Use of isozyme analyses and PCR based methods RAPD and RFLP for assessment of biochemical and genetic diversity of morphologically similar ectomycorrhizal Lactarius deliciosus from India


K. Mukesh Upadhyay *, A. K. Pandey and R. C. Rajak

The present study was undertaken to assess the genetic diversity and phylogenetic relationship among the morphologically similar and ecologically variant isolates of Lactarius deliciosus using polymerase chain reaction (PCR) based techniques random amplification of polymorphic DNA (RAPD), rDNA analysis and isozyme patterns. Two major groups were defined amongst the total isolates by cluster analyses. Four bands of amylase isozymes viz; A232.4, A342.7, A461.5, A672.0 and two bands each of pectinases (Pe364.18, Pe261.11), cellulases (C460.14, C356.12), glutamate dehydrogenases (GLD126.00, GLD229.18) and acid phosphatases (AP110.00, AP214.18) were common in all the morphologically similar isolates. The RAPD profile revealed that a total of 12 polymorphic bands were obtained, among these a locus of 550 bp was diversified in all strains except M221 and M222. Few common bands which is present in zymogram (isozymes) and electrogram (RAPD) exhibited genetic similarities amongst ecologically variant isolates. Amplifications of the ITS region resulted a single PCR product of approximately 560 bp for all the isolates. When amplified fragments were digested with restriction enzymes, identical banding patterns were observed, which showed that no inter-generic or intra-specific variation among the isolates.

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