| 基于AFLP技术的海南三亚野生和养殖牡蛎遗传多样性研究 |
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| 引用本文： | 高晓辉,王春生,吴月红,寿鹿,陈全震.基于AFLP技术的海南三亚野生和养殖牡蛎遗传多样性研究[J].海洋学研究,2012,30(1):1-10. |
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| 作者姓名： | 高晓辉 王春生 吴月红 寿鹿 陈全震 |
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| 作者单位： | 国家海洋局第二海洋研究所,国家海洋局海洋生态系统与生物地球化学重点实验室,浙江杭州310012 |
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| 基金项目： | “908”专项资助项目(908-ZC-I-02) |
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| 摘 要： | 2008年夏季,从海南三亚分别采集26份养殖牡蛎和32份野生牡蛎样品(1份样品代表1个体),开展扩增片断长度多态性(AFLP)和数量性状分析,采用POPGENE 32软件计算多态性位点百分率、遗传多样性指数、Hardy-Weinberg遗传偏离指数、群体遗传相似度和遗传距离。基于AFLP分析获得的遗传距离和线粒体细胞色素氧化酶I亚基(CO I)基因序列,采用邻接法构建养殖和野生牡蛎的系统发育树。结果表明,选择性引物对牡蛎品种具有较好的鉴别效率;相比养殖牡蛎,野生牡蛎多样性丰富、品种间同源性较低、亲缘关系相差较远、遗传基础较宽;通过CO I基因序列分析可知,养殖牡蛎均为香港巨牡蛎Crassostrea hongkongensis,而野生牡蛎属于囊牡蛎属Saccostrea,主要为僧帽牡蛎Saccostrea cucullata。目前海南三亚牡蛎种质资源多样性相对丰富,引种还未对三亚自然环境的牡蛎资源造成明显影响。
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| 关 键 词： | 牡蛎 扩增片断长度多态性 遗传多样性 COI基因 海南 三亚 |
AFLP analysis of genetic diversity of the wild and cultured oyster populations in Sanya, Hainan province |
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| GAO Xiao-hui
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WANG Chun-sheng
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WU Yue-hong
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SHOU Lu
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CHEN Quan-zhen.AFLP analysis of genetic diversity of the wild and cultured oyster populations in Sanya, Hainan province[J].Journal of Marine Sciences,2012,30(1):1-10. |
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| Authors: | GAO Xiao-hui WANG Chun-sheng WU Yue-hong SHOU Lu CHEN Quan-zhen |
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| Institution: | (Second Institute of Oceanography,SOA,Laboratory of Marine Ecosystem and Biogeochemistry,SOA,Hangzhou 310012,China) |
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| Abstract: | 32 wild and 26 cultured oysters were collected from Sanya,Hainan province in June 2008.Genetic diversity of the two sampled populations were studied with Amplified Fragment Length Polymorphism(AFLP) method,and statistical analysis were made on the AFLP electrophoretogram with 1 representing the presence of the AFLP bands and 0 representing the absence of AFLP bands to transfer the AFLP electrophoretogram into 0/1 binary character matrix.The percentage of polymorphic loci,number of effective alleles,the Shannon’s genetic diversity index,the Hardy-Weinberg deviation index,as well as Nei’ genetic identity and genetic distance were analyzed with the population analysis software POPGENE 32(version 1.31).Moreover,cytochrome oxidase subunit I(CO I) gene fragments of 35 samples were amplified for both wild and cultured populations and sequenced.The CO I gene sequence was read by Chromas software,complemented with annual correction,and sequence similarity analysis was made with BLASTn program and GenBank entry sequence.Clustalw 1.8 software package was used to make multi-sequence matching sequencing,in which the gaps formed were filled with neutral elements.Based on AFLP genetic distance and CO I gene sequence,dendrograms of the two oyster populations were constructed with MEGA 3.1 software and Neighbor-jointing method.The results show that the selective primer has a good efficiency for identifying oyster species.Two selective primers were used for the PCR amplification of both wild and cultured oysters and loci from 150 bp to 700 bp were taken for calculation.A total of 65 loci from the cultured oyster and 149 loci from the wild oyster were detected,in which 61 and 149 were polymorphic loci which account for 93.85% and 100% of the total respectively.The number of effective alleles,polymorphism information content of the marked polymorphic loci and Shannon’s genetic diversity index were 1.247 7 and 1.305 3,0.165 5 and 0.212 2,0.278 6 and 0.355 4 respectively,which shows that the genetic diversity of the wild population is higher than that of the cultured population.Nei’s genetic identity and genetic distance vary from 0.600 to 0.908,0.097 to 0.511 for the cultured population,from 0.503 to 0.879,0.129 to 0.687 for the wild population,which shows that the homology of the cultured species is high and the genetic base is narrow,while that of the wild species is relatively low and wide.The NJ tree based on AFLP data roughly shows that the pedigree of the cultured oysters is more concentrated while that of the wild oysters more differentiated,which indicates closer relationships between cultured individuals and more distant relationships between the wild individuals.Totally 35 CO I gene fragments(20 from the cultured oysters and 15 from the wild ones) were amplified via PCR,and about 620 bp nucleotide sequences were retrieved after the primer sequence removed.Homologous comparison shows small sequence difference in the groups,5 within the cultured population(4 transition and 1 transversion sites),11 within the wild population(8 transition and 3 transversion sites).There is significant sequence difference(147 variation loci) between the two groups.The NJ tree based on CO I gene sequences and comparison with known GeneBamk sequence demonstrates that the two populations have a quite distant phylogenetic relationship,and that the cultured oysters are Crassostrea hongkongensis,while the wild oysters are mainly Saccostrea cucullata.The results reveal that introduced species have not had distinct impacts on the oyster resources in Sanya.However,if the fact that after a long time cultivation,inbreeding makes the low level genetic diversity resulted from bottleneck effect further lowered will influence the yield and quality needs to be tracked for long time.The information of genetic diversities of different populations will give us theoretical guidance in breeding and genetic improvement. |
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| Keywords: | oyster AFLP genetic diversity CO I gene Sanya Hainan |
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