长柄石杉不同地理居群叶绿体DNA trnL-trnF序列变异与聚类分析

采用直接测序法,对石杉科广布种长柄石杉(Huperzia serrata var.longipetiolata)21个居群(福建省19个居群和江西省2个居群)共计126株个体的叶绿体DNA trnL-trnF序列进行测序,用Clustal X和MEGA5.0软件进行序列分析,UPGMA法构建聚类图,以期了解序列变异与地理分布的关系,为长柄石杉资源的保护及开发利用提供参考依据.扩增获得的序列全长在928-967 bp之间,经排序后两端切平,序列长925 bp,G+C平均含量为34.05%;21个长柄石杉居群的cpDNA trnL-trnF序列存在20个变异位点,5个信息位点.UPGMA聚类结果显示,福建省居群与江西居群存在较大遗传距离,分聚为不同分支;福建省内19个居群再分聚为3个分支,其中11个居群间的遗传距离为零.研究结果表明长柄石杉居群间存在较明显的遗传分化和一定强度的基因流,居群间的叶绿体DNA trnL-trnF序列变异与地理分布尤其是山脉形成的隔离和屏障有关,较高的遗传多样性和基因流水平有利于长柄石杉种群的生存与进化.     

麻疯树种质资源遗传多样性的ISSR分析

采用ISSR分子标记技术对大戟科属麻疯树9个自然居群的遗传多样性水平和遗传结构进行了研究.用10个引物对9个居群共135个样品进行了扩增,共获得169条清晰的扩增位点,其中多态性位点164个.POPGENE分析结果表明,麻疯树居群具有丰富的遗传多样水平(多态百分率PPB=97.04%,Neis遗传多样性H=0.2357,Shannon信息指数I=0.3760).AMOVA分析表明,9个自然居群间遗传多样性出现了较大的遗传分化(Gst=0.5398)可能与其有限的基因流(Nm=0.4667)和遗传漂变有关;而居群内有限的遗传分化可能是由于麻疯树自交、近交占主导方式的繁育方式有关.并进行了麻疯树居群的聚类分析.图3表4参19     

黄连木居群遗传多样性的SSR标记分析

为系统揭示黄连木天然居群在遗传多样性水平上的差异及遗传分化状况,采用SSR(Simple sequence repeats)分子标记对其8个居群进行遗传多样性分析.在建立良好的反应体系基础上,从已有的阿月浑子SSR引物中筛选出9对适合进行黄连木遗传分析的引物,在8个黄连木居群中共检测到43条等位基因,位点平均等位基因数为4.78,平均多态位点百分率达90.28%.各居群内平均有效等位基因数为2.08,平均期望杂合度(He)为0.472,说明各居群的遗传多样性处于中等水平.按检测到的有效等位基因数(Ne)和期望杂合度(He),各居群的遗传多样性由高至低依次为安康唐县顺平辉县略阳栾川林州涉县.居群间的遗传分化系数(FST)平均为0.319,说明居群内变异是黄连木变异的主要来源,并根据遗传距离将8个居群分为三大类.     

入侵植物加拿大一枝黄花和本地一枝黄花的遗传多样性比较

为了阐明加拿大一枝黄花成功入侵的机制,利用简单序列重复区间标记(ISSR)方法对加拿大一枝黄花和本地一枝黄花的遗传多样性进行比较研究。从100条引物中筛选出12条引物用于PCR扩增,利用POPGEN32软件对2种一枝黄花进行遗传多样性分析。结果表明,加拿大一枝黄花在物种水平上的多态位点百分率为95.19%,Nei’s基因多样性指数为0.308 5,Shannon’s信息指数为0.415 8;本地一枝黄花在物种水平上的多态位点百分率(89.80%)、Nei’s基因多样性指数(0.249 1)和Shannon’s信息指数(0.383 4)都比加拿大一枝黄花小。加拿大一枝黄花和本地一枝黄花居群间遗传分化系数分别为0.118 2和0.131 3,居群内变异分别为0.881 8和0.868 7,表明2个物种居群间的遗传分化不明显,遗传一致度高,且主要的遗传变异存在于居群内。入侵植物加拿大一枝黄花具有较高遗传多样性,且高于本地一枝黄花,这可能是加拿大一枝黄花成功入侵的原因之一。     

金花茶遗传多样性和遗传结构AFLP分析

利用扩增片段长度多态性(amplified fragment length polymorphism,AFLP)分子标记技术,对金花茶(Camellia nitidissima) 4个天然种群的遗传多样性和遗传结构进行研究,为金花茶资源的有效保护和高效利用提供科学依据。结果表明,8对引物共扩增出1 619个DNA位点,其中1 473个位点具多态性,多态性位点百分率为90. 98%。金花茶物种水平基因多样性指数和Shannon信息指数为0. 146和0. 246,种群水平基因多样性指数和Shannon信息指数为0. 131和0. 215; 4个金花茶种群的多态位点百分率、基因多样性指数和Shannon信息指数变化趋势一致,其中多态位点百分率介于56. 83%～72. 11%之间,基因多样性指数介于0. 122～0. 138之间,Shannon信息指数介于0. 197～0. 228之间。金花茶种群间的遗传分化系数介于0. 139～0. 289之间,遗传变异主要存在于种群内。分子变异分析(AMOVA)结果表明,金花茶种群内遗传变异为81. 52%,种群间遗传变异为18. 48%,遗传变异主要存在于种群内。非加权组平均法(UPGMA)聚类结果表明,种群间遗传变异与其地理距离无明显相关性。应加强现有金花茶种群的就地保护,促进种群自然更新,同时加强迁地保护。     

威海荣成鹿角菜野生种群遗传多样性及资源保护策略

利用随机扩增多态性DNA（RAPD）方法对威海荣成地区鹿角菜（Silvetiasiliquosa）野生种群共30个个体的遗传多样性水平及遗传结构进行研究。结果表明,利用21条随机引物共检测到112个多态性位点,多态性位点百分率为93．75％,Nei基因多样性指数为0．3515±0．1352,Shannon多样性指数为0．5046±0．1126。通过聚类分析而划为一类的3个亚种群间的遗传分化指数为0．1349～0．2189,基因流为1．7793。研究结果表明威海荣成地区的鹿角菜种群遗传多样性较高,种群内存在较为明显的遗传分化,种群遗传资源不受遗传漂变的影响。针对当地鹿角菜遗传资源的现状,应加强现存种群的就地保护,恢复种群规模;进行取样养殖保护时则需避免因近交而造成种群资源退化。     

缢蛏6个群体遗传结构的ISSR分析

采用ISSR分子标记对采自福建宁德(ND)、浙江台州(TZ)、上海崇明(CM)、山东海阳(HY)、天津汉沽(HG)和辽宁庄河(ZH)的缢蛏6个野生群体进行遗传多样性和遗传关系分析.筛选出10条ISSR引物对6个群体共216个样品进行扩增,共得到138个清晰的扩增位点,其中多态性位点135个,多态位点百分率为97.83%,在物种水平上的遗传多样性较为丰富.群体遗传多样性分析结果表明,6个群体的多态位点百分率在61.59%～72.46%之间,基因多样性指数和Shannon's信息指数分别在0.104 3～0.176 6和0.178 9～0.280 8之间,其中ND群体的多样性指数最低.群体间遗传分化、遗传距离和聚类结构显示,ND群体和TZ群体亲缘关系最近,首先聚为一支,之后依次与CM群体及HG群体和HY群体聚类,最后与ZH群体聚在一起,表明地理位置较远的群体,遗传距离也较远.     

大贵寺国家森林公园野生青檀居群的遗传多样性

利用ISSR分子标记技术对湖北省大贵寺国家森林公园野生青檀（Pteroceltis tatarinowii Maxim．）居群进行遗传多样性分析。用11条ISSR引物对6个青檀居群64个样品进行扩增,扩增片段长度在200bp至3000bp之间,检测到179个位点,其中多态位点165个,多态位点百分率（P）为92．14％。青檀在物种水平上的Nei基因多样性指数（胁）和Shannon信息指数∽分别为0．273和0．423,在居群水平上分别为0．225和0．343,显示出青檀有着较高的遗传多样性。居群间的遗传分化系数（Gsr）为0．192,表明青檀居群间存在着较大的遗传分化;UPGMA聚类分析表明遗传距离与海拔差距有一定的相关性,相邻海拔的青檀居群间遗传距离较近。不同海拔导致的异质生境可能是影响青檀居群遗传分化的主要原因。     

长江中下游及云贵高原芦苇居群的遗传变异特征

芦苇(Phragmites australis)为全球广泛分布的多型种,在水生生态系统,尤其是湿地生态系统中承担重要的生态功能,准确划分其生态型可为种质资源保护与利用提供理论依据,同时也为修复受损生态系统异地引种提供参考。利用叶绿体DNA rpl16基因序列,选取淡水湖泊较为集中且地理跨度较大的长江中下游和云贵高原,以地理位置为划分指标,探讨19个湖滨带芦苇居群的亲缘关系、遗传多样性和遗传结构,并分析了不同湖泊芦苇形态差异与遗传差异的异同,旨在分子水平为不同地理区域芦苇生态型的划分提供遗传分化方面的证据。结果表明:(1)rpl16基因序列分析显示:长江中下游芦苇居群基因水平上的变异程度较云贵高原的低;(2)基于rpl16基因序列重建的最简约系统发育树显示:以地理区域为划分指标,可将长江中下游和云贵高原的芦苇居群分为两个单系;(3)云贵高原芦苇居群的核苷酸多样性A=0.00047,单倍型多态性Hd=0.466,其遗传多样性显著高于长江中下游(Pi=0,Hd=0);(4)居群水平上的遗传分化系数F_(st)=0.535 37,其中53.54%的遗传变异来自于不同地理区域居群间,长江中下游芦苇居群的遗传差异指数F_(st)=0.537 24,基因流N_m=0.215 34,云贵高原芦苇居群的遗传差异指数F_(st)=0.532 78,基因流N_m=0.21923,居群间的遗传分化较强烈。分析结果证明了两种地理生态型芦苇具有遗传上的差异,且地理隔离可能是引起差异的主要原因。     

裸果木种群遗传多样性及其与土壤因子的关联性研究

为深入了解裸果木(Gymnocarpos przewalskii)遗传基础、斑块状分布的种群遗传结构及遗传多样性与土壤因子间的相关性,对分布于西北荒漠区的12个裸果木种群遗传多样性及土壤养分及进行分析,为裸果木遗传多样性的地理变异趋势预测及种质资源的科学管理和保育提供依据。结果表明,裸果木居群遗传多样性较高,其中,Nei’s基因多样性指数(He)变化范围为0.235~0.269,Shannom信息指数(I)变化范围为0.351~0.405,居群内具有丰富的遗传变异,基因分化系数(Gst)为0.279。基于遗传一致度对种群进行PCA分析,12个种群裸果木可分为4大类:甘肃肃北县种群为分布的核心区,单独为一类;以河西走廊中西部为中心分化2类,甘肃瓜州及甘肃柳园镇为河西走廊西部中心种群,与向西南和向西延伸的甘肃阿克塞县和新疆伊州区种群亲缘关系较近,聚为一类;甘肃金塔县、甘肃高台县、甘肃清泉乡、甘肃肃州区为中心的河西走廊东部分布聚为一类;向东和向东南延伸的内蒙古阿右旗、宁夏沙坡头区域和甘肃民勤县聚为一类。依据裸果木分布区土壤养分含量进行主成分聚类,将种群划分为3大类:宁夏沙坡头样地中有机质、磷、氮和水分含量较高,被单独划为一类;甘肃肃北县等种群因土壤养分含量处于中等水平,聚为一类;内蒙古阿右旗等种群因钾元素含量高,被划分为一类。将遗传特征参数与土壤养分含量进行相关性分析,土壤中氮含量及全盐含量对种群遗传多样性水平有一定影响,处于贫瘠土壤上种群机体能够更加有效地利用土壤中的限制性养分。     

我国主要花生品种的AFLP分析

对32个来源于中国不同产地的花生品种进行了AFLP指纹图谱及相似性聚类分析．结果表明：所有供试花生品种的遗传相似性为35％，在45％的相似性水平处分为3个群，表明我国的花生品种存在遗传多态性．AFLP指纹图谱分析技术能很好地反映出花生品种之间存在的遗传差异．图2表1参26     

Quaternary geographical sibling speciation and population structuring in the Eastern Atlantic skates (suborder Rajoidea) Raja clavata and R. straeleni

The European Raja clavata and the South African R. straeleni are marine skates which exhibit highly conserved morphological and ecological traits. Owing to this, taxonomic and evolutionary relationships between the two taxa have not yet fully elucidated. Here, we have tested the hypothesis that restricted gene flow and genetic divergence between these taxa might be associated with climatic/oceanographic discontinuities by surveying genetic variation in ten geographical samples at control region (CR) and amplified fragment length polymorphism (AFLP) loci. The clustering of CR haplotypes in two reciprocally monophyletic clades consistent with taxon zoogeography and the significant AFLP F values between the European and South African populations indicated the two taxa as recently diverged peripatric sibling species. Within each species, significant spatial genetic heterogeneity among samples at both markers revealed population structuring. We argued that structured populations and isolated sibling species might represent two stages of geographical speciation.     

Genetic structure of natural populations of California red abalone (<Emphasis Type="Italic">Haliotis rufescens</Emphasis>) using multiple genetic markers

Mitochondrial cytochrome oxidase subunit one (COI) sequence, nuclear microsatellites, and amplified fragment length polymorphisms (AFLPs) were used to evaluate connectivity among nine red abalone (Haliotis rufescens) populations sampled between August 1998 and November 2003 along approximately 1,300 km of California coastline from Crescent City (41°46′N, 124°12′W) to San Miguel Island (34°02′N, 120°22′W). COI sequences and microsatellite genotypes did not show significant genetic divergence among nine sampled populations. A subset of five populations spanning the geographic range of the study was scored for 163 polymorphic AFLP markers. Of these, 41 loci showed significant divergence (P < 0.001) among populations. Still, no AFLP markers were diagnostic for any of the study populations, and assignment tests did not consistently assign individuals to the correct population. Although the AFLP data are the first to suggest there is significant genetic differentiation among California red abalone populations, the discordance between the different genetic markers needs further study before unambiguous conclusions can be drawn with respect to connectivity among the populations. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Genetic diversity and relatedness of the mangrove Rhizophora mangle L. (Rhizophoraceae) using amplified fragment polymorphism (AFLP) among locations in Florida, USA and the Caribbean

Rhizophora mangle L. is a widespread mangrove species in the Western Hemisphere. Mangrove habitat loss and their importance to coastal and reef ecosystems make greater understanding of their genetic structure useful for conservation and management. An amplified fragment polymorphism (AFLP) analysis was performed on samples from Florida and the Caribbean to discover the genetic structure present. R. mangle had variable genetic diversity not related to latitude; P ranged 7 %–92 %. Some other factor, perhaps human impact, has caused low genetic diversity in some populations. Across Florida R. mangle populations varied in genetic diversity with less diversity (G_st?=?0.195) and greater gene flow on the Atlantic coast (Nm =2.07) than on the Gulf coast (G_st?=?0.717, Nm?=?0.197). Gene flow between Caribbean islands was low (Nm?=?0.386) compared to continental populations (Nm?=?1.40), indicating that long distance dispersal is not common between islands. Analysis of molecular variance (AMOVA) analysis showed significant deviations from Hardy-Weinberg expectations at the level of region among subpopulations and overall genetic difference among subpopulations for R. mangle. One implication for management is that small continental populations and island populations may be genetically isolated and distinct from each other.     

RAPD分子标记在鉴定香樟优选株和普通株中的应用

从85个随机引物中选10个随机引物对8个香樟样品的DNA进行了扩增，应用RAPD技术区别6个优选株和2个普通株，共获得78个DNA片断，把这些DNA片断作为遗传位点用UPCMA法计算出各材料间的遗传相似性系数，并作了聚类分析．在D＝0．738处8个样品分为两大类，而且普通株同优选株的亲缘关系很近．图2表2参9     

中国一年生野生大豆群体的遗传多样性研究

选用来自全国各地的一年生野生大豆２００ 余份材料,从形态性状、等位酶标记和细胞器ＤＮＡＲＦＬＰ标记的遗传丰富度和遗传离散度两方面分析了中国野生大豆群体的遗传多样性．结果表明：中国野生大豆天然群体存在着遗传分化,各地理生态群体间的遗传多样性水平不同,南方群体最高,黄淮海群体次之,东北群体最低．南方为一年生野生大豆的遗传多样性中心,也可能是起源中心．     

四川省部分豆科植物根瘤菌的遗传多样性

从四川省豆科植物鸡眼草属、合欢属、豇豆属、葛属、金合欢、杭子梢分离获得了103株根瘤菌,采用AFLP、BOXAIR-PCR研究了这些菌株的遗传多样性.结果表明,分离自四川省豆科植物6个属的根瘤菌存在明显的遗传多样性,AFLP分析中,全部供试菌株的相似性为21%,全部供试菌株可分为26个AFLP遗传群;BOXAIR-PCR聚类分析结果表明,103个菌株分在70%相似性水平处,供试菌株被分为18个BOX遗传群.除部分菌株外,多数根瘤菌按宿主类型聚群,同一宿主而地理来源不同的根瘤菌分布于不同的遗传群,表明宿主和地理环境对根瘤菌具有深刻影响.图3表1参9     

Genetic Identification of Spotted Owls, Barred Owls, and Their Hybrids: Legal Implications of Hybrid Identity

Abstract:  Recent population expansion of Barred Owls (  Strix varia ) into western North America has led to concern that they may compete with and further harm the Northern Spotted Owl (  S. occidentalis caurina ), which is already listed as threatened under the U.S. Endangered Species Act (ESA). Because they hybridize, there is a legal need under the ESA for forensic identification of both species and their hybrids. We used mitochondrial control-region DNA and amplified fragment-length polymorphism (AFLP) analyses to assess maternal and biparental gene flow in this hybridization process. Mitochondrial DNA sequences (524 base pairs) indicated large divergence between Barred and Spotted Owls (13.9%). Further, the species formed two distinct clades with no signs of previous introgression. Fourteen diagnostic AFLP bands also indicated extensive divergence between the species, including markers differentiating them. Principal coordinate analyses and assignment tests clearly supported this differentiation. We found that hybrids had unique genetic combinations, including AFLP markers from both parental species, and identified known hybrids as well as potential hybrids with unclear taxonomic status. Our analyses corroborated the findings of extensive field studies that most hybrids genetically sampled resulted from crosses between female Barred Owls and male Spotted Owls. These genetic markers make it possible to clearly identify these species as well as hybrids and can now be used for research, conservation, and law enforcement. Several legal avenues may facilitate future conservation of Spotted Owls and other ESA-listed species that hybridize, including the ESA similarity-of-appearance clause (section 4[e]) and the Migratory Bird Treaty Act. The Migratory Bird Treaty Act appears to be the most useful route at this time.     

Genetic evidence for the existence of cryptic species in an endangered clam <Emphasis Type="Italic">Coelomactra antiquata</Emphasis>

Coelomactra antiquata is a commercially important bivalve species, but has been suffering from severe population decline due to over-exploitation and the deterioration of environmental conditions. Previous genetic survey of C. antiquata conducted with allozymes combined with morphology revealed high levels of genetic differentiation between northern and southern populations which suggests a cryptic species might exist in C. antiquata. To test this hypothesis, amplified fragment length polymorphisms (AFLPs) and 16S rRNA gene sequence were used to re-evaluate the spatial genetic structure of six populations of C. antiquata along the coast of China. Both genetic markers display a sharp genetic break between the four northern populations (northern lineage) and two southern population (southern lineage). Large numbers of private alleles (AFLP) were found within the northern or southern populations and a deep divergence of about 6.5% in 16S rRNA gene sequence between the northern and southern lineages suggests the occurrence of potential cryptic or sibling species of C. antiquata. Applying previously published rates of mutation, divergence between the two lineages is estimated to have occurred approximately 3 million years ago and may be due to allopatric isolation during the middle Pliocene times. While no genetic differentiation was found within the northern or southern populations in both AFLP and 16S mtDNA markers, the results indicate that the northern and southern lineage should be managed separately and any translocation between the two areas should be avoided.     

Genetic Analysis of the Endangered Island Endemic Malacothamnus fasciculatus (Nutt.) Greene var. nesioticus (Rob.) Kearn. (Malvaceae)

Isozyme, randomly amplified polymorphic DNA, and ribosomal DNA analyses were used to evaluate the genetic distinctness and diversity of the endangered island endemic Malacothamnus fasciculatus var. nesioticus (Santa Cruz Island bush mallow). Analysis of amplified DNA profiles and electrophoretic data indicates that var. nesioticus is genetically distinct from other varieties within the species. No isozyme or amplified DNA variation was detected among individuals of nesioticus populations, but restriction site analysis of ribosomal DNA revealed the presence of three closely related genotypes within one of the nesioticus populations. Apparent clonal structuring was found in both nesioticus populations and in one mainland population, var. nuttallli . Results from these analyses provide the background necessary for the design of conservation efforts with recovery goals that insure the long-term survival of Malacothamnus fasciculatus var. nesioticus .