五种长臂虾遗传多样性和亲缘关系的RAPD分析
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摘要
本文应用随机扩增多态DNA(RAPD)技术,结合一定的形态学研究手段,对五种长臂虾的遗传多样性和亲缘关系进行了分析。
     从40个随机引物中筛选出33个对两种越南古洁虾进行RAPD扩增,共扩增出304个DNA片段。在越南古洁虾指名亚种和越南古洁虾广东亚种中分别扩增出279和296个DNA片段,其中多态性片段数分别为184和191,多态性片段的比例(P)分别为65.9%和64.5%。越南古洁虾指名亚种和越南古洁虾广东亚种的遗传多态度(H_o)分别为0.163和0.129,两种群的遗传变异H_(pop)/H_(sp)均值为0.942,而种群间的遗传变异(H_(sp)-H_(pop))/H_(sp)均值为0.058。未获得可区分两种越南古洁虾的RAPD标记。通过DPS(3.11专业版)种群遗传分析软件包、采用UPGMA方法分析,其聚类结果为广东亚种12个个体先相聚,然后与指名亚种的2个个体相聚,最后才与指名亚种的另4个个体相聚,这与形态学分析的结果不一致。两种群的遗传相似度(S)高达0.943,遗传距离(D)仅为0.057。
     从40个随机引物中筛选出31个对三种沼虾进行RAPD扩增,共扩增出450个DNA片段。在胖掌沼虾、海南沼虾和台湾沼虾中分别扩增出353、316和290个DNA片段,其中多态性片段数分别为228、147和132,多态性片段的比例(P)分别为64.6%、46.5%和45.5%。胖掌沼虾、海南沼虾和台湾沼虾的遗传多态度(H_o)分别为0.211、0.135和0.142,种群内的遗传变异H_(pop)/H_(sp)均值为0.682,而种群间的遗传变异(H_(sp)-H_(pop))/H_(sp)均值为0.318。获得33个RAPD标记,其片段长度在259-3333bp之间.通过DPS(3.11专业版)种群遗传分析软件包、采用UPGMA方法分析,其聚类结果为各种的个体先分别相聚,然后海南沼虾与台湾沼虾相聚,最后与胖掌沼虾相聚,这与形态学分析的结果一致。海南沼虾与台湾沼虾、海南沼虾与胖掌沼虾、台湾沼虾与胖掌沼虾的遗传相似度(S)分别为0.845、0.694和0.706,遗传距离(D)分别为0.155、0.306和0.294。
In this paper,random amplified polymorphic DNA (RAPD)technique,together with the method of morphology,was employed in analyzing the genetic diversity and the genetic relationship among five species of important Palaemoninae prawns.
    In this experiment,33 out of 40 random primers (10 bp) were selected to carry out the analysis of RAPD and 304 DNA fragments were amplified among two species of Genus Coutierella.The number of DNA fragments amplified of Coutierella tonkinensis tonkinensis and Coutierella tontdnensis guangdongensis was 279 and 296, respectively,and 184 and 191 of them were polymorphic,respectively,amounted to 65.9% and 64.5% respectively.The genetic diversity index was 0.163 and 0.129, respectively. Interpopulation and intrapopulation average genetic variation of two species Coutierella tonkinensis was 0.942 and 0.058,respectively.There was no RAPD marker amplified by 33 primers. The cluster analysis of the above two species was performed with UPGMA algorith by using DPS(3.11 professional version)software package.The result is not as same as the result of morphologic analysis.The genetic similarity index and the genetic distance among the two species of Coutierella tonkinensis prawns was 0.943 and 0.057,respectively.
    31 out of 40 random primers(10 bp) were selected to carry out the analysis of RAPD and 450 DNA fragments were amplified among three species of Genus Macrobrachium.The number of DNA fragments amplified of M.inflatum, M.hainanense and M.formosense was 353,316 and 270,respectively,and 228,147 and 61 of them were polymorphic,respectively,amounted to 64.6%,46.5% and 22.6%,respectively.The genetic diversity index was 0.211,0.135 and 0.142,respectively.Interpopulation and intrapopulation average genetic variation of the three species was 0.682 and 0.318,respectively.33 amplified fragments were distinctive among the three species with the molecular weight varying from 259 bp to 3333 bp. The cluster analysis of the above three species was performed with UPGMA algorith by using DPS(3.11 professional version)software package. The individuals of the same species clustered together before the different species did.The result was as same as the result of morphologic analysis. The genetic similarity index among M.hainanense and M.formosense,M.hainanense and M.inflatum, M.formosense and M.inflatum was 0.845,0.694 and 0.706,respectively.The genetic distance of them was 0.155,0.306 and 0.294,respectively
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