梧桐科翅子树属的亲缘关系

翅子树属传统上均作为梧桐科山芝麻族的成员,但也有少数人将它归到Dombeyeae族中。本文根据对翅子树属的外部形态及花粉形态特征的研究,并结合其解剖、染色体和胚胎发育的资料,对翅子树属的亲缘关系进行了分析。翅子树属与山芝麻族在外部形态上比较接近,但在解剖、花粉形态和染色体基数等特征上则差异显著;与Dombeyeae族在花粉形态上一致,但在外部形态(花结构)、染色体基数和瓦形细胞等特征方面则明显不同,反映了翅子树属在这两个族之间的过渡状态,不宜作为任何一族的成员,应该作为一个独立的族存在,其在梧桐科中的位置在Dombeyeae附近。     

根据ITS序列证据重建防己科蝙蝠葛族的系统发育

研究了国产防己科蝙蝠葛族tirb．Menispermeae9属20种和外类群青牛胆族trib．Tinosporeae 2属3种植物完整的ITS(包括5.8S rDNA)序列。trib．Menispermeae的ITS长527～601 bp,排序后长667bp。当gap处理为missing时具281个有信息位点。PAUP软件分析结果表明：①trib．Menispermeae是一个单系类群,该分支得到hootstrap l00％的支持;②确定了存疑种Pachygone valida的系统学位置,该种是Coc—culus属的成员;③Sinomenium和Menispermum两属有很近的系统学关系,组成族内稳定的一支,它们的ITS序列同源性极高,ITS1比族内其它属长41～73bp;④Stephania和Cyclea也是系统发育关系很近的两个类群。前者具两个主要分支,其IIS1、ITS2的G+C含量差异较大,在种类组成上,该两大支与传统上Stephania属内处理的2个亚属——千金藤亚属subgen．Stephania和山乌龟亚属subgen．Tuberiphania基本一致;Cyclea属内种间的ITS序列差异小,同源性极高。     

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。     

基于三个DNA片段讨论菖蒲科的分子系统学

菖蒲属长期以来被置于天南星科,但近年来分子系统学的研究表明菖蒲属与天南星科的关系较远,应成立一个单型科,即菖蒲科（Acoraceae)。本文选择菖蒲科3个种及1个变种即金钱蒲（Acorus gramineus),石菖蒲（A.ttrinowii)。金边菖蒲（A.tatarinowii),金边菖蒲（A.tatarinowii var.flawo-marginatus)和菖蒲（A.calamus)不同染色体倍性的4个居群进行了核糖体RNA内源转录间隔段（ITS）进行测序。并据此讨论了菖蒲属的属下系统关系,结果表明,菖蒲科为一个自然的单系类群,不同倍性的菖蒲构成一个自展数据支持率为100%的分支,无中肋的金钱蒲,石菖蒲和金边石菖蒲构成另一个自展数据支持率为100%的分支,通过比较分析菖蒲科与其相关类群之间18S和rbcL两段DNA序列,对菖蒲科的系统位置进行了讨论,在基于18S和rbcL的系统树中,菖蒲均处于单子叶植物中较为孤立的位置。     

最大似然法和贝叶斯推论与似然比检验探讨泽泻目系统发育关系

应用最大似然法(ML)、贝叶斯推论(BI)、邻接法(NJ)和似然比检验(hLRTs)进行泽泻目分子系统学研究。所用的rbcL基因序列代表了泽泻目14科46属以及作为外类群的6相关属。研究结果表明,*等级制似然比检验表明泽泻目rbcL序列最适合的DNA进化模型为GTR+I+G,最大似然法、贝叶斯法和邻接法构建的系统发育树拓扑结构相似,没有显著的差异,但贝叶斯树支持率较高;泽泻目为一单系类群,由两个主要谱系分支构成,深层分布格局由5个主要分支构成。基于分子系统发育树,文中对泽泻目科间、水鳖科+茨藻科、泽泻科+花蔺科+黄花蔺科、和"Cymodoeaceae complex"的系统发育关系进行了讨论。研究结果还表明,泽泻目系统发育关系可能还需要更多的证据进一步的澄清。     

锦葵科植物DNA条形码通用序列的筛选

对锦葵科植物样品的ITS、ITS2、rbcL、matK和psbA-trnH序列进行PCR扩增和测序,比较各序列的扩增效率、测序成功率、种内和种间变异的差异以及barcoding gap图,使用BLAST1和Nearest Distance方法评价不同序列的鉴定能力,进而从这些候选序列中筛选出较适合锦葵科植物鉴别的DNA条形码序列。结果表明,ITS序列在采集的锦葵科植物11个种26个样品中的扩增成功率较高,其种内、种间变异差异和barcoding gap较ITS2、psbA-trnH及rbcL序列具有更明显的优势,且纳入60个属316个种共1228个样品的网上数据后,其鉴定成功率可达89.9%。psbA-trnH序列的扩增和测序成功率最高,其鉴定成功率为63.2%,并能鉴别一些ITS序列无法鉴别的种。实验结果表明,ITS和psbA-trnH是较适合鉴别锦葵科植物的DNA条形码序列组合。     

rDNA片段的序列分析在苦苣苔亚科系统学研究中的应用

本文测定了苦苣苔亚科4族、5属、5种植物的核糖体DNA中的内转录间隔区(ITS)序列及5．8s rRNA基因的3′端序列。这几种苦苣苔亚科植物的ITS-1的长度范围为234～258 bp,ITS-2的长度范 围为218～246bp。Whytockia bijieensis的ITS-1(258bP)和ITS-2(218 bp)在长度、序列及GC含量上 均与其它几个种有较大差异,其代表的尖舌苣苔族可能很早就自苦苣苔亚科的祖先沿单独的一个分支 演化。以w．bijieensis作为功能性外类群,运用PAUP软件分析仅得到一个最简约树。在简约树上, Cyrtandra umbelliferm、Briggsia longipes和Anna mollifolia形成一个单系群,bootstrap分析对该分支的 支持强度达97％,Chirita crasslfolia位于该分支的基部。由于系统树上Cyrtandra umbellifera代表的 浆果苣苔族和Anna mollifolid代表的芒毛苣苔族均起源于长蒴苣苔族,结合这3个族在形态上存在过渡系列,建议将浆果苣苔族和芒毛苣苔族均并入长蒴苣苔族。     

rDNA片段的序列分析在苦苣苔亚科系统学研究中的应用

本文测定了苦苣苔亚科4族、5属、5种植物的核糖体DNA中的内转录间隔区(ITS)序列及5．8s rRNA基因的3′端序列。这几种苦苣苔亚科植物的ITS-1的长度范围为234～258 bp,ITS-2的长度范 围为218～246bp。Whytockia bijieensis的ITS-1(258bP)和ITS-2(218 bp)在长度、序列及GC含量上 均与其它几个种有较大差异,其代表的尖舌苣苔族可能很早就自苦苣苔亚科的祖先沿单独的一个分支 演化。以w．bijieensis作为功能性外类群,运用PAUP软件分析仅得到一个最简约树。在简约树上, Cyrtandra umbelliferm、Briggsia longipes和Anna mollifolia形成一个单系群,bootstrap分析对该分支的 支持强度达97％,Chirita crasslfolia位于该分支的基部。由于系统树上Cyrtandra umbellifera代表的 浆果苣苔族和Anna mollifolid代表的芒毛苣苔族均起源于长蒴苣苔族,结合这3个族在形态上存在过渡系列,建议将浆果苣苔族和芒毛苣苔族均并入长蒴苣苔族。     

DNA序列在悬钩子属植物分子系统学研究中的应用进展

悬钩子属植物种类繁多,类群复杂,而且多为多倍体和杂种。该文就近年来国内外有关DNA序列在悬钩子属植物分子系统学研究中的应用现状和进展进行了综述,并对中国悬钩子属植物系统发育研究进行了展望。研究认为:叶绿体DNA序列多应用非编码区,且多与ITS序列联合分析;核基因组中ITS序列应用最为广泛,主要用于研究悬钩子属空心莓组与木莓组的进化关系、栽培品种间亲缘关系及部分杂种和多倍体的起源等;在该属植物中发现了ITS个体内多态性,但未进行ITS假基因检测,其系统学应用价值需重新评价;低拷贝核基因只有GBSSI和LEAFY有相关应用。同时认为,悬钩子属植物系统学研究中应用的DNA序列及研究类群均较少,缺乏对整个悬钩子属全面而系统的研究。指出应进一步选择具有代表性的样本、筛选合适的DNA片段,并结合形态学、孢粉学和细胞学等手段对中国悬钩子属植物系统关系进行深入研究。     

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。     

Escaping introns in COI through cDNA barcoding of mushrooms: Pleurotus as a test case

DNA barcoding involves the use of one or more short, standardized DNA fragments for the rapid identification of species. A 648‐bp segment near the 5′ terminus of the mitochondrial cytochrome c oxidase subunit I (COI) gene has been adopted as the universal DNA barcode for members of the animal kingdom, but its utility in mushrooms is complicated by the frequent occurrence of large introns. As a consequence, ITS has been adopted as the standard DNA barcode marker for mushrooms despite several shortcomings. This study employed newly designed primers coupled with cDNA analysis to examine COI sequence diversity in six species of Pleurotus and compared these results with those for ITS. The ability of the COI gene to discriminate six species of Pleurotus, the commonly cultivated oyster mushroom, was examined by analysis of cDNA. The amplification success, sequence variation within and among species, and the ability to design effective primers was tested. We compared ITS sequences to their COI cDNA counterparts for all isolates. ITS discriminated between all six species, but some sequence results were uninterpretable, because of length variation among ITS copies. By comparison, a complete COI sequences were recovered from all but three individuals of Pleurotus giganteus where only the 5′ region was obtained. The COI sequences permitted the resolution of all species when partial data was excluded for P. giganteus. Our results suggest that COI can be a useful barcode marker for mushrooms when cDNA analysis is adopted, permitting identifications in cases where ITS cannot be recovered or where it offers higher resolution when fresh tissue is. The suitability of this approach remains to be confirmed for other mushrooms.     

Divergent and reticulate species relationships in Leucaena (Fabaceae) inferred from multiple data sources: insights into polyploid origins and nrDNA polymorphism

Previous analyses of species relationships and polyploid origins in the mimosoid legume genus Leucaena have used chloroplast DNA (cpDNA) restriction site data and morphology. Here we present an analysis of a new DNA sequence data set for the nuclear ribosomal DNA (nrDNA) 5.8S subunit and flanking ITS 1 and ITS 2 spacers, a simultaneous analysis of the morphology, ITS and cpDNA data sets for the diploid species, and a detailed comparison of the cpDNA and ITS gene trees, which include multiple accessions of all five tetraploid species. Significant new insights into species relationships and polyploid origins, including that of the economically important tropical forage tree L. leucocephala, are discussed. Heterogeneous ITS copy types, including 26 putative pseudogene sequences, were found within individuals of four of the five tetraploid and one diploid species. Potential pseudogenes were identified using two pairwise comparison approaches as well as a tree-based method that compares observed and expected proportions of total ITS variation contributed by the 5.8S subunit optimized onto branches of one of the ITS gene trees. Inclusion of putative pseudogene sequences in the analysis provided evidence that some pseudogenes in allopolyploid L. leucocephala are not the result of post-allopolyploidization gene silencing, but were inherited from its putative diploid maternal progenitor L. pulverulenta.     

Phylogeny of the core Malvales: evidence from ndhF sequence data

The monophyly of the group comprising the core malvalean families, Bombacaceae, Malvaceae, Sterculiaceae, and Tiliaceae, was recently confirmed by molecular studies, but the internal structure of this clade is poorly understood. In this study, we examined sequences of the chloroplast ndhF gene (aligned length 2226 bp) from 70 exemplars representing 35 of the 39 putative tribes of core Malvales. The monophyly of one traditional family, the Malvaceae, was supported in the trees resulting from these data, but the other three families, as traditionally circumscribed, are nonmonophyletic. In addition, the following relationships were well supported: (1) a clade, /Malvatheca, consisting of traditional Malvaceae and Bombacaceae (except some members of tribe Durioneae), plus Fremontodendron and Chiranthodendron, which are usually treated as Sterculiaceae; (2) a clade, /Malvadendrina, supported by a unique 21-bp (base pair) deletion and consisting of /Malvatheca, plus five additional subclades, including representatives of Sterculiaceae and Tiliaceae, and Durionieae; (3) a clade, /Byttneriina, with genera traditionally assigned to several tribes of Tiliaceae, plus exemplars of tribes Byttnerieae, Hermannieae, and Lasiopetaleae of Sterculiaceae. The most striking departures from traditional classifications are the following: Durio and relatives appear to be more closely related to Helicteres and Reevesia (Sterculiaceae) than to Bombacaceae; several genera traditionally considered as Bombacaceae (Camptostemon, Matisia, Phragmotheca, and Quararibea) or Sterculiaceae (Chiranthodendron and Fremontodendron) appear as sister lineages to the traditional Malvaceae; the traditional tribe Helictereae (Sterculiaceae) is polyphyletic; and Sterculiaceae and Tiliaceae, as traditionally circumscribed, represent polyphyletic groups that cannot sensibly be maintained with their traditional limits for purposes of classification. We discuss morphological characters and conclude that there has been extensive homoplasy in characters previously used to delineate major taxonomic groups in core Malvales. The topologies here also suggest that /Malvatheca do not have as a synapormophy monothecate anthers, as has been previously supposed but, instead, may be united by dithecate, transversely septate (polysporangiate) anthers, as found in basal members of both /Bombacoideae and /Malvoideae. Thus, “monothecate” anthers may have been derived at least twice, independently, within the /Bombacoideae (core Bombacaceae) and /Malvoideae (traditional Malvaceae).     

Molecular detection of Bartonella species infecting rodents in Slovenia

Rodents, collected in three zoogeographical regions across Slovenia, were tested for the presence of bartonellae using direct PCR-based amplification of 16S/23S rRNA gene intergenic spacer region (ITS) fragments from splenic DNA extracts. Bartonella DNA was detected in four species of rodents, Apodemus flavicollis, Apodemus sylvaticus, Apodemus agrarius and Clethrionomys glareolus, in all three zoogeographic regions at an overall prevalence of 40.4%. The prevalence of infection varied significantly between rodent species and zoogeographical regions. Comparison of ITS sequences obtained from bartonellae revealed six sequence variants. Four of these matched the ITS sequences of the previously recognized species, Bartonella taylorii, Bartonella grahamii, Bartonella doshiae and Bartonella birtlesii, but one was new. The identity of the bartonellae from which the novel ITS sequences was obtained were further assessed by sequence analysis of cell division protein-encoding gene (ftsZ) fragments. This analysis demonstrated that the strain is most likely a representative of possible new species within the genus.     

Its evolution in Platanus (Platanaceae): homoeologues, pseudogenes and ancient hybridization

BACKGROUND AND AIMS: Platanaceae is an old family of angiosperms extending back to the Early Cretaceous but consisting of a single extant genus, Platanus. Species of Platanus have long been known to hybridize, and the London plane, Platanus x hispanica, is a well-known example of a hybrid species that formed in historical times. In addition, morphological studies have suggested past interspecific or interlineage hybridization and reticulation as possibly important factors in the evolution of the genus. This study aims at unravelling the complex evolutionary information contained in internal transcribed spacer (ITS) sequences. METHODS: The ITS of the 35S nuclear ribosomal DNA are biparentally inherited, multi-copy markers with a high potential for resolving intrageneric relationships especially when ancient hybridization (reticulation) is involved. Phylogenetic trees, splits graphs and motif analysis are used to extract phylogenetic information from 223 cloned ITS sequences, representing ten species and varieties of Platanus. Non-pseudogenous and pseudogenous sequence motives are assessed to explain how different evolutionary modes contribute to possibly conflicting character state patterns in the ITS. KEY RESULTS: It was found that putative non-functional ITS copies ('pseudogenes') form distinct groups in phylograms and splits graphs, and that pseudogenous lineages reflect ancient hybridization events conserved in the ITS. Specifically, pseudogenous clones of an 'Atlantic' North American clade share sequence motives with non-pseudogenous clones of the western ('Pacific') North American P. racemosa species aggregate. In addition, evidence was found for recent lateral gene flow as a possible factor in the evolution of the central Mexican P. rzedowskii. CONCLUSIONS: Broad ITS data sets that cover intra- and interindividual variability reveal past and ongoing speciation processes in Platanus. Evolutionary pathways can be visualized with splits graphs, but not with bifurcating trees.     

9种梧桐科植物叶表皮特征的扫描电镜研究

用扫描电镜对9种梧桐科植物叶片进行观察研究以明确其叶表皮微形态特征差异.结果显示,绝大部分梧桐科植物的气孔类型和气孔形状相似,但其表皮的毛状体类型、形态以及表皮纹饰等方面有显著差异.结果表明9种梧桐科植物的叶表皮毛和表皮纹饰可作为梧桐科种间分类的特征,且研究结果支持将胖大海归属于胖大海属,将翅子树单独列为翅子树族的分类观点.     

Molecular systematics of Trilliaceae II. Phylogenetic analyses of Trillium and its allies using sequences of rbcL and matK genes of cpDNA and internal transcribed spacers of 18S–26S nrDNA

Coding regions of the rbcL and matK genes of cp DNA and internal transcribed spacers (ITS) of nuclear ribosomal DNA were sequenced to study phylogenetic relationships within and among all four genera of Trilliaceae: Trillium, Paris, Daiswa and Kinugasa . The rbcL gene has evolved much slower than matK and in particular ITS; hence the phylogenetic trees based on the rbcL gene show a much lower resolution than trees based on either matK or ITS. The general topology of phylogenetic trees resulting from separate parsimony analyses of the matK and ITS sequences are relatively congruent, with the exception of the placement of T. pusillum . Both matK and ITS phylogenies reveal that T. rivale diverges at the base of the trees. In both trees, Paris, Daiswa and Kinugasa form a relatively weakly supported group. Within this group, the allo-octaploid Kinugasa japonica is the sister group of Daiswa species. The Paris–Daiswa – Kinugasa group, the major Trillium group, and T. undulatum and T. govanianum showed a loosely related topology, but their affinities are not evident according to these two molecular markers. However, phylogenetic analysis of amino acid sequences derived from matK shows that T. rivale together with clades T. undulatum–T. govanianum, Daiswa–Kinugasa and Paris is basally diverged as a sister group to the remainder of Trillium .     

亚热带森林乔木树种DNA条形码研究——以哀牢山自然保护区为例

作为新一代植物志iFlora的重要组成部分,DNA条形码已经成为物种鉴定中重要且有效的方法。本研究以亚热带森林的乔木树种为研究对象,开展了DNA条形码的尝试性工作。为评估DNA条形码对鉴定亚热带森林树种的有效性,收集并研究了来自哀牢山自然保护区内5l科111属中204个树种的525个乔木个体。结果显示,所选4个DNA片段（rbcL,matK,trnH-psbA和ITS）的PCR扩增成功率都超过90％;测序成功率rbcL和matK最高,分别为90．7％和90．5％,trnH-psbA次之（83．6％）,ITS最低（73．5％）,表明4个片段在亚热带森林乔木中都具有较好的通用性。应用BLAST与NJ Tree两种方法,对物种和属水平的鉴别成功率进行统计,发现单片段中ITS最高,分别为68．4％-81．3％和99．0％～100％,核心条码rkL和matK组合的成功率是52．8％～60．2％和86．7％～90．5％,再与补充条码trnH-psbA和ITS联合,可以成功鉴别74．7％～79．6％哀牢山自然保护区亚热带森林中的乔木物种。由于ITS片段在亚热带森林部分重要树种类群（樟科和壳斗科等）中的测序成功率较差,所以对这些植物类群采用trnH-psbA作为DNA条形码是一个更好的选择。     

衣藻属的系统发育分析——基于形态形状和nrDNA ITS序列

通过实验分析莱茵衣藻 ( Chlamydomonas reinhardtii) 1个种和互连网获得衣藻属 1 5个种及丝藻属 1个种 ( Ulothrix zonata) ,共 1 7个种的 nr DNA ITS序列 ,并以 U.zonata为外类群 ,采用计算机分析软件包对其进行分析及构建分子系统发育树图。同时以 1 2个传统分类性状 ,对此 1 6种衣藻构建数据矩阵 ;以 U.zonata动孢子的相应性状为外类群原始性状 ,用Wagner法在计算机上对其进行分枝分析 ;然后比较并分析分子系统树和表征性状分支分析树的异同。初步尝试以 ITS分子序列系统发育分析作为传统性状分析的补充来研究衣藻种间的亲缘关系。