基于ITS序列西南地区不同居群薤白亲缘关系及其地理分布格局研究

该研究以西南地区分布的薤白(Allium macrostemon Bunge)为材料,对21个不同居群的薤白进行核糖体DNA的ITS序列测定,并结合GenBank相关序列,对27个不同居群薤白开展了系统分析,以探讨西南地区不同居群薤白的亲缘关系和生物地理分布格局。结果表明:(1)西南地区不同产区薤白nrDNA ITS序列长度介于607～638 bp之间,G+C含量为48%～52%,平均值为50%。(2)基于贝叶斯法和最大简约法构建的系统树均表明薤白为一单系群。(3)系统发育树分析结果显示,27个居群的薤白总体划分为a组和b组,其中位于西南地区东面的四川剑阁县、重庆巫溪县、贵州德江县和云南丘北县的薤白共同聚为a组,位于西南地区西面的四川汶川县、四川石棉县、四川江油市以及云南虎跳峡、云南昆明市、云南三坝的薤白归为b组。研究表明,nrDNA ITS序列可以鉴定不同地区薤白的亲缘关系。     

基于nrDNA ITS和cpDNA trnL-F序列探讨薤白的系统地位

薤白(Allium macrostemon)是葱属中重要的药食同源种类,其系统地位和分类等级迄今尚无统一定论,一直存有争议。为了探讨其系统位置及其与相关类群的亲缘关系,本研究利用nr DNA ITS和cp DNA trn L-F序列数据对该种及若干近缘类群进行了系统发育分析。基于贝叶斯法和最大简约法构建的系统树均表明薤白为一单系种,与多系的组Caerulea、Pallasia的部分种类有密切的系统发育关系,而与组Codonoprasum和Allium的种类关系疏远;结合总苞、花丝和子房等形态学特征,不支持将薤白划归Allium组或置于广义的Scorodon组。薤白可能代表一个新的组一级的类群,但尚待增加类群采样(尤其是中亚的种类)进一步研究。本文还在系统发育框架下探讨了中药薤白(Allii Macrostemonis Bulbus)的基源植物(薤白、藠头、棱叶韭和长梗韭)及Pallasia组部分物种的系统位置。     

不同种源何首乌的ITS序列分析及其亲缘关系研究

采用PCR直接测序法,测定了10个种源何首乌的核糖体DNA ITS区序列,并结合GenBank中相关植物的ITS序列(以萹蓄Polygonu maviculare为外类群),应用遗传距离与系统树分析法对不同种源何首乌的亲缘关系进行了分析。结果表明:(1)ITS1序列长度为195bp,G C含量为69.23%~72.31%,ITS2序列长度为189bp,G C含量为77.25%~80.95%,序列间遗传分化距离为0.00175~0.04945;(2)10个种源何首乌构成2个分支,广西田阳种源自成一支,与其它种源亲缘关系较远;(3)结合形态、分布与化学成分,支持将田阳何首乌作为何首乌的一个变种——棱枝何首乌。     

葱属12种植物的RAPD分析

采用随机引物扩增多态DNA技术对葱属部分植物进行了种间亲缘关系的研究。结果说明12种材料之间存在丰富的多态性,遗传距离变幅在0.2500-0.7887之间,聚类分析说明蒙古韭,山韭,野韭,韭菜（栽培韭）,野生韭菜,矮韭亲缘关系较近,聚为一支,其中韭菜与野生韭菜亲缘关系最近。天蒜,薤白,蒜聚为一支,葱,洋葱,红葱聚为一支,其中葱与洋葱,红葱的遗传分化较大。     

基于ITS序列分析豹子花属与5种百合的亲缘关系

以滇蜀豹子花和多斑豹子花为材料,采用PCR直接测序法测定其ITS序列,结合GenBank中其它3种豹子花和5种百合的ITS序列,构建了这10种植物的系统发育树.结果表明:(1)10种植物的ITS序列长度在625bp～627 bp之间,总G C含量在60.38%～61.12%之间,5.8S的G C含量除大理百合为45.4%外,其余9种植物为55.01%或54.60%,说明ITS序列在进化上保守性较强,同属不同种甚至不同属间的长度差异不明显;(2)NJ、MP、ME聚类树的分支趋势一致,都是豹子花属植物先聚在一起再和5种百合相聚,滇西豹子花和豹子花在3种聚类树中都以99%以上的支持率聚成一支,说明这2个种的亲缘关系最近;(3)在10种植物中,形态相似且分布海拔和区域重叠的种类先相聚,说明这些物种的亲缘关系密切.     

基于ITS与 trn L-F序列探讨槭树科的系统发育

报道了槭树科41种(其中槭属39种)植物的 trn L-F和ITS序列(其中部分种的ITS序列为重新测定),以期通过分子手段对槭树科内部尤其是复杂的槭属的系统发育关系进行重建.以无患子科和七叶树科为外类群,基于对57个种单独的ITS序列(包括从GenBank下载的16种的序列)、41种 trn L-F序列及41种两者序列的联合数据,分别采用最大简约法(Maximum Parsimony Method)和邻接法(Neighbor-Joining Method)对槭树科的系统发育进行了分析.结果显示,整个槭树科为一单系类群;金钱槭位于槭树科的基部;但由于云南金钱槭( Dipteronia dyerana )聚在了槭属内部,认为金钱槭属和槭属均可能是非单系类群;槭属内组间关系的支持率普遍较低,但多数组的组内关系得到了较好的支持.将两个片段结合比单独的ITS或 trn L-F分析能更好地解决槭属内部的系统关系,其中sect.Palmata 和sect.Microcarpa ,sect.Platanoidea 、sect.Lithocarpa 和sect.Macrophylla ,sect.Integrifolia 、sect.Trifoliata 和sect.Pentaphylla ,以及sect.Acer 、sect.Goniocarpa 和sect.Saccharina (sensu Ogata)的组间亲缘关系得到了一定的支持,但对其中部分组的划分可能应做进一步调整.重新评价了徐廷志系统中对sect.Rubra 和sect.Saccharodendron 的处理.     

基于ITS与trnL—F序列探讨槭树科的系统发育

报道了槭树科41种（其中槭属39种0植物的trnL-F和ITS序列（其中部分种的ITS序列为重新测定），以期通过分子手段对槭树科内部尤其是复杂的槭属的系统发育关系进行重建。以无患子科和七叶树科为外类群，基于对57个种单独的ITS序列（包括从GenBank下载的16种的序列），41种trnL-F序列及41种两序列的联合数据，分析采用最大简约法（Maximum Parsimony Method)和邻接法（Neighbor-Joining Method)对槭树科的系统发育进行了分析。结果显示，整个槭树科为一单系类群；金钱槭位于槭树科的基部；但由于云南金钱槭（Dipteronia dyerana)聚在槭属内部，认为金钱槭属和槭属均可能是非单系类群；槭属内组间关系的支持率普遍较低，但多数组的组内关系得到了较好的支持。将两个片段结合比单独的ITS或trnL-F分析能更好地解决槭属内部的系统关系，其中sect,Palmata和sect.Micrcarpa,sect,Platanoidea,sect,Lithocarpa和sect.Macrophylla,sect,Integrifolia.Trifoliata和sect Pentaphylla,以及sect.Acer,sect.Goniocarpa和sect.Saccharina(sensu Ogata）的组间亲缘关系得到了一定的支持，但对其中部分组的划分可能应做进一步调整。重新评价了徐廷志系统中对sect.Rubra和sect.Saccharodendron的处理。     

珍珠菜rDNA ITS序列扩增及分析

以贵州境内珍珠菜属植物为材料,对其rDNA转录间隔区(ITS)序列进行PCR扩增和序列测定。实验共得到7个种的ITS序列,它们分别是:过路黄(Lysi machia christinae,GenBank登录号FJ362382),矮桃(L.cle-throides,GenBank登录号FJ362383),叶头过路黄(L.phyllocephala,GenBank登录号FJ362386),临时救(L.con-gestiflora,GenBank登录号FJ362387),显苞过路黄(L.rubiginosa,GenBank登录号FJ362388),茂汶过路黄(L.stellarioides,GenBank登录号FJ362384),腺药珍珠菜(L.stenosepala,GenBank登录号FJ362385),其中后2个种是国际上首次得到的。采用Blast方法将测序结果进行同源搜索,采用邻接法构建与其相关植物的ITS序列系统发育树。结果表明,珍珠菜属7种植物ITS序列总长度为613~620 bp;ITS1区序列长度为234~239 bp,5.8SrDNA区序列长度163 bp,ITS2区序列长度216~219 bp,7种植物的ITS序列差异主要集中在ITS1与ITS2区。聚类分析将茂汶过路黄聚为一支,其它6种植物聚为一支,表明茂汶过路黄与其它6种植物的碱基差异较大,从分子水平上支持据形态特征把花辐射生长的茂汶过路黄另立一类。     

基于ITS序列的东亚当归属植物的分类学研究

采用PCR直接测序法,测定了东亚地区狭义当归属Angelica s.s.及其近缘共7属40种代表植物的核糖体DNA ITS序列,并结合GenBank中相关植物的ITS序列(含外类群3种),应用遗传距离与系统树分析法对东亚地区狭义当归属植物内部以及当归属与其近缘属植物之间的亲缘关系进行了分析。结果表明:(1)广义当归属中的狭义当归属、柳叶芹属Czernaevia和高山芹属Coelopleurum之间的亲缘关系较近,山芹属Ostericum与它们的亲缘关系较远,这与果实形态、化学分析的结果一致,建议将山芹属作为一个相对独立的分类群处理。(2)ITS序列分析结果支持狭义当归属不是单起源的自然分类群,而应该被分成若干组的观点。(3)ITS序列以及化学成分分析结果表明,前胡属Peucedanum与狭义当归属之间的亲缘关系很近。(4)形态、化学成分以及ITS等多方面分析结果显示,当归A.sinensis与狭义当归属的多数植物之间均有一定的差距,其归属问题值得商榷。(5)ITS序列与化学成分的分析结果均显示藁本属Ligusticum不是一个自然类群。     

基于ITS序列分析仲彬草属植物的亲缘关系

以旱雀麦为外类群,用PAUP 4.0b10软件并采用最大简约法和邻接法对11份仲彬草属物种的ITS区序列进行系统发育分析,两种方法得到的系统发育树基本一致。结果表明:(1)整个ITS序列长度变异范围为596~601 bp;G C含量在所有ITS中的变化范围为61.20%~62.44%;序列间的遗传分化距离为0.003~0.033,平均值为0.015;(2)疏花仲彬草和塔克拉干仲彬草2个物种聚为一支,位于系统发育树的底部,在最大简约法和邻接法分析中分别获得78%和82%的自展支持率,它们之间的亲缘关系较近;(3)形态相似、地理分布一致的物种有聚在一起的倾向,表现出较近的亲缘关系;(4)ITS区序列分析的结果与细胞学、形态学的研究结果基本一致,因此ITS区序列分析能反映仲彬草属种间关系。     

Systematic Position of Allium macrostemon Based on nrDNA ITS and cpDNA trnL-F Sequence Data

Allium macrostemon is an important medicinal and edible plant. Its systematic position and taxonmical classification remain controversial to date. To explore this issue, we reconstructed phylogenetic relationships among Amacrostemon and other related taxa using nrDNA ITS and cpDNA trnL F markers. The phylogenetic trees derived from Bayesian inference and maximum parsimony analysis showed that Amacrostemon is monophyletic, and has a close relationship with some species of polyphyletic sections Caerulea and Pallasia instead of sections Codonoprasum and Allium. The including of Amacrostemon within section Allium was not supported by both molecular data and morphological characters of spathe, filaments and ovary. Allium macrostemon should be included in a new section, however further studies using additional samples (especially those from Central Asia) is necessary. In addition, we also provided a discussion on the phylogenetic relationships among four original plants (Amacrostemon, Achinense, Acaeruleum and Aneriniflorum) of Allii Macrostemonis Bulbus and systematic position of partial species of section Pallasia.     

栽培种甘薯及其近缘野生种nrDNA ITS序列分析

采用核糖体DNA内转录间隔区(nrDNA ITS)序列比较分析了甘薯及其近缘野生种的遗传多样性及系统进化关系,首次报道了栽培种甘薯‘徐薯18’(Ipomoea batatas‘Xushu18’)及其近缘野生种I.triloba(DOM),I.cordatotriloba(MEX),I.nil(PER),I.nil(JPN),I.hederacea Jacq.(USA),I.hederacea Jacq.(HK)和种间杂交种67-1(I.batatas‘Xushu18’×I.hederacea Jacq.)及回交种(67-1×I.batatas‘Xushu18’)的nrDNA ITS序列。序列分析表明,栽培种甘薯及其近缘野生种nrDNA ITS序列长度为570~600bp。其中,ITS1序列为185~209 bp,GC含量为53.11%~61.83%;ITS2序列为214~226 bp,GC含量为61.21%~72.89%;5.8S序列均为165 bp,GC含量为54.55%~55.76%。此外,栽培种甘薯及其近缘野生种ITS序列信息位点均集中在ITS1和ITS2区;与其他甘薯属植物相比,I.wrightii ITS2的末端缺失了6~8个碱基。系统进化分析表明,栽培种甘薯‘徐薯18’(I.batatas‘Xushu18’)和野生种I.triloba、I.cordatotriloba、I.lacunosa、I.trifida的亲缘关系较近,与I.wrightii、I.pes-tigridis、I.grandifolia、I.nil、I.hederacea Jacq.、I.purpurea的亲缘关系较远;杂交后代与栽培种甘薯‘徐薯18’(I.batatas‘Xushu18’)亲缘关系较近,与野生种父本I.hederacea Jacq.的亲缘关系较远。     

Potential phylogenetic utility of the low-copy nuclear gene pistillata in dicotyledonous plants: comparison to nrDNA ITS and trnL intron in Sphaerocardamum and other Brassicaceae.

We report the potential phylogenetic utility of DNA sequence data from the last 700 bp of a ca. 1-kb intron of the MADS-box gene pistillata from a sampling of Sphaerocardamum species and other Brassicaceae. These results are compared with nrDNA ITS and the chloroplast trnL intron for the same taxa to demonstrate the potential phylogenetic utility of this pistillata intron and to identify potential historically independent sequences for an ongoing study of relationships within Sphaerocardamum. Analyses of the DNA sequence data for Brassicaceae indicated that pairwise divergences and potentially informative characters were higher in the pistillata intron (0.6-30.8%, 284 characters) and ITS (0-24%, 94 characters) than in the chloroplast trnL intron (0-4.2%, 17 characters). A comparison of Sphaerocardamum sequences identified low divergences and numbers of informative characters for trnL intron (0-2.4%, 1 character) and nrDNA ITS (0-2.5%, 2 characters) and substantially more variation among the pistillata sequences (0.15-3.7%, 19 characters). Phylogenetic analyses of these pistillata sequences fully resolve ingroup relationships without character conflict. Results of pistillata PCR amplifications from a broader dicot sample showed that some primers may be useful in amplifying orthologous pistillata sequences. Ultimately this pistillata intron may be a valuable source of phylogenetic characters at lower taxonomic levels.     

rDNA internal transcribed spacer sequence analysis of Craterellus tubaeformis from North America and Europe

The basidiomycete Craterellus tubaeformis (Fries) Quélet is an important widespread ectomycorrhizal basidiomycete found in the Northern Hemisphere. In this study, 12 samples of C. tubaeformis from North America and Europe were analyzed using internal transcribed spacer (ITS) sequences to reveal the correlation between ITS genotypes and geographic locations and to provide molecular evidence for the identification of C. tubaeformis from different habitats in North America and Europe. The analyses identified abundant sequence variations within C. tubaeformis. The length of the ITS region varied from 571 to 640 bp. The proportion of variable sites was 17.6%, and the proportion of parsimony information sites was 16.7%. Phylogenetic analysis showed some correlations between the ITS genotypes and geographic locations of C. tubaeformis; however, some discrepancies between geographical location and affinity were also found. The results indicated that C. tubaeformis from different habitats in North America and Europe underwent genetic drifting and evolved into 2 different species. nrDNA ITS could be a good markers for distinguishing among C. tubaeformis from different habitats, but rational affinity should be determined by associating the available ITS data with other information sources.     

A molecular phylogeny of the wild onions (Allium; Alliaceae) with a focus on the western North American center of diversity

Nuclear ribosomal DNA (ITS and ETS) sequences from 39 native Californian (USA) Allium species and congeners were combined with 154 ITS sequences available on GenBank to develop a global Allium phylogeny with the simultaneous goals of investigating the evolutionary history (monophyly) of Allium in the Californian center of diversity and exploring patterns of adaptation to serpentine soils. Phylogenies constructed with ITS alone or ITS in combination with ETS provided sufficient resolution for investigating evolutionary relationships among species. The ITS region alone was sufficient to resolve the deeper relationships in North American species. Addition of a second marker (ETS) further supports the phylogenetic placements of the North American species and adds resolution within subgenus Amerallium, a clade containing many Californian endemics. Within the global phylogeny, the native North American species were found to be monophyletic, with the exception of Allium tricoccum and Allium schoenoprasum. All native Californian species included in the analysis fell into a monophyletic subgenus Amerallium section Lophioprason, although endemic Californian species were not monophyletic due to the inclusion of species with ranges extending beyond the California Floristic Province. The molecular phylogeny strongly supports previous morphology-based taxonomic groupings. Based on our results, serpentine adaptation appears to have occurred multiple times within section Lophioprason, while the ancestor of the Californian center of diversity may not have been serpentine-adapted.     

樟属植物ITS序列多态性分析

对樟科樟属(Cinnamomum Schaeffer) 17个代表样本的核糖体DNA内转录间隔区(nrDNA ITS)进行克隆测序。对获得的87条不同ITS序列的长度变异、GC含量、5.8S区二级结构的稳定性、遗传距离、进化模式以及系统发育关系进行了相关分析。研究结果显示, ITS序列在樟属植物内存在明显的多态性, 87条序列中的22条序列被鉴定为假基因序列, 其余65条序列为功能基因序列; 假基因序列采用中性进化模式, 变异明显大于功能序列。ITS序列在樟属植物中出现一致性进化不完全和假基因现象也可能发生在樟科其它类群中, 这可能是导致樟科植物ITS序列直接测序方式成功率低的重要原因。     

Molecular authentication of Anthemis deserti Boiss. (Asteraceae) based on ITS2 region of nrDNA gene sequence

The dried plant material of medicinally important Anthemis deserti Boiss. (family: Asteraceae) especially when it remains in the powdered form often look similar to Anthemis melampodina Del.; and therefore, difficult to distinguish, finally lead to chances of adulteration. The adulteration in medicinal plants effects on the efficacy of the drugs. The molecular authentication of herbal plant materials such as based on the internal transcribed spacer 2 (ITS2) sequences of nuclear ribosomal DNA (nrDNA) is considered as more reliable method compared to other the biochemical or histological methods. The present study aims to molecular authentication ofA. deserti based on molecular phylogenetic analyses of ITS2 gene sequence of nrDNA region. The ITS2 region of nrDNA of A. deserti were sequenced, and the molecular phylogenetic analyses were performed together with the GenBank sequences. The Maximum Parsimony tree revealed the close relationships of A. deserti with A. melampodina; however, the Neighbor-Joining and Maximum Likelihood tree clearly revealed that A. deserti is distinct from A. melampodina, which is also supported by the differences in nucleotides at five diffident positions (i.e. 22, 28, 87, 175 and 198) in the DNA sequence alignment.     

Internal transcribed spacer region evolution in Larix and Pseudotsuga (Pinaceae)

The nuclear ribosomal DNA (nrDNA) internal transcribed spacer (ITS) region has been characterized in the sister genera Larix and Pseudotsuga (Pinaceae). Complete sequences were obtained for seven species of Larix from North America and Eurasia and five species of Pseudotsuga from western North America and eastern Asia. ITS region lengths ranged from 1759 to 1770 bp in Larix and from 1564 to 1571 bp in Pseudotsuga. In both genera, ITS1 is three times as long as the 5.8S plus ITS2 and contains subrepeats as observed in other genera of Pinaceae. Secondary structure models predicted that the subrepeats fold into terminal stem and loop domains. ITS polymorphism detected within individuals of Larix and Pseudotsuga suggests a slow rate of concerted evolution among nrDNA loci. Except for the placement of L. sibirica, phylogenetic analyses of the ITS region agreed with previously reported restriction site analyses of Larix and Pseudotsuga. The data were not consistent with phylogenetic hypotheses for Larix based primarily upon ovulate cone characters, failing to support a derivation of the North American L. laricina from a short-bracted Eurasian lineage. The phylogenetic hypothesis did not conflict with a stepping stone model of evolution for Pseudotsuga, but a basal lineage could not be inferred for either genus.     

Molecular phylogeny of selected Old World Astragalus (Fabaceae): incongruence among chloroplast trnL-F, ndhF and nuclear ribosomal DNA ITS sequences

This study reports maximum parsimony and Bayesian phylogenetic analyses of selected Old World Astragalus using two chloroplast fragments including trnL-F and ndhF and the nuclear ribosomal internal transcribed spacer (nrDNA ITS). A total of 52 taxa including 34 euploid Old World and New World Astragalus , one aneuploid species from the Neo-Astragalus clade as a representative and 14 other Astragalean taxa, plus Cheseneya astragalina and two species of Caragana as outgroups were analyzed for both trnL-F and nrDNA ITS regions. ndhF was analyzed in 30 taxa and the same number for the combination of these three datasets were examined. In general, the trnL-F dataset and the ndhF and nrDNA ITS datasets generated more or less the same clades within Astragalus . However, in the trnL-F and ndhF phylogenies, Astragalus species are not gathered in a single clade, the so-called Astragalus s.s., as indicated by the nrDNA ITS tree. Visual inspection of these three phylogenies revealed that they were inconsistent regarding the position and relationships of Astragalus hemsleyi , A. ophiocarpus , A. annularis–A. epiglottis / Astragalus pelecinus, A. echinatus and A. arizonicus . Incongruence length difference test suggested that the trnL-F , ndhF and nrDNA ITS datasets were incongruent. In spite of this, phylogenetic analyses of the combined datasets as one unit or as three partitions generated trees that were topologically similar as a mix of the cpDNA and the nrDNA ITS trees. However, the combined dataset provided more resolved and statistically supported clades. The recently described A. memoriosus appeared closely related to A. stocksii (both from sect. Caraganella ) based on both trnL-F and nrDNA ITS sequences.