG agar medium (for enumeration of enterobacteriaceae) and MRS agar medium (for enumeration and isolation of LAB). 2Research Group of Industrial Microbiology and Food Biotechnology. cabbage. and Weissella kimchi were found. WOUTERS2 and L. to purify and characterize these metabolites. carrots. DE VUYST2 1 Institute of Biology Bucharest. a stabilization of the end product. such as the bile salt resistance. Gram-positive. and cauliflower are used too. Romanian Academy. causing an acidification and hence. especially during the winter. independent of the vegetable type. of above 1400 KDa.5% (w/v) of bile salts.0. In the case of two strains. after 24 h of incubation showing viability rates of 109 CFU/ml and 1012 CFU/ml. Concerning the bile salt resistance. Lb. while 19 strains showed inhibition against one or all three Bacillus strains used as indicators. respectively. brevis 183 and Leuc.0 and pH 4. of about 20 g/L. 3. All six strains tested were resistant to pH 3. or to improve the shelf life of certain fermented foods. For instance. and Lb. citreum 52 have been shown to produce large amounts of EPS. After 2 h of exposure to 0.0. six selected strains were investigated for tolerance to acidity (pH values of 2. while the strains showing antibacterial activity could be used in a probiotic complex.3% (w/v). To evaluate the probiotic potential.0. and 4. The gel permeation chromatography-based screening revealed seven EPS-producing LAB strains. The two strains producing high amounts of high molecular mass EPS might be used to control the rheological properties of fermented foods. Lb.0. namely Leuc. All isolated EPS have a high molecular mass. plantarum/pentosus 358 were active against 5 of the indicator strains used. and a monomer composition dominated by the presence of glucose. brevis 308. four strains reached viability rates of at least 108 CFU/ml. respectively. the activity was maintained after neutralizing the pH of the cell free supernatant. all six strains showed a high resistance to a concentration of 0. probably.5%. None of the tested strains showed an antimicrobial activity against Staphylococcus aureus ATCC 25923. reaching a viability of 104 CFU/ml. Several strains showed antibacterial activity against more then one indicator strain. the viability of strains Lb. Eighty five of the tested strains showed a variable antimicrobial activity against Listeria monocytogenes ATCC 1911. 105 . citreum 177 and Leuc. reaching a cell viability of 105-108 CFU/ml after 24 h of treatment. mesenteroides 348. Romanian traditionally fermented vegetables can be regarded as important sources of LAB strains with functional properties.0 respectively) and to bile salts (concentrations of 0. Two of the positive strains.3% and 0. plantarum 307. All exept five isolates were shown to produce capsular polysaccharide and 31 developed mucoid colonies on sucrose-based media. while other 23 LAB were active against 3 indicator strains. w/v). such as production of EPS or bacteriocins. namely Leuc.LAB strains. with potential biotechnological application. being due. 35 of the strains showed a fade inhibition zone against Escherichia coli ATCC25922 and 26 strains against Salmonella enterica ATCC 14024. After 3 h of exposure to pH 2. brevis 183 survived even after 24 h of exposure to pH 2.0. reached about 106 CFU/ml. brevis 183. Moreover. citreum 344 and Lb. a value still adequate for the use of these strains as probiotics. to the production of bacteriocins. Lb. three Bacillus strains and four Gram-negative bacteria. Changing the carbon source was found to be directly correlated to the ability of synthesis of metabolites. reducing the spread of the pollutant. and emulsify the lipid constituents. the morphological and structural appearance of the bacterial cells growing under contamination stress should not be 106 . The researches have focused on the isolation and characterization of several bacterial strains from oil-contaminated areas and assessing both their tolerance to high concentrations of petroleum contaminants and their involvement in the bioremediation process. which possess the ability to metabolize the carbon that is found in the oil structures. this being accompanied by the production of metabolites such as polyhydroxyalkanoates (PHA) .also known as the biopolymers. Their intra. These bacterial products have the capacity to "isolate" the contaminated area.and extra-cellular biosynthetic capacity of the above mentioned metabolites was also assessed. Studies conducted so far have allowed us to select and identify groups of hydrocarbon-oxidizing microorganisms capable of surviving in extreme conditions of contamination stress. incidental or as a result of historical activities that have induced chronic pollution to deeper layers of the soil. Last but not least.M. the oxidative reactions. able to synthesize under contamination stress conditions. CÎRSTEA Institute of Biology Bucharest. even simultaneously. and extension of their characterization by electron microscopic studies approach. able to grow both in the presence and absence of pollutant. favoring thus. and rhamnolipids (RHL) . PHA and RHL. the bacteria acting either directly or complementary for the reduction of the contaminants. Among the tested microbial strains. but also a novelty as concerns demonstration of the versatile character of these microorganisms. Nevertheless. bioremediation appears to be a promising alternative for reducing harmful compounds that affect the ecosystem. biodegradable. by increasing the contact surface. the most representative turned out to be bacterial strains belonging to Pseudomonas genus. The current research is a continuation.STUDIES CONCERNING THE MORPHOLOGICAL AND FUNCTIONAL VARIATIONS OF SOME BACTERIAL CELLS UNDER CONTAMINATION STRESS IMPACT M. ŞTEFĂNESCU and D. represent a permanent challenge for finding solutions to protect the affected environment. Romanian Academy Areas contaminated by oil spills. The investigated microbial activity consisted in breaking down the complex compounds into simpler constituents.biosurfactants. Grown on the same mineral medium.e. modification and return of the cell to its initial shape. i. using electron microscopy techniques. Cultivation was performed on mineral media supplemented with carbon sources such as: octanoate. while the cell sections allow identification of endometabolite accumulations. the tolerance reaching concentrations up to 10%. The undertaken observations. 107 . The captured microscopic images are relevant in this respect. The paper submitted to your attention is the beginning of a research work aiming to argument the structural cellular versatility concept. such as PHA. Acumulation within the cell or as exo-products entails structural differentiations If the activity of rhamnolipids was found to be comparable to that of detergents. is a prerequisite for understanding the complex mechanisms of microbial decontamination of affected environments. depending on variation of the perturbing factors. but without carbon sources.overlooked. which result in changes in membrane ion pumps system. polyhydroxyalkanoates mode of action is less demonstrated in the direct bioremediation process. The intracytoplasmic metabolic accumulations and the extracellular synthesis products entail characteristic cellular changes that can only be highlighted by electron microscopic observations. nhexadecane. The formation of these metabolites of bio-plastic type involves the use of carbon within the contaminants structure for producing biodegradable polyesters. glucose. microorganisms did not develop. A more complete characterization of bacterial strains for potential bioremediation use. allowed visualization of the external structure of the selected cells. benzene. The bacterial strains to which we relate belong to Pseudomonas genera (figure 1). were less toxic for V. showing 99% similarity to V. The adaptation mechanisms. alginolyticus IBBCt2 showed a complex response of cells 60 min after solvent shock (i.ORGANIC SOLVENT RESISTANCE MECHANISMS IN Vibrio alginolyticus IBBCT2 M. modifications of the cell viability. alginolyticus strain J608 (JF836167. alginolyticus IBBCt2 cells to organic solvents had different effects on the lipolytic and proteolytic enzymes production. n-hexane. The high organic solvent resistance of V. Isolation of strain IBBCt2 from Constanta seawater was carried out on artificial seawater medium. The results from the present study suggest that the salt stress does not protect the bacterial cells against the toxic effects of 1% organic solvents (cyclohexane. Organic solvent tolerant bacteria are a relatively novel group of extremophilic microorganisms with novel tolerance mechanisms. changes in the membrane’s lipid and protein profile. The taxonomic affiliation of strain IBBCt2 was determined on the basis of phenotypic characteristics and the 16S rRNA gene sequence.e. n-decane. Romanian Academy Constanta harbor has been contaminated from shipping activities and accidental fuel spills with petroleum and petroleum products. C23DO) and transporter (HAE1. STANCU Institute of Biology Bucharest. mxylene. Exposure of V.. Moreover. todC1. alginolyticus IBBCt2. which enable them to overcome the toxic and destructive effects of organic solvents in solvent-saturated environments. with organic solvents as the sole carbon source. xylM.M. n-Hexane. ethylbenzene). alginolyticus IBBCt2 could be due to the presence of some catabolic (alkB. alginolyticus IBBCt2 and the mechanisms behind its resistance have been studied in saline stress conditions. compared with toluene. B 108 . The regions harboring catabolic and transporter genes are located on the chromosome in this bacterium. m-xylene. ndecane and cyclohexane with logarithm of partition coefficient in n-octanol and water (log POW) > 3. n-hexane. toluene. modifications of the genomic fingerprinting). acrAB) genes.35. which contain various types of toxic organic solvents.1). alginolyticus IBBCt2 cells to salt stress decreases the organic solvents tolerance of this bacterium. alkB/alkB1. The toxic effects of 1% organic solvents on V. The analysis of the 16S rRNA gene sequence located strain IBBCt2 (GenBank JN874640) within the genus Vibrio. and in the presence of salt the toxicity of the solvents seems to be intensified. and ethylbenzene resistance in V. toluene. the exposure for 60 min of V. underlying cyclohexane.17. m-xylene and ethylbenzene with log POW < 3. n-decane. using the enriched cultures method. The study was funded by project no. RO1567-IBB05/2012 from the Institute of Biology Bucharest of Romanian Academy. The author is grateful to Ana Dinu for technical support. 109 . Microscopic investigations revealed that most of the isolated strains were represented by Gram-negative rods. carboxymethyl cellulose and RBB-xylan revealed that all selected strains were able to degrade at least one of the tested substrates. research on extremophiles gained an increasing interest. TEODOSIU and M. they have an important role in the ecology of these extreme environments. They are categorized as slight.HALOPHILIC BACTERIA FROM DIFFERENT HYPERSALINE HABITATS IN SLANIC PRAHOVA. gelatin. based on the extent of their halotolerance. casein. Green Bath and Shepherd’s Bath lakes. representing a model of adaptation to frequent variations of the extracellular osmolarity. Tween 80. Bride’s Cave. for oil recovery. ENACHE Institute of Biology Bucharest. ROMANIA AND THEIR BIOTECHNOLOGICAL POTENTIAL R. The present study reports the isolation. in the biodegradation of organic pollutants and forensics. The investigated bacterial strains were isolated on MH selective medium containing 10% NaCl [2]. having oxidizing and catalytic properties and being able to produce indole and reduce nitrates to nitrites.5 – 2. NEAGU. being represented by Gram-negative or Grampositive bacteria. The presence of bacterial 16S rRNA genes in the case of all investigated strains. S. Qualitative determination of the capacity to extracellularly hydrolyze starch. demonstrated by PCR amplification [3] with bacterial specific primers 27 F and 1492 R [4] also proved the affiliation of the tested strains to halophilic bacteria. Romanian Academy Lately. Halophilic bacteria represent a category of extremophilic microorganisms remarkable by their ability to survive in environments with very high salt concentrations. moderate or extreme. The results showed a low diversity of microorganisms in the samples prelevated from the salt wall of the subterranean mine (around 3000 CFU/g rock salt). Samples were harvested from the wall of subterranean salt Mine Unirea (around 200 m underground).5M NaCl [1]. mainly due to their prospective applicative roles in various biotechnological fields. Because of their abundance in hypersaline habitats with different salt concentrations. COJOC. G. Romania. Microorganisms in this group are physiologically different. preliminary characterization and biotechnological potential of some halophilic bacterial strains isolated from hypersaline habitats in Prahova county. with an optimal growth in media containing 0. two water pounds inside Mine Unirea. olive oil. Specific inhibitors (sodium deoxicolate and chloramphenicol) testing allowed the separation of the halophilic bacterial strains from the haloarchaean ones. Some of the 110 . Florida.. Microbiol.. Boca Raton. The results showed that there was no strict correlation between the optimum conditions for the exopolysaccharides synthesis and the demands for optimum growth... Merciu S. 1988. Ventosa A.. 1998. Numerical Taxonomy of Moderately Halophilic Gram-negative Rods.. I. Polz M..F. 1. 64: 3724-3730. Physiology of halophilic Eubacteria. Greene R. Cojoc R.C.. Amplification of Bacterial 16S Ribosomal DNA with Polymerase Chain Reaction. Vol. Rodriguez-Valera F. 2. pharmaceutical industry). 58(4): 289-294. Dumitru L. Microbiol.. Extracellular Hydrolytic Enzymes of Halophilic Bacteria Isolated from a Subterranean Rock Salt Crystal.. Enache M.. Some of the selected halophilic bacterial strains harbored the capacity to synthesize extracellular polysaccharides [6].. Inc... 3. In this case.e. Romanian Biotechnological Letters.. Merciu S. Kamekura M. production of fermented food and food supplements. pp. 4.J. Blitchington R. 28: 19421946. Cavanaugh M. 1990.. Polish Journal of Microbiology.. Popescu G. Dumitru L. Gen. Wilson K. 14(5): 46584664.isolates even showed combined hydrolytic activities [5]. Highly Thermostable Exopolysaccharide Produced by the Moderately Halophilic Bacterium Isolated from a Man-Made young Salt Lake in Romania. 2009. Bias in Template-to-Product Ratios in Multitemplate PCR. both the carbon source and growth conditions influenced the dynamics of the culture development and the exopolymers synthesis. Oancea P. this characteristic recommending them as possible valuable candidates for use in biotechnological processes involving complex compound degradation in order to obtain desired final products (i. J. agriculture.. Clin. J.. Cojoc R.. 5. 6.B. (ed) Halophilic Bacteria.. Kamekura M. 109-138. Quesada E. Ramos-Cormenzana A. Enache M. CRC Press. Appl. In: RodriguezValera F. Pincu E. Environ. 111 ... Ruiz-Berraquero F.C. Microbiol. Kushner D. 128: 1959-1968. 1982..H. 2009.. 2National Institute for Laser. in dark as well as in light. with respect to bacterial growth. DAMIAN2. PASCU1.coli).3. culture collection cyanobacteria (Synechocystis PCC 6803) and cyanobacterial strains isolated and purified in our laboratory (single cells strains and filamentous ones). V. The cytotoxicity was tested. One working hypothesis to explain the results is based on :i) the ultrastructural differences between Gram-negative and Gram-positive bacteria and ii) cellular localization of main energetic processes in cyanobacteria. I. IORDACHE2 and D. I. photosynthesis only at intracellular membranes and intermediary metabolism within cytoplasme. is different in light as compared with the dark regime. The main results show that i) nanoparticles. Constaţa Cytotoxicity of gold. Magurele. APOSTOL2 . APOSTOL2 1 Institute of Biology Bucharest. aureus and E.thylakoides. M. 3Ovidius University. depending evidently on the ratio between nanoparticles and bacteria. 112 . all of them interacting very deeply. ARDELEAN1. (S. D.METALIC NANOPARTICLES CITOTOXICITY AGAINST PHOTOTROPHIC AND HETEROTROPHIC BACTERIA I. In cyanobacteria respiration occurs both at the cell membrane and at intracellular membranes. I.I. multiplication and intermediary metabolism associated with the reduction of artificial electron acceptors such as resazurine or MTT. SARCHIZIAN3 . CÎRNU1 . as well as the ability to reduce resazurine and ii) the citotoxycity. silver and aluminium was studied against heterotrophic bacteria. Plasma and Radiation Physics. especially silver ones. for quantification of resazurine reduction the readings were done simultaneously at 570nm and 600nm. Romanian Academy. affect bacterial growth and multiplication. The screening part of the work was done in micro plates. our investigation focused on determining the structural and functional properties of MS-l.Mol. E. POPA1. coli CPSase x-ray structure revealed a similar folding and the presence of all SYN domains and subdomains. Preliminary functional analysis showed an NH3-dependent CPSase activity for MS-l. C.R. Institute of Biology Bucharest. USA The genome of M. and c Department of Biochemistry and Molecular Biology. smithii GLN and MS-l with E. Mich.. 2012.. . smithii CPSase. Pair alignment of the primary structures of MSl and E. suggesting that this archaeal CPSase could be inhibited and activated by UMP and ornithine respectively. coli GLN and SYN subunits showed the conservation of active site residues. and of the contribution of the GLN subunit to this reaction are currently under way to elucidate the role of M. 2012). the predominant archaeal component of human gut. In addition. Mich. Detroit. 3-D molecular modeling of MS-l based on the E. Ypsilanti. Romanian Academy. Structural analysis indicated high sequence identity of both M. The gene coding for MS-l was cloned and expressed in E. the residues involved in allosteric regulation in E. EVANS2 and C.Purcarea.. 113 .R. coli and other bacterial and archaeal CPSase corresponding subunits. 2 Department of Chemistry. 22:287–299. D. smithii. In view of determining the role of this complex system involved in ammonia-metabolism enzyme that appears to be characteristic for some intestinal methanogens. smithii full-lenth enzyme and of the two-CPSase system in ammonia utilization by the human gut methanogen for arginine and pyrimidine nucleotides synthesis. PERERA2. N. Kibédi-Szabó. Perera. J. GUY-EVANS2. and the recombinant enzyme fused with a His-tag polypeptide was purified in one step by affinity chromatography. Eastern Michigan University.Guy-Evans. 1. PURCĂREA1 1 Department of Microbiology. Evans.kDa synthetase (SYN/MS-l) subunit. Popa. In addition to a reduced size enzyme (MS-s) that appears to be the smallest CPSase characterized so far (Popa et al. coli enzyme are conserved in MS-s. Further functional characterization of the reaction mechanism and enzyme allosteric regulation of the large M. H. The kinetic parameters were calculated from the saturation curves determined for all three substrates. Wayne State University School of Medicine.Z. H. coli.Microbiol Biotechnol. C. measured at 370C. N. contains genes coding for two carbamoyl phosphate synthetases (CPSase). D.STRUCTURE-FUNCTION INVESTIGATION OF Methanobrevibacter smithii FULL-LENGTH CARBAMOYL PHOSPHATE SYNTHETASE E. this methanogen also contains a putative full-length CPSase composed of a 40kDa glutaminase (GLN) subunit and a 118. Cultivation on 31 carbon sources was carried on using EcoPlates (BIOLOG). Cluj-Napoca. D. Romanian Academy. A. resulted from the different number and position of DNA amplicons. HILLEBRAND-VOICULESCU2. 3Ştefan cel Mare University. and the microbial diversity was investigated by culture-dependent and independent methods. and SSU-rRNA gene fragments were amplified by PCR using specific GC-clamp primers. DGGE analysis indicates for each sample a high variety of both bacterial and eukaryotic microorganisms. PERŞOIU3. liquid cultivation at 4°C and 15°C on five different rich and supplemented mineral media suggests the prevalence of heterotrophs in all ice samples. While up to date there are almost no reports on microorganisms’ biodiversity from underground ice sediments. T.M.CELL GROWTH AND DGGE ANALYSIS OF MICROORGANISMS FROM SCĂRIŞOARA CAVE ICE SEDIMENTS C. A. Bucharest. Total DNA was extracted from each ice sample. Ice samples of different ages were collected from the perennial ice block of the Great Hall and Little Reserve. Suceava. 4Emil Racoviţă Institute of Speleology. 2Emil Racoviţă Institute of Speleology. BRAD4 and C. Moreover. Each of the ice samples of 1. 400 and 900 years old showed a different substrate preference and selectivity. IŢCUŞ1. which is temperature dependent. Romania. PASCU1. consisting of (1) cell growth on various substrates at different temperatures. The microbial species isolated from recent and old perennial ice sediments will be further identified by SSU-rRNA 454 pyrosequencing. with a higher cell growth occurring at 15°C corresponding to psychrotolerant species predominance. and (2) DGGE analysis of 16S/18S-rRNA amplified genes from each sample. PURCĂREA2 Institute of Biology Bucharest. Romania 1 Cave glaciers represent an important environment for paleoclimate and exobiology studies. 114 . Romania. we are interested in characterizing the prokaryotic and eukaryotic microorganisms’ communities in different aged ice layers from Scărişoara Cave and determining their chronological distribution and climate biomarkers. PLANT AND ANIMAL CYTOBIOLOGY. BIOTECHNOLOGY POSTERS 115 . 116 . other were pyknotic. Strands of multistratified collagen fibers made evident by PSR stain. The EM observations showed the cytoplasm of the tumor cells full of relative uniform in shape mitochondria and numerous electron-light vesicles.2. The histological observations revealed that in the tumor territory the histologic hepatic structure was no more preserved. A possible relation between the iron overloading and the occurrence of this tumor represented a subject of interest. pH 7. in 1975. The last surviving mouse of the experimental group was sacrificed after seventeen months from the beginning of the experiment. Albino Swiss young adult male mice of 22±2 g weight were treated with Ferrum Hausmann. In the vicinity of the nucleus were often observed bundles of fine parallel fibrilles which seemed to originate from 117 . Small liver and tumor fragments were fixed in 4% formaldehyde in saline. Peters (2). respectively in cold 2. Romanian Academy A fibrolamellar carcinoma like tumor was found in the liver of the last survivor of an experimental group of iron overloaded mice.FIBROLAMELLAR LIVER CARCINOMA LIKE TUMOR IN AN EXPERIMENTALLY IRON LOADED MOUSE P. PRUNESCU and C. described for the first time by R. Some nuclei seemed to be polyploid. Glycogen and lipids were absent. Histological sections of 5 μm thickness were stained with Hemalum-Eosine (H-E). Among the packed mitochondria there were dilated profiles of smooth and rough endoplasmic reticulum and free ribosomes. They have abundant. L. The fibrolamellar carcinoma was constituted by short cellular cords. The uninvolved liver presented red-brown color.5% glutaraldehyde in 0. Ultrathin sections double contrasted with uranyl acetate and lead citrate were observed at a Zeiss 100 EM. The tumor cells presented polygonal or spindle shapes. to obtain the loading of 2 mg iron/g body weight of living animal. by intraperitoneal (ip) inoculations. Perls method to make evident the ferric ions and Picro-Sirius red (PSR) stain specific for collagen fibers.C. eosinophilic cytoplasm and nuclei with irregular outlines. An intra-hepatic spherical formation of about 8 mm diameter with white-yellow color was observed. Golgi complexes were not frequent. pseudoglandular structures like the cysts and fibrous stroma which invaded the tumor tissue. Small Golgi vesicles were packed with electron-dense material.1 M cacodylic buffer. surrounded the cellular pseudotrabeculae and formed branches into the tumor tissue. PRUNESCU Institute of Biology Bucharest. This tumor was studied in order to make a comparison with the fibrolamellar liver carcinoma (FLC) in man (1). In: Okuda K. 1991. Craig J. where they can be concentrated in the thicker fascicles of the fibrous stroma. must be considerate about the biological significance of the possible perturbation of the cellular divisions.. In the untransformed liver parenchyma the histological hepatic structure was relatively maintained. 28(3-4): 169-172.It is possible that the long time lapsed for the mouse iron overloaded liver to represent the major irritative condition which led to the tumorigenesis process.The main features of the fibrolamellar liver carcinoma (FLC) described in man were observed in the experimental fibrolamellar tumor occurred in mouse... 107-168.R. pp. 1. . Peters R. Pathology of hepatocellular carcinoma. New York. 3.. Rev. Ferritin accumulation in nuclei of hepatocytes in polymaltosed iron injected mice. sinusoids lining and Glisson’ capsule siderosomes agglomerations were observed. 118 .. Cancer. 46: 372-379. Peters R. 2. The great majority of the hepatocytes presented nuclear inclusions containing ferritin specifically stained in bright-green following Perls method (3). Prunescu C-C. . Roum.The mouse hepatocytes nuclei capacity to concentrate ferritin molecule as nuclear inclusions following the treatment of iron loading. Hepato Carcinoma.. Prunescu P. Biochim. and Masao Omata.L. In the portal spaces. .this zone..eds. Wiley. Peters R. Other similar bundles of fibrils run through the cytoplasm to get the intercellular space.A.L. 1980. hepatocytes cords.. Such nuclear ferritin aggregates with a conspicuous ultrastructure were also scattered in the hepatocytes cytoplasm. Edmonson H.L. Fibrolamellar carcinoma of the liver: A tumor of adolescents and young adults with distinctive clinico-pathologic features.. embryos. Cluj-Napoca. being common to Romania and Bulgaria also. this taxon was maintained in the collection during 4 years. the use of biotechnology can help the stop or diminishing of the plant biodiversity loss. VOICHITA. We used as explants. M. for this purpose being tested several compounds.E. HOLOBIUC. Romanian Academy In the last decade. MITOI and F. to obtain a moderate growth retardation or even a growth stopping without regeneration. ex situ preservation strategies gained more importance in the frame of the plants biodiversity conservation increased concern. botanical gardens). Borza Botanical Garden. C. in reduced space and protected against the biotic and abiotic agents. medium– term preservation using growth retardation methods and long-term conservation using cryopreservation procedures using plant propagules with morphogenic potential (apexes. HELEPCIUC Institute of Biology Bucharest. 119 . For medium-term maintenance it was ellaborated an efficient method to preserve plant material in minimal cultures with growth retardation. parviflorus AS EX SITU PRESERVATION TOOL I. field collection. In the first step an optimized micropropagation protocol was established.E. Normal plants were regenerated from this cultures and were biochemically characterized. It is possible to modulate in vitro conditions as if to obtain a limited growth with the maintenance of the regeneration capacity at high level. Besides traditional methods (seeds banks. The preservation based on in vitro protocols can be made on different duration: short time based on multiplication in active collection. stem fragments collected from a plant growing in the Al. The main advantage of this approach is the possibility to supply healthy plant material in short time. meristems).LONG-TERM CULTURES IN MINIMAL GROWTH CONDITIONS IN Dianthus trifasciculatus KIT SSP. Dianthus trifasciculatus ssp. Based on the elaborated medium-term preservation protocol. parviflorus is cited in the Red Book of Vascular Plants from Romania. The taxon taken into study. (ii) boiling. Romanian Academy. We tested three variants of callus raw extract: (i) grinding at room temperature. non-morphogenetic. HRISTEA2 1 Institute of Biology Bucharest. and its availability all year round represent additional advantages of plant in vitro systems. or uniformity represents a challenge in the nanotechnology field. bio-labeling. Our aim was to obtain AgNPs with useful characteristics. as a result of surface plasmon vibration in the metal nanoparticles. The development of environmental-friendly technologies for the production of nanoparticles with controllable size. Performances of AgNPs biosynthesis are dependent on the different biological organisms/systems used: bacteria. 2National Institute for Electrical EngineeringAdvanced Research.g. distribution. 120 . MITOI1 and G. optical receptors. cosmetics. fungi or plants. The shift of color in the reaction mixture from white to orange. The biological compounds involved in reducing AgNO3 to Ago are different for each species or even strain. COGĂLNICEANU1. using callus (non-differentiated.GREEN SYNTHESIS OF SILVER NANOPARTICLES USING PLANT CELL CULTURES G. yeast. medical and pharmaceutical applications. All three extracts rapidly mediated biosynthesis of AgNPs from AgNO3. shapes. proliferative cells) of strawberry (Fragaria x ananassa Duch. at light and 60oC conditions.). cytodifferentiation. secondary metabolism) by applying specific physical or chemical signals. was obtained in the range of minutes. morphogenesis.440 nm. proliferation.Bucharest.E. The maximum absorbance peaks were achieved at wavelengths between 420. The kinetics of AgNPs synthesis evidenced by the UV-Vis absorbance spectroscopy monitored every 30 min for 6 hrs was different for the three extracts. Silver nanoparticles (AgNPs) have many applications: non-linear optics. The absence of contaminants. the absorbance registered for grinding (i) and freezing (iii) extracts was higher compared to the boiled extract (ii) which did not increased anymore. catalysis. typical for AgNPs. INCDIE-CA. The boiling extract (ii) reduced AgNO3 faster than the other variants and the highest absorbance values were obtained in shorter time.SA. After 24 h of biosynthesis reaction in the same conditions. the stability of the plant tissue culture. spectrally selective coatings for solar energy absorption. M. and (iii) freezing. Plant cell cultures are closed systems that allow the possibility of manipulating important biological processes (e. Salmonella panama. Fourier-transform infrared spectroscopy (FTIR) analysis revealed that the biosynthetised Ag nanoparticles were in the size range of 10-25 nm. 121 . Salmonella enterica. Escherichia coli. Our method proves to be a simple and rapid biosynthesis of AgNPs from strawberry callus extract that is environmentally safe and has potential for various medical. We tested the growth inhibition effect of the synthesised AgNPs solutions against different bacteria strains (Staphilococcus aureus. licheniformis and the lowest effect was against E. An intermediary effect exhibited on B. coli and two types of Salmonella. To our knowledge this is the first report for the use of plant cell cultures to synthesize AgNPs. aerugionosa.Scanning electron microscopy (SEM). EDAX. aureus and P. X-ray diffraction (XRD). Pseudomonas aeruginosa and Bacillus licheniformis) with the agar disc-diffusion method. A similarly inhibition zone was observed for AgNO3 which suggest that the antimicrobial activity of AgNPs was due to Ag presence. cosmetic and industrial applications. The AgNPs were more effective against S. Bw. P14) were applied on the roots of two weeks old cucumber plantlets. activate the defense system in plants. having an important role in fungal pathogen inhibition. peroxidases (POX). Enzymatic defense mechanisms refer to different defense-related enzymes production.E. Bw1 and Pseudomonas sp. Also.DEFENSE-RELATED ENZYMES GENE EXPRESSION IN Cucumis sativus L. 3Departemnt of Biotechnology. CORNEA3 Institute of Biology Bucharest. with various catalytic activities. CIUCĂ2. involved in flavonoids and other phenolic derivatives synthesis. but also mixed treatment with bacteria and phytopathogenic fungi. systemic or local expression of lytic enzymes can be triggered by microorganisms. which contribute to reactive oxygen species (ROS) detoxification. M. BREZEANU1 and C. B40. Lytic enzymes like chitinases and glucanases are involved in breaking down polymers like chitin and glucan.P. Suspensions of Pythium debaryanum phytopathogen and antagonistic bacteria (Bacillus sp. using specific primers. Romanian Academy. plants use numerous defense mechanisms. National Agricultural Research and Development Institute. 2Department of Molecular Biology. M. indicating that simple treatment with antagonistic bacteria. which are a part of fungal cell wall. Reverse transcription-PCR (RT-PCR) method was used to determine alterations in defense-related genes expression of treated plants. Preliminary results indicate that differences in gene expression appear in plants exposed to bacteria and fungi comparing with non-treated plants. chemical and enzymatic (protein-based) defense mechanisms. which can be divided in structural. HELEPCIUC1. PLANTLETS INDUCED BY MICROORGANISMS F. Bucharest 1 As a response of interaction with microorganisms. catalases (CAT). Fundulea.E. Another important defense-related enzyme is phenylalanine ammonia lyase (PAL). 122 . MITOI1. The purpose of this study was to evaluate gene expression of defenserelated enzymes in cucumber plantlets treated with antagonistic bacteria and fungal phytopathogens. University of Agronomical Sciences and Veterinary Medicine. A. Plant-microbe interaction can induce the expression of antioxidant enzymes like superoxide dismutases (SOD). The recognized virological check for grapevine varieties from the Genofond of the Research-and-Practical Institute for Horticulture and Food Technologies Institute indicates that seedless genotypes demonstrated virus particles in 70-80% of analyzed samples. the application of in vitro techniques. Chisinau. with rapid multiplication in controlled environmental conditions resulting in healthy material. using the maternal and paternal forms plants infected with viruses degenerative complex: GFLV. Previous results indicate that the success of in vitro technique depends on the selected medium. S. hybrid combination and post-anthesis period (determines embryo age until the time of transfer onto culture medium). Republic of Moldova Viticulture is one of the most important branches of the agriculture complex in the Republic of Moldova. Arabis mosaic virus (ArMV). seedless x seed. Regeneration through somatic embryogenesis permits obtaining plants free from some harmful viruses and allows recovery of a higher proportion of seedless progenies. and Grapevine virus B (GVB). autopollination. ANDRONIC. it is not possible to obtain hybrid progeny in grape crosses involving seedless (‘stenospermocarpic’) female parents. The International Council for the Study of Virus and Virus-like Diseases of Grapevine recognizes over 70 infectious agents affecting grapevine. In conformity of the proposed objectives were made the crosses according to the scheme: seedless x seedless. Assessment of the phytosanitary status of referenced plants confirmed the widespread prevalence of the degenerative and leafroll complex of viruses (Grapevine fanleaf virus (GFLV). As biological 123 .and chemotherapy). Among the tested techniques (meristem culture aided with thermo. GRIGOROV Institute of Genetics and Plant Physiology of the ASM. GLRaV-1. offers the unique possibility of obtaining new forms of seedless grapevine vines. Modern biotechnology and especially. selfpollination. GLRaV-3 and ArMV. somatic embryogenesis seems to be a promising procedure for virus elimination. providing significant revenue for the national budget. Technologies associated with in vitro embryo development represent a single enabling method in a relatively short period of time to produce plants from new combinations of genotypes with different degree of seedlessness. Grapevine leafroll-associated virus (GLRaV -3).DIRECT SOMATIC EMBRYOGENESIS AS POSSIBLE WAY OF VIRUS ELIMINATION FROM SEEDLESS GRAPEVINE L. SMEREA and T. Using conventional breeding procedures. Grapevine seedless varieties have major advantages compared with varieties with seeds as they having a broader range of industrial and food application and command a larger demand in international markets. France) against GFLV.5 mg/l of IAA. Interlaikin. Two “recovery” medium were selected: Nitsch. The composition of mediums was the same: half strength of macroand microelements. Romulus.3mg/l of putrescine (NN2).2 mg/l of giberilinic acid (GA3) (NN1) and Nitsch. immuno-electron microscopy (IEM) or negative staining (NS). Nitsch (1969) with addition of 2.2 mg/l of GA3 and 0. Viruses were detected using serologically test enzyme-linked immunosorbent assay (ELISA). 100 mg/l of mio-inositol. depending on the cultivation mediums. The way of converting into plants the embryos recovered by “in ovulo” manipulations. ArMV. 3 mg/l of glicine and 6. especially with the seedless feature. It was find that. I5-58). Ovules were cultivated for 30 days in total darkness conditions at 25±20C. and then transferred in culture room in the same temperature regime and period of 16 hours of light. The berries resulted from free crosses of the genotypes were harvested 3540 days after anthesis. peculiarities of starting material. Apiren roz Basareabean. Centinales seedless. 0. Primary results of virological check of grapevine plantlets before ex vitro adaptation show that somatic embryogenesis proved to be effective technique in eradicating phloem-limited viruses. The number of required plants varied between 75-90% from viable embryos.2 % calcium hypochlorite and washed with sterilized distillated water. the reaction of immature embryos to the in vitro cultivation depends essentially on genotype. GLRaV-1 and GLRaV-3.8 g/l of agar. More descendants with desired characteristics.5 mg/l of IAA. as well as phytosanitary status of parental forms. must take into account the peculiarities of the varieties that were involved in the crosses and phytosanitary status. Every 30 days until seeds residuum were dissected to recover viable embryos. I-2-24. Considerable attention is paid to the selection of parental genitors. 1 mg/l of zeatin and 0. Apiren Negru de Grozeşti. the requirements during grape harvest and viable embryos excising. are obtained from combinations where both parents are seedless. 124 . The ovules were aseptically extracted and inoculated on cultivation medium in Petri dishes or pots. according to manufacturer instructions. genotypes used in crossings and the age of embryos at the time of transfer onto mediums. Nitsch (1969) supplemented with 1. Experimental data shows the results of virus eradication are variable depending on the number and type of viruses. For ELISA were used commercial antisera (Sediag. 1-15-15 and TAD VOG) and 6 paternal genotypes (Apiren roz. Collected berries were cold pretreated and then surface sterilized with 70 % ethylic alcohol followed by 5. Viable embryos were placed on “recovery” mediums.materials were used the 5 maternal forms (Apiren roz Basarabean. and vitamins after Murashige Skoog (1962) with 15 g/l of sugar. the long. total phenolic content and the evaluation of antioxidant potential by DPPH test. MIHAI.term callus extracts of Vitis vinifera cvs Isabelle. In the present study. This study was achieved by carrying out a quantitative analysis of the role of elicitors on resveratrol and total phenols production.ANTIOXIDANT ACTIVITY OF POLYPHENOLS FROM Vitis vinifera CVS. cancer. ethephon. The highest antioxidant activity was achieved using as elicitors treatment. the combination of mannitol (2mM) and jasmonic acid (40 µM). in different concentrations. but it has become clear that the mechanisms of action of polyphenols go beyond the modulation of oxidative stress. in the two-stage culture system. 125 . jasmonic acids. abscisic. were comparatively analyzed due to the influence of some biotic and abiotic elicitors in the long. F. For the first stage of the experiment. Polyphenolic compounds play an important role in adsorbing and neutralizing free radicals. compounds in general responsible for the antioxidant activities. Further subcultivation of the callus during the second stage of the experiment ensured the biosynthetic capacity of the callus using in the medium the addition of elicitors such as Fusarium oxysporum extract. MITOI Institute of Biology Bucharest. quenching singlet oxygen or decomposing peroxides. it has been established that oxidative stress is among the major causative factors in induction chronic and degenerative diseases such as: artherosclerosis. Romanian Academy In the last decade. in Vitis vinifera L callus culture. mannitol. Nowadays. Antioxidants can be effective for the suppression of such disorders and in prevention of the free radical formation. Parkinson`s disease and immune dysfunction.E. diabetes mellitus. HELEPCIUC and M.term callus was grown on the medium variants supplemented with mannitol in concentration 2mM to ensure the callus proliferation. The antioxidant properties of polyphenols have been widely studied.E. suggesting that this culture system could be used as a new method for improving natural antioxidants biosynthesis. a considerably increasing interest consists in finding naturally occurring antioxidants from plant sources used in food industry and in preventive medicine in order to replace synthetic antioxidants restricted due to their possible carcinogenic effect. isabelle LONG-TERM CALLUS CULTURE UNDER THE INFLUENCE OF SOME ELICITORS TREATMENT R. The elicitor treatment consisted in using eight medium variants in a two-stage experiment. Their synthesis and accumulation is generally stimulated in response to biotic/abiotic stresses. cvs Isabelle callus extract can be used as an easily accessible and rich source of natural antioxidants.The antioxidant activity may be attributed to the high content of phenolic compounds and resveratrol in the callus extract subject to the treatment of these combined elicitors. Moreover. this two-stage culture system for the Vitis vinifera L. 126 . The main conservation measures already taken. After 30 days of culture on basal media (MS) supplemented with different combination of auxins and cytokinins the best morphogenetic response for both type of explants. non-organogenic callus yield from roots explants could be important as ecdydisteroids source. Root explants yield non-embryogenic callus which proliferate when subcultured on fresh media supplemented with 1mg/l BAP and 0. Population are strongly fragmented and with low number of individuals. Likewise. were registered on MS media supplemented with 1mg/l 6-benzylaminopurine (BAP) and 0. The proposed method relied on indirect morphogenesis pathway followed by the morphogenetic competent somatic cells. Meristemoid–like structures and non-organogenic calli were induced from explants derived from aseptically growing seedlings (leaf fragments and roots). 127 . concerns only in situ approaches. the leaf epidermal and mesophyll cells showed embryogenic competence while root epidermal and parenchymal cells show callusogenic competence.1mg/l 1naphthylacetic acid (NAA). The results suggest that the response type was highly related with the explant source.AN EFFICIENT METHOD FOR SOMATIC EMBRYOGENESIS INDUCTION AND PLANT REGENERATION FOR Serratula bulgarica ACHTAROFF ET STOJ.1mg/l NAA. The method presented is efficient for plant regeneration and subsequent natural population recovery or ex situ conservation. EX SITU GERMPLASM CONSERVATION A. Onto media supplemented with 1mg/l Kinetin (Kn) plantlets derived from leaf explants elongate and rooted at optimal rate. with multiple applications in medicine and plant defense. Romanian Academy Serratula bulgarica is a Balkan near-endemic and critically endangered plant species growing in the herbaceous plant communities from wet meadows to steppes from North-Western Bulgaria and Eastern Romania. AIFTIMIE-PĂUNESCU Institute of Biology Bucharest. As a standard a sample from natural habitat individual was used. BANCIU and F. project number POSDRU/89/1. HELEPCIUC Institute of Biology Bucharest. 128 . The results showed the identity of the amplicons on all five samples proving that the regenerants kept their genotype and the in vitro culture conditions did not affect the original genetic pattern of the plant species.E. The genetic stability of the regenerants was evaluated using the RAPD method by amplifying the DNA of regenerants with non-specific plant polymorphic primers. RO1567-IBB06/2012 and from European Social Found through Sectoral Operational Progeamme Human Resources Development 2007-2013.GENETIC STABILITY ASSESSMENT OF Scilla autumnalis REGENERANTS C. pharmacological and ornamental applications. A number of 5 samples from individuals regenerated from a single plant were used to compare their amplicons. This work was cofinanced from the Romanian Academy project no. Romanian Academy Scilla autumnalis is a critically endangered plant species from Romanian flora of conservative.5/S/63258 “Postdoctoral school for zootechnical biodiversity and food biotehnology based on the eco-economy and the bioeconomy required by eco-san-genesys”. The protein samples were separated by two-dimensional gel electrophoresis and stained with Coomassie Brilliant Blue 250. Current study shows the differences between gametophytes and sporophytes of the mentioned species by comparing the proteomic profiles and determination of ploidy levels by flow cytometry. It is possible that the diploides origin of Athyrium filix-femina and Asplenium trichomanes gametophytes to appeare by a false meiosys.E. Ploidy level was determined using two step procedure with Otto I+II buffers and the fluorochrome DAPI. MITOI1. Our primary experiments were focused on developing an optimal in vitro multiplication method and medium term conservation for these species. S. Romanian Academy. DOMŻALSKA2. SPANIEL3.E. 4University of Pitesti. The differences in protein expression suggest that gametophyte and sporophyte have different metabolic pathways of protein synthesis. C. This study presents a preliminary analysis of proteomics profiles and compares the ploidy level for detection of further changes that may occur 129 . Polish Academy of Sciences. Although in previous studies using PAGE technique we have not observed significant differences between gametophyte and sporophyte.COMPARATIVE STUDIES BETWEEN FERNS GAMETOPHYTE AND SPOROPHYTE BY BIDIMENSIONAL ELECTROPHORESIS AND POLYPLOIDY ANALYSIS F. We used gametophytes and sporophytes from Athyrium filix-femina. 3Institute of Botany of Slovakian Academy of Sciences. The reactivity of each species has been evaluated on the basis of biometric (morfometric and gravimetric) and biochemical determinations. F. Polypodium vulgare and Asplenium trichomanes obtained by in vitro culture and multiplicated on MS 1/2 medium. Faculty of Science Athyrium filix-femina. Botanical Garden – Center for Biological Diversity Conservation. SOARE4 1 Institute of Biology Bucharest. the patterns obtained by two-dimensional gel electrophoresis showed the existence of distinct spots for both gametophyte and sporophyte extracts. ALDEA1. A. In case of Athyrium filix-femina and Asplenium trichomanes both sporophyte and gametophyte were diploid. 2Plant Biotechnology Department. L. Proteins were isolated using to a phenol-based extraction procedure (Hurkman and Tanaka 1986). HELEPCIUC1. while for Polypodium vulgare we detected that the gametophyte was haploid and sporophyte was diploid.C. BREZEANU1 and L. M. Polypodium vulgare and Asplenium trichomanes are three valuable ferns species due to their potential as ornamental plants and secondary metabolite production originate to Valsan Valley protected area. BANCIU1. Proteomic analysis allows the observation of translationally manifested cell physiological response. 130 .during the process of cryopreservation at the molecular level. (LICHENOPHYTA) SYMBIOTROPHIC FORMATIONS REGENERATED IN “IN VITRO” CULTURE D. These kinds of storage bodies are associated with the cytoplasm and may represent reserves of lipids. in order to elaborate an experimental system for lichens thalli resynthesis useful for both classical and modern cell biological researches as well as for biotechnological applications and for their conservation and repopulating strategies. Romanian Academy The aim of the paper is to realize a ample cytological study of the symbiotrophic formations which are developed in “in vitro” culture. Concentric bodies or ellipsoidal bodies are visible and are present in the cytoplasm like small formations. singly or in clusters with a core body surrounded by electron transparent halos. The cell wall also consist of the cell layers and the thickness of the outer layer. The most affected cellular substructures seem to be chloroplasts of the phycobiont cells which presents a small number of thylakoids and pyrenoglobules randomly spreaded.) MOTT. 131 .ELECTRONOMICROSCOPICAL ANALYSIS OF THE Usnea barbata (L. probably in relation with their composition. Possibly. and may represent adaptative modification. Another characteristic is the presence of so . VOICU and A. observations on their ultrastructure revealed some significant differences. Although symbiotrophic formations regenerated in “in vitro” culture seems to be similar to their natural correspondents. One of them presents electronodense content and in other one electron light content. BREZEANU Institute of Biology Bucharest. The fine structure of the mycobiont is from many points of views similar with that of the old mycobiont from the native thallus. varying according to the age. proteins and other metabolic substrates. Their presence may be determinated by inhibitory effect of the culture conditions especially on the thylakoid system development. Our investigations allowed us to appreciate that the experimental systems do not induce severe alterations of the inner structure of the cells and could represent an alternative methodology to obtain lichen biomass of the biotechnological interest namely secondary metabolites. Two categories of storage bodies were identified. As a peculiarity is that the outer layer consists of an amorphous electronodense material and a fibrilar matrix and the plasmalemma is highly folded.called “storage droplets”. they are involved in enabling of phycobiont to endure adverse environmental conditions. Copyright © Institutul de Biologie Bucureşti Material editat de Institutul de Biologie Bucureşti 11 decembrie 2012 132 .