Genetic characterisation of microsporidia infecting Indian tasar silkworm, Antheraea mylitta, using morphology and molecular tools

Abstract

The utility of inter simple sequence repeat-PCR (ISSR-PCR) assay in the genetic characterisation and elucidation of the phylogenetic relationship of different microsporidian isolates infecting tropical tasar silkworm, Antheraea mylitta Drury, is demonstrated. A total of 22 different microsporidians collected from the diseased tasar silkworms from Jharkhand state of India were analysed using morphological characters and ISSR-PCR. Observations spores under phase contrast microscope revealed oval to elongate in shape with length ranging from 3.8 μm to 5.1 μm and width from 2.6 μm to 3.3 μm. All the microsporidian isolates except MIJ-1gC showed gonadal infection and transovarial transmission in infected tasar silkworms. Fourteen out of 20 ISSR primers tested generated reproducible profiles and yielded a total of 281 fragments, of which 273 were polymorphic (97%). The degree of banding pattern was used to evaluate genetic distances and for phylogenetic analysis. The results demonstrated that ISSR analysis may be a useful and efficient tool for taxonomical grouping and phylogenetic classification of different microsporidians in general.