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Population-genetic monitoring of the Ukrainian population of buffaloes (Bubalus bubalis) using 11 microsatellite DNA loci
The problem of preservation of existing biodiversity on the Earth has acquired the global nature that has caused concern of the international community. A special place in biodiversity conservation is given to agriculture where the preservation of genetic resources of livestock breeding as a part of the biological community in the world has become actual.
Whereas in Ukraine for the preservation of genetic diversity of cattle there are programs designed for several years, the situation with buffaloes seems to be worse. The number of buffaloes in the world is constantly increasing. In 1961 the number of buffaloes was 88.32 million head, in 2011 there were already about 182 million heads, for 50 years the number of buffaloes increased to 93.68 million heads or by 206.1 %. Buffaloes are bred on all continents, but most of them – 95 % of the total livestock population – are in Asian countries. For 50 years (1961–2011) the trend of annual increase in the number of buffalo by 4.12 % has been observed.
Therefore, the study of genetic diversity of buffaloes is particularly important. A way of its studying is using molecular genetic markers, including sequences of DNA, the polymorphism of which is caused by differences in the nucleotide sequences of different alleles at one locus. One of these types of genetic markers is microsatellite DNA loci. Recent years genetic characteristics of buffaloes by using microsatellite acquired special distribution.
Despite a number of existing microsatellite loci used for the research of buffaloes genetic analysis by using microsatellite loci for cattle is very efficient.
11 microsatellite loci (BM1818, BM2113, BM1824, INRA023, ILST006, ETH10, ETH185, ETH225, SPS115, TGLA126, TGLA227) included in the list are recommended by ISAG for genotyping of cattle were selected for the analysis.
Genetic analysis of 64 buffaloes was conducted at the Laboratory of Molecular Genetics and Cytogenetics of animals of the Center of Biotechnology and Molecular Diagnostics of the State Scientific Institution Russian Research Institute of livestock. Sampling for animal studies was performed in vivo, by select in samples from ears (plucking tissue) for studies of DNA.
As a result of the research 73 allelic variants were identified and the frequency they met with was determined. The number of alleles by locus varied from 3 (BM2113) to 12 (TGLA227). The average number of alleles by locus was 6.55.
The actual heterozygote varied from 0.260 (BM2113) to 0.980 (INRA023), while the theoretically expected one varied from 0.291 (BM2113) to 0.753 (TGLA227).
The excess of the average value of the actual heterozygote theoretically expected indicates the surplus of heterozygous genotypes in the population. This is confirmed by the fixation index, which reflects the level of inbreeding of the individuals in relation to the population. On average it was 5.5 % for the eleven loci. Overall, half of the studied loci recorded a surplus of heterozygous genotypes with the highest level for the locus TGLA126 (34.2 %). The maximum deficit of heterozygotes was observed in BM1818 (27.3 %).
Loci with the value PIC>0.500 are highly polymorphic, PIC within 0.250–0.500 characterizes moderately polymorphic loci and if the value is PIC<0.250 markers are lowly polymorphic. On average after the studied loci the population was highly polymorphic (PIC=0.584). Exceptions were loci BM1818, BM2113 and ETH185, which recorded the average polymorphism. The locus TGLA227 was the most polymorphic in the studied population.
The analysis of probability of exclusion of casual coincidence of alleles allowed to assess the efficiency of using the microsatellite loci for the genetic examination of the origin. Despite the fact that the genetic study of the Ukrainian population of buffaloes was conducted on microsatellite loci recommended for studying cattle, the effectiveness of their use was extremely high and amounted to 99.99 %. The locus BM2113 was the least effective, while the efficiency of INRA 023 was 96 %.
As a result of the genetic studies of the Ukrainian population of buffaloes narrowing of genetic diversity is not fixed. Using microsatellite loci recommended for genetic analysis of cattle showed high level of polymorphism among buffaloes. The efficiency of using the mentioned list of microsatellites was extremely high, what indicating the suitability of their application for genetic analysis of buffaloes.
Key words: buffalo, genetic diversity, population, microsatellite loci, alleles.
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