which allow to amplify the ITS region of the fungal species.
The conditions for Polymerase Chain Reaction for ITSrDNA
amplification was 95 C for 5 min, 35 cycles of 94 C
for 45 s, 60 C for 30 s, 72 C for 45 s and final 72 C for
5 min. The product size was approximately 575 bp which was
visible in 1.2% agarose gel under UV light. The PCR product
of each isolate was purified using PCR purification kit (Fermentas,
Lithuania) as per the provided protocol. Sequencing
of the PCR product was done by a Sanger’s Dideoxy method
on applied Biosystem 3730XL (Bioloink, New Delhi, India).
The sequence of each isolate was subjected to BLAST search
(http://wwwncbinlmgov/BLAST) with NCBI database [1].
All the sequences were aligned with representative sequences
in the NCBI database using CLUSTAL W [48] and employing
MEGA 52 software, the phylogenetic analysis of the alignment
was performed with Maximum Likelihood Method.
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