Molecular characterization and identification of fungi causing stern canker of oilseed rape in Serbia

Abstract

During 2008-2010 the infected Brassica napus plant material was collected from nine localities: Karavukovo, Crvenka, Prigrevica, Subotica, Rimski Šančevi, Srbobran, Beška, Banatsko Karađorđevo, and Srpski Miletić. Infected tissue samples were taken from the root of both upper and basal stems, as well as from the leaf, flower and pods. Two reference isolates from Great Britain were used in these studies for the analysis of 119 isolates from Serbia, using polymerase chain reaction (PCR) and PCR-RFLP. Digestion of PCR products was performed with 5 selected endonucleases: BamHI, HaeIII, Rsal, EcoRII, and AluI. On the basis of PCR analysis, all isolates originating from Serbia belong to L. complex: 111 belong to species Leptosphaeria maculans, and 8 to species Leptosphaeria biglobosa NA1 ('Leptosphaeria biglobosa brassicae), respectively.

Kulture gljiva izolovane su iz obolelih biljnih delova uljane repice koji su prikupljeni iz devet lokaliteta (Karavukovo, Crvenka, Prigrevica, Subotica, Rimski šančevi, Srbobran, Beška, Banatsko Karađorđevo, Srpski Miletić) u periodu od 2008 do 2010. godine. Uzorci obolelog tkiva su uzeti sa korena, prizemnog stabla, gornjeg dela stabla i grana, kao i sa lista, cveta, ljuski i semena. Molekularna karakterizacija je sprovedena korišćenjem lančane reakcije polimeraze PCR i PCR-RFLP, a ukupno je ispitivano 119 izolata poreklom iz Srbije i dva referentna izolata iz Velike Britanije. Digestija PCR produkata je sprovedena sa 5 odabranih endonukleaza: BamHI, HaeIII, RsaI, EcoRII i AluI. Na osnovu PCR analize, svi izolati poreklom iz Srbije pripadaju L. kompleksu: 111 pripada vrsti Leptosphaeria maculans, a 8 vrsti Leptosphaeria biglobosa NA1 (Leptosphaeria biglobosa brassicae).