Genotypic and phenotypic characterization of common bunt agent (Tilletia foetida)

Common bunt is a wheat disease that causes important crop losses and extensively spreads in Turkey and World. Two fungi species that Tilletia foetida and Tilletia caries primary cause this disease in our country. It has been reported that in time pathogen have become resistant according as fungicide uses and environmental factors. In this study, for determination of genetic diversity of pathogen T. foetida using ISSR-PCR method, DNA isolation of 13 T. foetida isolates and 1 T. caries isolate as out-group was carried with classical CTAB method. Isolated DNAs were amplified with PCR method using 25 different ISSR primers. Amplified DNA fragments were run in 1.2% agarose gel and digitally photographed. Band analysis of PCR yields was carried using Phoretix 1d pro software, Jaccard similarity matrix was obtained with UPGMA method using data from binary method and phylogenetic tree showing genetic distance and similarity among common bunt isolates was formed. It was observed that isolates have high genetic varieties, close locations of T. foetida grouped together and T. caries departed from the other isolates as out-group. Phylogenetic analyses data of isolates were compared with pathogenicity rates against Heines VII wheat variety that haven't got resistance genes against common bunt. It was determined that 847 and 861 numbered primers among 25 ISSR primers used in study amplify species-specific bands and it was observed that this primers can use an markers for early detection of contaminated wheat seeds before sowing. Scanning electron microscope (SEM) analyses of isolates were carried and obtained phenotypic morphological data were compared with genotypic molecular analyses data. In addition, isolated mRNAs from T. foetida were converted to cDNAs by revers transcription and transferred to PGEM-Teasy plasmid vectors. Recombinant plasmid vectors including cDNAs were transferred to XLI-Blue hosts and at the first time a cDNA library of T. foetida pathogen was constructed for using in phenotypic characterization researches.