Determination of bean common mosaic virus found in Vojvodina region

Abstract

Our investigation was aimed towards determination of the presence and distribution of economically the most harmful bean virus in Vojvodina in 2006 (Bean common mosaic virus). Some 262 samples from Vojvodina grouped according to symptom types were collected during 2006: type A symptom: dark green nerves edgings; type B symptom: mosaic in the form of chlorotic and green areas slightly wrinkled; type C symptom mosaic in the form of green and yellowish smeared areas, and type D symptom: dark green cap shaped leaves. Viruses were identified using DAS ELISA test according to the instructuions given by the antiserum producer (LOEWE Biochemica GmbH, Germany). On the basis of the obtained results it was concluded that BCMV virus was found in 35% of tested plants. The greatest plant infection was found in Čonoplja (66%), and the most common symptom type was B. Chosen bean leaf samples were tested using method of reverse transcription and polymerase chain reaction (RT - Reverse Transcription and Polymerase Chain Reaction). Four sets of BCMV specific primers were used for each sample testing. Determination of BCMV virus as a member of Potyviridae family was comfired by PCR reaction, and by obtaining specific fragments of certain basic pairs. Set of primers specific for Russian and NL-3D strains were used to prove that determined viruses did not belong to the above mentiond strains.

Tokom 2006. godine prikupljeno je 262 uzorka, sa 22 lokaliteta, koji su grupisani po tipu simptoma: simptom tipa A: tamno zeleno obrubljivanje nerava; simptom tipa B: mozaik u vidu hlorotičnih i zelenih površina, sa blagom naboranošću; simptom tipa C: mozaik u vidu zelenih i žućkastih razlivenih površina i simptom tipa D: tamno zelena klobučavost listova. Serološka identifikacija virusa izvršena je DAS ELISA testom uz korišćenje komercijalnog seta antiseruma virusa BCMV, kompanije LOEWE Biochemica GmbH, Nemačka. BCMV konstatovan je u 35% testiranih biljaka. Najveći broj zaraženih biljaka zabeležena je u Čonoplji (66%) a najčešći tip simptoma bio je tip simptoma B. Odabrani izolati testirani su RT-PCR metodom. Svaki uzorak testiran je sa četiri seta BCMV specifičnih prajmera. PCR reakcijom, dobijanjem specifičnih fragmenata određenih baznih parova potvrdila se determinacija virusa BCMV, kao i pripadnost ovog virusa familiji Potyviridae. Sa setom prajmera specifičnim za sojeve Russian i NL-3D dokazalo se da determinisani virusi ne pripadaju pomenutim sojevima.