| 摘要: |
| 采用酚-氯仿和试剂盒两种提取法对三角帆蚌怀珠群与非怀珠群各6 个个体进行基因组DNA 的提取, 然后在优化RAPD(随机扩增多态性DNA)检测条件基础上, 从80 个随机引物中筛选出12 个扩增较好且多态性强的引物进行RAPD 扩增, 产物通过水平板琼脂糖凝胶电泳和垂直板聚丙烯酰胺凝胶电泳两种方法进行检验并对DNA 的多态性进行分析。结果显示在怀珠群和非怀珠群检测到的位点数、多态位点比例、平均Shannon 多态性信息指数、平均Nei’s 基因多样性指数、群体内个体间平均遗传相似率和平均遗传距离分别为10
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| 关键词: 三角帆蚌, 人工育珠, 随机扩增多态性DNA(RAPD), 遗传多样性 |
| DOI：10.11693/hyhz200905018018 |
| 分类号: |
| 基金项目:湖南省杰出青年基金资助, 041002 号; 国家星火计划资助, 2006EA770019 号; 湖南省“十一五”重点建设学科“动物学”资助, 07-A-2 号 |
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| GENOMIC DNA EXTRACTION AND RAPD ANALYSIS ON HYRIOPSIS CUMINGII DISTINGUISHED BY PEARL-BREED |
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YANG Pin-Hong1, WANG Zhi-Tao2
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1.College of Life Science, Hunan University of Arts and Science, Hunan Province Marine Products Engineering Technique Research Center, Dahu Aquaculture Co., Ltd.;2.College of Life Science, Hunan University of Arts and Science
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| Abstract: |
| Random amplified polymorphic DNA (RAPD) technique was applied to assess the genetic diversity of two populations of Hyriopsis cumingii distinguished by pearl-breed, six individuals from each population were randomly selected, followed by genomic DNA extraction using the SDS-phenol-chloroform method and KIT method. Of 80 10-nucleotide-long random primers used in the preliminary analysis, 12 primers produced well-amplified and reproducible band patterns. They were screened and used for further analysis. The amplified products were electrophoretically analyzed through horizontal agarose gel and vertical polyacrylamide gel methods. The results show that the number of loci detected, percentage of polymorphic fragments, Shannon’s index of phenotype diversity, Nei’s index of genediversity, average genetic distance and similarity estimated by Lynch’s index are 100, 33%, 0.1927, 0.1324, 0.904, 0.096 in pearl-breed population, and 94, 47.87%, 0.2711, 0.1879, 0.861, 0.139 in non-pearl-breed population, respectively. The average inter-population genetic distance between this two populations is 0.821, showing that the gene variability in pearl-breed
population is greater than that in non-pearl-breed one. Specific bands and variable bands were observed between this two groups, and these bands may be the effective genes associated with pearl-growth. The molecular phylogenetic trees constructed by UPGMA and NJ methods with MEGA4.0 clearly reveal that the all pearl-breed population are clustered into one group, while those of non-pearl-breed, another group.
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| Key words: Hyriopsis cumingii, Artificial pearl-breading, Random amplified polymorphic DNA (RAPD), Genetic diversity |
