大鸨(Otis tarda)两个亚种的遗传多样性与系统分化

大鸨(Otis tarda)为中国Ⅰ级重点保护动物,分为两个亚种,即指名亚种(Otis tarda tarda)和东方亚种(Otis tarda dybowskii)。研究从代表母系遗传特征的mtDNA控制区和代表双亲遗传特征的核微卫星两方面对两个亚种的遗传多样性与系统分化进行了分析。指名亚种mtDNA控制区3段序列(CtrIaL/CtrIIoH、L438/H772和LCR2a/HCR8)的单倍型数、Л、δ和К都明显高于东方亚种,更显著高于松辽平原西北部繁殖区。东方亚种3个微卫星(Otmic08、Otmic16和Otmic26)的等位基因数、Ho和He明显低于指名亚种。因此东方亚种的遗传多样性都明显低于指名亚种,甚至低于Madrid种群。两个亚种存在于不同的系统分支,证实了两个亚种的系统关系,欧洲指名亚种存在更多的系统分支。     

大鸨(Otis tarda)两个亚种的遗传多样性与系统分化

大鸨（Otistarda）为中国Ⅰ级重点保护动物,分为两个亚种,即指名亚种（Otis tarda tarda）和东方亚种（Otis tarda dybowskii）。研究从代表母系遗传特征的mtDNA控制区和代表双亲遗传特征的核微卫星两方面对两个亚种的遗传多样性与系统分化进行了分析。指名亚种mtDNA控制区3段序列（CtrIaL/CtrIIoH、L438/H772和LCR2a/HCR8）的单倍型数、Л、δ和К都明显高于东方亚种,更显著高于松辽平原西北部繁殖区。东方亚种3个微卫星（Otmic08、Otmic16和Otmic26）的等位基因数、Ho和He明显低于指名亚种。因此东方亚种的遗传多样性都明显低于指名亚种,甚至低于Madrid种群。两个亚种存在于不同的系统分支,证实了两个亚种的系统关系,欧洲指名亚种存在更多的系统分支。     

微卫星DNA与生化标记分析对长爪沙鼠群体遗传分析的比较

目的比较生化标记和微卫星DNA标记方法对长爪沙鼠群体遗传分析的可靠性。方法应用27个生化位点和13个微卫星DNA位点,采用已建立的生化标记和微卫星DNA标记分析方法对国内2个长爪沙鼠群体进行遗传分析,计算并比较两种方法测得的各群体遗传参数。结果生化基因位点中有13个位点在整体中呈现遗传多态性,多态率为48.1%;微卫星位点中有11个位点在整体中表现出多态性,多态率均为84.6%。两种方法测得的平均有效等位基因数趋于一致,微卫星DNA的多态位点百分率和平均杂合度均明显高于生化标记方法。但生化标记和微卫星DNA检测对两个长爪沙鼠群体的遗传多样性差异反映一致,所反映的群体平衡状况也基本一致。结论生化标记分析和微卫星DNA方法均可较好地反映长爪沙鼠群体遗传结构。     

勐养保护区亚洲象微卫星位点筛选及种群遗传多样性分析

本文以亚洲象肌肉样品提出的DNA为模板,从非洲象31个微卫星位点和5个已知亚洲象微卫星位点中筛选勐养亚洲象的微卫星位点,进而对在西双版纳勐养保护区3年采集到的191 份亚洲象粪便样品中提出的DNA进行特异性PCR扩增、基因分型、检测位点信息和遗传多样性分析.结果表明:36个位点中有14个位点能在亚洲象肌肉样品提出的DNA中成功扩增,且经测序证实为微卫星位点.其中9个多态位点能在185份粪便样品DNA中稳定扩增.勐养种群中,3个位点可能偏离Hardy Weubberg平衡,至少8个位点间无明显连锁存在,平均等位基因数3.78±1.72,平均期望杂和度0.32 ± 0.06,平均观察杂和度0.36±0.02,平均多态信息含量0.28 ,说明这9个位点适用于勐养亚洲象的遗传学研究.根据微卫星位点的杂合度和等位基因频率,相比于其他亚洲象种群,勐养亚洲象种群遗传多样性较低且等位基因频率具有特异性.     

基于转录组测序的大熊猫多态性微卫星标记筛选

结合已公布的大熊猫Ailuropoda melanoleuca基因组和本实验室所测6只大熊猫的转录组数据,筛选多态性微卫星位点并分析其组成及特征。结果显示:共获得326个多态性微卫星位点,其中二碱基多态性微卫星最多,共228个,占69.93%;三、四、五、六碱基所占比例分别为9.51%、14.11%、5.21%、1.22%。根据分析结果中缺失率与标准差2项指标以及位点序列长度,选取20个多态性二碱基微卫星位点,用于25只大熊猫个体血液DNA进行PCR验证并做后续分析。结果表明:不同位点的等位基因数为2~8,平均等位基因数为3.70,观测杂合度、期望杂合度分别为0~1.000、0.280~0.784,平均值分别为0.472和0.532。在Bonferroni校正后,证实4个位点显著偏离哈迪-温伯格平衡,所有位点未观察到显著连锁不平衡(P>0.01)。20个位点多态信息含量(PIC)在0.246~0.734,其中具有高度多态性的位点9个(PIC>0.50),11个位点呈中度多态性(0.25相似文献   

我国粘虫种群的微卫星位点筛选及遗传多样性分析

【目的】筛选适用于我国粘虫Mythimna separata种群遗传学研究的微卫星位点,从分子水平揭示粘虫种群的遗传多样性。【方法】利用已报道的微卫星标记及本实验室粘虫转录组测序的SSR序列,采用PCR产物荧光标记与自动扫描分型方法,分析各位点在我国河南、陕西、山西3个省份的7个粘虫地理种群200头试虫中的扩增稳定性和多态性。【结果】7个粘虫地理种群有7个位点能稳定扩增且具有较高的多态性。这7个位点等位基因丰富度(Ar)为4.167~12.402,观测杂合度(Ho)平均为0.640,期望杂合度(He)平均为0.752,多态信息含量(PIC)为0.547~0.884;各位点均存在无效等位基因且偏离哈迪-温伯格平衡,所有成对位点不存在显著连锁不平衡情况。【结论】从来自河南、陕西、山西的7个不同粘虫地理种群中成功筛选了7个能稳定扩增的SSR位点,且在这7个不同的粘虫地理种群中均具有较高的多态性,可用于我国粘虫种群遗传结构研究。粘虫不同地理种群间基因交流频繁,基因交流阻止了由遗传漂变引起的群体间分化,不同地理种群间遗传分化很低甚至不存在遗传分化。     

秦川母牛群体遗传特性的微卫星标记研究

为了从DNA分子水平揭示秦川牛群体遗传多态性和群体遗传结构,寻找可用于秦川牛的微卫星标记,本研究选择了12个普通牛（Bos taurus）微卫星标记检测了90头秦川母牛各微卫星位点的遗传变异及多态性。结果表明,在秦川母牛群体中,12个微卫星位点共检测到了247个等位基因,各位点的等位基因数在13（INRA005）～33个（HEL13）之间,平均每个微卫星位点的等位基因数为21个;总有效等位基因数和平均每个位点平均有效等位基因数（Ne）分别分为142．6229和11．8852。各位点平均基因频率取样方差（V（pij））为2．6036×10^-4。12个微卫星位点平均观察杂合度（Ho）和平均期望杂合度（He）在0．7842（INRA005）～0．9775（BM315）和0．7952（BM315）～0．9446（HEL13）之间。12个位点平均多态信息含量（PIC）在0．7653（INRA005）～0．9420（HEL13）之间,平均为0．8965．12个微卫星位点均属于高度多态位点,这表明秦川母牛群体中所检测各微卫星位点具有丰富的遗传多态性,具备较大的选择潜力。12个微卫星位点的平均固定指数（F）为-0．0076,即各位点杂合子的缺陷度不高,即偏离Hardy—Weinberg平衡的程度不大。     

塔城盆地大鸨迁徙前期种群分布与集群大小研究

2014年10月采用样线法调查了塔城地区大鸨指名亚种Otis tarda tarda迁徙前期种群分布与集群特征。调查面积约2000 km2。调查期间共记录大鸨54群374只,主要分布于阿不都拉乡、恰夏乡和168团3个区域。受食物可利用度影响,迁徙前期塔城大鸨主要在冬小麦和春小麦留茬地中采食。不同分布区域和栖息地类型没有影响大鸨的平均集群大小。农田生态系统食物资源分布均匀,导致迁徙前期大鸨多集小群,偶集大群,以避免集大群而带来的资源竞争。     

基于转录组平台的蛤仔微卫星标记筛选

以菲律宾蛤仔转录组测序所得拼接序列为基础,采用MISA软件进行微卫星分析,对其中的145个微卫星位点进行引物设计,得到具有清晰扩增条带的微卫星位点58个。对大连庄河野生蛤仔群体的扩增结果表明,18个位点显示单态性,40个位点表现为多态性。该群体40个多态性微卫星位点得到的等位基因数在2—6之间,平均等位基因数为3.4250±0.9718,观测杂合度和期望杂合度分别在0.0000—1.0000和0.0615—0.7996之间,平均值分别为0.2727±0.2272和0.4739±0.1902,群体平均Nei指数为0.4664±0.1872。多态信息含量(PIC)在0.0586—0.7529之间,平均值为0.4148±0.1707,其中16个微卫星位点的PIC值大于0.5,为高度多态性,15个位点0.25PIC0.5,为中度多态性,其余9个为低度多态性。经Sequential Bonferroni校正的Hardy-Weinberg平衡检验,有10个位点尚未偏离平衡。基于转录组平台筛选微卫星标记的方法,在很大程度上推动了DNA分子标记的开发。研究开发的微卫星标记可用于蛤仔群体遗传学、遗传连锁图谱构建及其他相关研究,为蛤仔分子标记辅助育种及群体种质保护等工作提供技术支持。     

恒河猴群微卫星DNA多态性的分析

目的　确立一种对恒河猴群个体的遗传物质进行准确可靠、快速简便的遗传检测方法。方法　利用聚合酶链反应 (PCR)扩增技术对 2 0只恒河猴群个体间进行了DNA多态性的分析。结果　筛选出 9个微卫星DNA位点具有显著多态性 ,4个微卫星DNA位点没有多态性 ,还有 2个位点等位基因数目较少。结论　利用这些多态性微卫星位点建立一种对恒河猴群个体进行有效、准备可靠、快捷简便的遗传背景监测方法。     

Isolation and characterization of microsatellite loci in the great bustard,Otis tarda

With the aim to study population genetics of the endangered great bustard, Otis tarda, dinucleotide microsatellite loci were isolated using an adapted hybrid‐capture enrichment protocol. This work reports the characterization of a set of six polymorphic microsatellite markers within the great bustard (n = 52). Results from cross‐species amplifications in several other members of the family Otididae demonstrate that five primer pairs also successfully amplified homologous loci outside the species Otis tarda.     

Isolation and characterization of 19 novel microsatellite loci in Rhododendron aureum and Rhododendron brachycarpum (Ericaceae)

Nineteen polymorphic microsatellite loci are described for Rhododendron aureum Georgi (Ericaceae), an endangered species in Korea, and the closely related Rhododendron brachycarpum. The sequences containing repeat motifs were identified using low-depth next generation sequencing and 148 microsatellite loci were examined for amplification success and the detection of polymorphisms. All 19 loci were polymorphic and the number of alleles for these markers varied from 4 to 25, with an average of 15 alleles per locus. Three R. aureum loci showed departure from Hardy–Weinberg equilibrium (HWE) and one R. brachycarpum locus deviated significantly from HWE. These microsatellite loci will be useful for investigating the genetic diversity and population structure of both species.     

Isolation and characterization of 17 microsatellite loci for the Chinese longsnout catfish (Leiocassis longirostris)

We describe the isolation and development of 17 polymorphic dinucleotide microsatellite loci for the Chinese longsnout catfish (Leiocassis longirostris). All loci were polymorphic in the 30 individuals tested. The number of alleles per variable locus ranged from 6 to 18, with a mean of 11.71. Observed and expected heterozygosities ranged from 0.467 to 1.000 and from 0.540 to 0.929, respectively. Eight loci were found to have deviated from HWE in the sampled population after Bonferroni correction. Linkage disequilibrium tests revealed significant linkage between two loci (LLW5 and LL27). These microsatellite loci will be useful for revealing population structure, genetic diversity, and phylogeography of the Chinese longsnout catfish.     

Characterization of 11 novel polymorphic microsatellite loci in the threatened Korean loach,Iksookimia koreensis,isolated using a next-generation sequencing method

To investigate the genetic diversity and population structure of Iksookimia koreensis, we characterized 11 novel polymorphic microsatellite markers developed using next-generation sequencing. The number of alleles per locus ranged from 4 to 10 (average = 6.26). Observed and expected heterozygosities ranged from 0.333 to 0.866 and from 0.375 to 0.866, respectively. No loci showed significant deviation from Hardy–Weinberg equilibrium (HWE). These loci were also used successfully to study the genetic diversity of a closely related species, Iksookimia longicorpa. Four of the 11 loci amplified in the two species showed different allele frequency and distribution, indicating deep genetic divergence between I. koreensis and I. longicorpa. The newly developed microsatellite markers reported here will provide a useful tool for examining gene flow, population genetic structure, and genetic diversity of these species.     

Isolation and characterization of 12 polymorphic microsatellite markers for the frog Pelophylax hubeiensis

Twelve polymorphic microsatellite loci were isolated from an (AC)n- and (AG)n-enriched DNA library for the endemic Chinese frog Pelophylax hubeiensis (Ranidae). The number of alleles per locus ranged from two to eight, with a mean of 5.17. The observed and expected heterozygosities ranged from 0.226 to 0.839 and from 0.204 to 0.826, with means of 0.568 and 0.656, respectively. No significant linkage disequilibrium was detected among these loci. However, two significant deviations from HWE were discovered at loci Pehu-11 and Pehu-12 (P<0.05). MICRO-CHECKER tests showed that null alleles could be present at locus Pehu-12. These polymorphic microsatellite loci can be employed for exploring mating mechanisms, population genetic structure and other relevant genetic investigations of P. hubeiensis.     

内蒙古图牧吉冬季大鸨调查初报

大鸨(Otis tarda)属大型草原鸟类,过去曾广泛分布于黑龙江省西部及内蒙古的东部,目前已经处于濒危状态。图牧吉自然保护区是大鸨的主要栖息地,繁殖数量约200多只。1998年开始记录到越冬大鸨个体,2003年冬季,本区越冬数量达到165只(其中保护区内记录到85只)。本文对图牧吉自然保护区大鸨的越冬数量分布进行了调查,并对大鸨越冬行为及食性进行了初步的观察和分析,对大鸨越冬地管理及越冬鸟类的保护提出了建议。     

越冬期大鸨粪便氨基酸组成与消化能力分析

为评估陕西黄河湿地大鸨越冬期营养状况,于2012年1～2月收集大鸨新鲜粪便并对粪便中的蛋白质、灰分和氨基酸含量等做了测定.结果显示,粪便中粗蛋白、粗灰分含量分别为40.25％、19.81％.谷氨酸含量最高,为0.55％,组氨酸含量最低,为0.10％.粪便中豆苗、豆瓣、麦苗、麦粒、玉米、杂物的干物质比重分别为0.47％、4.05％、84.80％、6.75％、2.86％、1.07％.结果表明,麦苗是粪便干物质的主要成分,但相对大鸨越冬食性而言,麦苗粗蛋白含量偏低.粪便中谷氨酸、甘氨酸、天门冬氨酸和丙氨酸含量较大,这对优化大鸨对食物的选择方面起一定作用.     

Development of eighteen tetranucleotide microsatellite markers in Tibetan Macaque (Macaca thibetana) and genetic diversity analysis of captive population

In this study, eighteen tetranucleotide microsatellite loci were isolated from AAAG-enriched and GATA-enriched libraries of the Tibetan macaque, Macaca thibetana. These loci were tested on 53 individuals of M. thibetana, and most loci were proved to be highly polymorphic. A total of 109 alleles were detected with an average of 6.06 alleles per locus. The PIC values of these loci ranged from 0.192 to 0.879, with an average of 0.624. The observed and expected heterozygosities ranged from 0.170 to 0.800 and from 0.217 to 0.898, with an average of 0.583 and 0.675, respectively. 5 loci significantly deviated from Hardy–Weinberg equilibrium (HWE). Significant linkage disequilibrium (LD) was found between 9 pairs of loci. The newly identified polymorphic markers would facilitate the study of M. thibetana on the population structure and genetic diversity.     

应用微卫星DNA标记对Wistar和SD大鼠封闭群的遗传学研究

封闭群大鼠的遗传质量对其医学生物学实验结果有重要影响,但目前缺乏遗传检测方法和标准.本研究应用6个微卫星标记及其荧光标记一半自动基因分型技术,对北京和上海2家单位分别提供的Wistar和Spague-Darley(SD)大鼠封闭群进行了遗传检测和评估.6个微卫星位点均具有高度多态性,在两大鼠群体共发现等位基因36个,每位点等位基因数5-8个,其多态信息含量(PIC)从0.5892(D11Mgh3)到0.8019(D6Mit1),平均为0.688l.6个位点在Wistar和SD大鼠分别发现25和26个等位基因,其平均期望杂合度分别为O.6260和0.6249.两群体的各组遗传多样性指数间无显著差异.群体间的不同微卫星位点Fst范围0.046l到0.4363.平均为0.2069,表明其遗传分化程度较大;Nei(1972)遗传距离和Nei(1978)无偏遗传距离分别为1.2862和1.2726,表明了2群体之间较大的遗传差异:Hardy-Weinberg平衡检验表明Wistar大鼠在所有检测的6个位点均非常显著偏离Haraly-WeinJaerg平衡,SD大鼠在2个位点(D6Mit1和D11Mgh3)处于遗传平衡状态,且偏离位点均表现为杂合子缺陷.因此研究表明,Wistar和SD大鼠封闭群均具有较好的遗传多样性,且两群体之间有较大的遗传差异和分化程度,分别具有各自不同的遗传特征,偏离Hardy-Weinberg遗传平衡是其繁育过程中较多存在的问题.本研究结果将为两品系大鼠遗传检测方法和标准的建立提供基础资料和依据.     

Isolation and characterization of 16 microsatellite loci in the Great Tit Parus major

Six dinucleotide, three trinucleotide and seven tetranucleotide microsatellite loci developed for the great tit Parus major are presented. Thirty individual birds were screened at each locus. Loci were polymorphic (four to 19 alleles per locus). These markers provide a system to study paternity, genetic diversity in natural populations, gene flow, dispersal and inbreeding.