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Ministry of Education and Research MINISTRY OF EDUCATION, YOUTH, RESEARCH AND SPORT UNIVERSITY OF AGRONOMICAL SCIENCES AND VETERINARY MEDICINE BUCHAREST SCIENTIFIC BULLETIN Serie F XIV 2010 BIOTECHNOLOGY ISSN 1224-7774 BUCHAREST 2010 Editorial board and coordination of the Scientific Bulletin Biotechnology: U.S.A.M.V. Bucuresti, Facultatea de Biotehnologii B-dul Mărăşti nr.59, Bucureşti, sectorul 1,România Tel: + 40 21 318 25 67 Fax: + 40 21 318 28 88 Web: http://www.biotehnologii.usamv.ro For the publications exchange please contact Prof Dr Petru Niculiţă, Dean of the Faculty of Biotechnology Tel: + 40 21 318 36 40; Mobile: + 40 740 11 44 01 e-mail: [email protected] Redactarea şi coordonarea Buletinului Științific Biotehnologii: U.S.A.M.V. Bucureşti, Facultatea de Biotehnologii B-dul Mărăşti nr.59, Bucureşti, sectorul 1,România Tel: + 40 21 318 25 67 Fax: + 40 21 318 28 88 Web: http://www.biotehnologii.usamv.ro Pentru schimb de publicaţii va rugam sa contactati Prof Dr Petru Niculiţă, Decanul Facultăţii de Biotehnologii Tel: + 40 21 318 36 40; Mobil: + 40 740 11 44 01 e-mail: [email protected] La rédaction et la coordination de Bulletin Scientifique Biotechnologie: U.S.A.M.V. Bucuresti, Facultatea de Biotehnologii B-dul Mărăşti nr.59, Bucureşti, sectorul 1,România Tel: + 40 21 318 25 67 Fax: + 40 21 318 28 88 Web: http://www.biotehnologii.usamv.ro Pour l‟échange des publications prier de contacter Prof Dr Petru Niculiţă, Doyenne de la Faculté de Biotechnologie Tel: + 40 21 318 36 40; Portable: + 40 740 11 44 01 e-mail: [email protected] 2 SCIENTIFIC COMMITTEE Petru NICULITA - Prof. Dr., Dean of the Faculty of Biotechnologies - USAMV bucharest, Member of the Academy of Agricultural and Forestry Sciences (ASAS), President of the Food Industry Department- ASAS - Prof. univ. dr., Decanul Facultatii de Biotehnologii - USAMV Bucuresti, Membru titular al Academiei de Stiinte Agricole si Silvice (ASAS), Presedinte al Sectiei de Industrie Alimentara - ASAS Acad. Cristian HERA - Prof. Dr., Doctor Honoris Causa, Member of the Romanian Academy, Honorary President of the Academy of Agricultural and Forestry Sciences - Prof. univ. dr., doctor Honoris Causa, Membru titular al Academiei Romane, Presedinte de onoare al Academiei de Stiinte Agricole si Silvice Acad. Ion Paun OTIMAN - Prof. Dr., Doctor Honoris Causa, Member of the Romanian Academy, General Secretary of the Romanian Academy - Prof. univ. dr., doctor Honoris Causa, Membru al Academiei Romane, Secretar General al Academiei Romane Constantin BANU - Prof. Dr., Member of the Academy of Agricultural and Forestry Sciences - Prof. univ. dr., Membru titular al Academiei de Stiinte Agricole si Silvice Mona Elena POPA - Prof. Dr.,Scientific Secretary of the Faculty of Biotechnologies - USAMV Bucharest - Prof. univ. dr., Secretar Stiintific al Facultatii de Biotehnologii - USAMV Bucuresti Calina Petruta CORNEA - Prof. Dr., Faculty of Biotechnologies, USAMV Bucharest - Prof. univ. dr., Facultatea de Biotehnologii - USAMV Bucuresti EDITORIAL BOARD Florentina MATEI - Faculty of Biotechnologies - USAMV Bucharest; Facultatea de Biotehnologii - USAMV Bucuresti; Amalia MITELUT - Faculty of Biotechnologies - USAMV Bucharest; Facultatea de Biotehnologii - USAMV Bucuresti Mira TURTOI - Faculty of Biotechnologies - USAMV Bucharest; Facultatea de Biotehnologii - USAMV Bucuresti Mihaela GHIDURUS - Faculty of Biotechnologies - USAMV Bucharest; Facultatea de Biotehnologii - USAMV Bucuresti 3 CONTENT CORNEA,C.P., MATEI,S., CIUCĂ,M., VOAIDES,C., MATEI,M., POPA,G., POP,A. - IN VITRO 5 BIOCONTROL ACTIVITY OF NEW TRICHODERMA SPP. ROMANIAN ISOLATES ON DIFFERENT PLANT PATHOGENIC FUNGI DINU LAURA-DORINA, BACH SUSAN, HARLTON COLLEEN - CONSTRUCTION OF TN7GFP- 16 TAGGED E. COLI O157:H7 TRANSFORMANT FOR LONG-TERM SURVIVAL INVESTIGATIONS MARINESCU GINA, ANTONETA STOICESCU, ANDREI BOLOCAN - COMPARATIVE STUDY 23 OF SPENT BREWER’S YEAST β- GLUCANS DETERMINATION METHODS NICOLAE GETUTA, IOAN DOPCEA, ANA-MARIA NICOLAE, PETRE STELIAN, FLORENTINA 31 MATEI - CONSERVATION METHODS USED FOR YEAST ISOLATED FROM VINEYARDS – LYOPHILISATION ADVANTAGES RAVICHANDRAN KAVITHA, NAY MIN MIN THAW SAW, HEIDI RIEDEL, CAI ZHEN ZHEN, 37 INGA MEWIS, IRYNA SMETANSKA - OBTAINING OF NATURAL COLOURANTS FROM RASPBERRIES MITELUT AMALIA, MONA POPA, ALINA CULETU, VERONIKA ABRAM - ESTABLISHING 44 PRODUCE ORIGIN THROUGH BIOCHEMICAL MARKERS IDENTIFICATION POPA, N.C., TAMBA–BEREHOIU, R., POPESCU, S., TAMBA, S - INVESTIGATIONS ON 50 PREDICTIVE MODELLING OF THE FARINOGRAPHIC PARAMETERS HÎNCU F.A., E.M. PRUTEANU, D.E. DUŢĂ, N. BELC, E. IORGA, A.C. MITELUŢ - STUDIES ON 58 THE IDENTIFICATION OF CAROTENOIDS TYPE BIOCHEMICAL MARKERS MIHAELA DRĂGHICI, ALEXANDRA POPA, MONA POPA, PETRU NICULITA, GEICU MIHAELA, 65 AMALIA MITELUT - ROMANIAN ECOLOGICAL FOOD MARKET: AN OVERVIEW PETRE STELIAN, ANGHEL LAURA, DIGUTA CAMELIA, STEFANA JURCOANE, GETUTA 71 NICOLAE - GROWTH OF PSEUDOMONAS AERUGINOSA IN PILOT BIOREACTOR AND ITS TESTING IN THE BIOREMEDIATION OF SOILS TREATED WITH PESTICIDES SUCH AS DINITROPHENOLS (ISHORT COMMUNICATION) 4 Scientific Bulletin Biotechnology, U.Ş.A.M.V.Bucharest, Serie F, Vol. XIV, 2010, p. 5 - 15 IN VITRO BIOCONTROL ACTIVITY OF NEW TRICHODERMA SPP. ROMANIAN ISOLATES ON DIFFERENT PLANT PATHOGENIC FUNGI CORNEA,C.P.1,2, MATEI,S.3, CIUCĂ,M.4, VOAIDES,C.1,2, MATEI,M.3, POPA,G.1, POP,A.2 1USAMV Bucharest, Faculty of Biotechnology - Bucharest, Romania 2BIOTEHNOL - Bucharest 3INCDPAPM Bucharest, Romania 4ICDA Fundulea, Romania ABSTRACT The genus Trichoderma comprises various fungal strains that can act as biological control agents against a large diversity of plant pathogens. A number of commercial products are available but the diversity of plant pathogens, and their increased resistance to the current control products (chemical or biological products) determined the search of new strains, potentially useful for biological control. New strains of Trichoderma spp. were isolated from Romanian soils and their antagonistic activity against Phytium spp and Rhizoctonia spp. was examined. The in vitro biocontrol activity of Trichoderma spp., as well as of other antagonistic fungi (Trichotechium roseum and Gliocladium spp.) on the phytopathogens was increased in the presence of FeCl3. RAPD and ITS- PCR were used to examine the genetic variability among the isolates. Key words: Trichoderma species, antagonist, Botrytis spp., Phythium spp., Rhizoctonia spp., molecular tests Plant diseases caused by soil-borne pathogens like Phytium, Botrytis, Rhizoctonia, Fusarium and Phytophtora play an important role in the destruction of natural resources in agriculture. Chemical pesticides have been extensively used for control fungal plant disease but their employment favored the selection of fungicides resistant strains as well as negative effect on non-target organisms and environment (Benitez et al., 2004). In this respect, the development of alternative methods for plant pathogens is of great interest not only for scientists but also for agriculture. Biological control agents are risk free both for environment and non- target organisms, and could reduce the use of chemical products. Several commercial biological products based on antagonistic microorganism are available now on the market (Strasser et al., 2007) but the interest for selection of new antagonists is not diminished (Siddique et al., 2007; Dubey and Suresh, 2006; Woo et al., 2006). The filamentous fungi Trichoderma (Ascomycetes, Hyprocreales) have attracted the attention because the inhibitory action against various plants pathogens and the diversity of mechanisms of action (Harman et al., 2004). The purpose of the present study was the screening of some new fungal Romanian isolates efficient in vitro against Pythium and Rhizoctonia. The 5 experiments were also conducted to molecular characterization of the fungal strains, using RAPD and ITS-PCR. 1. MATERIAL AND METHODS 1. Culture of Trichoderma spp. The fungal antagonists were isolated using dilution plate techniques on TSM medium (Ahmed et al., 1999) from soil samples provided by ICDPAPM Bucharest, Romania. The fungal isolates were purified and obtained single spore cultures and their identification was based on morphological characters. All cultures were maintained on potato dextrose agar at 4oC. 2. Cultures of other fungal strains In the experiments were also used strains of Pythium spp., Phytophtora spp. Rhizoctonia solani, Botrytis cinerea (Bc1 and Bc2) Penicillium chrysogenum A2, Gliocladium roseum and Eppicoccum purpurescens kindly provided by dr.M.Oprea, Institute of Plant Protection, Bucharest, Romania. 3. In vitro antagonism test was performed by dual cultures technique on Petri dishes containing PDA supplemented or not, with 100 μg/mL FeCl3. Petri dishes were inoculated with mycelial disc of 7-day-old culture of the pathogen and antagonistic strains at equal distance from the periphery. Inoculated plates were incubated for 6 days at 25oC and the radial growth of the pathogen was measured. From the zone of interaction between the antagonist and phytopathogen, the mycelial mats were gently removed with a needle and examined under microscope for hyphal interaction (Dubey and Suresh, 2006). Inhibition was observed as presence of inhibition zones prior to any mycelial contact. The percent RI was calculated as follows: RI= 100x (R2-R1)/R2. R1 was the distance between the inoculum of pathogen and the edge of the colony (after 6 days at 25oC) measured in the direction of the inoculum of inhibitory strain. R2 is the colony growth of pathogen measured in the direction
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