O. A. Koshkina, T. E. Deniskova, N. A. Zinovieva
Federal Research Center for Animal Husbandry named after Academy Member L.K. Ernst, pos. Dubrovitsy, 60, Podol’sk, Moskovskaya obl., 142132, Russian Federation
Abstract. An analysis of mitochondrial DNA (mtDNA) polymorphism is popular method for assessing the genetic diversity of livestock species. In sheep, the investigations of mtDNA have been mostly based on the analysis of the fragments (for example, cytochrome b gene or D-loop) that does not allow to obtain complete pattern on maternal heredity due to possible sequencing errors. The aim of this work is to develop a system for the analysis of complete mitochondrial genomes and to assess the phylogenetic relationships between representatives of Russian sheep breeds with different wool types. The test system is based on the amplification of three overlapping mtDNA fragments, which after purification and quality control of the libraries were sequenced using NGS technology. The selection of primers was carried out in accordance with the reference sequence of the domestic sheep complete mitochondrial genome (accession number NC_001941.1). The optimal conditions for the PCR amplification were selected experimentally. Sequencing of the obtained mtDNA amplicons was performed on a miSeq instrument (Illumina, USA). The first implementation of the developed test system for the analysis of complete mitochondrial genomes of sheep was performed on individuals from three Russian breeds including Altai Mountain (n=5), Groznensk (n=5) and Buubei (n=5). To assess the effectiveness of the developed test system, haplotype diversity (Hd) and nucleotide diversity (π) were calculated. All studied sheep samples were characterized by high haplotype (Hd = 0.990) and nucleotide diversity (π = 0.00496). A total of 14 haplotypes were identified and were belonged to two well-known haplogroups A (n=7), B (n=7) and C (n=1), which corresponded to sheep of European and Asian origin. Our study resulted in development and the first implementation of the test system for the analysis of complete mitochondrial genomes. New data on phylogenetic relationships of Russian sheep breeds were obtained.
Keywords: mitochondrial DNA, sheep, polymorphism, genetic diversity, phylogeny, haplotypes, haplogroups.
Author Details: O.A. Koshkina, post graduate student, junior research fellow (e-mail: Этот адрес электронной почты защищён от спам-ботов. У вас должен быть включен JavaScript для просмотра.); T. E. Deniskova, Cand. Sc. (Biol.), leading research fellow (e-mail: Этот адрес электронной почты защищён от спам-ботов. У вас должен быть включен JavaScript для просмотра.); N. A. Zinovieva, D. Sc. (Biol.), member of the RAS, director.
For citation: Koshkina OA, Deniskova TE, Zinovieva NA. [Development and implementation of test system for investigation of complete mitochondrial genomes of domestic sheep]. Dostizheniya nauki i tekhniki APK. 2022;36(10):60-5. Russian. doi: 10.53859/02352451_2022_36_10_60.