Maduna Simo N, Rossouw Charné, Roodt-Wilding Rouvay, Bester-van der Merwe Aletta E
Department of Genetics, Stellenbosch University, Private Bag X1, Matieland 7602, South Africa.
BMC Res Notes. 2014 Jun 10;7:352. doi: 10.1186/1756-0500-7-352.
Similarly to the rest of the world, southern Africa's diverse chondrichthyan fauna is currently experiencing high fishing pressures from direct and non-direct fisheries to satisfy market demands for shark products such as fins and meat. In this study, the development of microsatellite markers through cross-species amplification of primer sets previously developed for closely related species is reported as an alternative approach to de novo marker development. This included the design of four microsatellite multiplex assays and their cross-species utility in genetic diversity analysis of southern African elasmobranchs. As this study forms part of a larger project on the development of genetic resources for commercially important and endemic southern African species, Mustelus mustelus was used as a candidate species for testing these multiplex assays in down-stream applications.
Thirty five microsatellite primer sets previously developed for five elasmobranch species were selected from literature for testing cross-species amplification in 16 elasmobranch species occurring in southern Africa. Cross-species amplification success rates ranged from 28.6%-71.4%. From the successfully amplified microsatellites, 22 loci were selected and evaluated for levels of polymorphism, and four multiplex assays comprising of the 22 microsatellites were successfully constructed, optimised and characterised in a panel of 87 Mustelus mustelus individuals. A total of 125 alleles were observed across all loci, with the number of alleles ranging from 3-12 alleles. Cross-species amplification of the four optimised multiplex assays was further tested on 11 commercially important and endemic southern African elasmobranch species. Percentage of polymorphism ranged from 31.8%-95.5% in these species with polymorphic information content decreasing exponentially with evolutionary distance from the source species.
Cross-species amplification of the 35 microsatellites proved to be a time- and cost-effective approach to marker development in elasmobranchs and enabled the construction of four novel multiplex assays for characterising genetic diversity in a number of southern African elasmobranch species. This study successfully demonstrated the usefulness of these markers in down-stream applications such as genetic diversity assessment and species identification which could potentially aid in a more integrative, multidisciplinary approach to management and conservation of commercially important cosmopolitan and endemic elasmobranch species occurring in southern Africa.
与世界其他地区一样,南部非洲多样的软骨鱼类动物群目前正面临来自直接和非直接渔业的巨大捕捞压力,以满足市场对鱼翅和鱼肉等鲨鱼产品的需求。在本研究中,报告了通过对先前为密切相关物种开发的引物组进行跨物种扩增来开发微卫星标记,作为从头开发标记的替代方法。这包括设计四种微卫星多重检测方法及其在南部非洲板鳃亚纲动物遗传多样性分析中的跨物种应用。由于本研究是一个关于开发南部非洲重要商业和特有物种遗传资源的更大项目的一部分,条纹鼬鲨被用作候选物种,用于在下游应用中测试这些多重检测方法。
从文献中选择了先前为五个板鳃亚纲物种开发的35个微卫星引物组,用于测试在南部非洲出现的16种板鳃亚纲物种中的跨物种扩增。跨物种扩增成功率在28.6%-71.4%之间。从成功扩增的微卫星中,选择了22个位点并评估其多态性水平,并在一组87只条纹鼬鲨个体中成功构建、优化和表征了由这22个微卫星组成的四种多重检测方法。在所有位点共观察到125个等位基因,等位基因数量在3-12个之间。在11种南部非洲重要商业和特有板鳃亚纲物种上进一步测试了四种优化的多重检测方法的跨物种扩增。这些物种的多态性百分比在31.8%-95.5%之间,多态信息含量随着与源物种进化距离的增加而呈指数下降。
35个微卫星的跨物种扩增被证明是一种在板鳃亚纲动物中开发标记的省时且经济高效的方法,并能够构建四种新的多重检测方法,用于表征一些南部非洲板鳃亚纲物种的遗传多样性。本研究成功证明了这些标记在下游应用中的有用性,如遗传多样性评估和物种鉴定,这可能有助于采用更综合、多学科的方法来管理和保护南部非洲出现的重要商业世界性和特有板鳃亚纲物种。