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进口大菱鲆Scophthalmus maximus L.苗种的遗传结构分析
申雪艳1,2, 宫庆礼1, 雷霁霖2, 孔 杰2, 翟介明2, 李 波3
1.中国海洋大学生命科学与技术学部;2.农业部海洋渔业资源可持续利用重点开放实验室,中国水产科学研究院黄海水产研究所;3.莱州明波水产有限公司
摘要:
利用随机扩增多态性DNA(RAPD)和微卫星两种分子标记技术分析了从法国(F)、英国(E)、西班牙(S)引进我国的三个大菱鲆群体的遗传结构。20个RAPD随机引物对每个群体各20尾大菱鲆的基因组DNA进行扩增,共获得125个重复性好且谱带清晰的RAPD标记,片段大小在200-2500bp之间,三群体的多态位点比例在12.80%—20.00% 之间,Nei基因多样性指数在0.0142—0.0352之间,Shannon多样性指数在0.0423—0.0720之间。微卫星引物的分析结果与RAPD一致。总体上,三个群体的遗传多样性均较低,其中西班牙群体遗传多样性水平最高,英国群体次之,而法国群体最低。利用Shannon多样性指数和Nei多样性指数估算群体遗传变异的来源,两者得出一致的结论:群体问遗传分化很弱。AMOVA分析结果进一步证实了Shannon多样性指数和Nei基因多样性指数的分析结果:群体的遗传变异有97% 以上是由群体内个体问的遗传变异引起的,遗传变异主要来自于群体内个体间,显著性检验表明群体间的变异对总变异的影响不显著。实验结果证明我国进口大菱鲆群体种质较单一。
关键词:  大菱鲆,种群,RAPD,微卫星,遗传变异
DOI:
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基金项目:国家“863”计划资助项目,2001AA620207号
POPULATION GENETIC STRUCTURE ANALYSIS OF THE IMPORTED TURBOT SEEDLINGS SCOPHTHALMUS MAXIMUS L.USING RAPD AND MICROSATELLITE TECHNIQUE
SHEN Xue-Yan1,2, GONG Qing-Li1, LEI Ji-Lin2, KONG Jie2, ZHAI Jie-Ming2, LI Bo3
1.College of Marine Life Sciences and Technology,Ocean University of China;2.Key Lab of Sustainable Utilization of Marine Fisheries Resources,Ministry of Agriculture,Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences;3.Laizhoa Mingbo Aquatic Pr0ducts Co., Ltd.
Abstract:
Random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) molecular markers were used to detect genetic diversity, genetic structure and genetic divergency among three populations of Scophthalmus maximus L. imported from England, France and Spain(June 2002). Genetic DNA was extracted from muscle of the 60 S. maximus L. Inter-stock genetic similarity and intra-stock genetic similarity was then analyzed. Thirty random primers were used for analysis of nuclear DNA polymorphism of the three populations. Results showed that twenty primers were effective. 20 RAPD primers together produced 125 reproducible bands ranging from 200bp to 2500bp. The mean percentage of polymorphic loci of the three populations ranged from 12. 80% — 20.00%, while Nei’s gene diversity was between 0.0142 and 0.0352. The average Shannon’s indices of population phenotype ranged from 0.0423 to 0.0720. All these polymorphic indices revealed genetic diversity between the populations differing from each other. Shannon indices suggested that differentiation of S. maximus L. were divided into two parts, 1.05% from inter-populations and 98.95% from intra-populations. Hierarchy analysis of the population verified Shannon indices, 98. 8 1% of the variance was from intra-populations. Concurrently, results of Analysis of Molecular Variation(AMOVA)revealed that the variance of inter-population hadn’t significantly affected the total variance of S. maximus L. AMOVA (Analysis of Molecular Variation)analysis verified Shannon indices and Nei’s gene diversity, over 97% of the variance was from intra-populations. At the same time, results of AMOVA revealed that the variance of inter-populations did not affect the total variance of S. maximus L. Results of SSR analysis including mean percentage of polymorphic loci, Nei’s gene diversity and average Shannon’s indices of phenotypic were all consistent with the results of RAPD revealing low genetic diversity of S. maximus L. populations. The lowest genetic diversity was in French population while the highest was in Spanish. Genetic distance of the three populations by RAPD analysis was 0.0124 to 0.0192 consistent with results of SSR. By comparing the results of RAPD and those of SSR, SSR was found to be more sensitive than RAPD in revealing polymorphism. SSR can reflect a higher level of genetic diversity than RAPD. In terms genetic similarity and distance, differences were seen between the results obtained from each method. Genetic diversity revealed by SSR was higher than that obtained by RAPD. As a whole, the results for population genetic structure detected by RAPD and SSR were consistent, yet a slight difference appeared with in intra-population genetic diversity, mainly due to differences in genetic variance detection abilities of the two techniques. All results here suggest that genetic diversity of imported turbot populations was very low. Thus, measures must be taken to avoid inbreeding depression of S. maximus L. in domestic populations in China.
Key words:  Turbot Scophthalmus maximus L., Population, RAPD, SSR, Genetic diversity “
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