Genetic Diversity and Phylogenetic Relationships among Microsporidian Isolates from the Indian Tasar Silkworm, Antheraea mylitta, as Revealed by RAPD Fingerprinting Technique
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In this study, we investigated genetic diversity of 22 microsporidian isolates infecting tropicaltasar silkworm, Antheraea mylitta collected from various geographical forest locations inthe state of Jharkhand, India, using polymerase chain reaction (PCR)-based marker assay:random amplified polymorphic DNA (RAPD). A type species, NIK-1s_mys was used ascontrol for comparison. The shape of mature microsporidians was found to be oval toelongate, measuring 3.80 to 5.10 μm in length and 2.56 to 3.30 μm in width. Of the 20 RAPDprimers screened, 16 primers generated reproducible profiles with 298 polymorphic fragmentsdisplaying high degree of polymorphism (97%). A total of 14 RAPD primers produced 45unique putative genetic markers, which were used to differentiate the microsporidians. Calculation of genetic distance coefficients based on dice coefficient method and clusteringwith un-weighted pair group method using arithmetic average (UPGMA) analysis wasconducted to unravel the genetic diversity of microsporidians infecting tasar silkworm. Thesimilarity coefficients varied from 0.059 to 0.980. UPGMA analysis generated a dendrogramwith four microsporidian groups, which appear to be different from each other as well as fromNIK-1s_mys. Two-dimensional distribution based on Euclidean distance matrix also revealedconsiderable variability among different microsporidians identified from the tasar silkworms. Clustering of few microsporidian isolates was in accordance with the geographic origin. Theresults indicate that the RAPD profiles and specific/unique genetic markers can be used fordifferentiating as well as to identify different microsporidians with considerable accuracy.
Abstract
In this study, we investigated genetic diversity of 22 microsporidian isolates infecting tropicaltasar silkworm, Antheraea mylitta collected from various geographical forest locations inthe state of Jharkhand, India, using polymerase chain reaction (PCR)-based marker assay:random amplified polymorphic DNA (RAPD). A type species, NIK-1s_mys was used ascontrol for comparison. The shape of mature microsporidians was found to be oval toelongate, measuring 3.80 to 5.10 μm in length and 2.56 to 3.30 μm in width. Of the 20 RAPDprimers screened, 16 primers generated reproducible profiles with 298 polymorphic fragmentsdisplaying high degree of polymorphism (97%). A total of 14 RAPD primers produced 45unique putative genetic markers, which were used to differentiate the microsporidians. Calculation of genetic distance coefficients based on dice coefficient method and clusteringwith un-weighted pair group method using arithmetic average (UPGMA) analysis wasconducted to unravel the genetic diversity of microsporidians infecting tasar silkworm. Thesimilarity coefficients varied from 0.059 to 0.980. UPGMA analysis generated a dendrogramwith four microsporidian groups, which appear to be different from each other as well as fromNIK-1s_mys. Two-dimensional distribution based on Euclidean distance matrix also revealedconsiderable variability among different microsporidians identified from the tasar silkworms. Clustering of few microsporidian isolates was in accordance with the geographic origin. Theresults indicate that the RAPD profiles and specific/unique genetic markers can be used fordifferentiating as well as to identify different microsporidians with considerable accuracy.
참고문헌 (35)
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