Molecular identification of Fusarium graminearum, sorghum pathogen in Serbia

Abstract

A total of 39 samples of sorghum (Sorghum bicolor) with symptoms of stem and root rot were collected and analyzed during 2009-2011 in Bački Petrovac and Čantavir, Serbia. Monosporic cultures were isolated from stem tissue, their pathogenicity was confirmed by the development of symptoms on artificially inoculated sorghum plants, and they were identified on the basis of macroscopic and microscopic morphological features as Fusarium graminearum. Molecular identification was performed utilizing polimerase chain reaction (PCR) with primer pair ef1/ ef2 and by amplification of protein coding TEF 1-alpha gen. Sequence of TEF gene from the selected isolate 535- 10 (JF747146) showed 98-99% nucleotide identity with sequences of 63 Gibberella zeae isolates deposited in NCBI GenBank. Amplification of the barcoding region of F. graminearum genome of sorghum isolate, contributes to the fast and accurate identification and characterization of Fusarium species in Serbia.

U periodu 2009-2011. na lokalitetima u Bačkom Petrovcu i Čantaviru prikupljeno je i analizirano 39 uzoraka biljaka gajenog sirka (Sorghum bicolor) sa simptomima truleži prizemnog dela stabla. Iz biljnog tkiva izolovane su monosporne kulture, čija je patogenost potvrđena pojavom simptoma na veštački inokulisanim biljkama sirka, a na osnovu morfoloških makroskopskih i mikroskopskih osobina identifikovan je Fusarium graminearum. Molekularna identifikacija obavljena je primenom lančane reakcije polimeraze (PCR, Polymerase Chain Reaction) uz korišćenje prajmera ef1/ef2 i amplifikaciju kodirajućeg proteinskog gena TEF 1-alfa. Sekvenca TEF gena odabranog izolata 535-10 (JF747146) je pokazala 98% do 99% nukleotidne identičnosti sa sekvencama 63 izolata Gibberella zeae deponovanih u NCBI bazi podataka. Amplifikacijom barkoding dela genoma F. graminearum izolata iz sirka dat je doprinos bržoj i preciznijoj identifikaciji i karakterizaciji vrsta roda Fusarium u Srbiji.