【作者】 贺顺连；

【导师】 肖克宇； 郭照良；

【作者基本信息】 湖南农业大学 ， 养殖， 2003， 硕士

【摘要】 本文应用随机扩增多态DNA(RAPD)技术对黄鳝属(Monopterus)中产自缅甸的山黄鳝(M.cuchia)，印度尼两亚的穴黄鳝(M．fossorius)，及中国的黄鳝(M．albus)，共12个个体基因组DNA的遗传标记进行分析。先用12个个体的混合DNA对OPA、OPB和OPD三个系列40个随机引物进行筛选，有35个引物获得了扩增谱带，30个引物获得了重复性良好的谱带。本试验选用筛选出的30个引物分别对三种黄鳝的12个个体核DNA进行PCR扩增，其扩增产物经琼脂糖凝胶电泳分离后，共获得315个扩增片段，长度为250～5417bp，具有良好的重复性。每个引物在种间均能扩增出DNA多态性片段，每条扩增条带含有2～16个大小不等的片段。30个引物均可将1种、2种甚至3种黄鳝分别区别开来。检出三种黄鳝特异性片段87个，其中M．cuchia有25个，M．fossorius有33个，M．albus有29个。 从所检测到的位点中，三种黄鳝种群间的多态位点比例(P)为80.32％，遗传多样性指数(H)为0.1575；种群内的多态位点比例和遗传多样性指数在M．albus中为33.53％、0.0801；在M．cuchia中为48.61％、0.0645，在M．fossorius中为44.88％、0.1380。结果表明，种群间和种群内都具有较高的遗传变异，种群间的遗传变异高于种群内的。 通过DPS2003种群遗传分析软件包计算显示：M．cuchia个体间遗传距离(D)最小为0.4505、最大为0.8356，平均为0.5842：M．fossorius个体间遗传距离最小为0.1400、最大为0.9048，平均为0.5250；M．albus个体间遗传距离最小为0.2500、最大为0.7647，平均为0.5293。M．albus与M．fossorius的遗传距离最大，为0.7415；M．cuchia与M．fossorius的遗传距离次之，为0.7035：M．cuchia与M．albus的遗传距离最小，为0.6860。根据遗传距离指数，用UPGMA方法进行聚类分析，构建了三种黄鳝亲缘关系聚类图。图中所示三种黄鳝中M．cuchia与M．albus亲缘关系最近，M．fossorius与其余两种相对要远。更多还原

【Abstract】 The samples of three species of synbranchi form eels, Monopterus cuchia (Hamilton, 1822), M fossorius (Nayar, 1952) and M. albus (Zuiew,793) were collected from Yangon of Myanmar, Indonesia and YuanJiang County, Hunan Province,China respectively. Randomly amplified polymorphic DNA (RAPD) technique is usded to analyse the specific genetic markers and their genefic diversity of the wild grouper population of four individuals from each species.The reactions were performed through 94 #, pre-denaturing for 5 min, then 45 cycles of 94 # denaturing for 1 min, 36# annealing for 1 min and 72# polymerizing for 2 min; The complete extention was conducted at 72 # for 7 min. 40 primers of OPA, OPB and OPD were screened, 30 primers produced reproducible fingerprints with the molecular weight varying from 250 bp to 5417 bp. The number of RAPD markers amplified by each primer varied from 2 to 16. Those interpretable patterns generated by 30 primers were used as the genetic markers to discriminate the differentiation of the three species. 87 out of 315 amplified fragments were distinctive for the three species, 33 were from M. fossorius, 25 and 29 were from M. cuchia and M. albus respectively. The percentage of polymorphic ioci(P) among inter population of three species is 80.32%. The genetic diversity index (H) among inter population of three species is 0.157 5. Based on the RAPD data, the genetic distances among these three species of Monopterus, were computed by using DPS2003 software package. As result, the biggest genetic distance occurres between M. fossorius and M.albu with a value of 0.7415, while the smallest genetic distance with a value of 0.6860 comes about between M. cuchia and M. albus, and the genetic distance between M. cuchia and M. fossorius is 0.7035. The cluster analysis of the above three species was performed with UPGMA algorithm over their genetic distances. The dendrogram of these three species was constructed.更多还原