虹鳟倍性鉴定SSR标记筛选与应用研究  

Screening and Application of SSR Markers for Identification of Ploidy in Oncorhynchus mykiss

作　　者：马海兵 黄天晴 徐革锋[1] 刘恩慧 谷伟[1] 王高超 郭福元 董福霖 郑龙华 张黎黎 姜再胜 王炳谦[1] MA Haibing;HUANG Tianqing;XU Gefeng;LIU Enhui;GU Wei;WANG Gaochao;GUO Fuyuan;DONG Fulin;ZHENG Longhua;ZHANG Lili;JIANG Zaisheng;WANG Bingqian(Heilongjiang Fisheries Research Institute,Chinese Academy of Fishery Sciences,Cold Water Fish Industry Technology Innovation Strategic Alliance,Harbin 150070,China;Yantai Jinghai Marine Fishery Co.LTD,Yantai 264006,China;Anhui Tongcheng Aquatic Products Service Center,Tongcheng 231400,China)

机构地区：[1]中国水产科学研究院黑龙江水产研究所冷水鱼类产业技术创新战略联盟,黑龙江哈尔滨150070 [2]烟台经海海洋渔业有限公司,山东烟台264006 [3]安徽省桐城市水产服务中心,安徽桐城231400

出　　处：《渔业科学进展》2024年第4期43-52,共10页Progress in Fishery Sciences

基　　金：财政部和农业农村部:国家现代农业产业技术体系(CARS-46);中国水产科学研究院基本科研业务费项目(2020TD32)共同资助。

摘　　要：虹鳟(Oncorhynchusmykiss)是我国冷水性鱼类的主要养殖品种。与二倍体相比,虹鳟三倍体具有生长速度快、肉质好和不育等优点。目前,鉴定虹鳟倍性主要是通过流式细胞术进行DNA含量分析,但这种方法需要采集鱼类血液或组织样本,会对其造成伤害。为了实现利用微创方法收集样本鉴定虹鳟倍性,本研究利用PCR扩增以及电泳分离技术对153个微卫星标记(SSR标记)进行分析,其中139个SSR标记能够在二倍体和三倍体中成功扩增,筛选出132个SSR标记具有多态性,最终鉴定出7个标记在三倍体中表现出较高的变异性。通过在52个已知倍性水平的参考样本和48个未知倍性的样本上进行验证,进一步从中筛选出3个SSR标记(SSR1054、SSR1056和SSR1468)足以区分倍性水平,并且根据3个标记序列重新设计了3对具有良好稳定性的引物序列进行倍性分析应用。本研究所筛选的SSR标记解决了现有虹鳟倍性鉴定中存在的技术流程复杂、耗材昂贵的问题,并有助于不同倍性虹鳟的遗传多样性研究。Rainbow trout(Oncorhynchus mykiss)is one of the world's most widely farmed cold-water economic fish.It is also the primary cold-water fish species in China.Since the 1960s,the rainbow trout germplasm has been introduced from North Korea,the United States,Denmark,and other countries,and a systematic study has been carried out on germplasm preservation,identification,breeding,and variety breeding.Compared with rainbow trout diploid,the triploid has the advantages of a high feed conversion rate,fast growth rate,and good meat quality.Moreover,the triploid gonadal hypoplasia can mitigate risks associated with high mortality and meat quality decline during spawning and avert the ecological risk of invasive alien species caused by breeding escape.Therefore,breeding triploid rainbow trout has more significant market and ecological benefits.As diploid and triploid rainbow trout are very similar in terms of morphology,it is difficult to distinguish the ploidy of rainbow trout by morphology alone.Currently,DNA content analysis by flow cytometry is the main method used to identify the ploidy of rainbow trout,but this method necessitates the collection of fish blood or tissue samples,and the body length of the fish to collect the blood must be at least 5 cm.Additionally,these sampling operations may harm the fish.This method also involves a delicate operation,complicated technical process,and expensive consumables,thus,the popularization and application of triploid identification of rainbow trout are greatly restricted.Considering the limitations of the technical test for the ploidy of rainbow trout,a minimally invasive and economical method with only a small amount of sampling is needed to identify and analyze the ploidy of rainbow trout in batches.Microsatellite markers(SSR markers)are simple repeated sequences widely distributed in eukaryotic genomes and have been widely used in ploidy and pedigree analyses of fish.The advantages of microsatellite(SSR)analysis include high reproducibility,low sample requirements,and rapid

关 键 词：虹鳟 鲑科鱼类 微卫星 三倍体 倍性检测 

分 类 号：S961.6[农业科学—水产养殖]