基于cyt b基因的长江中上游大鳞马口鱼遗传多样性及谱系生物地理学过程分析

以线粒体cyt b基因为分子标记,对长江上游干流及汉江等9条支流13个地理种群的大鳞马口鱼Opsari-ichthys macrolepis Yang et Huang进行遗传多样性及种群历史动态分析,并探讨其谱系生物地理学过程.结果显示,414尾大鳞马口鱼样本中共检测到79个单倍型,整体的单倍型多样性(h=0.930...     

西藏雪鸡青海亚种的种群遗传结构和地理变异

本文采用分子系统地理学的研究方法,对我国西藏雪鸡地理分布格局的形成原因进行了探讨。从4个地理种群的51个样品中成功地扩增出了535bp的线粒体DNA细胞色素b(mtDNACytb)片段,在所有可比较的535bp的序列中,发现15个变异位点,没有插入/缺失,且很大程度上倾向于转换(Transition)或颠换(Transversion),Ti/Tv=3.4∶1,并且变异位点多发生在密码子的第三位点,而密码子的第二位点没有变异位点。在4个地理种群中,共有13种线粒体DNA单倍型,且在各地理种群中都具有较高的单倍型多态性。AMOVA分析结果表明,西藏雪鸡4个地理种群间的遗传结构差异并不明显(ΦST=0.19,P<0.01),遗传差异主要发生在地理种群内,占80.75%,而且种内大部分分子变异是由各地理种群之间单倍型的差异引起的,单倍型缺乏比较明显的地理分布格局,在系统发生树上各地理种群中的单倍型相互散布在不同的分布群中,从而形成今天的分布格局。     

基于线粒体细胞色素b基因的黑鳍鳈(Sarcocheilichthys nigripinnis)生物地理学过程分析

黑鳍鳈(Sarcocheilichthys nigripinnis)为广泛分布于东亚地区的小型淡水鱼类,是生物地理学研究的良好材料。该研究以线粒体细胞色素b(Cyt b)基因序列为分子标记,对中国8个水系20个采样点的黑鳍鳈共142尾个体进行遗传变异及生物地理学过程分析。遗传多样性分析结果表明,142条Cyt b基因序列共检测出56个单倍型,总体单倍型多样性较高(h=0.971),而核苷酸多样性较低(π=0.0212),平均遗传距离较小(2.2%)。分子系统发育树结果表明,黑鳍鳈种群分为7个谱系,以秦岭为界可分为南、北两大支系,北方支系分化时间较早(谱系Ⅰ)而南方支系分化程度较高(谱系Ⅱ~Ⅶ)。该研究结果进一步揭示了克氏鳈(S.czerskii)的物种形成过程,估算其祖先在较晚近时期(~1.03 Ma)由黄河水系的黑鳍鳈分化而来。谱系生物地理学分析显示,黑鳍鳈的各地理种群表现为由北向南逐渐演化的趋势,地理隔离可能是限制该物种扩散和基因交流的主要原因。分子钟分析显示,分化时间发生于0.95~3.92 Ma。种群历史动态结果则推测黑鳍鳈经历过种群扩张,且该过程可能与更新世冰期与间冰期的更迭相关。     

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。     

基于线粒体COI基因序列的大豆食心虫中国东北地理种群遗传多样性分析

【目的】大豆食心虫Leguminivora glycinivorella (Matsumura)是一种危害大豆的主要害虫,在中国北方地区危害较重。本研究旨在探讨大豆食心虫在中国东北不同地理种群间的遗传变异。【方法】测定了10个不同地理种群153个个体的线粒体细胞色素氧化酶亚基Ⅰ (mtCOI)基因的657 bp序列,利用DnaSP 5. 0和Arlequin 3. 5. 1. 2等软件对大豆食心虫种群间的遗传多样性、基因流水平和分子变异进行分析。【结果】结果表明：10个地理种群间的COI基因共有36个变异位点和17个单倍型,其中1个单倍型为10个种群所共享。总种群的单倍型多样性指数Hd为0.456,各地理种群单倍型多样度范围在0~0.634之间。总群体的固定系数Fst为0.12545,遗传分化系数Gst为0.06326,总基因流Nm为3.49,且各种群间的基因流均大于1,种群间基因交流的水平较高。【结论】大豆食心虫种群内遗传多样性水平处于中低等水平。总群体和各种群的Tajima’s D检验结果皆不显著,说明中国东北地区大豆食心虫在较近的历史时期内没有出现种群扩张现象。AMOVA分子变异分析结果表明,大豆食心虫的遗传分化主要来自种群内部,而种群间未发生明显的遗传分化。各地理种群的单倍型在系统发育树上和中介网络图上散布在不同的分布群中,缺乏明显的地理分布格局。各种群的遗传距离与地理距离之间没有显著线性相关性,种群间的基因交流并未受到地理距离的影响。     

昆明城郊中国树鼩群体线粒体DNA遗传多样性

由于树鼩是灵长类动物的近亲,且具有体型小、繁殖周期短、饲养管理成本低等优点,长期以来被认为有望替代灵长类动物用于人类疾病的动物模型研究.然而,目前对树鼩的群体遗传结构还知之甚少,这极大地限制了其在疾病动物模型研究的应用,也是其品系资源创制的瓶颈.本研究通过分析80只采自于云南省昆明周边地区的野生树鼩(Tupaia belangeri chinensis)线粒体DNA(mtDNA)多态性,结合国外报道的2个树鼩(Tupaia belangeri)序列比较后发现,在604 bp的mtDNA控制区片段中兵检测到29个核苷酸替代变异,这些变异共界定了13种单倍型,表现较高的群体遗传多样度.另外,昆明地区的树鼩与国外报道的2个树鼩间存在较大的遗传分化,mtDNA控制区单倍型之间的核苷酸替换数大于18个,远高于昆明地区树鼢群体内部不同单倍型之间的差异.选择含有代表性的mtDNA控制区单倍型的17个昆明地区树鼩个体进一步测定了细胞色素b基因片段(1134 bp),结合前人报道的数据分析,结果进一步支持mtDNA控制区数据反映的遗传格局及揭示的昆明地区树询与国外报道树鼩之间的明显差异.本研究结果提示,昆明地区树鼩与国外树鼩之间存在较大遗传差异,在将树鼩用于人类疾病动物模型研究中要注意这些遗传差别.昆明城郊的树鼩群体具有较高的遗传多样度,在开展近交系建立等工作时须考虑选取群体内部具有代表性的mtDNA世系.

Abstract：

Due to their special phylogenetic position in the Euarchontoglires and close affinity to primates, tree shrews have been proposed as an alternative experimental animal to primates in biomedical research. However, the population genetic structure of tree shrews has largely remained unknown and this has hindered the development of tree shrew breeding and selection. Here we sampled 80 Chinese tree shrews (Tupaia belangeri chinensis) in Kunming, China, and analyzed partial mtDNA control region sequence variation. Based on our samples and two published sequences from northern tree shrews (T. belangeri), we identified 29 substitutions in the mtDNA control region fragment (～ 604 bp)across 82 individuals and defined 13 hapiotypes. Seventeen samples were selected for sequencing of the cytochrome b (Cyt b; 1134 bp) gene based on control region sequence variation and were analyzed in combination with 34 published sequences to solidify the phylogenetic pattern obtained from control region data. Overall, tree shrews from Kunming have high genetic diversity and present a remarkable long genetic distance to the two reported northern tree shrews outside China. Our results provide some caution when using tree shrews to establish animal models because of this apparent genetic difference. In addition, the high genetic diversity of Chinese tree shrews inhabiting Kunming suggests that systematic genetic investigations should be conducted before establishing an inbred strain for medical and biological research.     

基于线粒体细胞色素b基因的中国大沙鼠系统地理格局

通过内蒙、新疆、甘肃的41个大沙鼠样品和1个伊朗撒拉克大沙鼠的mtDNA Cytb基因全序列的遗传分析,对我国大沙鼠(Rhombomys opimus)的分子系统地理学进行了初步探讨。结果表明,我国41个大沙鼠样品的Cytb基因包含了50个核苷酸变异位点(占全序列的4.39%),其中转换48个,颠换2个,共定义23个单倍型。在四个地理种群中,内蒙古中部半荒漠区和阿拉善荒漠区的单倍型多样性最高,甘新荒漠区的单倍型多样性最低;北疆荒漠区的核苷酸多样性最高,内蒙古中部半荒漠区的核苷酸多样性最低。分子变异分析(AMOVA)表明,种群间的遗传变异占51.68%,种群内的遗传变异占48.32%。FST统计结果表明,除内蒙古中部半荒漠区与阿拉善荒漠区地理种群之间差异显著外(P<0.05),其它地理种群间的差异均极显著(P<0.01)。基于单倍型的系统树显示,42只大沙鼠形成三支。其中,伊朗撒拉克地区大沙鼠和中国地区大沙鼠之间的亲缘关系比中国两支大沙鼠之间的亲缘关系远;分析表明,中国分布的大沙鼠两支之间分歧时间估计在0.093Ma前。嵌套支分析表明,大沙鼠历史种群曾发生过异域片段化、受阻碍基因流和持续种群扩张事件。种群扩张分析提示大沙鼠在0.0119Ma前曾经历过一次种群扩张事件,种群可能受到末次冰期波动的影响。     

黑眶蟾蜍、黑斑蛙、中国雨蛙不同地理居群的染色体多样性

本文研究温州地区的黑眶蟾蜍、黑斑蛙、中国雨蛙的核型,分析了三个地理居群的黑星期五蟾蜍、四个地理居群的黑斑蛙、三个地理居群的中国雨蛙核型。结果表明不同地理居群的同种蛙有相同的染色体数和核型模式。黑星期五蟾蜍为2n＝22,NF＝44,核模式6＋5;黑斑蛙为2n=26,NF=52,核模式5＋8;中国雨蛙为2n＝24,NF＝48,核模式6＋6。但同一种蛙的不同地理居群之间在SM数目和顺序、次缢痕或随体的位置等有所不同。说明不同地理居群的同种蛙的染色体具有丰富的多样性。     

基于线粒体Cyt b基因的雉鸡甘肃亚种的种群遗传结构

采用聚合酶链式反应(PCR)和直接测序法分析雉鸡甘肃亚种的种群遗传结构。测序获得了8个种群79个样本的长819bp的线粒体细胞色素b(Cytb)基因序列,发现7个变异位点,界定了6种单倍型(Haplotype),其中Hap-1是共享单倍型,占79.75%。在8个种群中,除ZJC和CHX种群外,其它种群的核苷酸多样性和单倍型多样性都很低,种群间遗传分化小,基因流大。ZJC种群由于亚种间的基因渗透,导致其遗传多样性最高。CHX种群受徽成盆地周边山地的森林隔离作用,单倍型最多。分子方差分析(AMOVA)表明,遗传变异主要发生在种群内,占86.44%。种群间和地理种群组间差异不显著。地理种群组的核苷酸多样性(π)同形态上的变异规律一致。FS值检测表明,雉鸡甘肃亚种曾发生过快速扩张。     

黑眶蟾蜍、黑斑蛙、中国雨蛙不同地理居群的染色体多样性

本文研究温州地区的黑眶蟾蜍、黑斑蛙、中国雨蛙的核型,分析了三个地理居群的黑眶蟾蜍、四个地理居群的黑斑蛙、三个地理居群的中国雨蛙核型.结果表明不同地理居群的同种蛙有相同的染色体数和核型模式.黑眶蟾蜍为2n=22,NF=44,核模式6+5;黑斑蛙为2n=26,NF=52,核模式5+8;中国雨蛙为2n=24,NF=48,核模式6+6.但同一种蛙的不同地理居群之间在SM数目和顺序、次缢痕或随体的位置等有所不同.说明不同地理居群的同种蛙的染色体具有丰富的多样性.     

Adaptive changes of gastro-entero-pancreatic system endocrine cells in the black-spotted pond frog Rana nigromaculata after fasting

Changes in distribution density,morphology and secretory content of endocrine cells in the gastro-entero-pancreatic system of black-spotted frogs Rana nigromaculata before and after fasting were investigated using immunohistochemistry and antisera to six gut hormones.Six types of endocrine cells were detected in the digestive tract of Rana nigromaculata,including 5-hydroxytryptamine(5-HT),gastrin(GAS),somatostatin(SOM),glucagon(GLU),pancreatic polypeptide(PP)and vasoactive intestinal polypeptide(VIP)cells.A...     

Breeding ecology and oviposition site selection of black-spotted pond frogs (Rana nigromaculata) in Ningbo, China

The black-spotted pond frog (Rana nigromacu-lata) is one of the most widely distributed species in China. However, there have been only a few qualitative descrip-tions of their breeding ecology and oviposition site selec-tion. From 2004 to 2006, we investigated the breeding ecology and oviposition site selection of K nigromaculata in Ningbo, China, quantitatively. Analyses of breeding ecology show that: (1) mean frog density in the breeding season was 0.0903 ± 0.0029 individuals per meter (n = 11) (mean ± SE); (2) R. nigromaculata was a sexually dimorphic species, with females significantly larger than males in both body weight and snout-vent length; (3) the clutch size averaged 4643.04± 235.96 eggs (range 1546-7897, n = 50); and (4) the egg size ranged from 1.50 to 1.74 mm in diameter, with an average egg size of 1.6050 ± 0.0046 mm (n = 226). Oviposition sites differed significantly from random sites in percentages of water, bare ground and vegetation cover, water depth (cm), water temperature (°C) and water turbidity. Rana nigro-maculata preferred microhabitats with higher percentages of water and vegetation cover, while it avoided microha-bitats with deeper water. The results suggest that micro-habitats with higher percentages of water and vegetation cover, but not deeper water, should be priorities for pro-tection and conservation of the breeding habitats of R. nigromaculata.     

Phylogeography and genetic diversity of the red squirrel (Sciurus vulgaris) in China: Implications for the species' postglacial expansion history

The geographical distribution of Sciurus vulgaris spans much of the Palearctic from western Europe and the UK eastward to the pacific coast of East Asia. S. vulgaris occurs in China only in the far northwest and the northeast. Understanding of the species’ postglacial expansion history in East Asia has been limited by the paucity of molecular data. In this study, we used partial D-loop and cytochrome b gene sequences to assess mitochondrial DNA variation in S. vulgaris in China. Our objectives were to (1) determine phylogeographical patterns of S. vulgaris in China; (2) understand the species’ postglacial expansion history in this region; and (3) quantify genetic diversity levels within S. vulgaris populations in China. We identified a supported phylogenetic group in S. vulgaris from China, and found no tendency for haplotypes to cluster by geographic region. Our analysis of S. vulgaris from China and other regions supports the hypothesis that the Calabria region of southern Italy is a glacial refugium for the species. We tentatively propose a postglacial expansion pattern for the squirrels: migrating from Calabria via Central and Eastern Europe to Russia and from there to China, and firstly to the northwest and then to northeast in China. We found high levels of genetic diversity in S. vulgaris populations across China as a whole, and discussed its influential factors.     

黄河上游花斑裸鲤Cyt b基因的序列变异和遗传多样性

花斑裸鲤（Gymncypris eckloni）主要分布于黄河上游高原宽谷河段深水缓流处或静水湖泊中,在高原淡水生态系统的食物链中具有重要的地位。本文获得了黄河上游花斑裸鲤5个种群共68个个体线粒体DNA（mtDNA）细胞色素b基因的全序列（1 140 bp）,分析了序列变异和种群遗传多样性。68个序列经比对后,发现30个（2.63%）多态性位点,共定义了18个单倍型。结果显示,花斑裸鲤种群单倍型多样度和核苷酸多样度均低于其它鲤科鱼类,这可能与青藏高原经所经历的地质变迁和古气候环境的改变有关,由此生活在高原水域中的鱼类或多或少经历过瓶颈效应。AMOVA分析结果显示,遗传差异主要发生在种群之内,而不是来自不同地理组群间或组群内种群间。单倍型网络图和系统发育分析均没有显示出单倍型与地理位置的对应关系,提示黄河上游花斑裸鲤自然种群未出现分化,应作为一个整体进行保护。单倍型歧点分布呈现为单峰以及中性检验Fu’s Fs（&#8722;15.3400, P<0.001）和Tajima’s D（&#8722;0.6254, P =0.3080）结果综合表明,花斑裸鲤可能经历过近期的种群扩张事件。     

中国黄颡鱼的线粒体DNA多样性及其分子系统学

基于体侧色斑、背鳍前部形态、吻长及尾柄长的差异, Ng和Kottelat(2007)将分布于中国的黄颡鱼群体划为两个物种: 北方群体为Pseudobagrus sinensis, 南方群体为P. fulvidraco。本研究通过对70个黄颡鱼标本相关形态特征的测量及对线粒体cyt b基因序列的分析, 探讨了P. sinensis物种的有效性问题。结果表明: 依据体侧色斑和背鳍前部形态的差异, 可将黄颡鱼分为对应于P. sinensis和P. fulvidraco的两种形态类型, 但对尾柄长、吻长的测量发现二者没有差异。对70条cyt b基因序列的分析结果为: 两种鱼类有1个共同的单倍型; 两种鱼类的单系性在系统发育分析中都没有得到重现, 而二者聚在一起形成获得100%支持率的单系群; 两种鱼类群体之间存在持续的基因交流(Nm = 4.7); 两种鱼类在单倍型的巢式支系分析(nested clade analysis, NCA)中没有形成各自独立的进化谱系, 所有的单倍型以不超过5步的突变全部被纳入同一个进化网络中。因此我们认为P. sinensis不是有效物种, 而应被视为黄颡鱼的一种形态类型。基于cyt b基因的序列变异, 本研究对黄颡鱼群体的遗传多样性和种群结构作了初步分析。群体的核苷酸不配对分布及Tajima’sD中性检验表明, 约在10.1-14.1万年前, 黄颡鱼在其分布范围内经历过群体扩张, 推测这可能是导致黄颡鱼群体单倍型多样度高(h = 0.857 ± 0.0014)而核苷酸多样度低( π = 0.0023 ± 0.0003)的主要原因。此外, 分析结果显示黄颡鱼群体缺乏明显的地理结构, 推测原因可能是历史上水系的连通促进了不同地理群体之间的基因交流。     

Mitochondrial DNA Sequence Variation Suggests the Lack of Genetic Heterogeneity in the Adriatic and Ionian Stocks of Sardina pilchardus

A genetic stock structure analysis of 11 sardine samples from the Adriatic Sea and Ionian neighboring area was carried out through sequence variation analysis of a 307-bp cytochrome b gene fragment in order to identify self-recruiting units in the Adriatic Sardina pilchardus stock. The overall lack of genetic subdivision among samples detected by analysis of molecular variance, pairwise Φ_st values, and the exact test of population differentiation indicates this sardine stock is part of a larger self-recruiting population whose boundaries are larger than the investigated area. This conclusion is in agreement with preliminary allozymic and mitochondrial DNA restriction fragment length polymorphism data, but contradicts the previous identification of 2 subpopulations of sardines in the Adriatic Sea argued on morphologic differences, which could be rather attributed to different hydrographic or ecologic conditions occurring in different areas of the Adriatic Sea. The reduced gene flow observed between Adriatic-Ionian and Spanish sardine geographic samples (P < 0.001) suggests that reproductively isolated populations of sardines may occur in the Mediterranean Sea.     

黑麂皖-浙分布中心种群的遗传多样性

本文基于黑麂线粒体控制区序列对皖浙分布中心的4个黑麂种群的遗传多样性和基因流进行了研究。结果显示:4个种群的33个个体中有14个变异位点,占分析序列长度的2·91 %,并由此定义了12个单倍型;遗传多样性检测结果显示4个种群中开化种群具有最高的遗传多样性,应予以优先保护;4个种群间尚存在着一定的基因流,但存在可能由于遗传漂变而产生分化的危险。从Tajima’s D和Fu and Li’s D值的估算结果来看,这4个黑麂种群相对于中性进化的歧异度并没有明显的偏离,具有较为稳定的种群结构(P>0·1),没有明显的证据显示这4个黑麂种群间存在很强的平衡选择。     

Effects of habitat fragmentation on genetic diversity and gene flow among the Homidia socia (Collembola: Entomobryidae) populations in the Thousand Island Lake

In the present study, partial sequences of the mitochondrial cytochrome oxidase subunit I (COI) gene of 22 island populations of the springtail Homidia socia in the Thousand Island Lake were sequenced. Across all sequences, 37 haplotypes were identified for the 510‐bp mitochondrial (mt) DNA COI gene. Haplotype 2 was the most common, and was distributed in the most of the 22 island populations. Haplotype diversity ranged from 0.065 to 0.733, and the total genetic diversity was 0.56216. The genetic characteristics of the 22 island populations were analyzed using the fixation index and gene flow, with values of 0.00043–0.94900 and 0.02703–703.72540, respectively. Comparison between (island area and isolations) with population genetic diversity revealed that there were no significant correlations between them, except for a significant correlation between the number of haplotypes and island area. Mantel tests showed that there was no significant correlation between geographic distance and genetic distance among various groups. All the results indicated that there were no obvious relationships between island characteristics and the genetic diversity of the springtails. We consider that the low dispersal capacity of springtails and the island patches surrounded by water in the Thousand Island Lake are the major factors affecting the genetic diversity of H. socia.