基于微卫星标记的中华蜜蜂5个自然群体遗传多样性分析
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摘要
【目的】分析中国四川阿坝、黄土高原、海南岛、浙闽丘陵、滇南地区等地的中华蜜蜂(Apis cerana cerana)自然群体遗传多样性。【方法】提取样品基因组DNA后进行PCR扩增,并用聚丙烯酰胺凝胶电泳检测。通过计算15对微卫星标记在5个中华蜜蜂自然群体中的等位基因数、多态信息含量、期望杂合度、观测杂合度、遗传距离、遗传相似系数和遗传分化系数,进而评估各中华蜜蜂种群遗传多样性和各种群间遗传分化。【结果】15个微卫星位点共检测到64个等位基因,每个位点的等位基因数平均为4.266 7个,单个位点的等位基因数为2～6个。各群体平均等位基因数为2.6～3.066 7个,平均观测杂合度为0.550 3～0.664 3,平均期望杂合度为0.524 4～0.566 7,平均多态信息量为0.410 3～0.492 7。四川阿坝群体与黄土高原、海南岛、浙闽丘陵、滇南地区等地群体在Acc001,Acc096,Acc131等3个位点上存在明显分化;黄土高原群体与海南岛、浙闽丘陵、滇南地区等地群体在Acc114,Acc121,Acc132等3个位点上表现出一定程度的遗传分化;海南岛群体与浙闽丘陵、滇南地区两地群体在Acc001,Acc002,Acc004,Acc131,Acc132等5个位点上均表现出较强的遗传分化;浙闽丘陵与滇南地区群体间在Acc002,Acc121,Acc131等3个位点上遗传分化极明显。5个中华蜜蜂自然群体之间的遗传距离在0.160 1～0.491 0之间。【结论】5个中华蜜蜂群体遗传多样性丰富,群体间遗传分化明显。
[Purposes]Genetic diversity of five Apis cerana cerana natural populations in Sichuan Aba,Loess Plateau,Hainan Island,Zhejiang-Fujian hilly,and Southern Yunnan Province was analyzed.[Methods]PCR implications were performed after genomic DNAs of samples were extracted,and then PAGEs were accomplished.By calculating several parameters including number of alleles,polymorphic information content,expected heterozygosity,observed heterozygosity,Nei's distance,genetic similarity coefficient and genetic differentiation coefficient of 15 pairs of simple sequence repeat markers in five Apis cerana cerana wild populations,the genetic diversity and the genetic differentiation of each population were evaluated.[Findings]A total of 64 alleles were detected at all loci for all the samples,with an average of 4.266 7 alleles per locus and 2～6 alleles per locus.The average allele number of each population was ranged from 2.6 to 3.066 7,the average observed heterozygosity of each population was ranging from 0.550 3 to0.664 3,the average expected heterozygosity of each population was 0.524 4～0.566 7,and the polymorphic information content of the 5 populations was 0.410 3～0.492 7.There were obvious differentiation between Sichuan Aba population and any of the other population at three allele loci(Acc001,Acc096,and Acc131).The Loess Plateau population showed a certain degree of genetic differentiation with the populations in Hainan Island,Zhejiang-Fujian hilly,and Southern Yunnan Province at three allele loci(Acc114,Acc121,and ACC132).The population of Hainan Island,Zhejiang-Fujian hilly,and Southern Yunnan Province showed strong genetic differentiation at five sites(Acc001,Acc002,Acc004,Acc131,and Acc132).The genetic differentiation between the Zhejiang-Fujian hilly,and Southern Yunnan Province populations was very obvious at three loci(Acc002,Acc121,and Acc131).The Nei's distance between the five natural populations ranged from 0.160 1 to 0.491 0.[Conclusions]In summary,the genetic diversity of five natural Apis cerana cerana populations was high,and the pairwise genetic differentiation among populations was obvious.
[Purposes]Genetic diversity of five Apis cerana cerana natural populations in Sichuan Aba,Loess Plateau,Hainan Island,Zhejiang-Fujian hilly,and Southern Yunnan Province was analyzed.[Methods]PCR implications were performed after genomic DNAs of samples were extracted,and then PAGEs were accomplished.By calculating several parameters including number of alleles,polymorphic information content,expected heterozygosity,observed heterozygosity,Nei's distance,genetic similarity coefficient and genetic differentiation coefficient of 15 pairs of simple sequence repeat markers in five Apis cerana cerana wild populations,the genetic diversity and the genetic differentiation of each population were evaluated.[Findings]A total of 64 alleles were detected at all loci for all the samples,with an average of 4.266 7 alleles per locus and 2～6 alleles per locus.The average allele number of each population was ranged from 2.6 to 3.066 7,the average observed heterozygosity of each population was ranging from 0.550 3 to0.664 3,the average expected heterozygosity of each population was 0.524 4～0.566 7,and the polymorphic information content of the 5 populations was 0.410 3～0.492 7.There were obvious differentiation between Sichuan Aba population and any of the other population at three allele loci(Acc001,Acc096,and Acc131).The Loess Plateau population showed a certain degree of genetic differentiation with the populations in Hainan Island,Zhejiang-Fujian hilly,and Southern Yunnan Province at three allele loci(Acc114,Acc121,and ACC132).The population of Hainan Island,Zhejiang-Fujian hilly,and Southern Yunnan Province showed strong genetic differentiation at five sites(Acc001,Acc002,Acc004,Acc131,and Acc132).The genetic differentiation between the Zhejiang-Fujian hilly,and Southern Yunnan Province populations was very obvious at three loci(Acc002,Acc121,and Acc131).The Nei's distance between the five natural populations ranged from 0.160 1 to 0.491 0.[Conclusions]In summary,the genetic diversity of five natural Apis cerana cerana populations was high,and the pairwise genetic differentiation among populations was obvious.
引文
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[14]郭慧萍,周姝婧,朱翔杰,等.秦巴山区中华蜜蜂种群微卫星DNA遗传分析[J].昆虫学报,2016,59(3):337-345.GUO H P,ZHOU S J,ZHU X J,et al.Population genetic analysis of Apis cerana cerana from the Qinling-Daba mountain areas based on microsatellite DNA[J].Acta Entomologica Sinica,2016,59(3):337-345.
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[17]沈浩,刘登义.遗传多样性概述[J].生物学杂志,2001,18(3):5-7.SHEN H,LIU D Y.Overview of genetic diversity[J].Journal of Biology,2001,18(3):5-7.
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[19]高雅,李生斌.样本量对等位基因检出数量的影响[J].中国优生与遗传杂志,2008(8):7-9.GAO Y,LI S B.The effect of sample size on the number of alleles detected[J].Journal of eugenics and genetics of China,2008(8):7-9.
[20]BOTSTEIN D,WHITE R,SKOLNICK M.Construction of a genetic linkage map in man using restriction fragment length polymorphisms[J].American Journal of Human Genetics,1980,32:314-331.
[21]WRIGHT S.The genetical structure of populations[J].Annals of Eugenics,1951,15(4):323-354.
[2]INSUAN S,DEOWANISH S,KLINBUNGA S,et al.Genetic differentiation of the giant honey bee(Apis dorsata)in Thailand analyzed by mitochondrial genes and microsatellites[J].Biochemical Genetics,2007,45(4):345-361.
[3]SIMONA S,PETER K,JANEZ P,et al.Molecular characterisation of indigenous Apis mellifera carnicain Slovenia[J].Apidologie,2004,35(6):623-636.
[4]De La RUA P,HERNANDEZ GARCIA R,PEDERSEN B V,et al.Molecular diversity of honeybee Apis mellifera iberica L.(Hymenoptera:Apidae)from western Andalusia[J].Archivos de Zootecnia,2004,53(202):195-203.
[5]De La RUA P,GALIAN J,SERRANO J,et al.Microsatellite analysis of non-migratory colonies of Apis mellifera ibericafrom south-eastern Spain[J].Journal of Zoological Systematics and Evolutionary Research,2002,40(3):164-168.
[6]于瀛龙,周姝婧,徐新建,等.长白山中华蜜蜂(Apis cerana cerana)遗传多样性分析[J].福建农林大学学报(自然科学版),2013,42(6):643-647.YU Y L,ZHOU S J,XU X J,et al.Analysis on genetic diversity of Apis cerana cerana in Changbai mountains[J].Journal of Fujian Agriculture and Forestry University(Natural Science Edition),2013,42(6):643-647.
[7]朱翔杰,周冰峰,吴显达,等.福建中华蜜蜂微卫星标记的遗传多样性分析[J].福建农林大学学报(自然科学版),2011,40(4):407-411.ZHU X J,ZHOU B F,WU X D,et al.Genetic diversity of Apis cerana ceranain Fujian based on microsatellite markers[J].Journal of Fujian Agriculture and Forestry University(Natural Science Edition),2011,40(4):407-411.
[8]刘敏,吉挺,殷玲,等.武夷山中华蜜蜂形态特征与微卫星DNA标记的相关分析[J].福建农林大学学报(自然科学版),2009,38(5):532-536.LIU M,JI T,YIN L,et al.Relationship between microsatellite DNA makers and morphology feature of Apis cerana cerana populations in Wuyi mountain[J].Journal of Fujian Agriculture and Forestry University(Natural Science Edition),2009,38(5):532-536.
[9]陈杨,宗超,余林生,等.皖南山区与皖西大别山区中华蜜蜂群体微卫星DNA遗传多样性研究[J].中国蜂业,2011,62(Z4):8-11.CHEN Y,ZONG C,YU L S,et al.Study on Microsatellite DNA Genetic Diversity of Apis cerana cerana in Wannan moutain area and Wanxi Da Bie moutain area[J].Apiculture of China,2011,62(Z4):8-11.
[10]TAKAHASHI J,SHIMIZU S,KOYAMA S,et al.Variable microsatellite loci isolated from the Asian honeybee,Apis cerana(Hymenoptera;Apidae)[J].Molecular Ecology Resources,2010,9(3):819-821.
[11]于瀛龙,周姝婧,徐新建,等.贵州省东方蜜蜂微卫星DNA遗传分化与遗传多样性分析[J].福建农林大学学报(自然科学版),2017,46(3):323-328.YU Y L,ZHOU S J,XU X J,et al.Genetic diversity and genetic differentiation of Apis ceranain Guizhou province of southwest China[J].Journal of Fujian Agriculture and Forestry University(Natural Science Edition),2017,46(3):323-328.
[12]徐新建,周姝婧,朱翔杰,等.海南岛中华蜜蜂遗传多样性的微卫星DNA分析[J].昆虫学报,2013,56(5):554-560.XU X J,ZHOU S J,ZHU X J,et al.Microsatellite DNA analysis of genetic diversity of Apis cerana ceranain Hainan island,southern China[J].Acta Entomologica Sinica,2013,56(5):554-560.
[13]徐新建,周姝婧,朱翔杰,等.黄土高原中华蜜蜂遗传多样性的微卫星DNA分析[J].福建农林大学学报(自然科学版),2013,42(6):638-642.XU X J,ZHOU S J,ZHU X J,et al.Microsatellite DNA genetic diversity of Apis cerana cerana from the Loess Plateau,Northwest China[J].Journal of Fujian Agriculture and Forestry University(Natural Science Edition),2013,42(6):638-642.
[14]郭慧萍,周姝婧,朱翔杰,等.秦巴山区中华蜜蜂种群微卫星DNA遗传分析[J].昆虫学报,2016,59(3):337-345.GUO H P,ZHOU S J,ZHU X J,et al.Population genetic analysis of Apis cerana cerana from the Qinling-Daba mountain areas based on microsatellite DNA[J].Acta Entomologica Sinica,2016,59(3):337-345.
[15]李婉玫.基于微卫星和线粒体DNA的陕西秦巴山区中华蜜蜂遗传多样性研究[D].西安:陕西师范大学,2017.LI W M.Genetic diversity of Chinese honeybees in Qinba Mountain,Shaanxi Province based on microsatellite and mitochondrial DNA[D].Xi’an:Shaanxi Normal University,2017.
[16]LIU L,QIN M Z,YANG L,et al.A genome-wide analysis of simple sequence repeats in Apis cerana and its development as polymorphism markers[J].Gene,2017,599:53-59.
[17]沈浩,刘登义.遗传多样性概述[J].生物学杂志,2001,18(3):5-7.SHEN H,LIU D Y.Overview of genetic diversity[J].Journal of Biology,2001,18(3):5-7.
[18]BARKER J S F.Squential gel electrophoretic analysis of esterase-2in two populations of Drosophila buzzatii[J].Genetica,1994,92(3):165-175.
[19]高雅,李生斌.样本量对等位基因检出数量的影响[J].中国优生与遗传杂志,2008(8):7-9.GAO Y,LI S B.The effect of sample size on the number of alleles detected[J].Journal of eugenics and genetics of China,2008(8):7-9.
[20]BOTSTEIN D,WHITE R,SKOLNICK M.Construction of a genetic linkage map in man using restriction fragment length polymorphisms[J].American Journal of Human Genetics,1980,32:314-331.
[21]WRIGHT S.The genetical structure of populations[J].Annals of Eugenics,1951,15(4):323-354.