濒危植物宽叶羌活天然居群cpDNA非编码区多态性分析

采用叶绿体DNA非编码区直接测序的方法,对青海、甘肃、四川3个省区内濒危植物宽叶羌活14个天然居群的遗传多样性和遗传结构进行了研究,以明确其遗传背景,为宽叶羌活的保护提供理论依据。结果表明:(1)14个居群138个个体的序列长度介于509~515bp;碱基组成A+T含量较高(平均67.6%)。(2)根据序列的核苷酸变异共鉴定出31个单倍型,其中9种单倍型(H5、H1、H9、H10、H16、H17、H19、H20、H22)为居群所共享,而且单倍型H5分布最广、在10个群体的67个个体中检测到,占总样品数的48.56%。(3)14个居群宽叶羌活具有高水平单倍型多样性(Hd=0.750 3)和较高水平核苷酸多样性(Pi=0.007 11),但31个单倍型没有按地理分布形成明显的族群,各地理单元中的单倍型相互混杂,没有明显的地理分化模式。(4)AMOVA分析、居群间分化度(FST=0.602 4)分析和基因流(Nm=0.330)分析的结果一致,表明宽叶羌活大部分遗传变异(60.24%)发生在居群间,居群间的基因流较低,群体间变异是宽叶羌活的主要变异来源。(5)14个居群的遗传距离在0.000~0.007,平均为0.003,表明居群间的亲缘关系较远,且居群间的遗传距离与地理距离之间无显著相关关系(P=0.143),说明宽叶羌活居群遗传变异的分布没有明显的地理趋势。研究认为,宽叶羌活居群间高的遗传变异可能是由基因流受阻、遗传漂变、地理隔离造成的,并提出了对该物种遗传多样性的保护策略。     

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。     

臭柏天然居群遗传多样性及演变历史分析

为了合理有效地保育天然臭柏(Juniperus sabina L.)种质资源,追溯和阐释其分布格局的历史成因,本文对我国内蒙古自治区、陕西省、甘肃省和青海省共10个天然臭柏居群388个个体的核糖体内转录间隔区(ITS)序列片段进行测序分析。结果显示:臭柏ITS序列总长度为1089 bp,共含有25个变异位点,定义32个单倍型,其中H4和H6单倍型为共有单倍型;分子变异分析(AMOVA)显示,臭柏居群变异主要来源于居群内,遗传变异为95.04%,而居群间遗传变异仅4.96%,居群间差异水平极显著(F ST=0.0496,P<0.001);Network单倍型网络分析表明,H4和H6为古老单倍型,其他单倍型是由他们衍生而来;遗传分化系数N ST(0.072)0.05)。推测臭柏起源于第三纪中新世(Miocene)中期约12.38 Mya,在第四纪冰期可能存在多个小型避难所。沙埋产生不定根的扩繁能力和较好的有性更新环境可能是沙地居群遗传多样性高于山地居群的决定性因素。     

濒危植物连香树居群的遗传多样性和遗传分化研究

利用ISSR分子标记技术对濒危植物连香树10个居群的遗传多样性和遗传变异进行了分析,结果表明:连香树物种水平遗传多样性较高,多态位点百分率(PPB)达到69.59%,Nei’s基因多样性指数(H)和Shannon信息指数(I)分别为0.231 3和0.351 4;而在居群水平上,多态位点百分率(PPB)为30.61%,Nei’s基因多样性指数(H)和Shannon信息指数(I)分别为0.115 6和0.173 3。遗传变异分析表明,居群间遗传分化程度高,遗传分化系数(GST)为0.500 3,居群间基因流Nm为0.527 3。Mantel检测,居群间的遗传距离和地理距离之间不存在显著的相关性。生境的片断化使居群间的基因流受阻,可能是导致居群间高遗传分化和居群水平低遗传多样性的主要原因。     

中国野生半夏的遗传多样性和遗传结构研究

第四纪气候波动以及地理和环境隔离深刻地影响了现代植物的遗传多样性、遗传结构和地理分布格局。该研究采用分子谱系地理学的研究方法对药用植物半夏19个居群共212个个体的3个叶绿体片段psbK-psbI、atpF-atpH和trnL-F进行分析,探究半夏的遗传多样性、遗传结构、地理分布格局模式及成因,并探讨其居群历史动态。结果表明：(1)半夏总单倍型多样性H_d为0.882,总核苷酸多样性π为1.23×10^-3,在物种水平上表现出较高的遗传多样性。(2)分子方差分析(AMOVA)结果显示,半夏遗传变异主要发生在居群间,显著的遗传分化(F_ST =0.909,P<0.001)和较低的种群内遗传多样性(H_S =0.134);种群间遗传分化系数N_ST=0.913>G_ST=0.855(0.01相似文献   

濒危植物长柄扁桃的遗传变异格局研究

该研究基于ITS1-ITS4序列对濒危植物长柄扁桃(Amygdalus pedunculata)自然分布区内的8个居群,105个个体的遗传多样性及遗传结构进行了分析,以明确长柄扁桃的地理分布及其遗传多样性特征。结果表明:(1)ITS1-ITS4序列长度583bp,变异位点13个,共定义了8个单倍型;居群间总的遗传多样性(hT)为0.727,居群内平均遗传多样性(hS)为0.564,各居群单倍型多样性、核酸多样性的变化范围分别为0.182~0.636、0.000 6~0.006 3。(2)AMOVA分析表明,33.65%的遗传变异来自居群间,而66.35%的遗传变异来自居群内;居群间平均基因流(Nm)为0.986,居群间遗传分化系数NSTGST(GST=0.225,NST=0.362,P0.05),表明长柄扁桃居群间存在不十分显著的遗传分化,但谱系地理结构显著;中性检验和错配分布曲线表明,居群自然分布区内的长柄扁桃没有经历明显的近期扩张。(3)单倍型网络和最大似然树都表明:内蒙古和陕西的长柄扁桃居群分别聚为2个明显的分支。     

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。     

青藏高原地区山生柳遗传多样性研究

利用叶绿体非编码区片段研究分布于青藏高原地区的山生柳居群遗传多样性,对未来山生柳生态环境和青藏高原地区物种丰富度的保护具有指导意义。该研究设计并筛选出cpDNA引物5'trnG2G-3'trnG(UUC)和5'rpS12-rpL20,用扩增出的片段和对应的联合片段进行后续的遗传多样性分析。结果表明:通过山生柳的联合片段检测到35种单倍型,单倍型多态性0.626,核苷酸多态性0.000 85。中性检验Tajima’s D(-2.286 70,P0.01)和Fu’s Fs(-5.298 05,P0.02)都是显著负值,推测山生柳个体数近期经历过扩张。AMOVA分析显示,居群内和居群间遗传变异分别为93.70%和6.30%,表明居群内的变异是山生柳遗传变异的主要来源。居群间遗传分化程度中等偏低(FST=0.063),基因流(N_m)为7.439,说明山生柳各居群的基因交流非常频繁,不同地理居群间存在一定的基因流动。遗传分化系数N_(ST)(0.075)大于GST(0.068)和基于遗传距离和单倍型的UPGMA聚类分析,表明山生柳12个居群分为4组且与居群的地理分布没有明显相关性。山生柳是进行有性繁殖还是无性繁殖主要受环境因素的影响,居群内变异是山生柳遗传变异性的主要来源,居群间基因交流频繁。     

广西地不容种质资源的ISSR分析

采用简单序列重复区间扩增(ISSR)分子标记技术对广西地不容3个野生居群和1个引种居群共92个个体进行了遗传多样性研究。10个引物共扩增出61条带,其中60条具多态性,多态性位点百分率为98.36%。4个居群多态性百分率在73.77%～86.89%。Nei’s基因多样性指数(H)为0.3379,Shannon信息多样性指数(Ⅰ)为0.5055。3个野生居群Nei’s遗传分化系数(Gst)表明:83.87%遗传变异分布在居群内,16.13%的遗传变异分布在居群间。引种居群与3个野生居群间的遗传一致度达0.8846。引种居群有效地保护了广西地不容的遗传多样性。     

基于线粒体COⅡ基因的大豆蚜不同地理种群遗传分化研究

【目的】大豆蚜Aphis glycines逐渐成为栽培大豆上的世界性重要害虫,为明确大豆蚜不同地理种群的遗传分化及遗传多样性,本研究探讨其在中国的种群遗传变异。【方法】测定了采自7个省共14个地理种群339头大豆蚜的线粒体COⅡ基因的673 bp序列,利用Dna SP 5.0、Arlequin 3.5.1.2、Network4.6.1.3等软件对地理种群间的大豆蚜的遗传多样性、遗传分化程度及分子变异进行分析,并建立了单倍型网络图及单倍型系统进化树。【结果】在所分析的339个COⅡ序列中,共检测出7个单倍型,其中H1为各种群所共享。种群内遗传多样性较低(Hd=0.479±0.030,Pi=0.00166±0.00018),种群内遗传分化相对较大(Fst=0.1985),基因流水平较高(Nm=2.019)。中性检验结果不显著(Tajima’s D=﹣0.931,P>0.10,Fu’s Fs=0.220,P>0.10),说明中国地区大豆蚜在较近的历史时期内没有出现种群扩张现象。分子变异分析(AMOVA)结果表明,大豆蚜遗传变异主要来自种群内部(80.15%),而种群间未发生明显的遗传分化。根据各地理种群的单倍型建立的系统发育树、单倍型网络图表明,各单倍型散布在不同的地理种群中,无明显的地理分布格局。【结论】大豆蚜不同地理种群的遗传多样性不高,各种群的遗传距离与地理距离之间没有显著线性相关性,各种群间的基因交流并未受到地理距离的影响。     

Higher genetic diversity in introduced than in native populations of the mussel Mytella charruana: evidence of population admixture at introduction sites

Aim  Levels of genetic diversity can be used to determine haplotype frequency, population size and patterns of invasive species distribution. In this study, we sought to investigate the genetic structure of the invasive marine mussel Mytella charruana and compare variation from invasive populations with variation found within three native populations.
Location  Invaded areas in the USA (Florida, Georgia); native areas in Ecuador, Colombia and Brazil.
Methods  We sequenced 722 bp of the mitochondrial COI gene from 83 M. charruana samples from four invasive populations (USA) and 71 samples from two natural populations (Ecuador, Columbia). In addition, we sequenced 31 individuals of a congeneric species, Mytella guyanensis , from Salvador, Brazil. We constructed the phylogenetic relationship among all haplotypes and compared diversity measures among all populations.
Results  We found significantly higher levels of nucleotide diversity in invasive populations than in native populations, although the number of haplotypes was greater in the native populations. Moreover, mismatch distribution analyses resulted in a pattern indicative of population admixture for the invasive populations. Conversely, mismatch distributions of native populations resulted in a pattern indicative of populations in static equilibrium.
Main conclusion  Our data present compelling evidence that the M. charruana invasion resulted from admixture of at least two populations, which combined to form higher levels of genetic diversity in invasive populations. Moreover, our data suggest that one of these populations originated from the Caribbean coast of South America. Overall, this study provides an analysis of genetic diversity within invasive populations and explores how that diversity may be influenced by the genetic structure of native populations and how mass dispersal may lead to invasion success.     

乔氏新银鱼基于细胞色素b序列的种群遗传结构和种群历史

为促进乔氏新银鱼(Neosalanx jordani)种质资源的保护, 采集了长江流域和淮河流域5个乔氏新银鱼地理种群计129个样本, 利用线粒体细胞色素b基因(Cyt b)全序列作为分子标记, 初步分析了乔氏新银鱼种群的遗传多样性、遗传结构及种群历史动态。研究结果共检测到18个Cyt b单倍型, 发现和其他鱼类相比, 乔氏新银鱼具有较高的单倍型多样性(h, 0.590±0.047), 但核苷酸多样性较低(π, 0.00088±0.00011)。分子变异分析(AMOVA)表明, 乔氏新银鱼5个地理种群内个体间和流域内种群间均存在显著的遗传差异, 而长江流域与淮河流域之间遗传分化不明显, 显示乔氏新银鱼遗传分化与当前水系的分布格局不吻合。结果表明乔氏新银鱼目前的遗传格局主要是由于长距离独立的建群事件、基因流限制以及种群的持续扩张的共同作用而形成的。种群历史动态分析结果显示乔氏新银鱼种群为近期扩张种群, 其大部分变异发生在1.897万年之内, 与最后一次冰期全面消退、海平面上升、长江流域及淮河流域中下游大量湖泊(适宜生境)形成的时间相吻合。建议对现存的乔氏新银鱼种群, 特别是遗传多样性较高的那些种群(如鄱阳湖、太湖及洪泽湖种群)分别保护。     

Chloroplast DNA study in wild populations and some cultivars of Prunus avium L.

The PCR-RFLP technique was used to detect chloroplast DNA diversity in wild populations of Prunus avium from five European deciduous forests and some cultivars. A study of 10.8% of the total chloroplast genome detected eight insertion-deletion (indel) mutations, distributed over 12 haplotypes. Six haplotypes (H1, H2, H3, H4, H5 and H6) were found in wild populations and eight (H2, H6, H7, H8, H9, H10, H11 and H12) in the cultivars. Only two haplotypes (H2 and H6) are shared by the wild populations and the cultivars. The most-abundant and frequent haplotype in wild populations is H2 (frequency=78%). The wider geographical distribution along with the high frequency reflects its ancient origin. Of the five populations, three are polymorphic. Populations GA (Scotland) and KE (Germany) have unique haplotypes. The total cpDNA diversity in wild populations is h_T=0.40, and a major portion of it is within populations (h_S=0.37). The genetic differentiation among populations was low (G_STC=0.08) and no genetic structure among wild populations was observed. A minimum-length spanning tree, demonstrating relationships among the haplotypes in wild populations, indicated two possible chloroplast lineages. The ten identified cultivars were represented by seven haplotypes; this result proposes the possible utilisation of the PCR-RFLP technique for the characterisation of sweet cherry cultivars. The cpDNA diversity in P. avium should be considered carefully for phylogenetic studies involving this species. Received: 10 July 2000 / Accepted: 19 October 2000     

基于核基因LEAFY的中国珍稀濒危植物中华水韭的遗传多样性分析

利用核基因LEAFY第二个内含子片段对中国现存中华水韭（Isoetes sinensis Palmer）的遗传多样性进行分析,探讨了中华水韭自然居群的遗传多样性结构及其形成机制。结果显示：现存中华水韭7个自然居群共105个样本中存在78个单倍型,单倍型多样性（H_d）为0.989,核苷酸多样性（π）为0.021,遗传差异主要存在于居群内（72%）,且存在较高的基因流（N_m=0.59）。同时,居群遗传学分析结果发现中华水韭居群不存在明显的谱系地理格局（G_ST>N_ST）;Mantel检验中Rxy值为-0.286,P（rxy-rand≥rxy-data）值为0.370,表明居群遗传距离和地理距离之间没有明显相关性;UPGMA聚类分析显示处于海拔较高位置的2个居群与其它5个居群遗传关系较远;中性检验（Taijima's D、Fu & Li's D^*和F^*）检测结果均为负值,基于稳定模型的失配分布检测显示为多峰。根据中华水韭居群的地理位置,推测中华水韭的遗传结构可能与水系、海拔分布及其杂交后代多倍化的物种形成过程相关。     

Genetic structure and phylogeography of a relict tree fern,Sphaeropteris brunoniana (Cyatheaceae) from China and Laos inferred from cpDNA sequence variations: Implications for conservation

In this study, we analyzed the genetic structure and phylogeography of Sphaeropteris brunoniana from China and Laos. Combining cpDNA trnL-trnF and atpB-rbcL sequence variations, five haplotypes were identified from the 10 investigated populations. Moderate haplotype diversity (h= 0.66580) and low nucleotide diversity (π= 0.23 × 10⁻³) were detected. The S. brunoniana in Yunnan region had much higher genetic diversity (h= 0.60195, π= 0.35 × 10⁻³) than that of Hainan–Laos (h= 0.00000, π= 0.00). A high level of genetic differentiation (94.74%) between the two regions was revealed by amova. Nested clade analysis identified two major clusters of the five haplotypes, one clade in the Yunnan region and the other in Hainan–Laos. The analysis indicated that restricted gene flow with isolation by distance and allopatric fragmentation were likely the major processes that shaped the spatial distribution of the haplotypes. The isolated distribution of clades implied the emergence of independent refugia of this species in each region during Quaternary glaciations. The Yunnan populations frequently contained an ancestral haplotype, and most of them harbored other descendent haplotypes. Based on the distribution pattern of haplotypes and the nested clade analysis results, the Yunnan region potentially had several refugia of this species during glacial periods, whereas the Hainan populations were probable new colonizations.     

Genetic diversity pattern of Stipa purpurea populations in the hinterland of Qinghai–Tibet Plateau

Stipa purpurea is among constructive endemic species in alpine meadow and steppe on the Qinghai–Tibet Plateau. To reveal the genetic diversity of this species and its relationship with geographic distribution pattern, we sampled eight populations across a northward transect with an increasing aridity gradient in the hinterland of Qinghai–Tibet Plateau. Their genetic diversity was quantified using eight intersimple sequence repeat (ISSR) primers. We found that S. purpurea had relatively low genetic diversity ( H _e = 0.135) but significant genetic differentiation among populations ( G _st = 0.391), indicating relatively more genetic diversity retained within populations. A Mantel test revealed a significant relationship between genetic and geographic distance in the S. purpurea populations. The genetic diversity tended to decrease with increasing latitude and longitude, while no significant relationship was found between genetic diversity and altitude, suggesting the possible influences of humidity and temperature on genetic diversity of alpine plant. We propose conservation measures for this plant on the plateau.     

入侵植物薇甘菊种群的遗传分化

利用简单重复序列区间(Inter simple sequence repeat, ISSR)分子标记技术分析了入侵植物薇甘菊(Mikania micrantha)8个种群的遗传多样性及遗传分化。12个引物共扩增出171个位点,其中多态位点有103个,多态位点百分率(P%)为60.23%,Shannon信息指数(I)为0.281 8,Nei指数(h)为0.184 9,薇甘菊在物种水平具有较高的遗传多样性。AMOVA显示薇甘菊具有较高的遗传分化,36.49%的变异发生在种群间,63.51%的变异发生于种群内,基因分化系数(G_ST)为0.352 4。种群间的基因流较高,为0.918 7。薇甘菊8个种群之间的遗传相似性很高,平均为0.915 5;遗传距离很小,平均为0.088 4。采用UPGMA法对8个种群进行聚类,可以将8个种群分为两大类群,即内伶仃岛为一个类群,而深圳与东莞内陆种群组成另一类群。薇甘菊现有遗传结构的形成与其生活史特性及入侵生态学特性有关。     

Genetic Diversity and Differentiation in Hedychium spicatum, a Valuable Medicinal Plant of Indian Himalaya

Hedychium spicatum, a perennial rhizomatous medicinal plant distributed in subtropical and temperate parts, is considered nearly endemic to the Himalayan region.In this study allozyme markers were utilized to assess genetic variations and relationships among 12 distinct populations of this species from the West Himalaya of India. A high level of genetic diversity was found among the populations. Of the 13 loci generated using eight enzymes, 12 (92%) were polymorphic. F-statistics suggested a prevalence of a high heterozygote excess among populations (F(IS) = -0.51). Gene flow estimates and geographic distances between populations did not correlate significantly (r = -0.0258, P < 0.3550). The excess heterozygosity may be attributed to high pollinator mobility and inbreeding coefficient within the subpopulation, relative to the total F(IS) value. High frequencies of several alleles at different loci probably reflect the breeding pattern, as the species propagates clonally as well as through seeds. We also discuss conservation implications for the target species.     

Microsatellite and mitochondrial haplotype diversity reveals population differentiation in the tiger shrimp (Penaeus monodon) in the Indo-Pacific region

The black tiger shrimp (Penaeus monodon) is an ecologically and economically important penaeid species and is widely distributed in the Indo-Pacific region. Here we investigated the genetic diversity of P. monodon (n = 355) from eight geographical regions by genotyping at 10 microsatellite loci. The average observed heterozygosity at various loci ranged from 0.638 to 0.743, indicating a high level of genetic variability in this region. Significant departures from Hardy-Weinberg equilibrium caused by heterozygote deficiency were recorded for most loci and populations. Pairwise F(ST) and R(ST) values revealed genetic differentiation among the populations. Evidence from the assignment test showed that the populations in the West Indian Ocean were unique, whereas other populations examined were partially admixed. In addition, the non-metric multidimensional scaling analysis indicated the presence of three geographic groups in the Indo-Pacific region, i.e. the African populations, a population from western Thailand and the remaining populations as a whole. We also sequenced and analysed the mitochondrial control region (mtCR) in these shrimp stocks to determine whether the nuclear and mitochondrial genomes show a similar pattern of genetic differentiation. A total of 262 haplotypes were identified, and nucleotide divergence among haplotypes ranged from 0.2% to 16.3%. Haplotype diversity was high in all populations, with a range from 0.969 to 1. Phylogenetic analysis using the mtCR data revealed that the West Indian Ocean populations were genetically differentiated from the West Pacific populations, consistent with the microsatellite data. These results should have implications for aquaculture management and conservation of aquatic diversity.