高原鼢鼠微卫星多态位点筛选及其通用性检测
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摘要
跨种扩增是微卫星DNA标记获取的快捷有效途径之一。为了筛选出高原鼢鼠微卫星多态性位点,从已开发微卫星信息的鼹形鼠科和甘肃鼢鼠物种中进行了高原鼢鼠微卫星多态性位点及其通用性研究,结果表明:在87对微卫星位点中,只有54对进行了成功扩增,多态位点仅有9个,27个鼹形鼠科物种的微卫星在高原鼢鼠中并没有多态性,9个多态位点全来自于同属的甘肃鼢鼠。基于获得的9个多态位点,检测到高原鼢鼠的等位基因数在4~12,其平均观测杂合度(Ho)为0.544,期望杂合度(He)为0.671,多态信息含量为0.551,研究为高原鼢鼠的微卫星标记应用奠定了基础。
Cross-amplification is one of the quickest and effective methods of obtaining microsatellite markers.In this study,we amplified microsatellite loci of Eospalax baileyi using published microsatellite DNA primers from related species,and tested both transferability and polymorphism.The results showed that 54 out of 87 loci were successfully amplified,meanwhile only nine polymorphic loci were obtained.27 loci fromSpalaxspecies showed only two alleles and nine polymorphic loci were all obtained from Gansu zokor.Genetic diversity of plateau zokor populations was relatively medium.The number of alleles(NA)ranged from 4 to 12(average5.5),PIC value was 0.551,and the ranges of observed(Ho)and expected(He)heterozygosities were 0.544 and 0.671,respectively.The results will facilitate the further studies in microsatellite markers of E.bailyei.
Cross-amplification is one of the quickest and effective methods of obtaining microsatellite markers.In this study,we amplified microsatellite loci of Eospalax baileyi using published microsatellite DNA primers from related species,and tested both transferability and polymorphism.The results showed that 54 out of 87 loci were successfully amplified,meanwhile only nine polymorphic loci were obtained.27 loci fromSpalaxspecies showed only two alleles and nine polymorphic loci were all obtained from Gansu zokor.Genetic diversity of plateau zokor populations was relatively medium.The number of alleles(NA)ranged from 4 to 12(average5.5),PIC value was 0.551,and the ranges of observed(Ho)and expected(He)heterozygosities were 0.544 and 0.671,respectively.The results will facilitate the further studies in microsatellite markers of E.bailyei.
引文
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[26]孙波,鲍毅新,张龙龙,等.大鼠及小鼠微卫星引物在社鼠中的跨种扩增[J].动物学杂志,2009,44(6):145-150.
[27]陈昌文,曹珂,王力荣,等.中国桃主要品种资源及其野生近缘种的分子身份证构建[J].中国农业科学,2011,44(10):2081-2093.
[2]Santana Q C,Coetzee M P A,Steenkamp E T,et al.Microsatellite discovery by deep sequencing of enriched genomic libraries[J].Biotechniques,2009,46(3):217.
[3]Selkoe K A,Toonen R J.Microsatellites for ecologists:a practical guide to using and evaluating microsatellite markers[J].Ecology Letters,2006,9(5):615-629.
[4]Zane L,Bargelloni L,Patarnello T.Strategies for microsatellite isolation:a review[J].Molecular Ecology,2002,11(1):1-16.
[5]林能锋,苏永全,丁少雄,等.大黄鱼微卫星标记引物在石首鱼科几个近缘种中的通用性研究[J].中国水产科学,2008,15(2):237-243.
[6]叶林江,王静,孙鹏,等.四种栲属植物微卫星标记在钩栲中的通用性检测[J].植物分类与资源学报,2014,36(4):443-448.
[7]邵伟伟,华攀玉,周善义,等.棕果蝠微卫星位点的筛选及其对近缘种的通用性[J].兽类学报,2007,27(4):385-388.
[8]李华.中国鼢鼠亚科的分类研究[J].首都师范大学学报(自然科学版),1995,16(l):75-80.
[9]苏军虎,纪维红,南志标,等.鼢鼠亚科Mysopalacinae动物系统学研究现状与展望[J].动物学杂志,2015,50(4):649-658.
[10]施银柱,边疆晖,王权业.高寒草甸地区小哺乳动物群落多样性的初步研究[J].兽类学报,1991,11(4):279-97.
[11]崔庆虎,蒋志刚,刘季科,等.青藏高原草地退化原因述评[J].草业科学,2007,24(5):20-26.
[12]刘锦上,张卫国,江小蕾,等.高原鼢鼠的洞道空间对高寒草甸植被性状的影响[J].草地学报,2011,19(6):927
[13]Zhang Y M,Zhang Z B,Liu J K.Burrowing rodents as ecosystem engineers:the ecology and management of plateau zokors Myospalax fontanierii in alpine meadow ecosystems on the Tibetan Plateau[J].Mammal Review,2003,33(3-4):284-294.
[14]钟文勤,樊乃昌.我国草地鼠害的发生原因及其生态治理对策[J].生物学通报,2002,37(7):1-4.
[15]苏军虎,南志标,纪维红.家畜放牧对草地啮齿动物影响的研究进展[J].草业学报,2016,25(11):136-148.
[16]王海芳,苏军虎,刘荣堂,等.基于线粒体ND4基因的鼢鼠系统进化关系[J].草原与草坪,2008,129(4):20-23.
[17]Begall S,Burda H,Schleich C E.Subterranean rodents:news from underground[M].Springer Berlin Heidelberg,2007.
[18]王静,纪维红,徐长林,等.达乌尔黄鼠线粒体Cyt b基因序列特征及其系统进化分析[J].草原与草坪,2016,36(3):17-22.
[19]Karanth K P,Avivi A,Beharav A,et al.Microsatellite diversity in populations of blind subterranean mole rats(Spalax ehrenbergi,superspecies)in Israel:speciation and adaptation[J].Biological Journal of the Linnean Society,2004,83(2):229-241.
[20]苏军虎.青藏高原东缘两类典型土著动物种群遗传结构分析[D].兰州:甘肃农业大学,2014.
[21]苏军虎,张艳萍,娄忠玉,等.甘肃金鳟AFLP体系建立及应用[J].中国农学通报,2011,27(7):400-404.
[22]Botstein D,White R L,Sckolnick M,et al.Construction of agenetic linkage map in man using restriction fragment lengthpolymorphism[J].Am J Hum Genetic,1980,32(3):314-331.
[23]Nei M.Genetic distance between populations[J].Am Nat,1972,106:283-292.
[24]Crow A J,Kimura M.Evolution in sexual and asexual population[J].Am Nat,1965,99:439-450.
[25]邹喻苹,葛颂,王晓东.系统与进化植物学中的分子标记[M].北京:科学出版社,2001.
[26]孙波,鲍毅新,张龙龙,等.大鼠及小鼠微卫星引物在社鼠中的跨种扩增[J].动物学杂志,2009,44(6):145-150.
[27]陈昌文,曹珂,王力荣,等.中国桃主要品种资源及其野生近缘种的分子身份证构建[J].中国农业科学,2011,44(10):2081-2093.