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短蛸微卫星多重PCR体系建立及交配模式分析
刘文芬,冯艳微,王卫军,陈建强,杨建敏
1.上海海洋大学 水产科学国家级实验教学示范中心 水产动物遗传育种中心上海市协同创新中心 水产种质资源发掘与利用教育部重点实验室 上海 201306;2.山东省海洋资源与环境研究院 山东省海洋生态修复重点实验室 烟台 264006
摘要:
本研究对实验室开发的12对短蛸(Amphioctopus fangsiao)高多态性微卫星标记进行优化组合,构建了4组三重PCR体系,对短蛸4个母本(♀1、♀2、♀3和♀4)、104个子代及19个候选父本进行亲权鉴定,分析其交配模式。结果显示,构建的4组三重PCR体系可有效鉴定短蛸的亲权关系,104个子代鉴定率为94.23%。♀1组样品鉴定出12个子代,父本数为3个;♀2组样品鉴定出40个子代,父本数为7个;♀3组样品鉴定出19个子代,父本数为2个;♀4组样品鉴定出27个子代,父本数为4个;并且♂2、♂3、♂7和♂10均与2个以上的母本发生了交配。由此分析得出,短蛸交配模式存在多雌多雄关系。研究结果为高效开展短蛸人工繁育、增养殖及放流工作提供了重要参考。
关键词:  短蛸  微卫星多重PCR  交配模式
DOI:
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基金项目:
Development of Microsatellite Multiplex PCR Sets and Its Application in the Reproduction Model Analysis of Amphioctopus fangsiao
LIU Wenfen1,2,3,4,5, FENG Yanwei5, WANG Weijun5, CHEN Jianqiang5, YANG Jianmin5
1.ational Demonstration Center for Experimental Fisheries Science Education;2.Shanghai Collaborative Innovation for Aquatic Animal Genetics and Breeding;3.Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education;4.Shanghai Ocean University, Shanghai 201306;5.Shandong Marine Resource and Environment Research Institute, Shandong Provincial Key Laboratory of Marine Ecological Restoration, Yantai 264006
Abstract:
Amphioctopus fangsiao, a synonym of Octopus ocellatus in China, is one of the most important economic species in the northern coast of China. Owing to overfishing and ever-increasing market demand, the wild resources of A. fangsiao are decreasing significantly, and therefore artificial breeding is imperative. The present study on the reproduction model of A. fangsiao would provide basic data for artificial breeding and germplasm conservation of the species. In the present study, 4 groups of triplex PCR sets were established using 12 high polymorphic microsatellite markers developed in our laboratory. Through the identification of paternity of 4 female parents (♀1, ♀2, ♀3, and ♀4), 104 offspring and 19 candidate male parents of the reproduction model A. fangsiao were analyzed. The results showed that the four groups of microsatellite multiplex PCR sets worked effectively in the paternity relationship analysis, and the identification rate of 104 offspring was 94.23%. Twelve offspring and 3 males were identified in ♀1 group, and the number of offspring and males identified in the groups ♀2, ♀3, and ♀4 were 40 and 7; 19 and 2; and 27 and 4, respectively, and ♂2, ♂3, ♂7, and ♂10 mate with two or more females, which indicates that A. fangsiao is polygynandry. Polygynandry is an adaptive mechanism by aquatic animals to environmental changes and human fishing pressure. It can effectively improve the genetic diversity of alleles and bring about genetic variation among offspring. In addition, polyandry plays a positive role in maintaining the stability of the population structure of A. fangsiao, and in improving the reproductive efficiency and offspring quality. The offspring from four groups assigned to ♂2 were as high as 71.43% (10/14), 60.98% (25/41), 85.71% (18/21), and 75% (21/28), respectively, which was higher than that assigned to the other male parents. This indicates that most of the sperm involved in fertilization were from ♂2; thus, ♂2 takes advantage of sperm competition and cryptic female choice. The present study provides important data for artificial breeding and proliferation, and also for the release of A. fangsiao.
Key words:  Amphioctopus fangsiao  Microsatellite multiplex PCR  Paternity