REP-PCR fingerprinting based genetic variability in Tilletia Indica

The repetitive sequence based polymerase chain reaction (rep-PCR) was evaluated as a tool to investigate variability among different isolates of Tilletia indica. DNA primers (BOX and ERIC) corresponding to conserved repetitive element motifs, originally described in prokaryotes, were used to generate genomic fingerprints of 18 isolates of T. indica from different locations of North Western Plain Zone of India. The amplified bands in BOX ranged in length from 600 to 3200 bp whereas for the ERIC, bands ranged from 400 to 3000 bp. A total of 72 bands were amplified of which 70 (97.2%) were polymorphic. The cluster analysis showed that most isolates from the same regions were grouped in the same cluster or a close cluster. However, the isolate, KBJ2 from Jammu was found to be the most distinct from rest of the isolates. The variability found within closely related isolates of T. indica demonstrated the effectiveness of rep-PCR marker in identifying genetic diversity among T. indica isolates. The results of Principal Component Analysis (PCA) were similar to those detected by cluster analysis. The first two major axis of differentiation (PC1 and PC2) explained 52.98% of the total variation.