用RAPD技术对5种经济海胆基因组DNA多态性的研究  

Genomic DNA polymorphism in five species of sea urchin by RAPD

作　　者：常亚青[1] 丁君[1] 邢荣莲[1] 宋林生[2] 倪多娇[2] 

机构地区：[1]大连水产学院农业部海洋水产增养殖学与生物技术重点开放实验室,辽宁大连116023 [2]中国科学院海洋研究所实验海洋生物学开放实验室,山东青岛266071

出　　处：《中国水产科学》2004年第2期129-134,共6页Journal of Fishery Sciences of China

基　　金：国家"863"高技术研究发展计划项目(2002AA628170);国家教育部骨干教师资助计划(No.16) ~~

摘　　要：利用随机扩增多态DNA技术对紫海胆(Anthocidariscrassispina)、海刺猬(Glyptocidariscrenularis)、马粪海胆(Hemicentrotuspulcherrimus)、光棘球海胆(Strongylocentrotusnudus)和中间球海胆(Strongylocentrotusintermedius)等我国最主要的5种经济类海胆基因组DNA的多态性进行初步研究。经20个随机引物扩增,共得到377条扩增片段,片段长度为200～1600bp。根据片段的共享度计算出平均遗传距离并采用UPGMA和NJ两种聚类分析方法进行处理得到系统树。从聚类图可得知长海胆科与疣海胆科有较近的亲缘关系,球海胆科的光棘球海胆与中间球海胆有较近的亲缘关系,其次是马粪海胆。本研究还讨论了RAPD技术在海胆的遗传学研究中的广阔应用前景以及从分子生物学方法上为海胆传统分类方法提供科学依据。RAPD technique was applied to study the polymorphism of genomic DNA from five species of sea urchins in different families of the same order. Anthocidaris crassispina, Glyptocidaris crenularis, Hemicentrotus pulcherrimus, Strongylocentrotus nudus and Strongylocentrotus intermedius are most important edible sea urchin in China. In this experiment, the samples of A. crassispina were from coast line along Zhanjiang, Guangdong Provice, and the samples of G.crenularis, H.pulcherrimus, S.nudus and S. intermedius were all from the coast line along Dalian, Liaoning Province. The samples were wild except S.intermedius individuals which were the offspring of the parents introduced by Dalian Fisheries University in 1989. The amplification with 20 random primers produced 377 reproducible amplified fragments ranging from 200 bp to 1 600 bp. The amplified fragments were scored as present(1)or absent(0)for each DNA sample and an index of band sharing(F )was calculated by Nei and Li's matching coefficient method. The value of(1-F ) was used to evaluate genetic distances between species. The phylogenetic trees were constructed with the methods of UPGMA and NJ on the basis of genetic distances. The results of the two methods of cluster analysis are in agreement, and the relationship expressed by the RAPD analysis revealed the difference between families, genus and species. S. nudus and S. intermedius shared the closest phylogenic relationship in the phylogenetic trees, and the result was in good agreement with the concept from Chinese scholar about the taxonomy in sea urchin. The prospect of RAPD application in the field of genetics and taxonomy of sea urchin is discussed in this paper.

关 键 词：海胆 基因组DNA RAPD 亲缘关系 

分 类 号：Q959.268[生物学—动物学]