Molecular characterization of the phytopathogen C. theae (Petch) – Causative of birds eye spot infection in Camellia sinensis host through DNA polymorphism and proteome analysis

Abstract

The pathogenic fungus Cercospora infecting plants causes high degree symptomatic disease in several important crops including tea (Camellia sinensis). Genetic diversity studies on plant pathogens are essential to tackle plant diseases, hence in the proposed study, Cercospora theae isolated from different regions was characterized using the molecular tool like RFLP of the rDNA regions amplified using ITS primers. Protein profiling for all the isolates was also done by optimizing the protein extraction using sodium dodecyl sulphate (SDS), urea and Trichloroacetic acid (TCA) protocols. Genetic relatedness with the identical size of molecular weight among the three C. theae isolates of different regions was studied from the 5.8S rDNA-ITS amplified regions. Digestion of the amplified regions with seven specific endonucleases exposed the same banding pattern among the isolates. The sequence data from the three isolates showed maximum homology with the Genbank sequence of Cercospora spp. (HQ450006, JQ754039, JQ753964, JN942274, JN942272, FJ460222 and EU581822). Further, an optimized protocol for protein profiling of C. theae was determined to extract the majority of the proteins. The results also revealed that the most suitable protocol for studying the proteomics of C. theae was the SDS extraction method.