Ibrahim Alek, Baliarti Endang, Budisatria I Gede Suparta, Artama Wayan Tunas, Widayanti Rini, Maharani Dyah, Tavares Luis, Margawati Endang Tri
Department of Animal Production, Faculty of Animal Science, Universitas Gadjah Mada, Yogyakarta, Indonesia, 55281.
Research Center for Applied Zoology, National Research and Innovation Agency (BRIN), Bogor, Indonesia, 16911.
J Genet Eng Biotechnol. 2023 Mar 17;21(1):34. doi: 10.1186/s43141-023-00491-z.
The cytochrome b (Cyt b) gene is one of the most studied mitochondrial DNA (mtDNA) genes to determine sheep's genetic profile. This study aimed to determine the genetic diversity and relationships of several Indonesian local sheep populations on Java Island, Indonesia, based on the mtDNA Cyt b gene sequences. Blood samples were collected from forty-one individual sheep in seven populations of Indonesia local sheep breeds on Java Island (Priangan = 6, Garut = 6, Batur = 7, Wonosobo = 5, Javanese Thin-Tailed/JTT = 7, Javanese Fat-Tailed/JFT = 5, and Sapudi = 5). DNA extraction was performed on blood samples, and the mtDNA Cyt b gene was amplified using specific primers (Alek-CBF: 5'-CAACCCCACCACTTACAA-3' and Alek-CBR: 5'-CCTTGAGTCTTAGGGAGGTT-3'). The polymerase chain reaction (PCR) products were then sequenced, and data were analyzed using the MEGA version 7.0, DNA SP version 6.0, and NTSYS-pc version 2.11 software.
A total of 1140 bp complete mtDNA Cyt b gene sequences in this study obtained 1134 monomorphic sites (I), six polymorphic sites (V), one segregation site (S), and five parsimony informative sites (P) with a nucleotide diversity (Pi), the average number of nucleotide differences (K), and sequence conservation (SC) were 0.00119, 1.35610, and 0.9947, respectively. There were six haplotypes consisting of two unique haplotypes and four shared haplotypes with a haplotype diversity (Hd) of 0.5415. The genetic distance within and between populations ranged from 0.0000 to 0.0016 and 0.0000 to 0.0020, respectively. Wonosobo, JFT, and Sapudi sheep have the closest relationship, and then these three breeds were close to JTT sheep, followed by Batur, Priangan, and Garut sheep. Two haplogroups have been found based on the Ovine haplogroup clustering. All Wonosobo, JTT, JFT, Sapudi sheep, and most Batur sheep were clustered into haplogroup B. In contrast, Garut sheep were mostly clustered into haplogroup A, while Priangan sheep were clustered into both haplogroups with the same percentage.
Seven Indonesian local sheep breeds on Java Island have a close relationship clustered into two haplogroups, namely haplogroups A and B. Most Indonesian local sheep breeds on Java Island in this study were clustered into haplogroup B, except for Garut and Priangan sheep.
细胞色素b(Cyt b)基因是研究最多的线粒体DNA(mtDNA)基因之一,用于确定绵羊的遗传特征。本研究旨在基于mtDNA Cyt b基因序列,确定印度尼西亚爪哇岛几个本地绵羊群体的遗传多样性及亲缘关系。从爪哇岛七个印度尼西亚本地绵羊品种的41只个体绵羊采集血样(普里安甘羊=6只、加鲁特羊=6只、巴图尔羊=7只、沃诺索博羊=5只、爪哇细尾羊/JTT=7只、爪哇肥尾羊/JFT=5只、萨普迪羊=5只)。对血样进行DNA提取,使用特异性引物(Alek - CBF:5'-CAACCCCACCACTTACAA-3'和Alek - CBR:5'-CCTTGAGTCTTAGGGAGGTT-3')扩增mtDNA Cyt b基因。然后对聚合酶链反应(PCR)产物进行测序,并使用MEGA 7.0版、DNA SP 6.0版和NTSYS - pc 2.11版软件分析数据。
本研究共获得1140 bp的完整mtDNA Cyt b基因序列,有1134个单态位点(I)、6个多态位点(V)、1个分离位点(S)和5个简约信息位点(P),核苷酸多样性(Pi)、平均核苷酸差异数(K)和序列保守性(SC)分别为0.00119、1.35610和0.9947。有6种单倍型,包括2种独特单倍型和4种共享单倍型,单倍型多样性(Hd)为0.5415。群体内和群体间的遗传距离分别为0.0000至0.0016和0.0000至0.0020。沃诺索博羊、JFT羊和萨普迪羊亲缘关系最近,这三个品种随后与JTT羊关系较近,其次是巴图尔羊、普里安甘羊和加鲁特羊。基于绵羊单倍群聚类发现了两个单倍群。所有沃诺索博羊、JTT羊、JFT羊、萨普迪羊以及大多数巴图尔羊聚类到单倍群B。相比之下,加鲁特羊大多聚类到单倍群A,而普里安甘羊以相同比例聚类到两个单倍群。
爪哇岛的七个印度尼西亚本地绵羊品种亲缘关系较近,聚类为两个单倍群,即单倍群A和B。本研究中爪哇岛的大多数印度尼西亚本地绵羊品种聚类到单倍群B,加鲁特羊和普里安甘羊除外。