浙江农业学报 ›› 2019, Vol. 31 ›› Issue (8): 1257-1271.DOI: 10.3969/j.issn.1004-1524.2019.08.06

• 动物科学 • 上一篇    下一篇

连续三代减数分裂雌核发育团头鲂的遗传多样性分析和RAPD鉴别方法的建立

唐首杰1,2,3, 毕详4, 张飞明4, 张友良4   

  1. 1.上海海洋大学 农业农村部淡水水产种质资源重点实验室,上海 201306;
    2.上海海洋大学 水产科学国家级实验教学示范中心,上海 201306;
    3.上海海洋大学 水产动物遗传育种中心上海市协同创新中心,上海 201306;
    4.上海市松江区水产良种场,上海 201616
  • 收稿日期:2019-01-31 出版日期:2019-08-25 发布日期:2019-08-30
  • 作者简介:唐首杰(1981-), 男, 江苏常州人, 博士, 讲师, 主要从事水产动物种质资源与种苗工程研究。E-mail: sjtang@shou.edu.cn
  • 基金资助:
    上海市种业发展项目(沪农科种字〔2017〕第1-9号); 上海市科技兴农重点攻关项目(沪农科攻字〔2013〕第2-3号); 国家自然科学基金(30630051)

Genetic diversity analysis and RAPD identification of three successive generations of meio-gynogenetic population in Megalobrama amblycephala

TANG Shoujie1,2,3, BI Xiang4, ZHANG Feiming4, ZHANG Youliang4   

  1. 1. Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai 201306, China;
    2. National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China;
    3. Shanghai Collaborative Innovation for Aquatic Animal Genetics and Breeding, Shanghai Ocean University, Shanghai 201306, China;
    4. Songjiang Aquatic Seed Breeding Farm, Shanghai 201616, China
  • Received:2019-01-31 Online:2019-08-25 Published:2019-08-30

摘要: 为评估连续三代减数分裂雌核发育团头鲂群体的遗传多样性和遗传纯合度,寻找区分不同团头鲂育种群体(团头鲂浦江1号选育系、连续三代减数分裂雌核发育团头鲂群体)的稳定的分子遗传标记,本研究以团头鲂(Megalobrama amblycephala)浦江1号选育系F9群体为对照组,利用39条多态性RAPD随机引物比较分析了团头鲂人工减数分裂雌核发育一代群体(G1)、二代群体(G2)和三代群体(G3)的遗传多样性和遗传结构,获得了用于鉴别不同团头鲂育种群体(F9、G1、G2、G3)的稳定的RAPD分子遗传标记,探讨了连续多代诱导减数分裂雌核发育对团头鲂基因纯化的效果。结果显示,39条RAPD随机引物在F9、G1、G2和G3群体中扩增条带总数分别为213条、202条、200条和190条,F9、G1、G2和G3群体的多态位点比例分别为36.15%、35.64%、27.00%和26.84%,F9、G1、G2和G3群体的Shannon信息指数分别为0.207 9、0.185 7、0.146 1和0.138 3。3个雌核发育群体的遗传多样性水平(多态位点比例、Shannon信息指数)均明显低于对照组F9群体,随着雌核发育世代数的增加,遗传多样性水平呈现逐代降低的趋势,即G1>G2>G3。4个群体的群体内个体间的平均遗传相似系数为0.828 5~0.906 0,3个雌核发育群体的群体内个体间平均遗传相似系数均明显高于对照组F9群体;群体内个体间的平均遗传相似系数呈现随雌核发育世代数的增加而升高的趋势,即G3>G2>G1。群体间成对FST值为0.269 2~0.419 5,经置换检验得到的FST值的P值为0.000 0~0.009 0,均达到极显著水平(P<0.01),表明4个群体间存在极显著的遗传分化。有5条随机引物在群体间产生了特异DNA片段,其中,4条随机引物(S3、S40、S58和S75)可用于区分G3群体和其他3个群体(F9、G1和G2),引物S3的鉴别可靠性最高;仅1条随机引物(S71)能用于区分G2群体和其他3个群体(F9、G1和G3)。本研究结果表明,连续多代的人工减数分裂雌核发育诱导已对团头鲂育种群体产生以下两方面的影响:一方面,遗传多样性明显降低,并呈现逐代降低的趋势;另一方面,遗传纯度明显升高,并呈现逐代升高的趋势。连续多代减数分裂雌核发育能显著加快团头鲂基因的纯合速度,雌核发育三代群体(G3)已经是一个遗传一致性较高的高纯品系。

关键词: 团头鲂, 雌核发育, 遗传多样性, RAPD标记

Abstract: To assess the genetic diversity and genetic homozygosity of the three generations of meiotic gynogenetic populations of blunt snout bream (Megalobrama amblycephala), and to find a stable molecular genetic marker that distinguishes different breeding populations of M. amblycephala(Pujiang No.1 genetically selected strains F9, three consecutive generations of meiotic gynogenetic populations), genetic diversity and genetic structure of meiotic gynogenetic populations meio-G1 (the first generation), meio-G2 (the second generation), meio-G3 (the third generation) and the control group (Pujiang No.1 genetically selected strains F9) of M. amblycephala was analysed using thirty-nine RAPD markers in this study. Several stable RAPD markers for distinguishing different breeding populations were obtained. And the efficiency to pure gene for successive artificial meiotic gynogenesis in M.amblycephala was assessed. The results showed that: the number of loci detected in the control group, meio-G1, meio-G2 and meio-G3 were 213, 202, 200 and 190, respectively. The percentage of polymorphic loci detected in the control group, meio-G1, meio-G2 and meio-G3 were 36.15%, 35.64%, 27.00% and 26.84%, respectively. Shannon's information index estimated within the control group, meio-G1, meio-G2 and meio-G3 were 0.207 9, 0.185 7, 0.146 1 and 0.138 3, respectively. The genetic diversity of meio-G1, meio-G2 and meio-G3 were much lower than that in the control group.The genetic diversity of three consecutive generations of meiotic gynogenetic populations decreased with the increasing of gynogenetic generations. Genetic identity between individuals within each population ranged from 0.828 5 to 0.906 0. The genetic identity between individuals within populations of meio-G1, meio-G2 and meio-G3 were much higher than that in the control group. The genetic identity between individuals within three consecutive generations of meiotic gynogenetic populations increased with the increasing of gynogenetic generations. Genetic differentiation index (pairwise FST values) between four populations ranged from 0.269 2 to 0.419 5. And the probabilities of the permutation test of genetic differentiation between four populations ranged from 0.000 0 to 0.009 0, indicating extremely significant genetic differentiation between four populations. Five primers (S3, S40, S58, S71 and S75) were observed to produce specific bands for discriminating different populations. Of these primers, four primers (S3, S40, S58 and S75) could be used as molecular markers for distinguish between meio-G3 and the other three populations (F9, meio-G1 and meio-G2). Primer S3 has the highest reliability for population identification. Only one primer (S71) could be used as molecular markers for distinguish between meio-G2 and the other three populations (F9, meio-G1 and meio-G3).The results of this study indicated that the continuous generations of artificial meiosis gynogenesis had affected the breeding populations of M. amblycephala in the following two aspects: on one hand, the genetic diversity significantly reduced with the increasing of gynogenetic generations. On the other hand, the genetic purity increased significantly with the increasing of gynogenetic generations. Continuous multi-generation meiotic gynogenesis could significantly accelerate the homozygous rate of the gene, and the meio-G3 population was already a high-purity line with high genetic consistency.

Key words: Megalobrama amblycephala, gynogenesis, genetic diversity, RAPD marker

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