云南中缅树鼩线粒体细胞色素b和D-loop区遗传多样性研究

对3个地方种群的49只样本的线粒体Cyt b基因全序列（1140 bp）及33只样本的控制区D-loop基因区段（745 bp）进行了序列测定。结果表明：Cyt b基因多态性位点有47个,其中单变异位点位点23个,简约信息位点24个。共定义了24个单倍型,其中种群间的共享单倍型有2个（8.33%）,其余均为某个种群所特有,单倍型多样度范围为0.80952（剑川种群）~0.91532（禄劝种群）,核苷酸多样度指数介于0.00326（禄劝种群）~0.00635（剑川种群）之间;D-loop基因多态性位点有18个,其中单变异位点8个,简约信息位点10个。共定义了16个单倍型,无种群间的共享单倍型,单倍型多样度范围为0.76615（禄劝种群）~0.93333（丽江种群）,核苷酸多样度指数介于0.00269（禄劝种群）~0.00583（丽江种群）之间。从各单倍型的TCS网络进化图显示横断山种群位于分支的末端,表现出中缅树鼩由南向北的扩散模式,支持＂岛屿起源＂假说。     

汉寿中华鳖线粒体细胞色素b基因克隆及其遗传多样性分析

本文以汉寿中华鳖为研究材料,比较分析中华鳖线粒体细胞色素b基因(Cytb)与瑞鳖、砂鳖等的差异,为探讨其进化与遗传分子标记奠定实验基础。采用PCR技术,克隆汉寿中华鳖Cytb基因的序列,并检测其在不同组织以及不同生长发育时期的表达模式,最后通过分子进化树分析中华鳖与砂鳖的亲缘关系。通过克隆发现中华鳖细胞色素b基因的mRNA开放阅读框含有1 140 bp,其碱基组成为A+T的含量(61. 1%)高于G+C(38. 9%),编码379个氨基酸残基的序列。与山瑞鳖和砂鳖氨基酸序列比较发现,中华鳖与瑞鳖和砂鳖均存在一定差异; Cytb基因在中华鳖不同组织和各个不同发育时期中均有表达,且在精巢组织中表达量最高,在不同时期阶段呈"低-高-低"曲线表达模式;分子进化树分析发现汉寿中华鳖与砂鳖具有较强的亲缘关系,但是在Cytb中仍存在差异,所以能够很好的把它们区分开来。此外汉寿中华鳖与哺乳动物、鱼类和爬行类均存在巨大差异。结果表明,Cytb基因在探讨汉寿中华鳖的进化与遗传分子标记中具有重要意义,可作为分析中华鳖种属进化系统发育特性和群体遗传多样性的标记基因。     

应用RAPD标记技术对树鼩遗传多样性的分析

目的建立树鼩RAPD（random amplified polymorphic DNA）遗传标记分析方法,了解中缅树鼩种群的遗传多样性。方法采用PCR技术对40条随机引物进行优化,筛选出能有效用于树鼩群体遗传分析的RAPD位点,对48只树鼩个体的基因组DNA进行PCR扩增,并应用Popgene 1.32与RAPDistance Package Version 1.04等软件进行数据处理,分析树鼩的群体遗传特性。结果20个RAPD引物共检测到113个位点,平均每个引物可扩增出5.65个位点,其中多态位点数为69个（占61.1%）。个体间的遗传相似系数平均为0.8307,个体间遗传距离在0.09-0.27之间,平均遗传距离为0.1693。雄性群体的遗传多态度（H0）（0.1864）略高于雌性群体（0.1470）,平均遗传多态度（Hpop）为0.1667;树鼩遗传多态度所占的比例在群体内为48.29%,而雌、雄群体间为51.71%。NJ法进行聚类分析得出T15、T33和T47号树鼩个体聚类成一大类,其余45只树鼩个体聚成另一大类,雌、雄个体呈相互交叉现象。结论实验所筛选的随机引物可有效用于中缅树鼩种群的遗传结构分析。本树鼩群体具有较好的遗传多样性。     

基于线粒体DNA细胞色素b基因和控制区序列分析西藏雅鲁藏布江黄斑褶鮡种群遗传多样性

采用线粒体DNA（mtDNA）Cyt b基因和D-loop控制区为分子标记,对分布于西藏雅鲁藏布江大峡谷以上里龙段和以下墨脱段2个群体的黄斑褶 （Pseudecheneis sulcata）共60个样本进行遗传多样性研究。获得联合基因有效序列长度为1 893 bp,包括Cyt b基因1 060 bp和D-loop控制区833 bp。结果显示,里龙和墨脱2个群体的单倍型多样性值（Hd）均较高（0.701和0.761）,核苷酸多样性值（π）均较低（0.001 00和0.001 09）;高频率的单倍型Hap1和Hap2为2个群体所共享,推测为祖先单倍型;同时,里龙和墨脱群体分别存在5个和6个特有单倍型,且在2个群体中不共享;分子方差分析（AMOVA）显示遗传变异主要来源于种群内部,群体间呈中度遗传分化水平（Fst = 0.090 44,P < 0.05）;中性检验（Tajima''s D、Fu''s Fs）和核苷酸不配对（SSD、Hir）分析结果揭示,黄斑褶 种群曾经历过种群扩张现象。本研究推测,黄斑褶 2个群体间的基因流动存在障碍,雅鲁藏布大峡谷的海拔落差及水文情势等生态屏障可能是阻碍黄斑褶 迁徙和交流的主要原因。     

浙江省金华市中华蜜蜂线粒体DNA遗传多样性研究

为了解浙江省金华市中华蜜蜂的遗传多样性及遗传分化现状,基于mtDNA tRNA1eu-COⅡ序列对金华市9个区县市共81个中蜂样本进行序列测定,分析了金华市中蜂群体的遗传多样性、遗传结构和遗传分化情况.结果共发现14个单倍型,其中主体单倍型JH1为9个地理群体所共享,与已报道的福建省中蜂主体单倍型以及浙江省丽水市的中蜂...     

长江中上游银[鱼句]线粒体DNA遗传多样性分析

银[鱼句]（Squalidus argentatus）是长江常见小型鱼类,分布广泛于干支流,资源量较大。本研究采用线粒体DNA（mtDNA）细胞色素b基因序列对长江中上游干流江津、荆州、监利、洪湖、黄石江段及支流赤水河、湘江等7个江段共217尾银[鱼句]样品的遗传结构和遗传多样性进行了分析。结果显示,银[鱼句]群体Cyt b基因序列分别检出了81个变异位点和97个单倍型,平均单倍型多样性（Hd）和核苷酸多样性（Pi）分别为0.966和0.008 6。分子方差分析（AMOVA）显示,长江中上游银[鱼句]群体存在显著遗传分化。群体间两两比较分析发现,赤水河群体与其他群体基因交流程度低,遗传分化显著,其他群体间没有显著遗传分化。中性检验结果不支持银 历史上发生过群体扩张。     

中国内蒙古鄂伦春族线粒体DNA高变区Ⅰ和高变区Ⅱ遗传多态性

收集50份鄂伦春族无关人群外周血样本, 用ABI PRISM377测序仪对其mtDNA HVRⅠ和HVRⅡ进行测序, 计算多态性位点数、单倍型数目、单倍型频率、平均核苷酸差异数目等多态性指标; 结合已发表的其他民族mtDNA遗传资料, 根据Nei法计算鄂伦春族与各群体之间的遗传距离, 进行聚类分析, 绘制系统发生树。鄂伦春族群体mtDNA两个高变区与CRS序列比对, 分别发现52和24个多态性位点, 分别界定了38和27种单倍型, 单倍型多态性分别为0.964±0.018和0.929±0.019; 平均核苷酸差异分别为7.379和2.408; 用HVRⅠ序列多态性数据计算Fst和dA两种遗传距离, 相关系数r为0.993(P<0.01); 基于HVRⅠ序列的系统树显示鄂伦春族与中国台湾、南方汉族和中国香港人群遗传距离较近, 与北方汉族、蒙古族及其国外人群遗传距离相对较远。我国鄂伦春族人群mtDNA具有相对独特的遗传特征, 其遗传多态性和个体识别力较高, 可用于民族起源、迁徙、法医学个体识别等领域研究。     

基于线粒体细胞色素b基因序列的版纳鱼螈四个种群遗传多样性与遗传结构研究

版纳鱼螈Ichthyophis bannanicus 是蚓螈类在我国分布的唯一物种,目前已知分布在云南、广西和广东的部分地区.本研究获得4个版纳鱼螈种群的87个个体的线粒体DNA细胞色素6基因1 000 bp序列,对序列变异、遗传多样性、种群结构和种群动态进行了分析.经比对后87个序列发现39个(3.9%)多态性位点,共定义了22个单倍型.序列分析结果显示版纳鱼螈种群单倍型多样性和核苷酸多样性与其它两栖动物接近,说明版纳鱼螈各种群仍保持一定的遗传多样性水平.单倍型网络关系图和系统发生分析均显示来自云南和两广地区的版纳鱼螈分歧明显,已形成了不同的进化谱系.种群遗传结构分析也显示来自云南的勐腊种群与两广各种群之间显著的遗传差异,这提示分布在云南与两广地区的版纳鱼螈应作为不同的管理单元(MU)进行保护.综合两种中性检验(Fu's Fs=9.44,P=0;Tajima's D=1.88,P=0.013)和种群增长指数(g=5415.03±297.55)分析的结果表明,两广地区的版纳鱼螈可能经历近期的种群扩张事件.     

贵州省东方蜜蜂Apis cerana线粒体DNA遗传多样性研究

为探明贵州省东方蜜蜂Apis cerana遗传多样性及遗传分化现状,对贵州省9个地级市79个样点采集的东方蜜蜂样本线粒体DNA (mt DNA) COI-COII序列进行扩增与测序,分析其遗传多样性、遗传结构及亲缘关系。序列结果显示扩增后COI-COII序列长度约为760 bp,A+T平均含量（82.3%）明显高于C+G（17.7%）,碱基偏移性明显。单倍型分析共发现33个单倍型,其中主体单倍型H1为9个地理群体共有。单倍型多样度分析表明平均单倍型多样度为0.802,其中黔南地区最高,遵义地区最低;平均核苷酸多样度为0.014,其中六盘水地区最高,黔西南及安顺地区最低。中性检验结果表明贵州省部分地区东方蜜蜂可能经历过群体扩张。9个地区样点的平均遗传距离介于0.0012~0.0044之间,其中毕节与黔东南地区间最大,为0.0044,安顺与黔南、遵义地区间最小,均为0.0012。在单倍型系统进化树中,79个样点东方蜜蜂可分为两个类群。研究结果为贵州省东方蜜蜂遗传分化研究、种质资源保护与开发利用提供参考依据。     

家养双峰驼线粒体DNA遗传多样性的研究

利用Ａｐａｌ、Ａｖａｌ、ＢａｍＨ、Ｂｃｌ１、Ｂｇｌ、、Ｂｇｌ２、Ｄｒａ１、ＷＥｃｏＲ１、ＥｃｏｒＶ、Ｈｉｎｄ３、Ｋｐｎ、、Ｐｕｖ２、Ｓａｅ１、Ｓｅａ１、Ｓｍａｌ、Ｘｂａ１、Ｘｈｏ１１８种限制制性内切酶的ｍｔＤＮＡ限制睛段长度多态（ＲＦＬＰ）分析,对我国双峰驼３个类群的遗传多样性及其亲缘关系进行了研究。结果表明,在双峰驼ｍｔＤＮＡ蝇未发现长度变异和个体内异质现象,从３个类群的８７个个体中,共检出了３     

Genetic diversity and population structure of Schizopygopsis younghusbandi Regan in the Yarlung Tsangpo River inferred from mitochondrial DNA sequence analysis

The genetic diversity and population structure of Schizopygopsis younghusbandi from six sites in the middle reach of the Yarlung Tsangpo River were examined based on mitochondrial DNA cytochrome b (Cyt b) gene and control region (D-loop) sequences. 1141 bp of complete Cyt b sequences and 737 bp of partial D-loop sequences for 153 individuals of S. younghusbandi were obtained by using PCR amplification and sequencing. The results showed that S. younghusbandi populations had high haplotype diversity and low nucleotide diversity, and the population genetic diversity of this species was at a low level. Analysis of molecular variance revealed that most genetic variation occurred within populations, and that genetic differentiation was at low or moderate levels. The network of haplotypes based on Cyt b gene showed that there were two groups within the examined populations. Neutrality tests and mismatch distribution analysis suggested that this species might have undergone a population expansion and the expansion time was estimated to be 0.25–0.46 Ma BP. All these results would be crucial to establish scientific strategies for effective conservation and sustainable exploitation of wild resources of S. younghusbandi.     

Genetic diversity and structure of the noxious alien grass Praxelis clematidea in southern China

Praxelis clematidea (Asteraceae), a plant species native to South America, is a noxious weed in southern China. We examined the genetic variation and population structure of 12 populations (76 individuals) of P. clematidea from Fujian, Guangdong, and Hainan Provinces in China using inter-simple sequence repeat (ISSR) analysis. From an initial set of 69 ISSR primers, 10 were selected which yielded 80 reproducible bands. Polymorphic bands (P) were 100%, Shannon's information index (I) was 0.4226, and Nei's gene diversity (H) was 0.2791. We infer that the high levels of genetic diversity exhibited by P. clematidea may have contributed to its invasiveness. Gene flow among populations was 2.4930, which has led to homogenization. The coefficient of population differentiation (Gst = 0.1671) indicated low levels of genetic variation among populations and high levels of genetic polymorphism within populations. There was a negative correlation between population elevation and genetic diversity, while there was a significant positive correlation between genetic distance and geographic distance based on a Mantel test (r = 0.5820, P < 0.01). Some populations from different provinces clustered together in principal coordinate and UPGMA analyses indicating that human-mediated events may have contributed to the dispersal of the species.     

基于粪便DNA的东北马鹿遗传多样性

东北马鹿(Cervus canadensis xanthopygus)为国家二级重点保护野生动物,近些年其种群数量急剧下降、分布区不断退缩、种群基因交流受阻,很多地区更是难觅其踪迹。亟需对其种群的遗传变化,特别是遗传多样性和近交衰退等种群遗传信息开展进一步评价,增强保护与管理的针对性。本研究在大、小兴安岭和长白山脉的6个重点研究区域,共收集409份疑似马鹿粪便样本。首先基于mtDNA Cyt b基因测序技术进行物种鉴定,并对鉴定为马鹿的阳性样本利用微卫星技术进行个体识别。结果共识别出172只东北马鹿个体;Cyt b基因序列共检测出14个变异位点和11个单倍型,单倍型多样性为0.849 (0.105～0.732),核苷酸多样性为0.678%(0.099%～0.775%)。10个微卫星位点检测出种群平均等位基因数为5.7 (5.2～7.2),有效等位基因数为3.3 (2.5～4.1),观测杂合度为0.687 (0.644～0.725),期望杂合度为0.619 (0.564～0.689),近交系数为-0.113 (-0.160～-0.037)。结果表明,东北马鹿种群遗传多样性处于中等水平,其...     

Genetic diversity of mitochondrial cytochrome b gene in Chinese native buffalo

The origins of the domestic water buffalo remain contentious. To better understand the origins of Chinese water buffalo, we sequenced the complete mitochondrial cytochrome b (MT-CYB) gene from 270 individuals representing 13 Chinese domestic swamp buffalo populations. We found genetic evidence of introgression of river buffalo into Chinese swamp buffalo herds. Swamp buffalo haplotypes can be divided into two highly divergent lineages (A and B), suggesting that Chinese native swamp buffalo have two maternal origins. We found that the A→G transition in the buffalo MT-CYB gene stop codon resulted in buffalo haplotypes being terminated by one of two stop codons: AGA or AGG. AGA is common to river buffalo and lineage A of swamp buffalo, while AGG is specific to lineage B of swamp buffalo. Lineage A appears to have been domesticated in China. Further genetic evidence is required to clarify the origins of lineage B.     

环境因子对新疆鹅喉羚群体遗传多样性的影响

环境因素在物种进化和遗传变异过程中起着非常重要的作用。为探讨新疆鹅喉羚遗传多样性与环境因子的关系,本研究采用聚合酶链式反应（PCR）和直接测序的方法,测定了新疆鹅喉羚11个群体84份样本的线粒体DNA Cyt b基因（1140 bp）和D-loop区（1100 bp）序列,分析各群体遗传多样性及环境因子对遗传多样性的影响。结果显示新疆鹅喉羚具有较高的单倍型多样性,较低的核苷酸多样性,表明其遗传多样性处于较低水平。环境因子与群体遗传多样性的相关性分析结果表明,海拔、年均降水量、年均气温、人口数量是影响新疆鹅喉羚遗传多样性的主要环境因子,其中海拔是最关键的环境因子。本研究结论为新疆鹅喉羚群体有效合理的保护与管理提供理论依据。     

黑龙江省张广才岭北部小飞鼠的遗传多样性与种群历史动态

小飞鼠 (Pteromys volans) 为树栖夜行滑行类啮齿动物,在森林种子传播和维持生态系统平衡等方面发挥着重要的生态学作用。本研究利用mtDNA Cytb、控制区和nDNA微卫星3种分子标记,对黑龙江省张广才岭北部的小飞鼠种群进行遗传多样性与历史动态分析。检测出Cytb全序列 (1 140 bp) 的平均单倍型多样性为0.909,平均核苷酸多样性为0.616%;控制区全序列 (1 066 bp) 的平均单倍型多样性为0.945,平均核苷酸多样性为1.698%;微卫星检测出种群平均等位基因数13.167个,观测杂合度0.727,期望杂合度0.864,近交系数0.159。结果表明,小飞鼠种群遗传多样性丰富,但存在一定程度的杂合度不足和近亲繁殖;未检测到种群近期遗传瓶颈效应,种群内无遗传分化。高比例的稀有单倍型 (≥ 60%) 、低频率等位基因与近亲繁殖,提示未来种群面临遗传多样性下降的风险,建议加大对该物种的关注和保护力度。基于Cyt b基因的系统进化关系结果表明,小飞鼠存在3个明显的遗传谱系：远东、欧亚大陆北部和日本北海道,本研究中张广才岭和大兴安岭的样本单倍型归属为远东谱系。     

Genetic diversity of natural populations of Larix principis-rupprechtii in Shanxi Province,China

Prince Rupprecht's Larch (Larix principis-rupprechtii Mayr.) is one of dominant components of middle and high elevation forests in North China. Shanxi Province is well known as “the Hometown of Prince Rupprecht's Larch” in China. In this study, six natural populations of this species across Shanxi were selected to investigate the genetic variation of the species using amplified fragment length polymorphism (AFLP) markers. Results showed that in comparison with some other species of Larix, higher genetic diversity was revealed at the species level for L. principis-rupprechtii (percentage of polymorphic loci PPL = 71.9%, Nei's gene diversity H_E = 0.225, Shannon information index I = 0.341). Most of genetic variation existed within populations (80.5%), while the genetic differentiation among populations was significant (p < 0.001) and higher (G_st = 0.194) than most other species of Larix. The differentiation can be attributed to the limited gene flow (N_m = 1.035) among populations, which could be due to the spatial isolation and habitat fragmentation. The six populations can be divided into three groups based on the Nei's genetic distances between populations (from 0.033 to 0.076). There was no significant correlation (r = 0.268, p > 0.05) between genetic distance and geographic distance among populations. The measures for in-situ or ex-situ conservation should be taken to preserve the genetic diversity of this species.     

Genetic assessment of captive Chinese water deer populations in Zhoushan Archipelago,China

Chinese water deer is a rare and vulnerable animal in China because of the poaching for medical use and the habitat loss. In this study, the genetic diversity and population genetic structure of 40 Chinese water deer from three populations in Zhoushan Archipelago were investigated with ten highly polymorphic microsatellite loci, including 4 screened from the nuclear DNA in the study, and 6 selected from the literature. According to the results, these captive populations had a higher genetic diversity than other rare cervid species, such as forest musk deer. No signs of inbreeding were detected. Low genetic differentiation among these populations was found. The probable reasons included the isolation by distance, the exchange among islands, or the supplement of the wild Chinese water deer. We proposed the deer raisers to strengthen the exchanges from different islands or mainland, and if possible, some deer would be returned to the wild to expand the wild population.     

Genetic diversity and structure of natural Pinus tabulaeformis populations in North China using amplified fragment length polymorphism (AFLP)

Chinese pine (Pinus tabulaeformis carr.), endemic to China, is a conifer species with extensive and fragmented distribution in North China. In this study, the genetic diversity and structure of 20 natural populations of this species were investigated using amplified fragment length polymorphism (AFLP) markers. A total of 445 fragments were revealed with 8 pairs of primers, 379 (85.17%) of which were polymorphic. A moderate level of genetic diversity was detected at the species level (Shannon's information index I = 0.356, Nei's gene diversity H_E = 0.271) and at the population level (I = 0.219, H_E = 0.206). Most of genetic variation was within populations while a considerable level of genetic differentiation was detected (G_ST = 0.352, Ф_ST = 0.304). The high differentiation could be attributed to the complex and fragmented habitats, and a limited gene flow among populations (N_m = 0.572). The Mantel test indicated that there was significant correlation (r = 0.455, P < 0.001) between Nei's genetic distance and geographical distance among all the populations. The results suggested that proper countermeasures should be taken to prevent the habitat further deterioration and maintain the genetic diversity of this species.