基于微卫星标记的大口黑鲈（<i>Micropterus salmoides</i>）原种和养殖群体遗传多样性和结构分析

doi:10.3785/j.issn.1008-9209.2020.04.031

浙江大学学报（农业与生命科学版）

2020, Vol. 46

Issue (6): 687-698 DOI: 10.3785/j.issn.1008-9209.2020.04.031

数量遗传与生物信息

基于微卫星标记的大口黑鲈（Micropterus salmoides）原种和养殖群体遗传多样性和结构分析

苏胜彦¹(

),张林兵²,李海洋³,郜灿²,贺鑫晋⁴,田灿⁵,李建林¹,王美垚¹,唐永凯¹(

)

^1.中国水产科学研究院淡水渔业研究中心，农业农村部淡水渔业与种质资源利用重点实验室，江苏无锡 214081
^2.安徽张林渔业有限公司，安徽铜陵 244000
^3.安徽省农业科学院水产研究所，合肥 230031
^4.山西农业大学动物科技学院，山西晋中 030800
^5.上海海洋大学水产科学国家级实验教学示范中心，上海 201306

Genetic diversity and structure analyses of largemouth bass (Micropterus salmoides) original and cultured populations based on microsatellite markers

Shengyan SU¹(

),Linbing ZHANG²,Haiyang LI³,Can GAO²,Xinjin HE⁴,Can TIAN⁵,Jianlin LI¹,Meiyao WANG¹,Yongkai TANG¹(

)

^1.Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture and Rural Affairs, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, Jiangsu, China
^2.Zhanglin Fisheries Co. , Ltd. , Tonglin 244000, Anhui, China
^3.Institute of Aquaculture, Anhui Academy of Agricultural Sciences, Hefei 230031, China
^4.College of Animal Science, Shanxi Agricultural University, Jinzhong 030800, Shanxi, China
^5.National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China

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摘要：

为了解大口黑鲈（Micropterus salmoides）的种质资源遗传背景，采用11个微卫星位点研究了大口黑鲈原种（加州原种）、从台湾省引进的台湾选育种（台湾加州鲈）和大陆本土化的养殖种（优鲈1号）的遗传多样性及遗传结构。结果显示：加州原种的等位基因数和有效等位基因数显著高于优鲈1号和台湾加州鲈；其中，加州原种的5个位点的观察杂合度和期望杂合度显著高于优鲈1号和台湾加州鲈，而后两者类似。从多态信息含量来看，加州原种群体中有11个位点处于高度多态水平，优鲈1号群体和台湾加州鲈群体分别有6和4个位点处于高度多态水平。在遗传平衡检验中发现：台湾加州鲈群体中偏离哈代-温伯格平衡的位点较多（P＜0.05），而加州原种存在3个连锁座位对的连锁不平衡状态（P＜0.05）。在双相突变模型中，无论是WILCOXON检验还是符号检验，加州原种群体均处于突变-漂移不平衡状态。分子方差分析（analysis of molecular variance, AMOVA）发现：大口黑鲈群体中15.57%的遗传变异来自群体间，25.05%的遗传变异来自群体内个体间，59.38%的遗传变异来自个体间（P＜0.01）。聚类分析发现：加州原种和优鲈1号之间的遗传距离最大，其次为加州原种和台湾加州鲈之间的遗传距离。当K为2时，加州原种与优鲈1号、台湾加州鲈的遗传结构有明显的差异。对个体的种群鉴定发现：加州原种群体被误判的比例最低，其次是优鲈1号，最后是台湾加州鲈。在判别准确率为85.00%的水平上，加州原种被判别正确的概率是67.70%，优鲈1号是53.30%，台湾加州鲈是20.00%。从遗传多样性、遗传距离、平衡分析均发现，加州原种多态性高，遗传距离较另外2个群体较远，并可能存在过遗传瓶颈，聚类和个体的种群鉴定进一步证明了此观点。这提示从美国引进的大口黑鲈原种应该扩群保种并系统地进行育种开发工作。

关键词： 大口黑鲈; 微卫星标记; 遗传多样性; 遗传结构

Abstract:

In order to examine the genetic background of Micropterus salmoides, eleven microsatellite loci were used to study the genetic diversity and structure of the introduced subspecies (California M. salmoides), selected subspecies from Taiwan Province (Taiwan-California M. salmoides) and cultured species (Youlu No. 1) in mainland. The results showed that both the number of alleles and effective alleles was higher in California M. salmoides than in Taiwan-California M. salmoides and Youlu No. 1. The observed heterozygosity and expected heterozygosity at five loci were significantly higher in California population than in Taiwan-California and Youlu No. 1 populations, which were similar. According to the polymorphic information content, eleven loci in the California population were at high polymorphic levels. Six loci and four loci were at highly polymorphic levels for Youlu No. 1 and Taiwan-California populations, respectively. In the genetic equilibrium test, it was found that there were more loci deviating from Hardy-Weinberg equilibrium in Taiwan-California population, and there were three linkage pairs in California population (P＜0.05). For the two-phased model of mutation (TPM), both WILCOXON and sign tests showed that California population was in a status of mutation-drift disequilibrium. For population structure, it was found by analysis of molecular variance (AMOVA) that 15.57% of the genetic variation came from among populations, 25.05% from within populations, and 59.38% from among individuals (P＜0.01). The genetic distance between California population and Youlu No. 1 population was the largest, followed by that between California population and Taiwan-California population. When the K was two, there were significant differences in genetic structures between California population and other two populations. Through individual population identification, California population had the lowest rate of misjudgment, followed by Youlu No. 1 and Taiwan-California populations. When the accuracy was 85.00%, the probabilities of correct discrimination of California, Youlu No. 1 and Taiwan-California populations were 67.70%, 53.30% and 20.00%, respectively. According to the analysis of genetic diversity, genetic distance and equilibrium test, it was found that the original species of California had high polymorphism, and the genetic distance was bigger than that of the other two populations, and there might be genetic bottleneck. It is suggested that the species of M. salmoides introduced from the United States should be expanded and cultivated systematically.

Key words: Micropterus salmoides microsatellite marker genetic diversity genetic structure

收稿日期: 2020-04-03 出版日期: 2020-12-31

CLC:

Q 75

基金资助: 中国水产科学研究院基本科研业务费(2020JBFJ02);物种品种资源保护费（水产新品种生产性能测试等第三方评价）

通讯作者: 唐永凯 E-mail: susy@ffrc.cn;tangyk@ffrc.cn

作者简介: 苏胜彦（https://orcid.org/0000-0001-8881-8410），Tel：+86-510-85555796，E-mail：susy@ffrc.cn

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引用本文:

苏胜彦,张林兵,李海洋,郜灿,贺鑫晋,田灿,李建林,王美垚,唐永凯. 基于微卫星标记的大口黑鲈（Micropterus salmoides）原种和养殖群体遗传多样性和结构分析[J]. 浙江大学学报（农业与生命科学版）, 2020, 46(6): 687-698.

Shengyan SU,Linbing ZHANG,Haiyang LI,Can GAO,Xinjin HE,Can TIAN,Jianlin LI,Meiyao WANG,Yongkai TANG. Genetic diversity and structure analyses of largemouth bass (Micropterus salmoides) original and cultured populations based on microsatellite markers. Journal of Zhejiang University (Agriculture and Life Sciences), 2020, 46(6): 687-698.

链接本文:

http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2020.04.031 或 http://www.zjujournals.com/agr/CN/Y2020/V46/I6/687

表1 微卫星引物信息

参量 Parameter	品种 Species	JZL114	MiSaTPW01	JZL37	Mdo6	MiSaTPW117	JZL108
等位基因数 Number of alleles	加州原种 California M. salmoides	8	3	4	6	6	10
	优鲈1号 Youlu No. 1	5	3	3	5	3	4
	台湾加州鲈 Taiwan-California M. salmoides	6	2	3	4	3	4
	总计 Total	11	5	6	9	6	11
有效等位基因数 Effective number of alleles	加州原种 California M. salmoides	4.89	1.81	1.77	1.73	2.78	3.42
	优鲈1号 Youlu No. 1	3.15	1.27	1.18	2.87	2.04	1.60
	台湾加州鲈 Taiwan-California M. salmoides	4.05	1.30	1.14	2.19	2.60	1.59
	总计 Total	4.79	1.81	1.31	2.36	2.92	2.06
观察杂合度 Observed heterozygosity	加州原种 California M. salmoides	0.53	0.50	0.09	0.29	0.56	0.56
	优鲈1号 Youlu No. 1	0.79	0.08	0.12	0.73	0.54	0.31
	台湾加州鲈 Taiwan-California M. salmoides	1.00	0.00	0.10	0.80	0.53	0.30
	总计 Total	0.83	0.13	0.10	0.61	0.54	0.38
期望杂合度 Expected heterozygosity	加州原种 California M. salmoides	0.82	0.46	0.44	0.43	0.66	0.72
	优鲈1号 Youlu No. 1	0.70	0.21	0.15	0.66	0.52	0.38
	台湾加州鲈 Taiwan-California M. salmoides	0.77	0.24	0.13	0.55	0.63	0.38
	总计 Total	0.80	0.45	0.24	0.58	0.66	0.52
参量 Parameter	品种 Species	JZL85	MiSaTPW123	MiSaTPW165	MiSaTPW96		Mdo7
等位基因数 Number of alleles	加州原种 California M. salmoides	5	3	4	9		6
	优鲈1号 Youlu No. 1	5	3	3	5		4
	台湾加州鲈 Taiwan-California M. salmoides	5	2	4	4		3
	总计 Total	8	3	5	12		8
有效等位基因数 Effective number of alleles	加州原种 California M. salmoides	2.00	1.42	2.43	4.86		3.23
	优鲈1号 Youlu No. 1	2.43	1.69	2.28	2.67		1.46
	台湾加州鲈 Taiwan-California M. salmoides	2.45	1.76	1.87	2.36		1.57
	总计 Total	3.05	1.65	2.22	3.47		1.94
观察杂合度 Observed heterozygosity	加州原种 California M. salmoides	0.64	0.35	0.62	0.82		0.48
	优鲈1号 Youlu No. 1	0.48	0.18	0.58	0.79		0.29
	台湾加州鲈 Taiwan-California M. salmoides	0.43	0.43	0.43	0.83		0.43
	总计 Total	0.51	0.32	0.52	0.82		0.40
期望杂合度 Expected heterozygosity	加州原种 California M. salmoides	0.51	0.30	0.61	0.82		0.70
	优鲈1号 Youlu No. 1	0.60	0.42	0.57	0.64		0.32
	台湾加州鲈 Taiwan-California M. salmoides	0.60	0.44	0.47	0.59		0.37
	总计 Total	0.68	0.40	0.55	0.72		0.49

表2 大口黑鲈群体的遗传多样性检测结果

表3 大口黑鲈群体的中性测试分析

表4 微卫星位点的哈代-温伯格平衡检验

表6 IAM、TPM和SMM检验下大口黑鲈杂合度过量期望或缺失的基因座位数

表8 大口黑鲈群体11个微卫星位点的F统计量

表7 大口黑鲈群体的遗传多样性检测结果

表9 大口黑鲈群体的AMOVA结果

表10 大口黑鲈群体的遗传相似度和相对遗传距离

图1 大口黑鲈群体的聚类和遗传结构分析A.邻接聚类结果（图中的数字代表线的相对长度）；B. 大口黑鲈群体的遗传结构分析。

表11 大口黑鲈群体被误判的个体数

表5 大口黑鲈群体微卫星基因座间的连锁不平衡检验

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