青藏高原特有植物青海当归的谱系地理学初探

为探讨第四纪冰期气候波动对青藏高原多年生草本植物青海当归地理分布格局形成的影响,检测了青海当归16个居群147个个体的核糖体内转录间隔区(ITS)的序列变异,共发现了7个单倍型.结果表明除松潘居群外其余15个居群都拥有单倍型H1,青藏高原边缘的居群拥有6种ITS单倍型,其中5种为私有单倍型.谱系关系分析显示单倍型H1为较古老的单倍型,AMOVA分析表明,居群间的变异占了总变异量的67％,居群遗传分化显著,并且存在较明显的谱系地理结构(NST＞GST,P＜0.05).我们认为青海当归在第四纪冰期最大一次冰期来临时退缩到青藏高原边缘的避难所,冰期后在扩张过程中经历了奠基者效应形成了如今的分布格局.     

横断山区高山绣线菊的谱系地理学研究

横断山区作为青藏高原东南部主要的一个冰期避难所,第四纪冰期气候的变化对该地区的植物地理分布和居群遗传结构都产生了重要的影响。为了揭示该地区物种分布的分子系统地理学结构,选取在该地区广泛分布的一种高山灌木-高山绣线菊的叶绿体trnL-trnF序列进行研究。采集了15个居群182个个体进行测序,共发现7个单倍型。总的遗传多样性较高(HT=0.809),但居群内遗传多样性较低(HS=0.236)。分子变异分析(AMOVA)结果表明分布区内高山绣线菊的遗传变异主要存在于居群间(84.48%),且居群间的遗传分化很高(GST=0.708,FST=0.84476,NST=0.863),有着显著的谱系地理学结构(NST>GST,P<0.01)和较低的居群间平均基因流(Nm=0.09)。单倍型的系统进化树和进化分支网络分析得到了相似的拓扑结构,7种单倍型都按照地理分布聚为三支:横断山区西部、横断山区东部以及两者的交接地带。本研究推测该物种在横断山区存在多个冰期避难所,而没有表现出大规模的种群集体扩张和迁移的现象。青藏高原隆升、第四纪气候的反复波动以及横断山区特殊的地理环境使得原来连续的居群片段化,并发生范围扩张,从而塑造了高山绣线菊的现代生物地理分布格局。     

横断山地区海仙报春的谱系地理学研究

由于横断山地区特殊的地理效应,第四纪冰期气候的反复变化对该地区植物的地理分布和居群遗传结构产生重要影响。为了揭示该地区分布的物种的谱系地理结构,检测了该地区特有的、生于湿地环境的植物海仙报春（Primula poissonii）的叶绿体trnL—F和trnT—L两个序列变异。共对13个居群167个个体的两个叶绿体片段进行测序,共发现11个单倍型。三种单倍型被共享,其他单倍型都只存在于单个居群内。分子变异分析（AMOVA）结果表明分布区内海仙报春的遗传变异主要存在于居群间（99．08％）,且居群间的遗传分化很高（GST=0．916,FST=0．99077,NST=0．973）,有着极显著的谱系地理学结构（NST〉GST,P〈0．01）和较低的居群间平均基因流（Nm=0．08）。结合嵌套分支分析（NCA）,本文的研究结果推测该物种在冰期可能存在多个避难所,并没有表现出大多数温带植物在第四纪冰期所经历过的种群集体扩张和迁移的现象,异域片段化被认为是该物种现有单倍型分布格局形成的主要原因。海仙报春的谱系地理结构是气候的反复波动和横断山特殊的地理环境互相作用的结果。     

高山植物条纹狭蕊龙胆的分子亲缘地理学研究

草本植物由于较短的生活周期以及对环境变化的敏感性,可能会更好地揭示第四纪冰期以来植物居群变化的历史过程.分子亲缘地理学研究是揭示动植物居群历史的有力工具,但到目前为止对青藏高原草本植物的分子亲缘地理学研究几乎是空白.因此,本文选择在青藏高原及邻近地区生长的一年生高山草本植物条纹狭蕊龙胆为研究对象,进行了13个居群155个个体的叶绿体基因组(cpDNA)非编码片段trnH(GuG)-psbA基因间区序列变异检测,共发现7种单倍型,其中单倍型H印A是分布最广的,而单倍型Hap E、Hap F和Hap G是拥有的居群所特有的.青藏高原东北部、东部及邻近地区的每个居群拥有的单倍型非常单一,而高原东南部横断山区的单倍型分布很集中,遗传多样性也相对较高.分子变异分析(AMOVA)结果表明整个分布区条纹狭蕊龙胆的遗传变异主要存在于居群间(73.05%),且居群问的遗传分化很高(GsT=0.805,FsT=0.731,NsT=0.859),有着显著的亲缘地理学结构(NsT>Gsr,P<0.05)和较低的居群间的平均基因流(Nm=0.184).结合巢式支系法分析(NCA),根据本文的研究结果推测青藏高原东南部横断山区是该植物第四纪冰期时可能的避难所,而且在间冰期或冰期后.伴随着异域片段化和过去片段化从避难所发生范围扩张而形成当前单倍型及居群的分布格局.     

唐古特虎耳草谱系地理学研究

利用叶绿体基因(trnL-trnF和rpl16)对青藏高原地区的18个唐古特虎耳草(Saxifraga tangutica Engl.)居群(209个个体)进行谱系地理学研究,以揭示唐古特虎耳草的现有遗传结构及其历史演化过程。结果表明:(1)从209个个体中共检测到74个单倍型,且只有单倍型H5在居群中广泛分布,71.62%的单倍型为居群特有单倍型。(2)分子变异分析(AMOVA)显示,91.85%的遗传变异来源于居群内,居群间遗传分化不明显(F_(ST)=0.081);遗传分化系数N_(ST)(0.109)大于G_(ST)(0.097,P0.05)但不显著,表明唐古特虎耳草在其整个分布范围内没有明显的谱系地理结构。(3)中性检验表明,Tajima’s D(-2.045 07,P0.05)和FuLi’D*值(-3.629 27,P0.05)均为显著的负值,结合单峰的错配分布曲线,表明该物种经历过近期扩张。研究推测,唐古特虎耳草在第四纪冰期时可能存在多个微型避难所,由于第四纪冰期气候的反复波动,使得原来连续的居群片段化,避难所内的居群单独进化,从而形成了大量特有单倍型;唐古特虎耳草居群对第四纪冰期气候波动的反应可能更多的表现为垂直海拔高度的迁移,而非大规模的水平迁移。     

基于叶绿体DNA变异研究高山植物偏花报春的种内谱系地理结构

横断山地区是许多温带植物的冰期避难所。为揭示该地区分布物种的亲缘地理结构, 检测了该地区特有、分布相对较为普遍的偏花报春Primula secundiflora的叶绿体trnL-trnF和rps16区序列变异。研究了11个居群109个个体, 一共发现了15种单倍型。只有一种单倍型为3个居群所共有, 其他单倍型都只存在于单个居群内。总的遗传多样性较高(HT=0.966), 但居群内遗传多样性较低(HS=0.178)。尽管种内形态十分一致, 居群间却存在高水平的遗传分化(FST=0.976)。NST (0.982)显著高于GST (0.816), 表明偏花报春在居群间存在明显的亲缘地理结构。单倍型聚成四个主要的分支: 三个分支的单倍型分布在北部, 而另一分支的单倍型分布在南部。四个分支的隔离分布表明该物种在冰期存在多个避难所。未发现在其他温带物种中广泛存在的间冰期或者冰期后物种分布范围的统一扩张现象。但是, 在气候变迁过程中由于居群增长-缩小反复发生, 多数居群的遗传多样性降低。这些推断也被巢式分支分析所证实, 距离隔离而导致的限制性基因流以及异域片断化被认为是该物种现有单倍型分布格局形成的主要原因。这种独特的谱系地理结构主要是由于气候变迁与该地区复杂的地质环境相结合造成的。     

青藏高原东北部黄缨菊的谱系地理学

黄缨菊（Xanthopappus subacaulis）是青藏高原地区一种特有的高山草甸药用植物。为探讨第四纪冰期气候波动对黄缨菊居群遗传结构和空间分布格局的影响,对黄缨菊20个居群、123个个体的叶绿体DNA片段（psbA-trnH、rbcL和psbI-psbK）进行测序和数据分析。结果表明：黄缨菊居群共检测到6个单倍型,其中H1为古老单倍型,除居群P7外其余居群均具有单倍型H1,H3、H5和H6为特有单倍型,单倍型H3为居群P7的私有单倍型,单倍型H5和H6只存在于居群P18,单倍型H2和H4主要存在于青海湖流域的居群;总的遗传多样性（H_e）和核苷酸多样性（π）分别为3.101和0.008 903;居群间遗传变异（68.98%）大于居群内遗传变异（31.02%）,居群间遗传分化较高（F_ST=0.689 85,P<0.01）;居群遗传分化系数N_ST（0.727）大于G_ST（0.656）（P>0.05）,表明黄缨菊在分布区域内不存在明显的谱系地理结构;错配分布和中性检验结果显示,黄缨菊居群可能经历过近期扩张。据此,推测第四纪冰期黄缨菊可能在青海湖流域和甘肃临潭地区存在微型避难所,认为第四纪气候变迁及青藏高原隆升塑造了黄缨菊的现代地理分布格局。     

基于叶绿体基因序列rbcL-accD研究莛子藨的分子遗传多样性和谱系地理结构

莛子藨是莛子藨属的一种多年生草本植物,主要分布在青藏高原东北部及邻近高山地区和日本,为东亚特有种。本文基于叶绿体DNA基因间隔区rbcL-accD序列,对莛子藨物种水平的序列特征、分子遗传多样性和谱系地理结构进行研究。分析结果发现该区域莛子藨种群的遗传变异主要来自于居群间（72.28%）,优势单倍型（H2）广泛分布,特有单倍型分散在多个居群。居群间遗传分化系数N_STST,没有检测到显著谱系地理结构,而且居群间基因流较低（N_m=0.096）,青藏高原及其邻近地区特殊地形和生态环境造成的地理隔离或者生境异质化可能是现有单倍型分布格局形成的原因。种群动态历史分析显示莛子藨经历过不显著的种群选择或者扩张事件,综合分析推测莛子藨居群应该是在间冰期或者冰期后从不同避难所扩散形成现在的分布格局,具体避难所还需进一步深入研究才能确认。     

南方红豆杉谱系地理学研究

通过对南方红豆杉18个居群499个个体叶绿体基因组的trnL-F片段进行RFLP分析,共检测到6个叶绿体DNA单倍型,其中除台湾居群外的17个居群都包含有单倍型H1和H2;黄山及临安居群除单倍型H1、H2外,还分别各有1个特有单倍型。谱系关系分析显示,单倍型H1、H2为较古老的祖先单倍型。AMOVA分析显示,变异主要存在于居群内,居群间的遗传分化程度较低(26.39%)。嵌套分支分析(NCA)显示,单倍型分布与地理分布存在轻度的地理相关性,南方红豆杉现在的遗传结构可能源于居群的片段化效应,台湾居群可能是由于曾经的居群片段化而隔离或长距离扩散所致;在第四纪冰期,中国东、南部地区可能存在2个以上南方红豆杉的避难所。     

基于叶绿体DNA trnL-F序列研究肋果沙棘的谱系地理学

由于青藏高原的地理效应,第四纪冰期气候的反复变化应对现今该地区生物的地理分布及其居群遗传结构产生重大影响。肋果沙棘Hippophae neurocarpa是青藏高原地区的一个特有种,根据叶上的附属物（星状鳞毛或者鳞片状鳞毛）分为两个亚种：肋果沙棘亚种subsp.neurocarpa和密毛肋果沙棘亚种subsp.stellatopilosa。依据母系遗传的叶绿体DNA片段对该物种谱系地理学进行研究不仅能阐明该物种冰期避难所,而且对于理解两个亚种的母系分化也具有重要意义。共对两个亚种14居群的70个个体的trnL-F序列进行了测序,共发现8种单倍型,其中5种单倍型出现在肋果沙棘亚种中,4种出现在密毛肋果沙棘亚种中,两个亚种共享一种单倍型。种内谱系分化与两个亚种形态上的分化不一致。嵌套进化分支把8种单倍型分为三支：一支为肋果沙棘亚种,其他两支中两个亚种单倍型嵌套组成,且肋果沙棘亚种处于进化末端。我们的研究结果还表明肋果沙棘在冰期可能在高海拔地区存在多个避难所,并且存在来自避难所的邻域扩张。     

Phylogeographical Study on Eriophyton wallichii (Labiatae) from Alpine Scree of Qinghai Tibetan Plateau

In this study, we examined the nuclear ribosomal DNA internal transcribed spacer (ITS) sequence variation of Eriophyton wallichii, a perennial endemic to alpine scree of the Qinghai Tibetan Plateau. Nineteen haplotypes were detected by analyzing 187 individuals from 20 populations of E.wallichii. Most populations hold the unique haplotype, which showed different population with different haplotype. An analysis of molecular variance (AMOVA) for populations showed that the genetic variation mainly resided among populations (89.54%), the level of differentiation among populations was very high (GST=0.863; NST=0.957) but didn’t show a significant phylogeographical structure. It is different from foregoing other species reported in Qinghai Tibetan Plateau that the haplotypes of E.wallichii showed a distribution pattern that almost one population hold one unique haplotype, hardly share same halotype among populations.We hypothesized the unique genetic structure of E.wallichii might result from allopatric fragmentation in the “islands of alpine” due to the extremely isolated alpine scree and rough topography of the region, through the Quaternary climate oscillation dramatically caused population expanding or shrinking.     

Intraspecific differentiation of Allium wallichii (Amaryllidaceae) inferred from chloroplast DNA and internal transcribed spacer fragments

In the present study, we used two chloroplast DNA (cpDNA) fragments (trnL-F and rps16) and the nuclear ribosomal internal transcribed spacer (ITS) sequence data to examine intraspecific differentiation and phylogeographical history of Allium wallichii. Based on wide scale sampling (28 populations and 174 individuals) across the entire distribution range of this species, 33 cpDNA haplotypes and 25 ITS ribotypes were detected in our investigation. These cpDNA haplotypes were divided into three major lineages, which was further supported by the ITS phylogenetic results. High haplotype/ribotype diversity and population differentiation, together with most of the haplotypes/ribotypes being exclusive to single populations, implied restricted gene flow among populations and significant geographical isolation. Nearly all populations with high haplotype/ribotype diversity were found in the Hengduan Mountain Region (HMR), whereas the populations of the Himalayas and Nanling Mountains showed a lower level, suggesting the HMR might serve as a potential divergence center for A. wallichii. The main lineages of A. wallichii diverged from each other between Mid–Late Pliocene and Early Quaternary based on two sets of molecular markers, indicating that the Quaternary climatic fluctuation could not have contributed greatly to the divergence of the main lineages of A. wallichii. Instead, the intricate topography and heterogeneous habitats resulting from the drastic uplift of the Qinghai–Tibet Plateau from the Late Pliocene could be responsible for the intraspecific differentiation of A. wallichii. The present study further highlights the importance of geographic isolation and habitat heterogeneity in shaping and maintaining high species diversity within the HMR.     

基于AFLP分子标记技术的黑花韭与多星韭遗传多样性与亲缘关系分析

采用AFLP分子标记技术分析了分布在滇中、滇西、滇西北的3个黑花韭居群、3个二倍体多星韭居群和6个四倍体多星韭居群的遗传多样性与亲缘关系.结果表明,12个居群在DNA水平上已发生明显分化.居群间的基因分化系数(GST)为0.444.由GST计算的居群间基因流(Nm)为0.625,表明居群间部分基因流动受阻.12个居群间...     

中国小菜蛾群体遗传变异的时空动态

【目的】小菜蛾 Plutella xylostella 是十字花科蔬菜最重要的迁移性害虫之一,在中国越冬区(OR)的群体和季节性繁殖区(SBR)的群体之间具有“源-汇”关系和复合群体的特征。本研究旨在揭示中国小菜蛾群体遗传变异的时空动态,以期进一步阐明该虫的迁移特性及其对区域灾变的影响。【方法】基于已经发表的4组不同采样时间中国小菜蛾 COI 序列(共1 567条)数据,将每个数据集细分为3组:OR群体、SBR群体以及特定采样时间(ST)的群体。对不同组(群体)进行遗传多样性、遗传分化、单倍型相似性和群体演化史的分析。【结果】基于 COI 序列的小菜蛾群体遗传变异分析结果表明,同一采样时间内,OR群体和SBR群体遗传多样性水平相当。虽然SBR群体在单倍型频率上不稳定,导致包含SBR的群体间遗传分化较大,但SBR群体单倍型均能够在同一采样时间的OR群体中找到,或SBR单倍型仅与OR单倍型相差2个碱基以内;SBR群体中单倍型频率在不同年份间变化大,遗传分化最大,而ST群体之间遗传分化的变异水平最低。不同采样时间样品 COI 序列的碱基错配分布图呈现相同的单峰状,且不同采样时间样本主要是由几种相同的单倍型构成。【结论】这些结果表明:迁移是影响中国季节性繁殖区小菜蛾群体遗传变异的主要因素;中国不同区域的小菜蛾群体遗传变异在时间尺度上具有稳定性和相关性。小菜蛾复合群体特征为阐明其季节性迁移特性提供了重要的生态学依据。     

云南宾川县鸡足山地区多星韭的生态型

根据生态学和遗传学的综合研究,鸡足山地区的８个多星韭种群可划分为６个生态型：１个柳叶韭生态型,１个岩生二倍体生态型和４个四体倍生态型即林生四倍体生态型,岩生四倍体生态型,草生白花四倍体生态型和草生紫花四倍体生态型。     

Reproductive biology of a mycoheterotrophic species, Burmannia wallichii (Burmanniaceae)

The breeding system of a mycoheterotrophic species, Bumannia wallichii (Miers) Hook. f. (Burmanniaceae), is assessed using field observations, floral anatomy, pollen histochemistry and isozyme electrophoresis. The structure of the flower effectively prevents pollinators from accessing the stamens, and no pollinators were observed visiting natural populations in Hong Kong. The pollen is starch-rich but lipid-poor. Analysis of isozyme variation revealed a small proportion of polymorphic loci, a low level of total heterozygosity ( H _T= 0.1972), and a very high overall fixation index of populations ( F _IT= 0.6602). These results strongly suggest that B. wallichii is predominantly self-pollinated. The coefficient of gene differentiation ( G _ST) is low (0.05), although it is suggested that this is due to genetic homogenization of populations consonant with the formation of numerous tiny 'diaspora' seeds that are wind dispersed.     

我国辣椒种质资源eIF4E基因多态性分析

利用国家蔬菜种质资源库的1904份辣椒资源材料,采用测序技术获得eIF4E(eukaryotic translation initiation 4E)基因exon1序列,研究e IF4E基因多样性及我国辣椒种质资源群体多样性。结果表明:在1904份材料中共发现17个单倍型,14个有义多态性位点,其中9个为新的位点,位点大多集中在eIF4E蛋白表面环上;8个地理群体的平均单倍型多样性(Hd)和平均核苷酸多样性(Pi)分别为0.519和0.00210;群体间分化指数(Fst值)及基因流(Nm)表明不同群体间表现差异的分化程度;AMOVA分析表明总变异主要来源于群体内个体间的变异(97.23%),只有2.77%变异发生在群体间。本研究将有助于了解我国辣椒eIF4E基因多样性,为抗PVY育种提供更多抗源材料。     

Chloroplast DNA variation within prairie cordgrass (Spartina pectinata Link) populations in the U.S.

Chloroplast DNA (cpDNA) is most often maternally inherited and highly conserved leading to previous observation of little to no sequence variation. Comparing cpDNA haplotypes have provided valuable insight into the establishment and migration of polyploid populations. However, to use chloroplast haplotypes to their full potential intrapopulational variation needs to be addressed. In this study, cpDNA haplotype variation was surveyed within 16 natural populations of prairie cordgrass (Spartina pectinata Link) located east of the 100th west meridian and north of the 35th north parallel in the U.S.A. using two non-coding, polymorphic chloroplast regions. Two main clades were defined with subclades as follows: haplotype 1 and haplotype 2A and 2B. It was discovered that seven populations showed intrapopulational chloroplast genome variation. Of the total amount of variation, 95.5% occurred within the octoploid populations and 4.5% occurred within the tetraploid populations. Both variant haplotypes, 2A and 2B, were found in a larger sampling of one of the natural populations, but no variation was found in a mixed ploidy population. The intrapopulational cpDNA variation we found in this study cannot directly be related to mechanisms of introduction of the non-native populations into native populations. Therefore, this cpDNA variation could be novel natural variation that has been fixed as the octoploid populations were established and moved northwest. This analysis provides insight into determining the usefulness of indels and single nucleotide polymorphisms for population identification and may provide information in regards to the origin of chloroplast variation and its subsequent fixation and establishment in natural prairie cordgrass populations.     

Genetic variation of mitochondrial DNA in Chinese populations of Pectinophora gossypiella (Lepidoptera: Gelechiidae)

The pink bollworm Pectinophora gossypiella is an invasive pest insect that has successfully established populations in many cotton growing regions around the world. In this study, the genetic diversity and population structure of Chinese populations of P. gossypiella were evaluated using mitochondrial DNA sequence data (COII and Nad4). For comparison, individuals of Pakistan and America were also sequenced at the same two mtDNA regions. Extremely low genetic variation was observed in the two mitochondrial regions among all populations examined. Most of the populations harbored only one to two haplotypes. Although the Nad4 region showed relatively high haplotype diversity and nucleotide variation, ranging from 0.363 to 0.591 and from 0.00078 to 0.00140, respectively, there were only three haplotypes observed in this region. COII and Nad4 haplotype networks shaved one or two common haplotype(s) forming the center of a star-shaped phylogeny. Pairwise tests showed that most of the populations were not significantly differentiated from each other. The Chinese populations were differentiated from the Pakistani and American populations in the Nad4 region. The low level of population genetic variation of P. gossypiella is attributed to invasion bottlenecks, which may have been subsequently strengthened by its nonmigratory biology and the mosaic pattern of agricultural activities.     

三个地理种群的董氏须鳅遗传多样性及种群历史动态

研究采用线粒体DNA Cyt b和D-loop基因分析西辽河、松花江和穆棱河3条水系董氏须鳅(Barbatula toni)的遗传多样性和种群结构, 并探讨其种群演变历史。结果表明: 3条水系119个董氏须鳅样本共检测出57个单倍型, 各群体间不存在共享单倍型; 3个董氏须鳅地理群体均呈现出较高的单倍型多样性(0.805—0.926)和较低的核苷酸多样性(0.00095—0.00458); AMOVA分子方差分析、群体间分化指数(F_ST)、系统发育树及单倍型网络图均表明3个地理种群已经出现高度分化, 群体间遗传变异占总变异的79.45%, 遗传分化显著(P<0.01)。中性检验和错配分析表明, 松花江和穆棱河群体历史上发生了群体快速扩张。参考鳅科鱼类Cyt b基因0.68—0.84%/Ma的进化速率, 估算3个地理群体的分歧时间为1.082—0.669 Ma前, 松花江和穆棱河群体扩张时间为0.071—0.047 Ma前, 推测我国董氏须鳅在中更新世期随冰期迁移至不同避难所, 形成各地理群体, 并于晚更新世早期经历了群体扩张事件。