Use of ISSR markers for genotyping an experimental group of largemouth bass Micropterus salmoides (LACEPEDE, 1802), reared in ponds of Polissia of Ukraine

Abstract

The aim of this study was to investigate the genetic structure of the experimental group of largemouth bass reared in the ponds of Polissia of Ukraine using ISSR markers. To accomplish these tasks, ISSR genotyping of the genetic structure of largemouth bass was performed using four fragments of trinucleotide loci. The genetic structure of the experimental group of largemouth bass of the pond fsh farm "Nyvka" was characterised using 4 primers B – (GAG)6C; C – (AGC)6G; D – (ACC)6G and E – (AGC)6C. Fins fragments were used for the study. In the course of the work, the optimal conditions for ISSR-PCR analysis were selected. The study revealed a number of factors that affect the efciency of these markers: DNA concentration, number of amplifcation cycles. For 4 markers, 80 alleles with a molecular weight of 160–1320 bp were identifed. The ranges of amplicons for the selected markers were determined: marker B – from 150 to 1186 bp; marker C – from 640 to 200 bp; D – from 1320 to 225 bp; and E – within 630–160 bp. The most polymorphic marker is marker B – 26 alleles, the least polymorphic marker is marker E – 15 alleles. In the studied experimental group of largemouth bass, the effective number of alleles varied from 10.2 for marker E to 12.2 for marker C, D. The indicators of genetic variability were determined by calculating allelic frequencies, the maximum level of available heterozygosity is 0.918 for marker C, D, and the lowest for marker E is up to 0.902. A method has been proposed that makes it possible to analyse the genetic structure of the experimental group of largemouth bass using these primers and to implement genetic information at different stages of the selection process. Keywords: ISSR-PCR, genetic structure, largemouth bass, DNA– markers, genotype, amplicons, genetic polymorphism, molecular genetic marker.