罗非鱼“粤闽1号”及其繁育群体的遗传多样性和遗传关系分析
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摘要
罗非鱼"粤闽1号"是用尼罗罗非鱼(Oreochromisniloticus)和奥利亚罗非鱼(O.aureus)培育的全雄罗非鱼新品种。采用同工酶电泳技术和线粒体DNA控制区序列分析方法,对罗非鱼"粤闽1号"及其繁育群体的遗传多样性、遗传结构及群体之间的遗传关系进行研究。同工酶电泳结果显示,酯酶(EST)和乳酸脱氢酶(LDH)的酶谱具有组织和群体特异性。肌肉和脾脏中的EST表达量极低,而肝脏中的EST表达量较高,LDH在3种组织中均有较高表达,但酶谱存在差异。在罗非鱼"粤闽1号"及其繁育群体中,共检测到10条EST酶带和5条LDH酶带,多态性位点比例(P)为12.50%～71.43%,平均观测杂合度(Ho)为0.0417～0.6143, Hardy-Weinberg遗传偏离指数(D)为-0.2347～0.9072。线粒体DNA控制区序列分析结果显示,mtDNA D-loop区的碱基组成无显著差异,均呈现出A+T碱基偏向性(63.39%)。在罗非鱼"粤闽1号"及其繁育群体中,共发现20种单倍型,核苷酸多样性指数(Pi)为0～0.0519,奥尼罗非鱼雌鱼(WY1)、尼罗罗非鱼雌鱼(XX)、尼罗罗非鱼雄鱼(XY)和罗非鱼"粤闽1号"(XY2)群体的Pi值(0.0440～0.0519)明显高于超雄尼罗罗非鱼(YY1)、超雄奥尼罗非鱼(YY2)和奥利亚罗非鱼雌鱼(WZ)群体的Pi值(0～0.0009)。各群体之间的遗传分化指数(FST)为-0.0115～0.9963,XY2与WY1群体之间、XX与XY群体之间以及WZ和YY2群体之间的遗传分化不显著(F_(ST)<0.05,P>0.05)。XY2群体与尼罗罗非鱼群体(XX群体和XY群体)和奥利亚罗非鱼群体(WZ群体)之间的平均遗传距离分别为0.0639和0.0695。在NJ系统进化树中,XX、XY、YY1和XY2群体聚为一支,WY1、WZ和YY2群体聚为另一支。本研究揭示了罗非鱼"粤闽1号"的生化和分子遗传特征,为其种质鉴定、繁育群体的构建和优良性状的稳定遗传提供理论基础。
"Yuemin No.1" is a novel holandric species of tilapia, which was bred with Oreochromis niloticus and O. aureus. In order to reveal the genetic characteristics of tilapia "Yuemin No.1," the genetic diversity, genetic structure, and genetic relationships among groups of tilapia "Yuemin No.1" and its breeding populations were studied by isozyme electrophoresis and sequence analysis of the D-loop region of mtDNA. The results of isozyme electrophoresis showed that the zymogram of esterase(EST) and lactate dehydrogenase(LDH) were population-and tissue-specific in tilapia. The expression level of EST in the muscle and spleen was extremely low, whereas that in the liver was high. LDH was highly expressed in all the tissues, as revealed by the different zymograms. Ten bands of EST and five of LDH were detected in tilapia "Yuemin No.1" and its breeding populations. The proportion of polymorphic loci(P) ranged from 12.50% to 71.43%, the average observed heterozygosity(Ho) ranged from 0.0417 to 0.6143, and the Hardy-Weinberg genetic deviation index(D) ranged from-0.2347 to 0.9072. The result of sequence analysis of the D-loop region of mtDNA revealed that no significant difference occurred in base composition, and that A+T content(63.39%) bias was widespread in all the populations. There were 20 haplotypes in tilapia "Yuemin No.1" and its breeding populations. The nucleotide diversity index(Pi) ranged from 0 to 0.0519. The Pi value of WY1, XX, XY2, and XY populations(0.0440～0.0519) were significantly higher than that of YY1, YY2, and WZ populations(0～0.0009). The genetic differentiation index(F_(ST)) among populations ranged from-0.0115 to 0.9963. There was no significant genetic differentiation(F_(ST)<0.05, P>0.01) between the XY2 and WY1, XX and XY, and WZ and YY2 populations. The average genetic distance between XY2 population and O. niloticus(XX and XY populations) was 0.0639, whereas that between the XY2 population and O. aureus(WZ population) was 0.0695. Cluster analysis revealed two clusters; one cluster included XY, XX, YY1, and XY2 populations, and the other cluster included WY1, WZ, and YY2 populations. This study revealed the biochemical and molecular genetic features of tilapia "Yuemin No.1," and provide research data for germplasm identification, establishment of breeding populations, and maintainence of stable inheritance of desirable traits.
"Yuemin No.1" is a novel holandric species of tilapia, which was bred with Oreochromis niloticus and O. aureus. In order to reveal the genetic characteristics of tilapia "Yuemin No.1," the genetic diversity, genetic structure, and genetic relationships among groups of tilapia "Yuemin No.1" and its breeding populations were studied by isozyme electrophoresis and sequence analysis of the D-loop region of mtDNA. The results of isozyme electrophoresis showed that the zymogram of esterase(EST) and lactate dehydrogenase(LDH) were population-and tissue-specific in tilapia. The expression level of EST in the muscle and spleen was extremely low, whereas that in the liver was high. LDH was highly expressed in all the tissues, as revealed by the different zymograms. Ten bands of EST and five of LDH were detected in tilapia "Yuemin No.1" and its breeding populations. The proportion of polymorphic loci(P) ranged from 12.50% to 71.43%, the average observed heterozygosity(Ho) ranged from 0.0417 to 0.6143, and the Hardy-Weinberg genetic deviation index(D) ranged from-0.2347 to 0.9072. The result of sequence analysis of the D-loop region of mtDNA revealed that no significant difference occurred in base composition, and that A+T content(63.39%) bias was widespread in all the populations. There were 20 haplotypes in tilapia "Yuemin No.1" and its breeding populations. The nucleotide diversity index(Pi) ranged from 0 to 0.0519. The Pi value of WY1, XX, XY2, and XY populations(0.0440～0.0519) were significantly higher than that of YY1, YY2, and WZ populations(0～0.0009). The genetic differentiation index(F_(ST)) among populations ranged from-0.0115 to 0.9963. There was no significant genetic differentiation(F_(ST)<0.05, P>0.01) between the XY2 and WY1, XX and XY, and WZ and YY2 populations. The average genetic distance between XY2 population and O. niloticus(XX and XY populations) was 0.0639, whereas that between the XY2 population and O. aureus(WZ population) was 0.0695. Cluster analysis revealed two clusters; one cluster included XY, XX, YY1, and XY2 populations, and the other cluster included WY1, WZ, and YY2 populations. This study revealed the biochemical and molecular genetic features of tilapia "Yuemin No.1," and provide research data for germplasm identification, establishment of breeding populations, and maintainence of stable inheritance of desirable traits.
引文
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Yang J,He AY,He XJ,et al.Genetic diversity and relationships of mitochondrial DNA control region from the eight domesticated populations of Nile tilapia.Journal of Fishery Sciences of China,2014,21(4):693-699[杨洁,何安元,何学军,等.尼罗罗非鱼8个养殖群体线粒体控制区遗传多样性和遗传关系分析.中国水产科学,2014,21(4):693-699]
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Yang YH,Zhou JS,Li L,et al.Preliminary analysis on isozymes in different tissues and population genetic structure of Silurus lanzhouensis.Freshwater Fisheries,2015,45(1):25-29[杨元昊,周继术,李蕾,等.兰州鲇不同组织同工酶及群体遗传结构初步分析.淡水渔业,2015,45(1):25-29]
Zhang JD,He GQ,Chen G,et al.The tissue-specificities of isozymes and the genetic structure in three species of Theraponidae.Journal of Natural Science of Hunan Normal University,2015,38(5):27-34[张健东,何国清,陈刚,等.三种科鱼类同工酶组织特异性及群体遗传结构分析.湖南师范大学自然科学学报,2015,38(5):27-34]
Zhu LF.Gradient gel electrophoresis on polyacrylamide of isozymes and proteins of fishes.Acta Hydrobiologica Sinica,1992,16(2):183-185[朱蓝菲.鱼类同工酶和蛋白质的聚丙烯酰胺梯度凝胶电泳法.水生生物学报,1992,16(2):183-185]
Ding JQ,Liu P,Li J,et al.Genetic variation analysis of four geographic populations of Charybdis japonica by isozyme.Progress in Fishery Sciences,2013,34(2):82-89[丁金强,刘萍,李健,等.日本蟳4个地理群体遗传变异的同工酶分析.渔业科学进展,2013,34(2):82-89]
Gyllensten U.The genetic structure of fish:Differences in the intraspecific distribution of biochemical genetic variation between marine,anadromous,and freshwater species.Journal of Fish Biology,1985,26(6):691-699
Han Q,Zhao DH.Comparative study on three isozymes in different tissues of catfish(Silnrus asotus)with different color in Dongting Lake.Guangdong Agricultural Sciences,2015(2):104-107,113[韩庆,赵东海.洞庭湖不同体色鲇不同组织同工酶的比较研究.广东农业科学,2015(2):104-107,113]
Hu YT,Hu W,Jiang H,et al.Sequence and phylogenetic analysis of the complete mitochondrial genome of Chuzhou Crucian Carp(Carassius auratus).Progress in Fishery Sciences,2015,36(5):63-70[胡玉婷,胡王,江河,等.滁州鲫(Carassius auratus)线粒体全基因组序列分析及系统进化.渔业科学进展,2015,36(5):63-70]
Janko K,Flaj?hans H,Choleva L,et al.Diversity of European spined loaches(genus Cobitis L.):An update of the geographic distribution of the Cobitis taenia hybrid complex with a description of new molecular tools for species and hybrid determination.Journal of Fish Biology,2007,71(Suppl C):387-408
Li SF,Cai WQ.Introgression in hatchery stocks of Tilapia nilotica and Tilapia aurea in China.Journal of Fisheries of China,1995,19(2):105-111[李思发,蔡完其.我国尼罗罗非鱼和奥利亚罗非鱼养殖群体的遗传渐渗.水产学报,1995,19(2):105-111]
Li SF.Genetical characterization of major freshwater culture fishes in China.Shanghai:Shanghai Scientific and Technical Publishers,1998,189-193[李思发.中国淡水主要养殖鱼类种质研究.上海:上海科技出版社,1998,189-193]
Liu H,Zhang HQ,Cai SL,et al.Analysis of the genetic diversity of six populations of cultured Litopenaeus vannamei based on the mitochondrial DNA control region.Progress in Fishery Sciences,2016,37(1):63-73[刘红,张海强,蔡生力,等.基于线粒体DNA控制区序列的6个凡纳滨对虾(Litopenaeus vannamei)养殖群体的遗传多样性分析.渔业科学进展,2016,37(1):63-73]
Slatkin M.Gene flow and the geographic structure of natural populations.Science,1987,236(4803):787-792
Teletchea F.Molecular identification methods of fish species:Reassessment and possible applications.Reviews in Fish Biology and Fisheries,2009,19(3):265-293
Wei YZ,Zhang GR,Huo B,et al.A comparative study on lactate dehydrogenase isozymes in six species of Schizothoracinae.Freshwater Fisheries,2017,47(5):3-8[魏玉众,张桂蓉,霍斌,等.雅鲁藏布江中游6种裂腹鱼乳酸脱氢酶同工酶的比较研究.淡水渔业,2017,47(5):3-8]
Wright S.Evolution and the genetics of populations:Variability within and among natural populations.Chicago:University of Chicago Press,1978,580
Wu CJ,Zou ZY,Yang H,et al.Structure of the mitochondrial DNA D-Loop region and analysis of genetic diversity in different strains of tilapia(Oreochromis).Chinese Journal of Zoology,2010,45(5):121-128[吴长敬,邹芝英,杨弘,等.罗非鱼mt DNA D-loop区部分序列结构和种群遗传多样性分析.动物学杂志,2010,45(5):121-128]
Wu YY,Li LH,Han JL.Studies on four isozyme from several tissues of Jifu tilapia.Biotechnology Bulletin,2008(Suppl),319-323[吴燕燕,李来好,韩君莉.吉富罗非鱼不同组织中4种同工酶研究.生物技术通报,2008(增刊):319-323]
Xie XY,Li SF.Comparison of base sequence diversity of Cytb and D-loop gene of Nile tilapia.Genomics and Applied Biology,2014,33(5):982-985[颉晓勇,李思发.罗非鱼选育群体Cytb与D-loop序列变异信息对比分析.基因组学与应用生物学,2014,33(5):982-985]
Xiong QM.Isozymes analysis of fishes(Vol.1).Hereditas(Beijing),1992,14(2):41-44,48[熊全沫.鱼类同工酶谱分析(上).遗传,1992,14(2):41-44,48]
Xu C,Wang KL,You F,et al.Biochemical genetics of Lateolabrax japonicus populationⅠbiochemical genetic analysis of isozymes.Oceanologia et Limnologia Sinica,2001,32(1):42-49[徐成,王可玲,尤锋,等.鲈鱼群体生化遗传学研究Ⅰ同工酶的生化遗传分析.海洋与湖沼,2001,32(1):42-49]
Yang J,He AY,He XJ,et al.Genetic diversity and relationships of mitochondrial DNA control region from the eight domesticated populations of Nile tilapia.Journal of Fishery Sciences of China,2014,21(4):693-699[杨洁,何安元,何学军,等.尼罗罗非鱼8个养殖群体线粒体控制区遗传多样性和遗传关系分析.中国水产科学,2014,21(4):693-699]
Yang L,Lu H,Liu YQ,et al.Study on the tissue specificity of 8isozymes in Cyprinus carpio from Dongping Lake.Chinese Agricultural Science Bulletin,2014,30(14):26-32[杨玲,卢红,刘羽清,等.东平湖鲤8种同工酶的组织特性研究.中国农学通报,2014,30(14):26-32]
Yang S,Huang ZH,Ye X,et al.Preliminary study on three isozymes of Oreochromis hornorum and Oreochromis mossambicus.Journal of Dalian Fisheries University,2006,21(2):122-126[杨淞,黄樟翰,叶星,等.荷那龙罗非鱼和莫桑比克罗非鱼3种同工酶的分析.大连水产学院学报,2006,21(2):122-126]
Yang YH,Zhou JS,Li L,et al.Preliminary analysis on isozymes in different tissues and population genetic structure of Silurus lanzhouensis.Freshwater Fisheries,2015,45(1):25-29[杨元昊,周继术,李蕾,等.兰州鲇不同组织同工酶及群体遗传结构初步分析.淡水渔业,2015,45(1):25-29]
Zhang JD,He GQ,Chen G,et al.The tissue-specificities of isozymes and the genetic structure in three species of Theraponidae.Journal of Natural Science of Hunan Normal University,2015,38(5):27-34[张健东,何国清,陈刚,等.三种科鱼类同工酶组织特异性及群体遗传结构分析.湖南师范大学自然科学学报,2015,38(5):27-34]
Zhu LF.Gradient gel electrophoresis on polyacrylamide of isozymes and proteins of fishes.Acta Hydrobiologica Sinica,1992,16(2):183-185[朱蓝菲.鱼类同工酶和蛋白质的聚丙烯酰胺梯度凝胶电泳法.水生生物学报,1992,16(2):183-185]