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。     

11种金花茶植物的RAPD分析及其系统学意义

利用随机扩增多态性DNA(RAPD)技术分析了11种金花茶植物,即:龙州金花茶Camellia longzhouensis J.Y．Luo,弄岗金花茶C．longgangensis C.F.Liang et S．L. Mo,大样金花茶 C．grandis(C. F. Liang et S．L．Mo)H．T,Chang et S．Y.Liang,扶绥金花茶 C.fusuiensis S．Y. Liang et X．J.Dong,中东金花茶C．achrysantha H．T．Chang et S．Y．Liang,小花金花茶C. micrantha S．Y. Liang et Y．C．Zhong,薄叶金花茶C．chrysanthoides H．T．Chang,毛籽金花茶C．ptilosperma S．Y．Liang et Q．D．Chen,陇瑞金花茶C．longruiensis S．Y．Liang et X．J．Dong,凹脉金花茶C．impressinervis H．T．Chang et S．Y．Liang和夏石金花茶C．xiashiensis S. Y．Liang et C．Z. Deng。选用14个引物(10 bp)扩增出201个DNA片段,用于种间遗传相似性分析,构建系统发育树图。分析结果表明:(1)小花金花茶、扶绥金花茶与中东金花茶之间的关系较近;(2)大样金花茶、薄叶金花茶和夏石金花茶之间的关系较近;(3)龙州金花茶、毛籽金花茶、陇瑞金花茶和弄岗金花茶之间的关系较近;(4)凹脉金花茶与其他各种关系相对较远。     

滇南桫椤的系统位置:来自叶绿体trnL内含子和DNA trnL-F间隔区序列的证据

为了探讨滇南桫椤Alsophilaaustroyunnanensis的系统位置 ,我们对该植物与中华桫椤Alsophilacostularis的叶绿体DNAtrnL内含子和trnL F间隔区序列进行PCR扩增和序列测定。滇南桫椤的trnL内含子、trnL F间隔区序列的长度分别为 5 70bp ,36 2bp ;中华桫椤的trnL内含子、trnL F间隔区序列的长度分别为 5 72bp ,36 1bp。结合已经发表的其他桫椤科植物的相应序列进行系统发育分析 ,用简约法和邻接法构建的系统发育树基本一致。结果表明 :1)所分析的桫椤科植物构成一个单系群 ;2 )滇南桫椤与Gymnosphaerapodophylla、Gymnosphaerapectinata、Gymnosphaerapseudogigantea、Gymnosphaeratinganensis、Gymnosphaeragigantea关系较近 ,聚成一支系 ;3)中华桫椤与Cyatheatsangii、Alsophilaspinulosa关系较近 ,聚成一支系。本文系统发育分析的结果支持将滇南桫椤归入黑桫椤属Gymnosphaera。     

基于叶绿体四个DNA片段联合分析探讨山茶属长柄山茶组、金花茶组和超长柄茶组的系统位置与亲缘关系

为探讨山茶属茶亚属(Camellia subgen.Thea)中长柄山茶组(sect.Longipedicellata)、金花茶组(sect.Chrysantha)和超长柄茶组(sect.Longissima)的系统位置和亲缘关系,本研究选取了该属4个亚属11组28个种及2个外类群的材料,对这些材料的叶绿体4个DNA片段(rp/16、psbA-trnH、trnL-F和rp/32-trnL)进行了测序,运用邻接法(neighbor-joining)、最大简约法(maximum-parsimony)和贝叶斯推断(Bayesian inference)对获得的序列进行了联合矩阵分析.并构建基因树.基因树的拓扑结构显示:1)金花茶组包括3个平行的支系,并且长柄山茶组的模式种长柄山茶(Camellia longipedicellata)嵌于其中一个支系,因而金花茶组可能是一个并系或多系类群;2)长柄山茶与越南分布的金花茶组种类在分子系统树上构成一个单系支,暗示了长柄山茶组和金花茶组之间可能具有紧密的亲缘关系;3)超长柄茶组不是一个单系类群,该组的河口超长柄茶(C.hekouensis)位于系统树的基部,与山茶属其余种构成姐妹群.由于缺乏更广泛取样的分析,超长柄茶在山茶属中的系统位置仍然不明确,超长柄茶组与长柄山茶组的亲缘关系问题也没有得到解决.     

基于叶绿体四个DNA 片段联合分析探讨山茶属长柄山茶组、金花茶组和超长柄茶组的系统位置与亲缘关系

为探讨山茶属茶亚属 (Camellia subgenThea) 中长柄山茶组 (sectLongipedicellata)、金花茶组 (sectChrysantha)和超长柄茶组 (sectLongissima) 的系统位置和亲缘关系,本研究选取了该属4个亚属11组28个种及2个外类群的材料,对这些材料的叶绿体4个DNA片段 (rpl16、psbA trnH、trnL F和rpl32 trnL) 进行了测序,运用邻接法 (neighbor joining)、最大简约法 (maximum parsimony) 和贝叶斯推断 (Bayesian inference) 对获得的序列进行了联合矩阵分析,并构建基因树。基因树的拓扑结构显示：1) 金花茶组包括3个平行的支系,并且长柄山茶组的模式种长柄山茶 (Camellia longipedicellata) 嵌于其中一个支系,因而金花茶组可能是一个并系或多系类群;2) 长柄山茶与越南分布的金花茶组种类在分子系统树上构成一个单系支,暗示了长柄山茶组和金花茶组之间可能具有紧密的亲缘关系;3) 超长柄茶组不是一个单系类群,该组的河口超长柄茶 (C. hekouensis) 位于系统树的基部,与山茶属其余种构成姐妹群。由于缺乏更广泛取样的分析,超长柄茶在山茶属中的系统位置仍然不明确,超长柄茶组与长柄山茶组的亲缘关系问题也没有得到解决。     

珍稀濒危植物越南小花金花茶的叶绿体基因组特征分析

越南小花金花茶(Camellia minima)是一种珍稀濒危的金花茶组植物,目前尚未有其叶绿体基因组的相关研究。该研究利用Illumina HiSeq 2000平台对越南小花金花茶进行了叶绿体基因组序列的测序、组装、注释和分析。结果表明：(1)越南小花金花茶叶绿体基因组全长156 961 bp,为典型的四分体结构,共注释到136个基因,其中包含87个蛋白编码基因、41个tRNA基因和8个rRNA基因。(2)分析鉴定出66个SSR位点和39个重复序列。(3)密码子偏好使用以A/U结尾的密码子,综合有效密码数(ENC)绘图、PR2-plot和中性分析推测自然选择是影响密码子使用模式的主导因素。(4)边界分析显示ycf1基因的长度和位置在不同金花茶组植物间存在差异。(5)对已发表的金花茶组植物叶绿体基因组的系统发育分析发现越南小花金花茶与小花金花茶聚为一支,亲缘关系最近。该研究结果为探索物种进化和提高外源基因表达提供了重要的参考信息,同时为后续的金花茶组植物的保护和利用提供了理论基础。     

山茶属金花茶组金花茶系的AFLP分析

应用扩增片段长度多态性(AFLP)分子标记方法,对分布于我国的金花茶组金花茶系的35个样品进行了分析,样品包括了《中国植物志》收录的该类群的15种2变种、2个未收录的种和4个已归并但在分类处理上存在分歧的种。4对引物组合：E—ACG／M—CAG,E—ACG／M—CTG,E—AGG／M—CTG,E—AGG／M—CAT用于选择性扩增,其中EcoRI引物的5′端用荧光染料进行标记。这4个引物组合共得到298条扩增带,其中278条带是多态性的。计算了样品间的Nei和Li(1979)相似性系数,基于这一相似性系数应用UPGMA法进行聚类分析,得到了树状分枝图。分析结果表明：(1)贵州金花茶是一个好种;(2)支持将毛籽金花茶、陇瑞金花茶、弄岗金花茶和大样弄岗金花茶归并到淡黄金花茶的观点;(3)薄叶金花茶、小花金花茶、夏石金花茶和小瓣金花茶之间的亲缘关系较近。     

基于ITS序列探讨珍珠菜属过路黄组的系统关系

测定了紫脉过路黄、临时救和过路黄的nrDNA ITS序列,并分析了珍珠菜属过路黄组17个物种的遗传距离及亲缘关系。结果表明,过路黄组植物的ITS序列长度在620～628间,一致性高达90.59%,种间遗传距离为0.002～0.199。系统发育树表明:(1)紫脉过路黄、临时救和小茄亲缘关系较近;(2)大叶过路黄、落地梅和过路黄亲缘关系较近;(3)山萝过路黄、贯叶过路黄、管茎过路黄、叶头过路黄、峨眉过路黄及三角叶过路黄亲缘关系较近。ITS序列分析结果为组内植物的鉴定、分类及系统进化提供了新的参考。     

基于rDNAITS序列分析Alternaria与相似属的关系及A.leucanthemi的分类地位

对Alternaria,Stemphylium,Ulocladium,Nimbya,Embellisia等五个形态相似属的代表性种进行了5.8S/ITS区段序列测定,连同从GenBank下载的相关有性型Pleospora,Lewia的同项资料,由Neighbor-joining方法构建系统发育树。序列对比结果显示,所有参试的菌株/种的5.8SrDNA序列保守程度较高,ITS序列变化较大,ITS1比ITS2变化更大。由5.8S/ITS构建的系统发育树可以足够将供试菌区分到属;系统发育树显示Alternaria,Ulocladium,Nimbya和Embellisia系统学关系较近,又存在一定的分化;Stemphylium聚在相对较远的分支上,与上述四属关系较远,相对独立进化;有性型的Pleospora与无性型的Stemphylium被聚到一个分支,这种不同阶段菌株在遗传基础上的一致,充分证明所用方法及所选测序的rDNA区段对本类群真菌的分类很有价值。Alternarialeucanthemi位于系统发育树的外侧,较为独立进化,与Alternaria的其它种亲缘关系较远,却与Stemphyliumspp.和Bipolarisspp.关系较近,对其目前的分类地位提出质疑。     

Phylogenetic relationships of sect.Nummularia (Lysimachia):evidence from ITS sequences

测定了紫脉过路黄、临时救和过路黄的nrDNA ITS序列,并分析了珍珠菜属过路黄组17个物种的遗传距离及亲缘关系.结果表明,过路黄组植物的ITS序列长度在620～628间,一致性高达90.59%,种间遗传距离为0.002～0.199.系统发育树表明:(1)紫脉过路黄、临时救和小茄亲缘关系较近;(2)大叶过路黄、落地梅和过路黄亲缘关系较近;(3)山萝过路黄、贯叶过路黄、管茎过路黄、叶头过路黄、峨眉过路黄及三角叶过路黄亲缘关系较近.ITS序列分析结果为组内植物的鉴定、分类及系统进化提供了新的参考.     

Phylogeny of Camellia sects Longipedicellata,Chrysantha and Longissima (Theaceae) Based on Sequence Data of Four Chloroplast DNA Loci

We focused on the systematic positions and relationships of three sections of Camellia subgenThea of Theaceae, i.e., sect.Longipedicellata, sect.Chrysantha (golden camellias) and sect.Longissima by using four chloroplast DNA regions (rpl16, psbA trnH, trnL F & rpl32 trnL). We sampled 28 species representing four subgenus, 11 sections in Camellia and two outgroups. Combined analyses of chloroplast DNA sequence data sets are performed with the neighbor joining, maximum parsimony and Bayesian inference methods, and the gene trees are constructed. The topologies of gene trees revealed that: 1) sect. Chrysantha is paraphyletic or polyphyletic, containing three parallel lineages and Camellia longipedicellata (type of sectLongipedicellata) nested inside; 2) all four species from Vietnam, together with C. longipedicellata, forming a well supported monophyletic clade, which implies the close relationship between sect. Longipedicellata and sect.Chrysantha; and 3) sectLongissima is not monophyletic because C.hekouensis is the sister to the rest of Camellia species. The systematic position of C.longissima and the relationship between sect.Longissima and sect.Longipedicellata are unsolved.     

Complex biogeographic patterns in Androsace (Primulaceae) and related genera: evidence from phylogenetic analyses of nuclear internal transcribed spacer and plastid trnL-F sequences

We conducted phylogenetic analyses of Androsace and the closely related genera Douglasia, Pomatosace, and Vitaliana using DNA sequences of the nuclear internal transcribed spacer (ITS) and the plastid trnL-F region. Analyses using maximum parsimony and Bayesian inference yield congruent relationships among several major lineages found. These lineages largely disagree with previously recognized taxonomic groups. Most notably, (1) Androsace sect. Andraspis, comprising the short-lived taxa, is highly polyphyletic; (2) Pomatosace constitutes a separate phylogenetic lineage within Androsace; and (3) Douglasia and Vitaliana nest within Androsace sect. Aretia. Our results suggest multiple origins of the short-lived lifeform and a possible reversal from annual or biennial to perennial habit at the base of a group that now contains mostly perennial high mountain or arctic taxa. The group containing Androsace sect. Aretia, Douglasia, and Vitaliana includes predominantly high alpine and arctic taxa with an arctic-alpine distribution, but is not found in the European and northeastern American Arctic or in Central and East Asia. This group probably originated in Europe in the Pliocene, from where it reached the amphi-Beringian region in the Pleistocene or late Pliocene.     

Infrageneric Phylogeny of the Genus Viola (Violaceae) Based on trnL trnF,psbA trnH,rpL16, ITS Sequences,Cytological and Morphological Data

The sequences of chloroplast trnL trnF (20 species), psbA trnH(17 species), rpL16 (17 species) and ITS(one species) regions of the genus Viola were analyzed for phylogenetic relationships in the genus with the maximum parsimony (MP) and Bayesian inference (BI) analyses. Hybanthus enneaspermus, Scyphellandra pierrei and Rinorea bengalensis were used as outgroups. Additional sequences corresponding to the same DNA regions for the other taxa in Viola were downloaded from GenBank. The phylogenetic tree indicated that subgenViola is not monophyletic. Moreover, some intrageneric relationships in Viola were clarified. Combining sequences with morphological characters and chromosome number, the inference of morphological evolution in Viola was put forward. It was inferred that: 1) erect stems might well be more primitive than stolons and rosettes; 2) long fimbriate margin in stipules and stipules with 1/2-3/4 adnate part might represent distinctive evolutionary trends of sectTrigonocarpae and of sectAdnatae respectively; 3) species with stigmatic beaks might have been derived from ancestors without beaks; simple structure stigmas transformed into complex structure stigmas, and then transformed back to simple stucture patterns.     

基于rDNA ITS序列的油茶叶片内生真菌多样性研究(英文)

油茶是我国热带和亚热带地区广泛栽培的一类重要的经济树种,其种子可生产优质食用油(茶油)。为了比较全面地了解油茶的内生真菌多样性,采用基于rDNA ITS的免培养法从油茶叶片中提取了总DNA,再从总DNA样本中直接扩增了内生真菌核糖体RNA基因内转录间隔区序列(ITS),进而构建了ITS克隆文库。共计从50个克隆子中获得了22种不同的克隆序列,其中3个为嵌合体,7个归属为植物,其余克隆序列根据序列相似性和系统发育分析归为12个不同的分类操作单元(OTUs),全部为子囊菌,分属于4纲4目,其中Aspergillus属2种,Mycosphaerellaceae科1种,Sordariales目4种,Helotiales目5种,Aspergillus属真菌是优势菌。结果表明油茶叶片内生真菌的种类分布较广。     

堇菜属组间的系统发育与亲缘关系：基于trnL-trnF、psbA trnH、rpL16、ITS序列，细胞学以及形态学证据

以鼠鞭草(Hybanthus enneaspermus)、鳞隔堇(Scyphellandra pierrei)、雷诺木(Rinorea bengalensis)作为外类群,对堇菜属(Viola)20个类群的trnL trnF序列,17个类群的psbA trnH、rpL16序列以及1个类群的nrDNA ITS序列进行了测定,并从GenBank下载相应的序列,运用最大简约法以及贝叶斯推论法进行系统分析,构建系统发育树。结果表明：堇菜亚属(subgen.Viola)不是一个单系类群,并明确了堇菜属部分组间类群的亲缘关系。本文还结合形态与细胞学证据对堇菜属进行性状演化的推测。结果表明：1）直立茎较匍匐茎、莲座状茎（叶基生）原始;2）托叶边缘长流苏状与托叶1/2~3/4合生分别是鸟嘴柱头堇菜组（sect.Trigonocarpae）和合生托叶组（sect.Adnatae）演化路线的重要性状标志;3）花柱样式从柱头无喙演化至柱头有喙,并由柱头简单演化至柱头复杂,再趋向于柱头简化。     

Molecular systematics of the genus Megoura (Hemiptera: Aphididae) using mitochondrial and nuclear DNA sequences

To construct the molecular systematics of the genus Megoura (Hemiptera: Aphididae), DNA based-identifi-cation was performed using four mitochondrial and three nuclear DNA regions: partial cytochrome c oxidase I (COI), partial tRNA-leucine + cytochrome c oxidase II (tRNA/COII), cytochrome b (CytB), partial 12S rRNA + tRNA-valine + 16S rRNA (12S/16S), elongation factor-1 alpha (EF1 alpha), and the internal transcribed spacers 1 and 2 (ITS1, ITS2). Pairwise sequence divergences between taxa were compared, and phylogenetic analyses were performed based on each DNA region separately, and the combined datasets. COI, CytB, EF1 alpha, ITS1, and ITS2 were relatively effective in determining species and resolving their relationships. By contrast, the sequences of tRNA/COII and 12S/16S were not able to separate the closely related species. CytB and EF1alpha gave better resolution with higher average sequence divergences (4.7% for CytB, 5.2% for EF1 alpha). The sequence divergence of COI (3.0%) was moderate, and those of the two ITS regions (1.8% for ITS1, 2.0% for ITS2) were very low. Phylogenetic trees were constructed by minimum evolution, maximum parsimony, maximum likelihood, and Bayesian phylogenetic analyses. The results indicated that the phylogenetic relationships between Megoura species were associated with their host preferences. Megoura brevipilosa and M. lespedezae living on Lespedeza were closely related, and M. nigra, monophagous on Vicia venosa, was rather different from M. crassicauda, M. litoralis, and M. viciae, which are oligophagous on Lathyrus and Vicia. The three populations of M. crassicauda formed a clade separated from M. litoralis and M. viciae. Nevertheless M. litoralis and M. viciae, which are morphologically similar, were not separated due to negligible sequence divergence. We discuss the phylogenetic relationships of the Megoura, and the usefulness of the seven DNA regions for determining the species level phylogeny of aphids.     

Phylogenetic position of endosymbiotic green algae in Paramecium bursaria Ehrenberg from Japan

Endosymbiotic green algae of Japanese Paramecium bursaria were phylogenetically analyzed based on DNA sequences from the ribosomal DNA operon (18S rDNA, ITS1, 5.8S rDNA, and ITS2). Phylogenetic trees constructed using 18S rDNA sequences showed that the symbionts belong to the Chlorella sensu stricto (Trebouxiophyceae) group. They are genetically closer to the C. vulgaris Beijerinck group than to C. kessleri Fott et Nováková as proposed previously. Branching order in C. vulgaris group was unresolved in 18S rDNA trees. Compared heterogeneities of 18S rDNA, ITS1, 5.8S r, and ITS2 among symbionts and two Chlorella species, indicated that the ITS2 region (and probably also ITS1) is better able to resolve phylogenetic problems in such closely related taxa. All six symbiotic sequences obtained here (approximately 4000-bp sequences of 18S rDNA, ITS1, 5.8S rDNA, and ITS2) were completely identical in each, strongly suggesting a common origin.     

ITS1 Sequences of Nuclear Ribosomal DNA in Wild Rices and Cultivated Rices of China and Their Phylogenetic Implications

he first internal transcribed spacer (ITS1) of nuclear ribosomal DNA of three wild rice species and two subspecies of cultivated rice, which are distributed in China, was amplified using PCR technique and sequenced with automated fluorescent sequencing. The sequences of ITS1 ranged from 193 bp to 218 bp in size and G/C content varied from 69.3%to 72.7%. In pairwise comparison among the five taxa, sequence site divergence ranged from 1.5 % to 10.6%. Phylogenetic analysis of ITS1 sequences using Wagner parsimony generated a single well-resolved tree, which revealed that Oryza rufipogon was much more closely related to cultivated rice species than to the other two wild species. Oryza granulata was less closely related to either cultivated rice species or the other two wild species, and might be a unique and isolated taxon in the genus Oryza. The phylogenetic relationships of the three wild rice species and two cultivated rice subspecies inferred from ITS1 sequences is highly concordant with those based on the molecular evidence from isozyme, chloroplast DNA (cpDNA), mitochondrial DNA (mtDNA) and nuclear DNA (nDNA) of the genus Oryza.