四川裂腹鱼乌江种群mtDNA控制区序列的遗传多样性分析

采用PCR扩增和直接测序法分析了四川裂腹鱼乌江种群32个个体mtDNA控制区的序列差异和遗传多样性.在该种群mtDNA控制区长度为464 bp的同源序列中,共计有9个变异位点,占总位点数的1.94%.32个个体共定义了9种单倍型,单倍型间平均遗传距离(P)为0.0060,单倍型多样度(Hd)为0.768.核苷酸多样性(π)为0.00324,平均核苷酸差异数(K)为1.500.用单倍型间遗传距离构建的NJ分子系统树聚为两个分支.结果提示四川裂腹鱼乌江种群遗传多样性匮乏,保护四川裂腹鱼乌江种群刻不容缓.     

线粒体DNA全序列揭示罗布人母系遗传结构

[目的]探知罗布人mtDNA遗传结构与遗传多样性,构建人群mtDNA遗传数据库。[方法]运用二代测序技术对164位罗布人mtDNA全序列进行检测、分型。[结果]共检出315处突变位点包含298处多态性位点,其中A263G、A750G、A4769G、A8860G、A15326G位点突变率为100%,核苷酸多样性为0.0084,平均核苷酸配对差异数目为9.4695;划分出27个单倍型,高频单倍型有:B5b2c/31.71%、H6a1b2/18.90%、D4c2a/13.41%、H9a/9.76%,单倍型多样性为0.8360,个体识别力为0.8309;主成分和邻接系统树分析显示:罗布人与维吾尔族、蒙古族、中亚人群遗传关系近,特别与维吾尔族亲缘关系最近。[结论]现代罗布人mtDNA遗传结构具有亚欧混合现象,遗传多样性丰富;单倍型主要集中在B(39.03%)、H(29.88%)、D(15.85%);mtDNA遗传结构数据可为人群在遗传进化、族源鉴定、个体识别等方面提供依据。     

都柳江鲤、鲫和草鱼种群mtDNA控制区及遗传多样性分析

采用PCR和DNA测序技术对贵州都柳江鲤(Cyprinus carpio)、鲫(Carassius auratus)和草鱼(Ctenopharyngodon idellus)种群的mtDNA控制区序列及遗传多样性进行了研究。获得了都柳江鲤、鲫和草鱼mtDNA控制区长度分别为899~901 bp、787 bp和901~905 bp的序列。该3种鱼类控制区碱基A、T含量较高,G含量最低。识别了该3种鱼类mtDNA控制区终止序列区、中央保守区和保守序列区等保守序列。其中,除CSB-2和CSB-3碱基组成相同外,其余核心序列碱基组成存在着差异。都柳江鲤、鲫和草鱼种群mtDNA控制区分别有24、24和11个多态位点,分属12、17和8个单倍型。都柳江鲤、鲫种群遗传多样性较高,草鱼种群遗传多样性较低。因此,有必要开展都柳江草鱼种群遗传多样性的保护。     

陇东黄土高原石鸡的分子系统地理结构

本文运用聚合链式反应和直接测序的方法测定陇东黄土高原 8个石鸡 (Alectorischukar)地理种群 78个样本的mtDNA控制区 4 91bp ,建立其分子系统地理结构。 78个样本共发现 2 4个变异位点 ,2 5种单倍型。 8个地理种群共享单倍型C1 ,6个种群共享单倍型C2 ,种群间有一定的基因交流。 8个地理种群的 2 5种单倍型在NJ树中相互混杂 ,没有形成独立的地理结构 ,但聚成两个分支。单倍型网络图显示 2 5种单倍型聚成两个星状的集群 ,分别以单倍型C1和C2为中心。这两个群间的遗传差异显著。陇东黄土高原石鸡的系统地理结构属于“系统发生连续 ,具有部分空间隔离”的地理格局。这种地理格局是更新世冰川、泥石流和人类活动共同作用的结果     

斑翅草螽线粒体基因组序列测定与分析

已经测定的昆虫线粒体基因组中, 直翅目草螽亚科的疑钩额螽Ruspolia dubia线粒体控制区长度最短, 仅70 bp。为此, 本研究采用L-PCR结合二次PCR扩增策略对另一种草螽亚科昆虫斑翅草螽Conocephalus maculates线粒体基因组序列进行了测定。序列注释发现: 斑翅草螽线粒体基因组序列全长15 898 bp, A+T含量为72.05%, 基因排列与典型的节肢动物线粒体基因组一致。全部蛋白质编码基因以典型的ATN作为起始密码子, 9个蛋白质编码基因具有完整的终止密码子, 其余4个以不完整的T作为终止信号。除trnS^AGN外, 其余21个tRNAs均可折叠形成典型的三叶草结构, 依照Steinberg等(1997)线粒体特殊tRNA结构类型-9, trnS^AGN的DHU臂形成一个7 nt环, 反密码子臂则长达9 bp, 含1个突起碱基, 而不是正常的5 bp。斑翅草螽与其他直翅目昆虫线粒体基因组的主要区别在于, 在trnS^UCN和nad1, nad1和trnL^CUN基因间各存在一段罕见的、大段的基因间隔序列, 长度分别为78 bp和360 bp。其中, 位于nad1和trnL^CUN之间的基因间隔序列N链可形成一个包含完整起始、终止密码子(ATT/TAA)、编码103个氨基酸的未知开放阅读框。同义密码子使用偏好与线粒体基因组编码的tRNA反密码子匹配情况无关, 但与密码子第3位点的碱基组成紧密相关; 相对密码子使用频率(relative synonymous codon usage, RSCU)大于1的密码子, 其第3位点全部是A或T。在已经测定的直翅目昆虫线粒体基因组tRNAs中, 均存在一定数量的碱基错配, 且以G-U弱配对为主, 表明G-U配对在线粒体基因组中可能是一种正常的碱基配对形式。本研究测定的斑翅草螽线粒体基因组序列, 和先前已经测定的直翅目线粒体基因组序列一起, 可以为重建直翅目的进化历史提供数据资源。     

汉族人线粒体DNA单倍型与散发型帕金森病关系的研究

目的：研究汉族人线粒体DNA（mtDNA）单倍型与散发性帕金森病（PD）的关系。方法：用酚／氯仿法从65例PD患者和50名健康者（对照组）的外周静脉血中提取基因组DNA,通过斑点杂交方法对PD患者和对照组的10个单核苷酸多态性（SNP）位点（G1719A,G4580A,C7028T,G8251A,G9055A,A10398G,A12308G,G13366A,C13708T,G16391A）和单倍型进行检测。结果：PD组G1719A突变5例、G9055A突变2例、C13708T突变4例,对照组G1719A突变1例,2组的突变率无显著性差异（P〉0.05）。PD组H单倍型54例（83.1％）,其他11例（16.9％）;对照组H单倍型49例（98.0％）,其他1例（2.0％）。结论：10398G不是汉族人散发性PD发病的易感因子,也没有保护作用;不宜用H、I、J、K、T、U、V、w和x等9个mtDNA单倍型来评估汉族人散发性PD发病风险。     

异刺草螽线粒体基因组序列分析

本研究采用高通量测序技术对异刺草螽的基因组进行测序,并组装得到了完整的线粒体基因组序列。结果显示:异刺草螽线粒体基因组序列全长16 038 bp,包含13个蛋白质编码基因、22个t RNA基因、2个r RNA基因和1个控制区。异刺草螽线粒体基因组的总碱基组成如下:A为37.3%、C为15.4%、G为10.3%、T为36.9%,A+T含量较高,为74.2%。异刺草螽线粒体基因的排列与祖先序列相同,该线粒体基因组序列为直翅目螽斯科的系统发生和进化研究提供了重要的分子基础。     

四种草螽雄性鸣声的研究

对分布于我国的长瓣草螽Conocephalus(Anisoptera)gladiatus,中华草螽C.(Amurocephalus)chinensis,悦鸣草螽 C.(Anisoptera)melaenus和斑翅草螽C.(Anisoptera)maculatus雄性鸣声进行了分析.研究结果表明,这4种草螽雄性鸣声的时域波形较简单,鸣声均由1种类型的脉冲组序列构成.长瓣草螽雄性鸣声每个脉冲组持续时间约0.13 s,脉冲组间隔约0.12 s,每个脉冲组通常由8脉冲构成,鸣声的频率范围5～20 kHz.中华草螽雄性的鸣声脉冲组由2个脉冲构成,每次鸣叫持续时间约为3.3 s,两次连续鸣叫间隔约10.5 s,鸣声频率范围为20 Hz～20 kHz.斑翅草螽雄性鸣声的脉冲组由10～13个脉冲构成,脉冲组持续时间2.1～2.5 s,两次连续鸣叫间隔时间约为3 s;鸣声频率从5.5 kHz到高于20 kHz.悦鸣草螽雄性鸣声由单一规则的重复脉冲组序列构成,每个鸣声脉冲组持续时间约0.035 s,脉冲组间隔约0.023 s,每个脉冲组由3个脉冲构成,脉冲组重复率20/s,鸣声频率6.0～20.0 kHz.     

小口白甲鱼都柳江种群mtDNA D环的序列变异及遗传多样性

采用PCR结合DNA测序技术,测定分析了易危鱼类小口白甲鱼(Onychostoma lini)都柳江种群36个个体mtDNA D环约470bp序列的变异及遗传多样性。结果表明,在36个个体中,该序列的长度为469～475bp,其碱基组成为A+T的平均含量(68.4%)高于G+C(31.6%)。共检测到25个多态位点,其中转换19个、颠换6个。核苷酸多样性(π)为0.00575,平均核苷酸差异数(K)为2.695。36个个体分属5个单倍型,单倍型多样度(Hd)为0.260,单倍型间的平均遗传距离(P)为0.026。5个单倍型构建的UPGMA系统树聚为2个分支。目前小口白甲鱼都柳江种群mtDNA D环序列存在着较丰富的变异和遗传多样性。     

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 structure of Oryza rufipogon Griff. in China

Oryza rufipogon Griff. (common wild rice; CWR) is the ancestor of Asian cultivated rice (Oryza sativa L.). Investigation of the genetic structure and diversity of CWR in China will provide information about the origin of cultivated rice and the grain quality and yield. In this study, we used 36 simple sequence repeat (SSR) markers to assay 889 accessions, which were highly representative of whole germplasm in China. The analysis revealed a hierarchical genetic structure within CWR. First, CWR has diverged into two ecotypic populations, a south subtropical population (SSP) and a middle subtropical population (MSP), probably owing to natural selection by the different climates. The distribution of specific alleles and haplotypes indicated that Chinese CWR had both indica-like and japonica-like variations; the SSP was an indica-like type, whereas the MSP was more japonica-like. The SSP and MSP further diverged into five (HN, GD-GX1, GX2, FJ and YN) and two (JX-HuN1 and HuN2) geographical populations, respectively. The genetic data suggest the isolation by distance, although water systems also appear to play an important role in the formation of homogenous populations, and occasionally landscape was also involved. The population GD-GX1, which grew widely in Guangdong and Guangxi provinces, was the largest geographical population in China. It had a high level of genetic diversity (GD) and the closest genetic relationship with other inferred populations. The population HN, with the smallest SSR molecular weights and the highest level of GD, may be the most ancestral population.     

Chloroplast phylogeography of a desert shrub,Calligonum calliphysa (Calligonum,Polygonaceae), in arid Northwest China

In order to investigate the influence of Pleistocene climate oscillations and paleogeographic events on the evolutionary history of xerophytic plants in arid Northwest China, the phylogeography of a well adapted desert shrub, Calligonum calliphysa (Calligonum, Polygonaceae), was studied. A total of 11 natural populations and 90 individuals were sampled. Two cpDNA intergenic spacer regions (rpl32-trnL and ycf6-psbM) were sequenced and 13 haplotypes were identified. Significant genetic differentiation was found among populations and groups, suggesting that short distance seed transfer and geographic isolation have restricted gene flow. Based on SAMOVA analysis, the 11 populations were divided into 3 clades with no haplotypes were shared between these. The time of divergence within C. calliphysa was estimated at between 2.36 and 0.18 Ma, during the early to middle Pleistocene, and for Clade 3, in the range of 0.88–0.29 Ma. The largest number of populations (7) and haplotypes (9) were found in Clade 3. We found fragmentation of genetic variation, with most unique haplotypes among populations located at the edges of the Gurbantunggut Desert, which may have occurred because vegetation there was sensitive to habitat variation caused by climate change, and we detected that a demographic expansion event followed the expansion of this desert.     

Genetic diversity and maternal origin of Northeast African and South American donkey populations

To investigate the mtDNA variation and origin of maternal lineages in South American donkeys and to reassess the domestication of donkeys in northeast Africa, we analyzed sequences (489 bp of the D‐loop) from 323 domestic donkeys sampled from Peru, Brazil, Ethiopia and Egypt. Altogether, the 323 sequences displayed 53 different haplotypes (45 in Ethiopia, 14 in Egypt, eight in Peru and six in Brazil). Among the four populations, Egyptian donkeys possessed the highest haplotype diversity (0.910 ± 0.032), followed by Brazilian donkeys (0.879 ± 0.060). The Clade I haplotypes dominated in Peruvian donkeys (65%), whereas Clade II haplotypes dominated in Brazilian donkeys (67%). Estimates of F_ST values showed a high genetic differentiation between Peruvian and Brazilian donkey populations (F_ST = 0.4066), which could be explained by the complex introduction history of South American donkeys. Phylogeographic analysis indicates that northeast Africa could be the most probable domestication center for Clade I donkeys. Analysis of molecular variance confirmed a weak genetic structure in domestic donkey populations among four continents (Europe, Asia, Africa and South America).     

Genetic structure and differentiation of four lake populations of Coilia ectenes (Clupeiformes: Engraulidae) based on mtDNA control region sequences

To investigate genetic structure and differentiation of four lake populations of Coilia ectenes, a total of eighty-one individuals from four largest freshwater lakes (Dongting [DT], Poyang [PY], Chaohu [CH], Taihu [TH]) were analyzed based on mtDNA control region sequences. Seventy-five haplotypes were detected, and four to seven 38 bp tandem repeats were found. The indexes of nucleotide diversity (π) in these four populations were DT 0.78%, PY 1.09%, CH 0.81%, and TH 0.62%. Genetic distances within populations were between 0.63 and 1.14%, and from 0.76 to 1.60% among populations. A Neighbor–Joining (NJ) tree consisted of three major clades. Clade 1 consisted of 88.89, 26.09, and 4.55% individuals of DT, PY, and CH. Analysis of molecular variance (AMOVA) showed that most variances occurred within populations, suggesting that this is the main source of total variance. Our results suggest that the four lake populations of C. ectenes have not developed significant genetic structure.     

Assessment of genetic diversity and population structure of swamp eel Monopterus albus in China

Swamp eel has become one of the most economically important fish in China. However, the wild swamp eel is facing the serious challenge of declining population and germplasm degeneration because most of farming swamp eel fingerlings was collected by fishing wild individuals. In this study, the genetic variation of Monopterus albus in six dominant farming regions was investigated based on the mitochondrial DNA D-Loop of 1008 bp in length. 180 individuals from 6 populations were examined and 74 haplotypes were observed. The overall genetic diversity was abundant and which its SD population was highest but CQ population was lowest. There was obvious genetic differentiation among investigated populations. Phylogenetic analysis revealed that these individuals were divided into four distinct genetic clades, clade A, B, C, and D. Clade A should be the most common ancestor clade. AH and CQ populations might originate from one single ancestor in maternal clade A. Clade C should be a native important clade in China. Though the genetic diversity did not suffered obvious decreasing, it is still imperative to take effective conservation measurements and establish an efficient selective breeding program.     

Identifying Source Populations and Genetic Structure for Savannah Elephants in Human-Dominated Landscapes and Protected Areas in the Kenya-Tanzania Borderlands

We investigated the genetic metapopulation structure of elephants across the trans Rift Valley region of Kenya and Tanzania, one of the remaining strongholds for savannah elephants (Loxodonata africana) in East Africa, using microsatellite and mitochondrial DNA (mtDNA) markers. We then examined this population structure to determine the source population for a recent colonization event of savannah elephants on community-owned land within the trans rift valley region. Four of the five sampled populations showed significant genetic differentiation (p<0.05) as measured with both mtDNA haplotypes and microsatellites. Only the samples from the adjacent Maasai Mara and Serengeti ecosystems showed no significant differentiation. A phylogenetic neighbour-joining tree constructed from mtDNA haplotypes detected four clades. Clade four corresponds to the F clade of previous mtDNA studies that reported to have originated in forest elephants (Loxodonta cyclotis) but to also be present in some savannah elephant populations. The split between clade four and the other three clades corresponded strongly to the geographic distribution of mtDNA haplotypes across the rift valley in the study area. Clade four was the dominant clade detected on the west side of the rift valley with rare occurrences on the east side. Finally, the strong patterns of population differentiation clearly indicated that the recent colonists to the community-owned land in Kenya came from the west side of the rift valley. Our results indicate strong female philopatry within the isolated populations of the trans rift valley region, with gene flow primarily mediated via male movements. The recent colonization event from Maasai Mara or Serengeti suggests there is hope for maintaining connectivity and population viability outside formal protected areas in the region.     

Multisource noninvasive genetics of brown bears (Ursus arctos) in Greece reveals a highly structured population and a new matrilineal contact zone in southern Europe

In human‐dominated landscapes, connectivity is crucial for maintaining demographically stable mammalian populations. Here, we provide a comprehensive noninvasive genetic study for the brown bear population in the Hellenic Peninsula. We analyze its population structuring and connectivity, estimate its population size throughout its distribution, and describe its phylogeography in detail for the first time. Our results, based on 150 multilocus genotypes and on 244‐bp sequences of the mtDNA control region, show the population is comprised by three highly differentiated genetic clusters, consistent with geographical populations of Pindos, Peristeri, and Rhodope. By detecting two male bears with Rhodopean ancestry in the western demes, we provide strong evidence for the ongoing genetic connectivity of the geographically fragmented eastern and western distributions, which suggests connectivity of the larger East Balkan and Pindos‐Dinara populations. Total effective population size (N _e) was estimated to be 199 individuals, and total combined population size (N _C) was 499, with each cluster showing a relatively high level of genetic variability, suggesting that migration has been sufficient to counteract genetic erosion. The mtNDA results were congruent with the microsatellite data, and the three genetic clusters were matched predominantly with an equal number of mtDNA haplotypes that belong to the brown bear Western mitochondrial lineage (Clade 1), with two haplotypes being globally new and endemic. The detection of a fourth haplotype that belongs to the Eastern lineage (Clade 3a1) in three bears from the western distribution places the southernmost secondary contact zone between the Eastern and Western lineages in Greece and generates new hypotheses about postglacial maxima migration routes. This work indicates that the genetic composition and diversity of Europe''s low‐latitude fringe population are the outcome of ancient and historical events and highlight its importance for the connectivity and long‐term persistence of the species in the Balkans.     

Population genetic diversity of Chinese sea bass (Lateolabrax maculatus) from southeast coastal regions of China based on mitochondrial COI gene sequences

Chinese sea bass (Lateolabrax maculatus), an important commercial and recreational fishery species in China, whose natural resources have decreased dramatically for decades due to overfishing and environmental changes. To provide guidelines for the source conservation and management, genetic diversity and population structure of L. maculatus were analyzed based on the mitochondrial cytochrome oxidase subunit I (COI) gene. Totally 192 individuals were sampled from five locations: Qingdao (QD), Chongming (CM), Dongtou (DT), Lieyu (LY) and Fangcheng (FC). After sequencing of a 586 bp fragment of COI gene, 20 haplotypes were defined. H4 (haplotype 4) and H8 were the dominant haplotypes and existed in all populations. Haplotype diversity (Hd) and nucleotide diversity (π) of each population ranged from 0.572 to 0.721 and from 0.00129 to 0.00271, respectively. The highest Hd and π were found in DT and LY populations, while the lowest value of Hd and π were in the QD population. Genetic distance ranged from 0.0031 to 0.0027 within populations and from 0.0018 to 0.0035 between populations. Analysis of molecular variance (AMOVA) revealed that significant genetic divergence was found in QD and FC populations. In addition, neutrality tests and mismatch distribution analysis indicated that this species experienced potential population expansion events.     

基于cpDNA trnL-F序列的胡黄连保护遗传学研究

胡黄连为特产中国-喜马拉雅特有高山植物,作为常用中、藏药材,受到灭绝性采挖,作为濒危和二级保护植物亟待科学的保护。该研究以云南和西藏7个野生居群91个个体为材料,基于 cpDNA trnL-F 非编码序列测序分析胡黄连的遗传多样性和遗传结构,分析显著进化单元,确立优先保护居群并提出科学的保护策略。结果表明：胡黄连 trnL-F 序列长度为871~876 bp,根据序列的核苷酸变异共鉴定出5个单倍型,西藏占有2个单倍型,云南占有3个单倍型,西藏和云南2个地区的所有单倍型均不共享。胡黄连具有较低的单倍型多样性(Hd ＝0.43419)和核苷酸多样性(Dij ＝0.00466)。种群间分化度(Fst ＝0.864520)和基因流(Nm ＝0.04)、居群间的遗传分化水平(GST ＝0.916)、AMOVA 分析(0.78％的遗传变异发生在居群内,60.97％的遗传变异发生在地区内居群间,38.25％的遗传变异发生在地区间)均表明,胡黄连居群间存在明显遗传分化。多数一致性树将胡黄连划分为3个进化分支(Ⅰ、Ⅱ、Ⅲ),这3个分支均与地理相关,分支Ⅰ分布于横断山区的4个居群,分支Ⅱ是分布于东喜马拉雅的一个居群,分支Ⅲ是分布于喜马拉雅中段的2个居群。3个分支分属于3个“进化显著单元(ESU)”。这3个 ESU 中白马雪山、茨中、定日、波密、聂拉木五个居群都需要保护,建议现阶段应优先保护的居群是云南白马雪山和西藏波密居群,以就地保护为主。     

New insights into the origin and the genetic status of the Balkan donkey from Serbia

The Balkan donkey (Equus asinus L.) is commonly regarded as a large‐sized, unselected, unstructured and traditionally managed donkey breed. We assessed the current genetic status of the three largest E. asinus populations in the central Balkans (Serbia) by analysing the variability of nuclear microsatellites and the mitochondrial (mtDNA) control region of 77 and 49 individuals respectively. We further analysed our mtDNA dataset along with 209 published mtDNA sequences of ancient and modern individuals from 19 European and African populations to provide new insights into the origin and the history of the Balkan donkey. Serbian donkey populations are highly genetically diverse at both the nuclear and mtDNA levels despite severe population decline. Traditional Balkan donkeys in Serbia are rather heterogeneous; we found two groups of individuals with similar phenotypic features, somewhat distinct nuclear backgrounds and different proportions of mtDNA haplotypes belonging to matrilineal Clades 1 and 2. Another group, characterized by larger body size, different coat colour, distinct nuclear gene pool and predominantly Clade 2 haplotypes, was delineated as the Banat donkey breed. The maternal landscape of the large Balkan donkey population is highly heterogeneous and more complex than previously thought. Given the two independent domestication events in donkeys, multiple waves of introductions into the Balkans from Greece are hypothesized. Clade 2 donkeys probably appeared in Greece prior to those belonging to Clade 1, whereas expansion and diversification of Clade 1 donkeys within the Balkans predated that of Clade 2 donkeys.