Genetic structure of the Ukrainian water buffalo population by ISSR-PCR markers

Abstract

The study of intraspecific genetic diversity of cattle, including species of the genus Bubalus from the subfamily Bull, is important because of the reduction of biodiversity of farm animals. The basis of genetic diversity is its genetic component. The loss of native species and breeds of cattle is a real threat to the biosphere, as the resilience of natural ecosystems and agroecosystems is directly linked to their genetic ability to adapt to environmental conditions. Polymorphism of ISSR-markers of the Ukrainian population of water buffaloes (Bubalus bubalis) from the farm of “TASBIO” LLC (Chernihiv region) was analyzed in 66 animals selected for the study. Genomic DNA samples were isolated from venous blood with a standard set of reagents. Genotyping was performed using specific ISSR primers: (ACC)6G, (GAG)6C, (AG)9C, (CTC)6C, (AG)8CA, (AG)8CGand (GA)6CC. We determined the difference in spectra both by the number of amplicons, their lengths (number of nucleotides) and by their polymorphism. As a result of the study, all primers showed polymorphism of buffalo DNA regions. Amplicons were defined in the range from 200 bp up to 4000 bp. Analysis of ISSR spectra revealed 87 loci, of which 71 were polymorphic. (AG)8CA-marker was the least polymorphic (PIC=0.234), (CTC)6C the most polymorphic (PIC=0.389). Conservative loci were found in four ISSR markers: 6 in (AG)8CA marker, 5 in (AG)8CG, 4 in (GA)6CC, and 1 in (AG)9C. 67 species-specific loci were identified for the Ukrainian water buffalo population: 10 for (AG)9C, 3 loci for (ACC)6G, 4 for (GAG)6C, 7 for (CTC)6C, 15 for (AG)8CA, and 14 in (AG)8CG and (GA)6CC. The ISSR primers used are recommended for molecular genetic analysis of buffalo DNA polymorphism.