rDNA sequence analysis

The PCR amplified internal transcribed (ITS) rDNA region from each of the isolates was purified using Favorgen PCR purification kit (FAVORGEN Biotech Corp, Taiwan), as per the manufacturer's instructions. The purified ITS amplicons were sequenced by Agrigenome, Bengaluru, India and then the sequences were analyzed in GenBank (http://www.ncbi.nlm.nih.gov/) by using the Mega BLAST sequence analysis tool. The full length ITS sequences were used as queries for establishing kinship with the published sequences and search result with maximum homology and highest score were marked for further analysis. All the ITS sequences obtained from the isolates used in the present study were submitted to GenBank. ClustalW program of the Bioedit sequence alignment editor (Thompson et al., 1994; Hall, 1999) was used to align all the obtained sequences. The resulting multiple-alignment file was used for phylogenetic analyses which were performed using Molecular Evolutionary Genetics Analysis (MEGA 7.0) with Neighbour-Joining method (Kumar et al., 2016).