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引用本文:李碧瀚,于洋,刘桂嘉,罗正,李富花.基于三代测序技术的凡纳滨对虾(Litopenaeus vannamei)高通量候选基因关联分析方法的建立.海洋与湖沼,2021,52(5):1255-1264.
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基于三代测序技术的凡纳滨对虾(Litopenaeus vannamei)高通量候选基因关联分析方法的建立
李碧瀚1,2, 于洋1, 刘桂嘉1, 罗正1,2, 李富花1
1.中国科学院海洋研究所实验海洋生物学重点实验室 青岛 266071;2.中国科学院大学 北京 100049
摘要:
凡纳滨对虾(Litopenaeus vannamei)是世界也是我国的主导对虾养殖品种,产业的发展离不开良种的支撑,分子育种被认为是加快良种选育的最有效途径,而目标性状相关分子标记的开发是发展分子育种的基础。研究主要目的是建立一种适用于凡纳滨对虾等水产经济物种的高通量候选基因关联分析方法,并在抗弧菌相关标记筛选中进行应用。首次将三代靶向测序技术用于对虾候选基因的基因分型,在抗弧菌性状候选基因LvPI3K的全长序列上发掘到91个SNP位点,通过关联分析鉴定到21个与抗弧菌性状显著相关的SNP标记(P<0.05),利用Sanger测序证实了三代靶向测序技术分型结果准确可靠。所建立的基于三代测序的靶向分型方法为凡纳滨对虾等水产动物提供了一种高效、低成本的基因分型方法,所发掘的抗弧菌性状相关位点对开展凡纳滨对虾抗弧菌性状分子育种具有一定指导意义。
关键词:  三代靶向测序  基因分型  抗弧菌性状  关联分析  凡纳滨对虾
DOI:10.11693/hyhz20210300071
分类号:S917.4
基金项目:国家重点研发计划,2018YFD0901301号,2018YFD0900303号;中国科学院战略性先导专项,XDA24030105号。
附件
A HIGH-THROUGHPUT METHOD FOR CANDIDATE GENE ASSOCIATION ANALYSIS IN LITOPENAEUS VANNAMEI BASED ON THE THIRD-GENERATION SEQUENCING TECHNOLOGY
LI Bi-Han1,2, YU Yang1, LIU Gui-Jia1, LUO Zheng1,2, LI Fu-Hua1
1.Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China;2.University of Chinese Academy of Sciences, Beijing 100049, China
Abstract:
The Pacific white shrimp (Litopenaeus vannamei) is a dominant aquaculture shrimp species in China. Development of the shrimp industry depends heavily on the improved broodstocks, and molecular breeding is considered as the most effective approach to accelerate the selective breeding. Development of molecular markers related to interested traits is the basis of molecular breeding. To establish a high-throughput candidate gene association analysis method suitable for L. vannamei and other aquaculture species, the third-generation target sequencing technology was applied. A total of 91 SNPs were discovered in LvPI3K, and 21 SNPs were significantly (P<0.05) correlated with disease-resistance trait against Vibrio in L. vannamei. The results were approved accurate and reliable after being validated in an independent population using the Sanger sequencing technology. This approach provides an efficient and low-cost genotyping method for aquaculture species, and the identified markers will be useful for the marker-assisted breeding of disease resistance in L. vannamei.
Key words:  third-generation target sequencing  genotyping  disease-resistant trait  association analysis  Litopenaeus vannamei
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