To study morphological characteristics of this pathogen, well
separated mycelia specimens from pure culture of the isolate were
stained with cotton blue and observed under light microscope
(10, 40 and 100).
Identification of one of the representative isolated pathogen
VBAM1 was carried out using D1/D2 region of LSU (Large SubUnit:
28S rDNA) based molecular technique. Forward and reverse DNA
sequencing reactions were carried out with DF and DR primers
using BDT v3.1 Cycle sequencing kit on ABI 3730xl Genetic Analyzer.
Consensus sequence of D1/D2 region of 28S rDNA gene
was generated from forward and reverse sequence data using
aligner software. The D1/D2 region of LSU gene sequence was used
to carry out BLAST with the nrdatabase of NCBI GenBank database.
Based on maximum identity score first ten sequences were
selected and the phylogenetic tree was constructed using MEGA
4 (Tamura et al., 2007).
2.3. Study of pectinase activity of isolated pathogens