基于RAPD和RFLP的淡水泥鳅遗传多样性评价

Md. Faruque Miah, S. Anwar, Md. Hazrat Ali, M. NiamulNaser, Kawser Ahmed
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引用次数: 1

摘要

背景:库奇鳝(Monopterus cuchia)是孟加拉国一种重要的经济鳗鱼,由于绝大多数鱼种来自自然栖息地,人工养殖尚未建立,因此正面临种群减少和种质退化的挑战。材料与方法:采用8种十聚体引物对孟加拉东北部自然种群30只褐毛鼠进行分析,用2种限制性内切酶对谷氨酰胺合成酶基因进行酶切。结果:经RAPD检测,30个个体共检测到735条条带,多态性位点228个,所有引物多态性均为100%。利用双相似性指数计算个体间的遗传距离,共测得36组遗传多样性,最低遗传距离为0.58,最高遗传距离为0.97,平均遗传多样性为0.81。Nei’s遗传相似值为0.06 ~ 0.6,平均为0.301746,遗传距离显著。UPGMA聚类结果显示,11个进化支共产生6个聚类,涉及22个个体,而其余样本则与具有特定连接距离的进化支有关联。利用+CfrI和+Hpy178III两种限制性内切酶分析了谷氨酰胺合成酶基因的遗传多样性,这两种限制性内切酶均能酶切541 bp的基因片段,并检测了野生型纯合子、多态性纯合子和杂合子的多态性。利用RFLP波段分析观察了谷氨酰胺合成酶基因的遗传多样性,鉴定出4个不同群体,p值分别为0、0.033、0.05和1,但群体间存在距离差异,表明实验个体间存在多态性。结论:RAPD和RFLP分析表明,该鱼在实验生态生境中具有丰富的遗传多样性,这将有助于种质资源多样性的保护,并为该鱼的可持续繁殖计划提供支持。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Assessment of Genetic Diversity of Freshwater Mud Eel (Monopterus cuchia) Using RAPD and RFLP Markers
Background: Monopterus cuchia, an economically important eel of Bangladesh, is confronting the challenge of population reduction and germplasm degeneration since the vast majority of fingerlings are collected from natural habitats and artificial cultivation of the fish is yet to be established. Materials and Methods: 30 individuals of M. cuchia from a natural population of Northern-East part of Bangladesh were analyzed using 8 decamer primers and glutamine synthetase gene was digested with 2 restriction enzymes. Results: A total of 735 bands with 228 polymorphic loci were detected among the selected 30 individuals by using RAPD assay while 100% polymorphism was revealed by all the primers. The genetic distance among the individuals was calculated by using the data from pair-wise similarity index where 36 groups of genetic diversities were measured while the lowest and highest genetic distance were found 0.58 and 0.97 respectively with an average genetic diversity of 0.81. The Nei’s genetic similarity values were found from 0.06 to 0.6 where average value was recorded 0.301746 which was found significant regarding genetic distance. Phylogenetic relationships using UPGMA clustering revealed linkage distance ranged from 3.6 to 6.24 and generated 6 clusters by 11 clades with the involvement of 22 individuals while the rest of the samples were connected to those clades with specific linkage distances. Genetic diversity of glutamine synthetase gene was analyzed with two restriction enzymes, e.g. +CfrI and +Hpy178III while both the enzymes digested the gene fragment at a length of 541 bp and polymorphism was detected in terms of wild type homogygotes, polymorphic homozygote and heterozygosity. The genetic diversity was observed by using the RFLP band analysis and four different groups of individuals were identified with the p-values of 0, 0.033, 0.05 and 1 respectively whereas different distances were found among the groups indicating the polymorphism among experimental individuals of glutamine synthetase gene. Conclusion: Herein, RAPD and RFLP analysis indicating the rich genetic diversity of this fish in the experimental ecological habitat and this would come helpful for the conservation of germplasm diversity and to support the sustainable breeding program of M. cuchia.
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