两个不同的人工雌核发育草鱼群体基因组DNA的RAPD分析
RAPD ANALYSIS ON GENOMIC DNA OF TWO ARTIFICIAL GYNOGENETIC GROUPS OF GRASS CARP,CTENOPHARYNGODON IDELLU
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摘要: 采用随机扩增多态性DNA(RAPD)技术对连续两代人工诱导雌核发育草鱼群体和一代人工诱导雌核发育草鱼群体的群体内的遗传相似度、遗传距离以及群体多样性进行了分析,并用一个随机取样的普通草鱼群体作比较.用26个多态性引物在3个群体中共检测到了291个扩增位点;在3个群体中检测到的位点数分别为265、22和282.其中多态性位点数分别为15、19和81.遗传学统计分析结果表明:3个群体的多态位点比例分别为5.66%、6.99%、28.2%;香农表型多样性指数分别为0.102、0.3169和0.8450;按照Nei指数统计的三个群体的遗传相似度分别为0.9851、0.9820、0.9114.这些结果表明:两个雌核发育草鱼群体的遗传多样性远低于普通草鱼群体,而其群体的基因组DNA同质性远高于普通草鱼.而在两个雌核发育草鱼群体中,两代雌核发育群体的遗传多样性要低于一代雌核发育群体的遗传多样性;而群体的遗传纯合度则前者高于后者.Abstract: Genetic similarity,genetic distance and Shannon's genetic diversity of two different artificial gynogenetic grass carp groups and a control common grass carp group were analyzed in the methods of RAPD (Random Amplified Polymorphism DNA).One of the two different gynogenetic groups was a meiotic-mito-gynogenetic grass carp group that was produced by artificial gynogenesis for two generations. Another.gynogenetic group was a mito-gynogenetic grass carp that was generated by artificial gynogenesis only one generation.The common grass carp group was randomly collected from the markets in Changsha. Total 291 loci were detected in all of the three groups through the RAPD-PCR with 26 polymorphic random primers. The loci number in the three groups,meio- mito-gynogenetic group,the mito-gynogenetic group and the common grass carp group,were 265,272 and 282 and the number of polymorphic loci were 15,19 and 81 respectively. Genetic statistical analysis showed that percentage of polymorphic loci of the three groups of grass carp,were 5.66%,6.99%,28.23%;and the genetic similarity within the three groups of grass carp estimated by Nei’s index were 0.9851,0.9820,0.9114,respectively. The genetic diversity indices within the three groups of grass carp estimated by Shannon index of phenotypic diversity were 0.1722,0.3169 and 0.8450 respectively. Generally,the genetic diversity within two artificial gynogenetic groups of grass carp is far lower than that of common grass carp,while the genetic diversity in meiotic-mito-gynogenetic grass carp group is much lower than that of mito-gynogenetic grass carp group. The relatively low level of genetic diversity in each of the two groups of gynogenetic grass carp revealed that the population purity in each of the two gynogenetic grass carp groups is higher than that of common grass carp group.
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Keywords:
- Gynogenetic grass carp /
- RAPD /
- Genetic similarity /
- Genetic distance /
- Genetic diversity
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PCR技术实验指南[M].北京:科学出版社. 2000][21] Apostol B L,Black IV W C,Reiter P,et al. population genetics with RAPD-PCR markers:the breeding structure of Aedes aegypti in Puerto Rico [J]. Heredity,1996,76:325-334[22] Nei M.Li W H. Mathematical model for studying genetic variation in terms of restriction endonucleases[J].Proc. Natl. Acad. Sci. USA, 1979,76:5269-5273[23] Wachira F N, Waugh R, Hackett C A. et al. Detection on genetic diversity in tea (Camellia sinensis) using RAPD markers [J]. Genome,1995,38:201-210[24] Streisinger G, Walker C, Dower N, et al. Production of homozygous diploid zebra fish [J].Nature,1981,291:293-296[25] Kuhnlein U.Assessment of inbreeding by DNA fingerprinting:Development of a calibration curve using defined strains of chickens [J].Genetics,1990,125:161-165 Wu Q J,Chen R D,Ye Y Z.Investigation on the carp gynogenesis with reference to establishing a pure Line [J]. Acta Genetica Sinica, 1981,8(1):50-55. [吴清江,陈荣德,叶玉珍. 鲤鱼人工雌核发育及其作为建立近交系新途径的研究. 遗传学报,1981,8(1):50-55][2] Williams J G K,Kubelik A R,livak K J,et al. DNA polymorphism amplified by arbitrary primers are useful as genetic markers[J]. Nucleic Acids Research,1990,18:6531-6535[3] Welsh J,McClelland M,. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res, 1990,18:7213-7218[4] Wu Q J, Gui J F. Fish Genetics and Breeding Engineering[M]. Shanghai:Shanghai Scientific Technical Publishes. 1999. [吴清江,桂建芳. 鱼类遗传育种工程. 上海:上海科学和技术出版社.1999][5] Zou S M, LI S F,Cai W Q,et al. Establishing gynogenetic groups of genetic improved Megalobrama amblycephala and its genetic analysis. [J]. Journal of Fisheries of China,2001,25(4):311-316.[邹署明,李思发,蔡完其,等.团头鲂良种雌核发育群体的建立及其遗传变异. 水产学报,2001,25(4):311-316][6] Lu R,Rank G H. Use of RAPD population genetic parameters in the alfalfa leafcutting bee,Megachile rotundata[J]. Genome,1996,39:63-70[7] Xia D Q, Cao Y, Wu T T, et al. Study on lineages of Protosalanx chinensis,Neosalanx taihuensis and N. oligodontis in Taihu Lake with RAPD technique[J]. Journal of Fisheries of China,2000,7(1):12-15.[夏德全,曹莹,吴婷婷,等. 用RAPD方法分析太湖大银鱼、太湖新银鱼和寡齿新银鱼的亲缘关系. 中国水产科学,2000,7(1):12-15][8] Marillia E F,Scoles G J. The use of RAPD markers inHordeum Phylogeny [J].Genome,1996,39:646-654[9] Sun X W,Liang L Q. A genetic linkage map of common carp [J]. Journal of Fishery Sciences of China,2000,7(1):1-5[孙效文,梁利群. 鲤鱼的遗传连锁图谱.中国水产科学,2000,7(1):1-5][10] Lin J Z,Ritland K. Construction of a genetic linkage map in the wild plant Mimulus using RAPD and isozyme markers[J]. Genome,1996,39:63-70[11] Zhou L,Fan L C, Gui J F. RAPD analysis of incorporation of heterologousgenetic materisls in multiple species of Silver Crucian carp[J]. Acta Hydrobiologica Sinica,1998,22(4):301-303. [周莉,樊连春,桂建芳. 银鲫复合种外源遗传物质整入的RAPD分析. 水生生物学报,1998,22(4):301-303][12] Iturra P,Medrano J F, Bagley M. Identification of sex chromosome molecular markers using RAPD sand fluorescentin situ hybridization in rainbow trout[J]. Genetica,1998,101:209-213[13] Zhang S M, Deng H, Wang D Q et al population structure and diversity of silver carp and grass carp from populations of Yangtze River system revealed by RAPD[J]. Acta Hydrobiologica Sinica, 2001,25(4):324-330. [张四明,邓怀,汪登强,等.长江水系鲢和草鱼遗传结构及变异性的RAPD分析. 水生生物学报,2001,25(4):324-330][14] 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):168-173.[章怀云,刘荣宗,张大明,等.草鱼和鲤鱼群体遗传变异的RAPD指纹分析.水生生物学报,1998,22(2):168-173][15] Stanley J G.Morphology of androgenetic and gynogenetic grass carp,Ctenopharyngodon idella(Valenciennes)[J].J. Fish Biol,1976,9:523-528[16] Luo C,Liu Y.Studies on production of gynogenetics grass carp and crucian carp[J]. Acta Sci Univ Norm Hunan,1991,14(2):154-159.[罗琛,刘筠.人工诱导草鱼和鲫鱼雌核发育的研究.湖南师范大学自然科学学报,1991,14(2):154-159][17] Li B X,Luo C.Cytological observations on induction of mitogynogenesis by heat-shocking of the eggs in grass carp,Ctenopharyngodon idellu[J].Acta Hydrobiologica Sinic,2003,27(2):155-160. [李冰霞,罗琛.热休克法抑制第一次卵裂实现草鱼雌核发育的细胞学观察[J].水生生物学报,2003,27(2):155-160][18] Tajima F. Evolutionary relationship of DNA sequence in finite populations[J]. Genetics,1983,105:437-445[19] Huang M M,Chen J H,Luo C et al.screening of polymorphic primers and polymorphic locus for grass carp genomic RAPD analysis[J].Science Technology and Engineering,2004,4(2):91～96[黄明敏,陈金辉,罗琛等.草鱼基因组随机引物扩增多态性引物及多态位点的筛选.科学技术与工程,2004,4(2):91～96][20] Dieffenbach C W,Dveksler G S.(translated by Huang P T,et al). PCR Primer:A Laboratory Manual[M]. Beijing:Science Press.2000.[C.W.迪芬巴赫,G.S.德维克斯勒著,(黄培堂等译). PCR技术实验指南[M].北京:科学出版社. 2000][21] Apostol B L,Black IV W C,Reiter P,et al. population genetics with RAPD-PCR markers:the breeding structure of Aedes aegypti in Puerto Rico [J]. Heredity,1996,76:325-334[22] Nei M.Li W H. Mathematical model for studying genetic variation in terms of restriction endonucleases[J].Proc. Natl. Acad. Sci. USA, 1979,76:5269-5273[23] Wachira F N, Waugh R, Hackett C A. et al. Detection on genetic diversity in tea (Camellia sinensis) using RAPD markers [J]. Genome,1995,38:201-210[24] Streisinger G, Walker C, Dower N, et al. Production of homozygous diploid zebra fish [J].Nature,1981,291:293-296[25] Kuhnlein U.Assessment of inbreeding by DNA fingerprinting:Development of a calibration curve using defined strains of chickens [J].Genetics,1990,125:161-165
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