北江大刺鳅(Mastacembelus armatus)的核型分析及线粒体Cyt b基因和D-loop的遗传多样性
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摘要
采用胸腔注射植物凝集素(PHA)、秋水仙素,以大刺鳅(Mastacembelus armatus)头肾组织为材料,低渗处理,空气干燥法进行染色体标本制备,分析其染色体核型。结果显示:大刺鳅的二倍体染色体数目为2n=48,其核型公式为2n=14m+34t, NF=62。对24尾大刺鳅线粒体Cyt b基因和D-loop序列进行测序分析,结果表明:(1)大刺鳅Cytb基因、D-loop序列长度分别为1438bp、895bp,两个基因的单倍型多样性分别为0.656和0.541,核苷酸多样性分别为0.00539和0.00817,平均核苷酸差异数分别为5.938和6.805,说明该种群的遗传多样性丰富。(2)两个基因的每个个体的遗传距离分别为0.000—0.018和0.000—0.026,说明该种群个体间的遗传距离小。(3)中性检验的Tajima’sD检验及Fu's Fs检验的结果说明该种群没有发生过扩张。结果说明北江大刺鳅的遗传多样性水平较高,可用于人工繁育。
To study and protect the resource of fish Mastacembelus armatus in the Beijiang River, Guangdong, South China, 24 individuals were sampled in the river reaches and their chromosomes karyotypes were analyzed. PHA(phytohemagglutinin) and colchicines were injected in vivo, and the kidney cells were collected and hypotonically air-dried.The results show that, the diploid chromosome number of M. armatus is 48 and the karyotype formula is 2 n=14 m+34 t,NF=62. In addition, the sequences of Cyt b gene and D-loop were analyzed using PCR and DNA sequencing, from which a1438-bp Cyt b gene and a 895-bp D-loop were obtained. The haplotype diversity of the two genes is 0.656 and 0.541, the nucleotide diversity 0.00539 and 0.00817, and the average number of nucleotide differences 5.938 and 6.805, respectively,indicating an obvious population genetic diversity. The genetic distance of each individual of the two genes are0.000—0.018 and 0.000—0.026, respectively, showing a close genetic distance. Moreover, the population size has not changed as shown in Tajima's D and Fu's Fs tests. Therefore, the genetic diversity of the M. armatus population in Beijiang River regions is intermediate, and the species can be used for artificial breeding.
To study and protect the resource of fish Mastacembelus armatus in the Beijiang River, Guangdong, South China, 24 individuals were sampled in the river reaches and their chromosomes karyotypes were analyzed. PHA(phytohemagglutinin) and colchicines were injected in vivo, and the kidney cells were collected and hypotonically air-dried.The results show that, the diploid chromosome number of M. armatus is 48 and the karyotype formula is 2 n=14 m+34 t,NF=62. In addition, the sequences of Cyt b gene and D-loop were analyzed using PCR and DNA sequencing, from which a1438-bp Cyt b gene and a 895-bp D-loop were obtained. The haplotype diversity of the two genes is 0.656 and 0.541, the nucleotide diversity 0.00539 and 0.00817, and the average number of nucleotide differences 5.938 and 6.805, respectively,indicating an obvious population genetic diversity. The genetic distance of each individual of the two genes are0.000—0.018 and 0.000—0.026, respectively, showing a close genetic distance. Moreover, the population size has not changed as shown in Tajima's D and Fu's Fs tests. Therefore, the genetic diversity of the M. armatus population in Beijiang River regions is intermediate, and the species can be used for artificial breeding.
引文
于旭蓉,仇雪梅,柳晓瑜等,2011.线粒体DNA多态性在海洋动物群体遗传结构研究中的应用.生物技术通报,(10):49-54
王方,黄小林,拉达等,2012.中国南部地区大刺鳅种群遗传和亲缘地理研究.见:中国鱼类学会.中国海洋湖沼学会鱼类学分会,中国动物学会鱼类学分会2012年学术研讨会论文集.兰州:中国海洋湖沼学会,中国鱼类学会,23
生物系细胞遗传学实验室,1985.四川广西两省59种鱼的染色体研究初报.武汉大学学报(自然科学版),(1):124-126
成庆泰,郑葆珊,1987.中国鱼类系统检索.北京:科学出版社,459
成美玲,田永胜,吴玉萍等,2018.云纹石斑鱼(♀)×鞍带石斑鱼(♂)杂交后代染色体核型分析.渔业科学进展,39(2):67-75
吕凤义,左艳玲,范月明等,2008.4种鲿科鱼类基于线粒体DNA控制区序列的比较研究.华南师范大学学报(自然科学版),(2):105-110
伍远安,梁志强,李传武等,2010.两种刺鳅肌肉营养成分分析及评价.营养学报,32(5):499-502
刘伟,代应贵,袁振兴等,2016.都柳江粗唇种群线粒体DNA Cyt b基因和D-loop序列组成及遗传多样性.淡水渔业,46(3):10-15,51
刘朋朋,钟立强,潘建林等,2013.基于线粒体D-loop区分析黄颡鱼(Pelteobagrus fulvidraco)五个淡水湖泊群体的遗传多样性.海洋与湖沼,44(3):728-733
杨华强,李强,舒琥等,2016.华南及邻近地区大刺鳅遗传多样性的ISSR分析.水生生物学报,40(1):63-70
吴仲庆,1998.水产生物遗传育种学.厦门:厦门大学出版社,7-2
沈朕,关洪斌,郑风荣等,2017.基于cytb和D-loop的4个大泷六线鱼群体遗传多样性分析.海洋科学进展,35(4):524-534
房祖业,陈晓东,吴咏诗等,2018.大刺鳅(Mastacembelus armatus)二、三、四碱基重复微卫星标记的筛选和特征分析.海洋与湖沼,49(1):174-182
钟立强,刘朋朋,潘建林等,2013.长江中下游5个湖泊黄颡鱼(Pelteobagrus fulvidraco)种群线粒体细胞色素b基因的遗传变异分析.湖泊科学,25(2):302-308
郭立,李隽,王忠锁等,2011.基于四个线粒体基因片段的银鱼科鱼类系统发育.水生生物学报,35(3):449-459
姬南京,常亚青,赵冲等,2014.3个地理群体仿刺参D-loop序列的变异及系统发生分析.水产学报,38(4):477-482
舒琥,江小璐,杨华强等,2017.华南地区7个大刺鳅野生群体的形态差异分析.广州大学学报(自然科学版),16(3):8-14
舒琥,蔡晓阅,刘锋等,2010.鲀形目3种鱼的染色体组型分析.动物学杂志,45(2):101-106
曾庆祥,方园,曾学平等,2016.大刺鳅的生物学特性与人工繁养殖技术.中国水产,(3):70-73
樊海平,邱曼丽,钟全福等,2018.不同生长阶段野生和养殖大刺鳅营养成分的比较.安徽农业科学,46(8):92-96
Grant W A S,Bowen B W,1998.Shallow population histories in deep evolutionary lineages of marine fishes:Insights from sardines and anchovies and lessons for conservation.Journal of Heredity,89(5):415-426
Levan A,Fredga K,Sandberg A A,1964.Nomenclature for centromeric position on chromosomes.Hereditas,52(2):201-220
Lynch M,Crease T J,1990.The analysis of population survey data on DNA sequence variation.Molecular Biology and Evolution,7(4):377-394
Nelson J S,2006.Fishes of the World.4th ed.New York:John Wiley and Sons,Inc.
Shaklee J B,Tamaru C S,Waples R S,1982.Speciation and evolution of marine fishes studied by the electrophoretic analysis of proteins.Pacific Science,36(2):141-157
Yang Y H,Kim K I,Cothran E G et al,2002.Genetic diversity of Cheju Horses(Equus caballus)determined by using mitochondrial DNA D-loop polymorphism.Biochemical Genetics,40(5-6):175-186
王方,黄小林,拉达等,2012.中国南部地区大刺鳅种群遗传和亲缘地理研究.见:中国鱼类学会.中国海洋湖沼学会鱼类学分会,中国动物学会鱼类学分会2012年学术研讨会论文集.兰州:中国海洋湖沼学会,中国鱼类学会,23
生物系细胞遗传学实验室,1985.四川广西两省59种鱼的染色体研究初报.武汉大学学报(自然科学版),(1):124-126
成庆泰,郑葆珊,1987.中国鱼类系统检索.北京:科学出版社,459
成美玲,田永胜,吴玉萍等,2018.云纹石斑鱼(♀)×鞍带石斑鱼(♂)杂交后代染色体核型分析.渔业科学进展,39(2):67-75
吕凤义,左艳玲,范月明等,2008.4种鲿科鱼类基于线粒体DNA控制区序列的比较研究.华南师范大学学报(自然科学版),(2):105-110
伍远安,梁志强,李传武等,2010.两种刺鳅肌肉营养成分分析及评价.营养学报,32(5):499-502
刘伟,代应贵,袁振兴等,2016.都柳江粗唇种群线粒体DNA Cyt b基因和D-loop序列组成及遗传多样性.淡水渔业,46(3):10-15,51
刘朋朋,钟立强,潘建林等,2013.基于线粒体D-loop区分析黄颡鱼(Pelteobagrus fulvidraco)五个淡水湖泊群体的遗传多样性.海洋与湖沼,44(3):728-733
杨华强,李强,舒琥等,2016.华南及邻近地区大刺鳅遗传多样性的ISSR分析.水生生物学报,40(1):63-70
吴仲庆,1998.水产生物遗传育种学.厦门:厦门大学出版社,7-2
沈朕,关洪斌,郑风荣等,2017.基于cytb和D-loop的4个大泷六线鱼群体遗传多样性分析.海洋科学进展,35(4):524-534
房祖业,陈晓东,吴咏诗等,2018.大刺鳅(Mastacembelus armatus)二、三、四碱基重复微卫星标记的筛选和特征分析.海洋与湖沼,49(1):174-182
钟立强,刘朋朋,潘建林等,2013.长江中下游5个湖泊黄颡鱼(Pelteobagrus fulvidraco)种群线粒体细胞色素b基因的遗传变异分析.湖泊科学,25(2):302-308
郭立,李隽,王忠锁等,2011.基于四个线粒体基因片段的银鱼科鱼类系统发育.水生生物学报,35(3):449-459
姬南京,常亚青,赵冲等,2014.3个地理群体仿刺参D-loop序列的变异及系统发生分析.水产学报,38(4):477-482
舒琥,江小璐,杨华强等,2017.华南地区7个大刺鳅野生群体的形态差异分析.广州大学学报(自然科学版),16(3):8-14
舒琥,蔡晓阅,刘锋等,2010.鲀形目3种鱼的染色体组型分析.动物学杂志,45(2):101-106
曾庆祥,方园,曾学平等,2016.大刺鳅的生物学特性与人工繁养殖技术.中国水产,(3):70-73
樊海平,邱曼丽,钟全福等,2018.不同生长阶段野生和养殖大刺鳅营养成分的比较.安徽农业科学,46(8):92-96
Grant W A S,Bowen B W,1998.Shallow population histories in deep evolutionary lineages of marine fishes:Insights from sardines and anchovies and lessons for conservation.Journal of Heredity,89(5):415-426
Levan A,Fredga K,Sandberg A A,1964.Nomenclature for centromeric position on chromosomes.Hereditas,52(2):201-220
Lynch M,Crease T J,1990.The analysis of population survey data on DNA sequence variation.Molecular Biology and Evolution,7(4):377-394
Nelson J S,2006.Fishes of the World.4th ed.New York:John Wiley and Sons,Inc.
Shaklee J B,Tamaru C S,Waples R S,1982.Speciation and evolution of marine fishes studied by the electrophoretic analysis of proteins.Pacific Science,36(2):141-157
Yang Y H,Kim K I,Cothran E G et al,2002.Genetic diversity of Cheju Horses(Equus caballus)determined by using mitochondrial DNA D-loop polymorphism.Biochemical Genetics,40(5-6):175-186
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