Masih Prachi, Luhariya Rupesh K, Das Rakhi, Gupta Arti, Mohindra Vindhya, Singh Rajeev K, Srivastava Rohit, Chauhan U K, Jena J K, Lal Kuldeep K
National Bureau of Fish Genetic Resources (ICAR), Canal Ring Road, P.O. Dilkusha, Lucknow, 226 002, Uttar Pradesh, India.
Mol Biol Rep. 2014 Aug;41(8):5187-97. doi: 10.1007/s11033-014-3386-0. Epub 2014 May 4.
This study is aimed to identify polymorphic microsatellite markers and establish their potential for population genetics studies in three carp (family cyprinidae; subfamily cyprininae) species, Labeo rohita, Catla catla and Cirrhinus mrigala through use of cyprinid primers. These species have high commercial value and knowledge of genetic variation is important for management of farmed and wild populations. We tested 108 microsatellite primers from 11 species belonging to three different cyprinid subfamilies, Cyprininae, Barbinae and Leuciscinae out of which 63 primers (58.33%) successfully amplified orthologous loci in three focal species. Forty-two loci generated from 29 primers were polymorphic in these three carp species. Sequencing of amplified product confirmed the presence of SSRs in these 42 loci and orthologous nature of the loci. To validate potential of these 42 polymorphic loci in determining the genetic variation, we analyzed 486 samples of three focal species collected from Indus, Ganges and Brahmaputra river systems. Results indicated significant genetic variation, with mean number of alleles per locus ranging from 6.80 to 14.40 and observed heterozygosity ranging from 0.50 to 0.74 in the three focal species. Highly significant (P < 0.00001) allelic homogeneity values revealed that the identified loci can be efficiently used in population genetics analysis of these carp species. Further, thirty-two loci from 19 primers were useful for genotyping in more than one species. The data from the present study was compiled with cross-species amplification data from previous results on eight species of subfamily cyprininae to compare cross-transferability of microsatellite loci. It was revealed that out of 226 heterologous loci amplified, 152 loci that originated from 77 loci exhibited polymorphism and 45 primers were of multispecies utility, common for 2-7 species.
本研究旨在通过使用鲤科引物,鉴定多态微卫星标记,并确定其在三种鲤鱼(鲤科;鲤亚科),即印度野鲮、印度鲃和露斯塔野鲮群体遗传学研究中的潜力。这些物种具有很高的商业价值,了解其遗传变异对于养殖和野生种群的管理至关重要。我们测试了来自鲤科三个不同亚科,即鲤亚科、鲃亚科和雅罗鱼亚科的11个物种的108个微卫星引物,其中63个引物(58.33%)在三个目标物种中成功扩增出直系同源位点。由29个引物产生的42个位点在这三种鲤鱼物种中具有多态性。扩增产物的测序证实了这42个位点中存在简单序列重复(SSR)以及这些位点的直系同源性质。为了验证这42个多态位点在确定遗传变异方面的潜力,我们分析了从印度河、恒河和雅鲁藏布江流域采集的三种目标物种的486个样本。结果表明存在显著的遗传变异,三个目标物种中每个位点的平均等位基因数在6.80至14.40之间,观察到的杂合度在0.50至0.74之间。高度显著(P < 0.00001)的等位基因同质性值表明,所鉴定的位点可有效地用于这些鲤鱼物种的群体遗传学分析。此外,来自19个引物的32个位点可用于一个以上物种的基因分型。本研究的数据与先前关于鲤亚科八个物种的跨物种扩增数据进行了汇总,以比较微卫星位点的交叉转移性。结果显示,在扩增的226个异源位点中,源自77个位点的152个位点表现出多态性,45个引物具有多物种通用性,适用于2至7个物种。