捞刀河浏阳段河蚬的遗传多样性及生殖特征研究
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摘要
研究以线粒体细胞色素氧化酶Ⅰ(COⅠ)部分序列(614 bp)为分子标记,对捞刀河浏阳段河蚬Corbicula fluminea(n=40)的种群遗传多样性进行评价,并分别从性腺组织学和精子形态学两方面分析其生殖特征,以期丰富河蚬的繁殖生物学信息,为开展其人工繁殖及资源保护提供参考。结果显示,40条河蚬COⅠ基因序列共检出4种单倍型,17个变异位点,平均单倍型多样性、平均核苷酸多样性和平均核苷酸差异数分别为0.664±0.042、0.014±0.006和8.595。捞刀河浏阳段河蚬存在雌雄同体和雌雄异体2种性别系统,雌雄同体、雄性和雌性的性比约为6﹕3﹕1。雌雄同体个体生殖滤泡存在滤泡混合型和滤泡并存型2种类型。23个雌雄同体和8个雄性个体的精子均为双鞭毛。结果表明,捞刀河浏阳段河蚬种群的遗传多样性相对较低,生殖方式多样且以雄核生殖为主。
This study attempted to theoretically support and comprehensively perceive genetic diversity and reproduction of Corbicula fluminea from the Laodao River in Liuyang city was investigated. Partial sequences of the cytochrome oxidase c subunit 1(COⅠ) of mitochondrial genes were utilized to evaluate the genetic diversity, and reproductive characteristics were observed by gonadal histology and sperm morphology using light microscopy. Among examined 40 individuals, only 4 haplotypes and 17 variable sites were detected in the partial sequences of the COⅠ, with an average haplotype diversity, nucleotide diversity, and number of variable sites equal to 0.664±0.042, 0.014±0.006,and 8.595, respectively. Hermaphrodite was a predominant reproduction in the population studied, and the hybrid hermaphrodite-sex ratio was 6﹕3﹕1. Furthermore, there were two types of reproductive follicles in hermaphroditic individuals, including sperm and egg cells in the same and in different follicular cells. The observation of sperm morphology revealed that biflagellate spermatozoa were discovered in both hermaphrodite(n=23) and male(n=8) cases. The results also showed that Corbicula fluminea samples from the Laodao River of Liuyang city contained a relatively low genetic diversity, but with multiple reproduction modes, and the predominant mode was androgenesis. This study attempted to present significant biological reproduction information required for Corbicula fluminea protection in China.
This study attempted to theoretically support and comprehensively perceive genetic diversity and reproduction of Corbicula fluminea from the Laodao River in Liuyang city was investigated. Partial sequences of the cytochrome oxidase c subunit 1(COⅠ) of mitochondrial genes were utilized to evaluate the genetic diversity, and reproductive characteristics were observed by gonadal histology and sperm morphology using light microscopy. Among examined 40 individuals, only 4 haplotypes and 17 variable sites were detected in the partial sequences of the COⅠ, with an average haplotype diversity, nucleotide diversity, and number of variable sites equal to 0.664±0.042, 0.014±0.006,and 8.595, respectively. Hermaphrodite was a predominant reproduction in the population studied, and the hybrid hermaphrodite-sex ratio was 6﹕3﹕1. Furthermore, there were two types of reproductive follicles in hermaphroditic individuals, including sperm and egg cells in the same and in different follicular cells. The observation of sperm morphology revealed that biflagellate spermatozoa were discovered in both hermaphrodite(n=23) and male(n=8) cases. The results also showed that Corbicula fluminea samples from the Laodao River of Liuyang city contained a relatively low genetic diversity, but with multiple reproduction modes, and the predominant mode was androgenesis. This study attempted to present significant biological reproduction information required for Corbicula fluminea protection in China.
引文
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[9]Zhu C,Li J,Xie S,et al.Genetic diversity analysis of Asian clam Corbicula fluminea in the Hongze Lake based on mitochondrial cytochrome b gene[J].Mitochondrial DNA Part A DNA Mapping Sequencing&Analysis,2017,29(3):419-426
[10]Li D M,Zhang T Q,Tang S K,et al.Genetic diversity of wild Corbicula fluminea population in the Hongze Lake analyzed by mitochondrial DNA COⅠgene sequence[J].Progress in Fishery Sciences,2015,36(5):81-86[李大命,张彤晴,唐晟凯,等.洪泽湖野生河蚬(Corbicula fluminea)线粒体COⅠ基因序列的遗传多样性分析.渔业科学进展,2015,36(5):81-86]
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[12]Pigneur L M,Etoundi E,Aldridge D C,et al.Genetic uniformity and long-distance clonal dispersal in the invasive androgenetic Corbicula clams[J].Molecular Ecology,2015,23(20):5102-5116
[13]Pigneur L M,Hedtke S M,Etoundi E,et al.Androgenesis:a review through the study of the selfish shellfish Corbicula spp[J].Heredity,2012,108(6):581-591
[14]Xu X,Qian L M,Li J Y,et al.The reproductive phase of gonad in Mollusca(Corbicula fluminea Müller)in Shanghai Diansan Lake[J].Journal of Animal Science(Current Zoology),1988,34(4):320-324[徐信,钱玲妹,李建英,等.淀山湖河蚬性腺发育分期的研究.动物学报,1988,34(4):320-324]
[15]Wang Q Z,Chang Y Q.Reproductive biology of Asian clam Corbicula fluminea in Dayang River in Liaoning Province[J].Journal of Dalian Ocean University,2010,25(1):8-13[王庆志,常亚青.大洋河河蚬的繁殖生物学研究.大连海洋大学学报,2010,25(1):8-13]
[16]Zhao X L,Zheng X D,Sun T Q,et al.The reproductive biology of Corbicula fluminea from the Yellow River Delta[J].Oceanologia et Limnologia Sinica,2012,43(5):1008-1015[赵雪琳,郑小东,孙同秋,等.黄河三角洲河蚬(Corbicula fluminea)的繁殖生物学研究.海洋与湖沼,2012,43(5):1008-1015]
[17]Bi T T.Reproductive biology and resource assessment for Corbicula fluminea in the Hongze Lake[D].University of Chinese Academy of Sciences.2013[毕婷婷.洪泽湖河蚬繁殖生物学特征与资源状况评价.中国科学院大学.2013]
[18]Zhang J.Annual production and reproductive characteristics of Corbicula fluminea in Datong Lake in Hunan Province[D].Hunan Agricultural University.2016[张健.大通湖河蚬的周年生产量及繁殖特性研究.湖南农业大学.2016]
[19]Folmer O,Black M,Hoeh W,et al.DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates[J].Molecular Marine Biology and Biotechnology,1994,3(5):294
[20]Wheeler D L,Barrett T,Benson D A,et al.Database resources of the National Center for Biotechnology Information.Nucleic Acids Res.33 Database Issue:D39-D45[J].Nucleic Acids Research,2007,35(Database issue):D5-D12
[21]Kearse M,Moir R,Wilson A,et al.Geneious Basic:An integrated and extendable desktop software platform for the organization and analysis of sequence data[J].Bioinformatics,2012,28(12):1647
[22]Tamura K,Dudley J,Nei M,et al.MEGA4:Molecular Evolutionary Genetics Analysis(MEGA)Software Version 4.0[J].Journal of Medical Molecular Biology,2007,24(24):1596-1599
[23]Thompson J D,Gibson T J,Plewniak F,et al.The Clustal-X windows interface:flexible strategies for multiple sequence alignment aided by quality analysis tools[J].Nucleic Acids Research,1997,25(25):4876-4882
[24]Rozas J,Sánchezdelbarrio J C,Messeguer X,et al.Dna SP,DNA polymorphism analyses by the coalescent and other methods[J].Bioinformatics,2003,19(18):2496-2497
[25]Avise,John C.Phylogeography:the History and Formation of Species[M].Cambridge:Harvard University Press.2000,134-135
[26]Houki S,Yamada M,Honda T,et al.Origin and possible role of males in hermaphroditic androgenetic Corbicula clams[J].Zoological Science,2011,28(7):526-531
[27]Grant W,Bowen B.Shallow population histories in deep evolutionary lineages of marine fishes:insights from sardines and anchovies and lessons for conservation[J].Journal of Heredity,1998,89(5):415-426
[28]Wu X P.River clam Corbicula fluminea(Müller)and river clam C.fluminalis(Müller)population ecology research:China institute of zoology,shellfish learning branch of China Marine lacustrine bog institute council of the second and third academic symposium proceedings[C].1986[吴小平.河蚬Corbicula fluminea(Müller)和江蚬C.fluminalis(Müller)的种群生态学研究:中国动物学会、中国海洋湖沼学会贝类学分会第二次代表会暨第三次学术讨论会论文集.1986]
[29]Morton B.The sexuality of Corbicula fluminea(Müller)in lentic and lotic waters in Hong Kong[J].Journal of Molluscan Studies,1983,49(1):81-83
[30]Renard E,Bachmann V,Cariou M L,et al.Morphological and molecular differentiation of invasive freshwater species of the genus Corbicula(Bivalvia,Corbiculidea)suggest the presence of three taxa in French rivers[J].Molecular Ecology,2000,9(12):2009-2016
[31]Ishibashi R,Komaru A.Abortive second meiosis detected in cytochalasin-treated eggs in androgenetic diploid Corbicula fluminea[J].Development Growth&Differentiation,2006,48(4):277-282
[32]Komaru A,Kawagishi T,Konishi K.Cytological evidence of spontaneous androgenesis in the freshwater clam Corbicula leana Prime[J].Development Genes&Evolution,1998,208(1):46-50
[2]Liao N,Chen S,Ye X,et al.Antioxidant and anti-tumor activity of a polysaccharide from freshwater clam,Corbicula fluminea[J].Food&Function,2013,4(4):539
[3]Zhou H,Liu C Q,Yan H,et al.The shape of Corbicula fluminea(Müller,1774),and its adaptability to the environment[J].Ecology Journal,2011,30(7):1497-1503[周会,刘丛强,闫慧,等.中国主要流域河蚬Corbicula fluminea(Müller,1774)形态及其对环境的适应性.生态学杂志,2011,30(7):1497-1503]
[4]Huang T K.Compendium of Materia Medica[M].Beijing:China Medical Science Press.2011,2195[黄泰康.现代本草纲目.北京:中国医药科技出版社.2001,2195]
[5]Cai R X.Animal Studies in Zhejiang.Software Animal[M].Hangzhou:Zhejiang Science and Technology Press.1991,235[蔡如星.浙江动物志.软体动物.杭州:浙江科学技术出版社.1991,235]
[6]Liu Y Y.Chinese Journal of Economic Animals:Freshwater Mollusks[M].Beijing:Science Press.1979,119-123[刘月英.中国经济动物志:淡水软体动物.北京:科学出版社.1979,119-123]
[7]Liu Y S,Zhang T Q,Tang S K,et al.Development and application of growth equations for Corbicula fluminea population in Hongze Lake[J].Journal of Water Ecology,2017,38(4):82-89[刘燕山,张彤晴,唐晟凯,等.洪泽湖河蚬种群生长方程估算及其应用.水生态学杂志,2017,38(4):82-89]
[8]Bi T T,Hou G,Zhang S Y,et al.Study on age structure,growth and mortality of Corbicula fluminea in Hongze Lake,China,through shell length-frequency data analysis[J].Acta Hydrobiologica Sinica,2014,38(4):797-800[毕婷婷,侯刚,张胜宇,等.基于壳长频率分析的洪泽湖河蚬渔获物年龄结构、生长与死亡参数估计.水生生物学报,2014,38(4):797-800]
[9]Zhu C,Li J,Xie S,et al.Genetic diversity analysis of Asian clam Corbicula fluminea in the Hongze Lake based on mitochondrial cytochrome b gene[J].Mitochondrial DNA Part A DNA Mapping Sequencing&Analysis,2017,29(3):419-426
[10]Li D M,Zhang T Q,Tang S K,et al.Genetic diversity of wild Corbicula fluminea population in the Hongze Lake analyzed by mitochondrial DNA COⅠgene sequence[J].Progress in Fishery Sciences,2015,36(5):81-86[李大命,张彤晴,唐晟凯,等.洪泽湖野生河蚬(Corbicula fluminea)线粒体COⅠ基因序列的遗传多样性分析.渔业科学进展,2015,36(5):81-86]
[11]Li D M,Zhang T Q,Tang S K,et al.Analysis of genetic diveristy and population structure of Corbicula flumineain Hongze Lake in mitochondrial COⅠgene sequence[J]Oceanologia et Limnologia Sinica,2015,46(6)1339-1346[李大命,张彤晴,唐晟凯,等.基于线粒体COⅠ序列的洪泽湖河蚬(Corbicula fluminea)遗传多样性和种群结构分析.海洋与湖沼,2015,46(6)1339-1346]
[12]Pigneur L M,Etoundi E,Aldridge D C,et al.Genetic uniformity and long-distance clonal dispersal in the invasive androgenetic Corbicula clams[J].Molecular Ecology,2015,23(20):5102-5116
[13]Pigneur L M,Hedtke S M,Etoundi E,et al.Androgenesis:a review through the study of the selfish shellfish Corbicula spp[J].Heredity,2012,108(6):581-591
[14]Xu X,Qian L M,Li J Y,et al.The reproductive phase of gonad in Mollusca(Corbicula fluminea Müller)in Shanghai Diansan Lake[J].Journal of Animal Science(Current Zoology),1988,34(4):320-324[徐信,钱玲妹,李建英,等.淀山湖河蚬性腺发育分期的研究.动物学报,1988,34(4):320-324]
[15]Wang Q Z,Chang Y Q.Reproductive biology of Asian clam Corbicula fluminea in Dayang River in Liaoning Province[J].Journal of Dalian Ocean University,2010,25(1):8-13[王庆志,常亚青.大洋河河蚬的繁殖生物学研究.大连海洋大学学报,2010,25(1):8-13]
[16]Zhao X L,Zheng X D,Sun T Q,et al.The reproductive biology of Corbicula fluminea from the Yellow River Delta[J].Oceanologia et Limnologia Sinica,2012,43(5):1008-1015[赵雪琳,郑小东,孙同秋,等.黄河三角洲河蚬(Corbicula fluminea)的繁殖生物学研究.海洋与湖沼,2012,43(5):1008-1015]
[17]Bi T T.Reproductive biology and resource assessment for Corbicula fluminea in the Hongze Lake[D].University of Chinese Academy of Sciences.2013[毕婷婷.洪泽湖河蚬繁殖生物学特征与资源状况评价.中国科学院大学.2013]
[18]Zhang J.Annual production and reproductive characteristics of Corbicula fluminea in Datong Lake in Hunan Province[D].Hunan Agricultural University.2016[张健.大通湖河蚬的周年生产量及繁殖特性研究.湖南农业大学.2016]
[19]Folmer O,Black M,Hoeh W,et al.DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates[J].Molecular Marine Biology and Biotechnology,1994,3(5):294
[20]Wheeler D L,Barrett T,Benson D A,et al.Database resources of the National Center for Biotechnology Information.Nucleic Acids Res.33 Database Issue:D39-D45[J].Nucleic Acids Research,2007,35(Database issue):D5-D12
[21]Kearse M,Moir R,Wilson A,et al.Geneious Basic:An integrated and extendable desktop software platform for the organization and analysis of sequence data[J].Bioinformatics,2012,28(12):1647
[22]Tamura K,Dudley J,Nei M,et al.MEGA4:Molecular Evolutionary Genetics Analysis(MEGA)Software Version 4.0[J].Journal of Medical Molecular Biology,2007,24(24):1596-1599
[23]Thompson J D,Gibson T J,Plewniak F,et al.The Clustal-X windows interface:flexible strategies for multiple sequence alignment aided by quality analysis tools[J].Nucleic Acids Research,1997,25(25):4876-4882
[24]Rozas J,Sánchezdelbarrio J C,Messeguer X,et al.Dna SP,DNA polymorphism analyses by the coalescent and other methods[J].Bioinformatics,2003,19(18):2496-2497
[25]Avise,John C.Phylogeography:the History and Formation of Species[M].Cambridge:Harvard University Press.2000,134-135
[26]Houki S,Yamada M,Honda T,et al.Origin and possible role of males in hermaphroditic androgenetic Corbicula clams[J].Zoological Science,2011,28(7):526-531
[27]Grant W,Bowen B.Shallow population histories in deep evolutionary lineages of marine fishes:insights from sardines and anchovies and lessons for conservation[J].Journal of Heredity,1998,89(5):415-426
[28]Wu X P.River clam Corbicula fluminea(Müller)and river clam C.fluminalis(Müller)population ecology research:China institute of zoology,shellfish learning branch of China Marine lacustrine bog institute council of the second and third academic symposium proceedings[C].1986[吴小平.河蚬Corbicula fluminea(Müller)和江蚬C.fluminalis(Müller)的种群生态学研究:中国动物学会、中国海洋湖沼学会贝类学分会第二次代表会暨第三次学术讨论会论文集.1986]
[29]Morton B.The sexuality of Corbicula fluminea(Müller)in lentic and lotic waters in Hong Kong[J].Journal of Molluscan Studies,1983,49(1):81-83
[30]Renard E,Bachmann V,Cariou M L,et al.Morphological and molecular differentiation of invasive freshwater species of the genus Corbicula(Bivalvia,Corbiculidea)suggest the presence of three taxa in French rivers[J].Molecular Ecology,2000,9(12):2009-2016
[31]Ishibashi R,Komaru A.Abortive second meiosis detected in cytochalasin-treated eggs in androgenetic diploid Corbicula fluminea[J].Development Growth&Differentiation,2006,48(4):277-282
[32]Komaru A,Kawagishi T,Konishi K.Cytological evidence of spontaneous androgenesis in the freshwater clam Corbicula leana Prime[J].Development Genes&Evolution,1998,208(1):46-50
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