中国胭脂鱼种群的遗传分析
GENETIC STRUCTURE OF CHINESE SUCKER(MYXOCYPRINUS ASIATICUS)BY mtDNA AND RAPD MARKERS
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摘要: 采用RAPD和PCR-RFLP技术,分析了长江中游两个中国胭脂鱼群体的遗传结构.0个随机引物进行RAPD分析,有3个引物显示了多态,宜昌、金口群体内个体之间的遗传相似度分别为0.9274、0.9313,群体之间遗传相似度为0.9000.12个限制性内切酶分析了两群体线粒体DNA ND-/6基因的限制性片段长度多态性,仅内切酶Ncil的酶切图谱显示了多态,基因型间的核苷酸序列歧化距离为0.23%,核苷酸多样性为0.004.分析表明,长江中游两个中国胭脂鱼群体遗传结构较为单一,群体之间表现了较为明显的遗传分化.Abstract: Two molecular techniques, RAPD and PCR RFLP,were used to detect population genetic structure of Chinese sucker (Myxocyprinus asiaticus) from the middle reaches of Yangtze River, Yichang section (Y population),and Jinkou section (J population). Total genome DNA was isolated from fin tissue by standard phenol chloro form extraction. Four kits of primers for RAPD analysis were from Operon technologies (oph, opl, opk, opq).Nei′s genetic distance from RAPD pattern was calculated for population analysis. Of 50 RAPD primers, only 3 produced polymorphism.The genetic similarity within Yichang and Jinkou population is 0.9274?0.9313 respectively. The genetic similarity between them is about 0.9000.4μL aliquots of the PCR products of mitochondrial ND-5/6 genes were digested by 12 restriction enzymes. Estimation of mtDNA haplotype diversity followed Nei and Tajima's nucleotide diversity indices. Of 12 restriction enzymes types,11 are monomorphic except Ncil enzyme. Only two kinds of genotypes were found. The nucleotide diversity was 0.004, and the pair wise sequence divergence was about 0.235% . Meanwhile, We found differentiations between the two populations to some extent. From Ncil enzyme cleavage patterns,the restriction pattern A was predominant (about 78%) in Y population, while in J population B was the major pattern (about 88%).In RAPD patterns amplified by primer Opq02, the 1.0 kb fragments appear mainly in Y population (about 78%),while the majority of the 0.8 kb fragments appear in J population (about 88%). Because of the booming pollution, and overexploitation, and because of the longer development and the sex mature period, Chinese sucker undertook a suddenly and sharply decreasing. Bottlenecks and genetic drift came into being. On the other hand,the Gezhou Dam has been the great barrier for the gene exchange of different populations. Therefore, the genetic structure of Chinese sucker is considerably simple and there was a trend of the differentiation of gene materials among different populations.
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Keywords:
- Chinese sucker /
- Genetic diversity /
- RAPD /
- PCR RFLP /
- ND-5/6 gene
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Genetic variation and local differentiation in the Pacific cod Gadus macrocephalus around Japan revealed by mtDNA and RAPD markers[J].Fisheries Science,1998, 64(5):673-880［13］Wirgin Isaac, Oppermann Tim, Stabile Joseph. Genetic divergence of Robust redhorse Moxostoma rubustum (Cypriniformes Catostomidae) from the Oconee River and the Savannah River based on MtDNA control region sequences[J].Copeia,2001,2:526-530[14] Pascale Lafontaine and J.J. Dodson. Intraspecific genetic structure of white sucker (catostomus commersoni)in northeastern North America as revealed by MtDNA polymorphism[J].Can.J.Fish.Aqua.Sci,1997,54:555-565[15] Yu Z T, Deng Z L, Zhou C S, et al. Prognosis of the effects of the Gezhou Damhydroelectric project on fish resources of the Yangtze River[C].In:The Chinese Ichthyological Society,Transactions of the Chinese Ichthyological Society. Beijing:Science Press,1985,(4):193-205[余志堂,邓中(米),周春生,等. 长江葛洲坝水利枢纽兴建后鱼类资源变化预测. 中国鱼类学会编,鱼类学论文集,北京:科学出版社,1985,第四集:193-205] Liu L H, Wu G X,Wang Z L,et al. A effect of the Gezhou Dam key water control project on sexul gland development and natural spawning of Chinese sucker (Myxocyprinus asiaticus) in the downstream below the Dam[J].Journal of Fisheries of China, 1992,16(4):346-456[刘乐和,吴国犀,王志玲. 葛洲坝水利工程对胭脂鱼性腺发育及自然繁殖的影响. 水产学报,1992,16(4):346-456][2] Wu G X, Liu L H, Wang Z L,et al. The age and growth of myxocyprinus asiaticus at Yichang reaches of Yangtze River below the Gezhou Dam[J].Freshwater Fisheries,1990,2:3-7[吴国犀,刘乐和,王志玲,等. 葛洲坝下宜昌胭脂鱼年龄与生长. 淡水渔业,1990,2:3-7][3] Zhang C G, Zhao Y H .Early development of Chinese sucker [J].Acta Zoological Sinica,2000,46(4):438-447[张春光,赵亚辉. 胭脂鱼的早期发育. 动物学报,2000,46(4):438-447][4] Sun Y H,Liu S Y, Wang W,et al. Genetics diversity analysis of mitochondrial D-Loop region of Chinese sucker (Myxocyprinus asiaticus)[J].Acta Genetica Sinica, 2002,29(2):787-790[孙玉华,刘思阳,王伟,等. 中国胭脂鱼线粒体控制区遗传多样性分析. 遗传学报,2002,29(2):787-790][5] Song P,Hu JR,Hu YC,et al.Development of DNA markers for Megalobrama amblycephala Yih with RAPD-derived PCR primers[J].J.Wuhan Univ.(Nat.Sci.Ed.),2001,4(4):493-497[宋平,胡珈瑞,胡隐昌,等. 依据RAPD片段克隆而建立的团头鲂PCR鉴定法. 武汉大学学报(理学版),2001,4(4):493-497][6] Lynch M. The similarity index and DNA fingerprinting[J].Mol Biol Evol,1990,7:478-486[7] Park L K,Brainard M A, Dightman D A. Low level of intraspecific variation in the MtDNA of chum salmon (Oncorhynchus keta)[J].Mol.Mar.Biol.Biotech,1993,2:362-370[8] McElroy D,Moran P E, Bermingham E, et al .REAP: An integrated environment for the manipulation and phylogenetic analysis of restriction data[J].Journal of Heredity,1992,83:157-158[9] Guoqing Lu, Sifa Li, Bernatchez L.MtDNA diversity,population structure, and conservation genetics of four native carps within the Yangtze River, China[J].Can.J.Fish.Aqua.Sci.1997,54:47-56[10] Zhang H Y, Liu R Z, Zhang X W et al. Assessment of population genetic variation of grass carp and common carp using RAPD finger-prints[J].Acta Hydrobiologica Sinica, 1998, 22(2):168-175[章怀云,刘荣宗,张学文 ,等. 草鱼和鲤鱼群体遗传变异的RAPD分析. 水生生物学报,1998,22(2):168-175][11] Bielawski J P, Pumo D E. RAPD analysis of Atlantic coast stripped bass[J].Heredity,1997,78:32-40[12] Saitoh. Kenji. Genetic variation and local differentiation in the Pacific cod Gadus macrocephalus around Japan revealed by mtDNA and RAPD markers[J].Fisheries Science,1998, 64(5):673-880［13］Wirgin Isaac, Oppermann Tim, Stabile Joseph. Genetic divergence of Robust redhorse Moxostoma rubustum (Cypriniformes Catostomidae) from the Oconee River and the Savannah River based on MtDNA control region sequences[J].Copeia,2001,2:526-530[14] Pascale Lafontaine and J.J. Dodson. Intraspecific genetic structure of white sucker (catostomus commersoni)in northeastern North America as revealed by MtDNA polymorphism[J].Can.J.Fish.Aqua.Sci,1997,54:555-565[15] Yu Z T, Deng Z L, Zhou C S, et al. Prognosis of the effects of the Gezhou Damhydroelectric project on fish resources of the Yangtze River[C].In:The Chinese Ichthyological Society,Transactions of the Chinese Ichthyological Society. Beijing:Science Press,1985,(4):193-205[余志堂,邓中(米),周春生,等. 长江葛洲坝水利枢纽兴建后鱼类资源变化预测. 中国鱼类学会编,鱼类学论文集,北京:科学出版社,1985,第四集:193-205]
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