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利用高通量测序开发的熊本牡蛎微卫星标记评价野生和养殖群体的遗传多样性及通用性
引用本文:黄飘逸,马海涛,喻子牛,张跃环,高红梅,彭建军.利用高通量测序开发的熊本牡蛎微卫星标记评价野生和养殖群体的遗传多样性及通用性[J].水产学报,2020,44(3):368-377.
作者姓名:黄飘逸  马海涛  喻子牛  张跃环  高红梅  彭建军
作者单位:重庆师范大学生命科学学院,重庆 401331;中国科学院南海海洋研究所,广东广州 510220
基金项目:国家自然科学基金(31702340,31470570);重庆市科委自然科学基金(cstc2014jcyjA80013);重庆市教育委员会自然科学基金;(kj1400534);广东省自然科学基金(2017A030310442)
摘    要:利用高通量测序的方法,从熊本牡蛎基因组中开发了20对具有多态性的微卫星标记,通过微卫星标记位点比较了野生群体和养殖群体的遗传多样性。野生群体中,所有位点共扩增出330个等位基因,等位基因数(N_a)范围为6~39,平均等位基因数为16.500 0;有效等位基因数(N_e)范围为1.352 9~33.361 7,平均值9.517 2;观测杂合度(H_o)范围为0.200 0~1.000 0,平均值0.671 5;期望杂合度(H_e)范围为0.265 6~0.987 7,平均值0.832 1;ShannonWeiner指数(Ⅰ)范围为0.648 3~3.585 8,平均值2.276 9;多态信息含量(PIC)范围为0.254 5~0.969 2,平均值0.803 5,共有16个位点符合Hardy-Weinberg平衡。养殖群体中,N_a平均值为10.250 0,N_e平均值为5.843 4,H_o平均值为0.639 1,H_e平均值为0.763 6,I平均值为1.791 4,PIC平均值为0.720 7。结果显示,熊本牡蛎养殖群体的遗传多样性低于野生群体,但仍然维持在高度多态水平。研究表明,在熊本牡蛎人工繁育过程中,使用大数量的亲本进行繁育,可有效防止选育群体的遗传多样性降低,但人工选育对选育群体的遗传多样性也产生了一定的影响。另外,分析了这些引物在近缘种葡萄牙牡蛎、长牡蛎、香港牡蛎、有明牡蛎、僧帽牡蛎、咬齿牡蛎以及舌骨牡蛎中的通用性情况,发现XB1-6、XB1-39和XB1-45 3个位点在8个物种中均能扩增出目的条带,XB1-41仅能在熊本牡蛎中扩增出目的条带。

关 键 词:熊本牡蛎  微卫星标记  野生群体  养殖群体  通用性
收稿时间:2019/3/9 0:00:00
修稿时间:2019/6/6 0:00:00

Evaluation of genetic diversity and versatility of wild and cultured populations based on high-throughput sequencing of Crassostrea sikamea microsatellite markers
HUANG Piaoyi,MA Haitao,YU Ziniu,ZHANG Yuehuan,GAO Hongmei and PENG Jianjun.Evaluation of genetic diversity and versatility of wild and cultured populations based on high-throughput sequencing of Crassostrea sikamea microsatellite markers[J].Journal of Fisheries of China,2020,44(3):368-377.
Authors:HUANG Piaoyi  MA Haitao  YU Ziniu  ZHANG Yuehuan  GAO Hongmei and PENG Jianjun
Institution:College of Life Sciences of Chongqing Nonmal Univercity, Chongqing 401331, China,South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510220, China,South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510220, China,South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510220, China,College of Life Sciences of Chongqing Nonmal Univercity, Chongqing 401331, China and College of Life Sciences of Chongqing Nonmal Univercity, Chongqing 401331, China
Abstract:Using high-throughput sequencing,we characterized 20 pairs of polymorphic microsatellite primers from Crassostrea sikamea genome, and we examined the markers in a wild population. A total of 330 alleles were found in 25 microsatellites.The observed number of alleles (Na) ranged 6-39 in average of 16.500 0,and the effective number of alleles(Ne) ranged 1.352 9-33.361 7. The observed and expected heterozygosity values range 0.200 0-1.000 0 and 0.265 6-0.987 7, respectively.The Shannon Weiner index ranged 0.648 3-3.585 8 and Polymorphic information content (PIC) ranged 0.254 5-0.969 2. 16 microsatellite markers accord with Hardy-Weinberg equilibrium. Using these loci to analyze the genetic diversity of a cultured population, the average number of alleles was 10.25; the average number of Ne was 5.843 4. The observed heterozygosity (Ho) average was 0.639 1; the expected heterozygosity (He) average was 0.763 6; the Shannon-wiener index (I) average was 1.791 4; and the polymorphic information content (PIC) average was 0.720 7. It was found that the genetic diversity of C. sikamea cultured population was lower than that of the wild population, but it maintained a high-genetic diversity. The results of this study indicate that in the artificial breeding process of C. sikamea, the use of a large number of parents for breeding can effectively prevent the decline of genetic diversity of the cultured population, but artificial breeding also has a certain impact on the genetic diversity of the cultured population. The 20 loci were surveyed the cross-species proportions in C. angulate, C. gigas, C. hongkongensis, C. ariakensis, Saccostrea cucullata, Ostrea mordax and Hyotissa hyotis. Loci XB1-6, XB1-39, and XB1-45 can amplify the target band in 8 species, and the XB1-41 can only amplify the target band in C. sikamea.
Keywords:Crassostrea sikamea  microsatellite markers  wild population  cultured population  universality
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