基于nrDNA ITS序列数据的兰属系统发育关系的初步分析

现存的兰属分类系统是基于宏观形态学性状、尤其是花粉块的数目以及唇瓣与蕊柱的愈合程度而建立的。兰属因此而划分为3个亚属：兰亚属（subgenus Cymbidium),大花亚属（subgenus Cyperorchis)和建兰亚属（subgenus Jensoa）。本文运用PCR扩增和直接测序的方法分析兰属（Cymbidium）27种、3个栽培品种以及3个外类群的核DNA ITS区段序列,通过最简约性分析产生的ITS系统发育树表明,兰属的3个亚属均可能为不自然的类群,大花亚属表现为一复系群,兰亚属的冬凤兰（C.dayanum）隐藏于基点;建兰亚属为一并系群,它的成员之一兔耳兰（C.lancifolium）偏离出去而成为兰属一最基部的分支;兰亚属为一复系群,它分为几支而分别与另两个亚属组合在一起,由于兰属ITS序列位点变异率较低,最简约性分析产生的几支主要分支均得不到Bootstrap分析的高度支持,各亚属内组之间的关系也不明确,研究兰属的系统发育关系还需要新的数据。     

基于nrDNAITS序列数据的兰属系统发育关系的初步分析(英)

现存的兰属分类系统是基于宏观形态学性状、尤其是花粉块的数目以及唇瓣与蕊柱的愈合程度而建立的。兰属因此而划分为 3个亚属 :兰亚属 (subgenusCymbidium) ,大花亚属 (subgenusCyperorchis)和建兰亚属 (subgenusJensoa)。本文运用PCR扩增和直接测序的方法分析兰属 (Cymbidium) 2 7种、3个栽培品种以及 3个外类群的核DNAITS区段序列。通过最简约性分析产生的ITS系统发育树表明 ,兰属的 3个亚属均可能为不自然的类群。大花亚属表现为一复系群 ,兰亚属的冬凤兰 (C .dayanum )隐藏于其中 ;建兰亚属为一并系群 ,它的成员之一兔耳兰(C .lancifolium)偏离出去而成为兰属一最基部的分支 ;兰亚属为一复系群 ,它分为几支而分别与另两个亚属组合在一起。由于兰属ITS序列位点变异率较低 ,最简约性分析产生的几支主要分支均得不到Bootstrap分析的高度支持 ,各亚属内组之间的关系也不明确。研究兰属的系统发育关系还需要新的数据。     

基于ITS序列对中国簇毛黄耆亚属(黄耆属)系统学问题的初步研究

测定了簇毛黄耆亚属(Pogonophace)4组8种和外类群Caragana roborovskyi的ITS序列,从GenBank中调出相关12属47种的ITS序列,组成数据距阵,应用PAUP程序中的最大简约法构建了系统发育树状图.扁荚组(Sect. Sesbanella)与亚属其余类群在系统树上处于不同的分支,亲缘关系较远,这个亚属不是一个单系类群;膨果组(Sect. Bibracteola),背扁组(Sect. Phyllolobium)和袋果组(Sect. Trichostylus)作为一个单系类群能得到ITS序列的支持,但与鱼鳔槐亚族比与黄耆属其他类群的关系更近; Astragatus complanatus和A. tribulifolius可能为一对替代种;亚属下的分组以及膨果组下系的划分都得不到ITS序列分析的支持.     

基于ITS序列对中国簇毛黄耆亚属（黄耆属）系统学问题的初步研究

测定了簇毛黄耆亚属（Pogonophace)4组8种和外类群Caragana roborovskyi的ITS序列,从GenBank中调出相关12属47种的ITS序列,组成数据距阵,应用PAUP程序中的最大简约法构建了系统发育树状图。扁荚组（Sect.Sesbanella)与亚属其余类群在系统树上处于不同的分支,亲缘关系较远这个亚属不是一个单系类群;膨果组（Sect.Bibracteola)。背扁组（Sect.Phyllolobium)和袋果组（Sect.Trichostylus)作为一个单系类群能得到ITS序列的支持,但与分组以及膨果组下系的划分都得不到ITS序列分析的支持。     

基于ITS序列探讨杜鹃属的亚属和组间系统关系

首次报道了15种杜鹃属(Rhododendron)植物、1种杜香属(Ledum)植物和Cassiope fastigiata的内转录间隔区(ITS) (包括5.8S)序列.加上从GenBank下载的13种杜鹃属植物和Bajiaria racemosa的ITS序列,以C. fastigiata和B. racemosa为外类群,用最大简约法对杜鹃属的亚属和组间的系统关系进行了分析.结果表明: 1)杜鹃属是一个单系类群,叶状苞亚属为杜鹃属的基部类群; 2)杜香属确应归并到杜鹃属中,且与有鳞杜鹃亚属有较近的亲缘关系; 3)有鳞杜鹃亚属和杜香构成一个单系分支,该分支是其余无鳞杜鹃花的姐妹群; 4)由无鳞杜鹃花组成的一个分支的内部支持率较低,其中常绿杜鹃亚属和映山红亚属均为内部支持率很高的单系类群,而羊踯躅亚属和马银花亚属均为多系类群; 5)在马银花亚属中,长蕊杜鹃组和马银花组均分别得到强烈支持,马银花组与异蕊杜鹃亚属可能构成姐妹群关系,异蕊杜鹃亚属和马银花组组成的一个分支可能与映山红亚属构成姐妹群关系.     

基于ITS序列探讨杜鹃属的亚属和组间系统关系

首次报道了 15种杜鹃属 (Rhododendron)植物、1种杜香属 (Ledum)植物和Cassiopefastigiata的内转录间隔区(ITS) (包括 5 .8S)序列。加上从GenBank下载的 13种杜鹃属植物和Bajiariaracemosa的ITS序列 ,以C .fastigiata和B .racemosa为外类群 ,用最大简约法对杜鹃属的亚属和组间的系统关系进行了分析。结果表明 :1)杜鹃属是一个单系类群 ,叶状苞亚属为杜鹃属的基部类群 ;2 )杜香属确应归并到杜鹃属中 ,且与有鳞杜鹃亚属有较近的亲缘关系 ;3)有鳞杜鹃亚属和杜香构成一个单系分支 ,该分支是其余无鳞杜鹃花的姐妹群 ;4 )由无鳞杜鹃花组成的一个分支的内部支持率较低 ,其中常绿杜鹃亚属和映山红亚属均为内部支持率很高的单系类群 ,而羊踯躅亚属和马银花亚属均为多系类群 ;5 )在马银花亚属中 ,长蕊杜鹃组和马银花组均分别得到强烈支持 ,马银花组与异蕊杜鹃亚属可能构成姐妹群关系 ,异蕊杜鹃亚属和马银花组组成的一个分支可能与映山红亚属构成姐妹群关系。     

紫乌头复合体nrDNA的ITS序列与系统发育分析

对乌头属 Aconitum L. 27个类群的nrDNA ITS序列进行了简约法与邻接法分析,两种方法得到的系统发育树基本一致。乌头亚属subgen. Aconitum 的蔓乌头系 ser. Volubilia 不是一个单系类群,显柱乌头系 ser. Stylosa 与兴安乌头系ser. Ambigua 各自作为单系类群也没有得到支持。特产于云南西北部横断山区的一些种之间存在非常近的系统发育关系,说明这些种可能是近期物种形成的产物。紫乌头 A. delavayi 复合体的不同类群在系统发育树上位     

中国紫金牛属的分支分类学研究

基于60个形态学性状,对中国广义报春花科（Primulaceae s.l.）紫金牛属（Ardisia）90个分类群的系统发育关系运用分支分析方法进行了分析。采用最简约性分析得到了100个同等简约树。50%多数规则一致树的分支结构与以前建立的紫金牛亚属划分系统基本一致。外类群酸藤子属、铁仔属、密花树属聚在分支树的最基部,紫金牛属为一单系类群。形态分支树的一致性指数和保持性指数和各分支内部支持率均较低,只在种与变种或亚种之间获得较高的支持率。高木亚属、腋序亚属、短序亚属、顶序亚属处于分支树较为基部的位置,推测这四个亚属的类群在紫金牛属中较为原始;圆齿亚属和锯齿亚属共同组成一大支,二者亲缘关系紧密,推测这两个亚属为该属中最为进化的类群。结合形态学对属内系统发育关系进行了讨论和推测了一些性状的演化趋势,以期为分类修订提供依据。     

基于叶绿体rps16基因和核基因ITS片段研究肉苁蓉属系统位置

在前人对列当科系统发育研究的基础上,追加了肉苁蓉属(Cistanche)的基因序列数据,运用最大简约法、最大似然法和贝叶斯推断方法探讨了其在列当科中的系统位置及列当科中属间关系.基于rps16基因序列及rps16+ ITS联合序列建立了列当科系统发育树,结果显示,肉苁蓉属、列当属(Orobanche)以及草苁蓉属(Boschniakia)聚在同一进化枝内,肉苁蓉属和列当属表现出最近亲缘关系;列当科中的全寄生类群、半寄生类群和非寄生类群分属在3个不同分支中.     

基于ITS序列探讨忍冬属的系统发育关系

以七子花(Heptacodium miconioides)为外类群,运用MEGA软件对20种忍冬属植物进行系统发育分析,采用邻接法(NJ)和最大简约法(MP)构建系统发育树,从分子系统学角度探讨忍冬属下的亲缘关系.结果表明:(1)在NJ和MP系统树中,没有形成系统树的基部分支,忍冬亚属(Subg.Chamaecerasus)和轮花亚属(Subg.Lonicera)没有形成姐妹群关系.(2)在各系统树中,囊管组内的各种没有聚为一支,故认为对囊管组的划分应进一步探讨.(3)忍冬属ITS区(ITS1+ITS2)的信息位点达到11.0%,信息位点比较丰富,证明ITS序列可以为解决忍冬属植物的系统发育问题提供较强的证据.     

利用RAPD技术分析云南兰属部分种的种间关系

采用RAPD技术对云南兰属的11个种和3个变种中的39个样本进行了相似性分析。在相似系数0．58水平上,大花亚属的虎头兰、西藏虎头兰、长叶兰和碧玉兰聚为一支;建兰亚属的寒兰、墨兰、蜜蜂兰、蕙兰、春兰、豆瓣兰、春剑、莲瓣兰和送春聚为另一支,兔耳兰与大花亚属关系更近。春剑和莲瓣兰相似性更高,它们与春兰的关系较远,不支持送春作为蕙兰下的变种。这些结果可为开展兰属育种提供重要参考价值。     

利用RAPD 技术分析云南兰属部分种的种间关系

采用RAPD 技术对云南兰属的11 个种和3 个变种中的39 个样本进行了相似性分析。在相似系数0. 58 水平上, 大花亚属的虎头兰、西藏虎头兰、长叶兰和碧玉兰聚为一支; 建兰亚属的寒兰、墨兰、蜜蜂兰、蕙兰、春兰、豆瓣兰、春剑、莲瓣兰和送春聚为另一支, 兔耳兰与大花亚属关系更近。春剑和莲瓣兰相似性更高, 它们与春兰的关系较远, 不支持送春作为蕙兰下的变种。这些结果可为开展兰属育种提供重要参考价值。     

四照花亚属的nrDNA ITS致同进化不完全

四照花亚属(Cornus subg.Syncarpea)隶属于山茱萸科山茱萸属(Cornus),我国该亚属共有5种8亚种。为探讨四照花亚属nrDNA ITS序列的致同进化不完全现象及假基因产生的可能原因,分析了该亚属4种(每种1~2个居群)共21个个体的nrDNA ITS序列。结果表明,这些类群的nrDNA ITS存在多态性,通过分析这些nrDNA ITS克隆序列的G+C含量、5.8S保守基序和二级结构最小自由能,推测其可能存在假基因。系统发育研究结果显示所有nrDNA ITS序列分成5个分支,同一个体的不同拷贝被分别置于两个甚至多个分支中,且不同分支显示了不同种间关系。四照花亚属物种个体内部存在nrDNA ITS不完全致同进化,可能归咎于不完全的世系分选(incomplete lineage sorting)、种间杂交或多倍化等进化事件,从而导致基因组内nrITS区序列出现多态性,同时也导致难以通过外部形态来划分亚属内种间界限。     

Phylogenetic analysis of Allium subg. Melanocrommyum infers cryptic species and demands a new sectional classification

Allium subgenus Melanocrommyum (Alliaceae) from Eurasia comprises about 150 mostly diploid species adapted to arid conditions. The group is taxonomically complicated with different and contradictory taxonomic treatments, and was thought to include a considerable number of hybrid species, as the taxa show an admixture of assumed morphological key characters. We studied the phylogeny of the subgenus, covering all existing taxonomic groups and their entire geographic distribution. We analyzed sequences of the nuclear rDNA internal transcribed spacer region (ITS) for multiple individuals of more than 100 species. Phylogenetic analyses of cloned and directly sequenced PCR products confirmed the monophyly of the subgenus, while most sections were either para- or polyphyletic. The splits of the large sections are supported by differences in the anatomy of flower nectaries. ITS data (i) demand a new treatment at sectional level, (ii) do not support the hypotheses of frequent gene flow among species, (iii) indicate that multiple rapid radiations occurred within different monophyletic groups of the subgenus, and (iv) detected separately evolving lineages within three morphologically clearly defined species (cryptic species). In two cases these lineages were close relatives, while in Allium darwasicum they fall in quite different clades in the phylogenetic tree. Fingerprint markers show that this result is not due to ongoing introgression of rDNA (ITS capture) but that genome-wide differences between both lineages exist. Thus, we report one of the rare cases in plants where morphologically indistinguishable diploid species occurring in mixed populations are non-sister cryptic species.     

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。     

Phylogeny of the tribe Menispermeae (Menispermaceae) reconstructed by ITS sequence data

The phylogeny of the tribe Menispermeae (Menispermaceae) represented by 20 species of 9 genera in China, was reconstructed based on sequence analysis of the internal transcribed spacers (ITS) (including ITS1, ITS2, and 5.8S rRNA gene ) of nuclear ribosomal DNA. Three species of two genera in the tribe Tinosporeae were designated as outgroups. Direct PCR sequencing method was used in the study, The sizes of ITS within trib. Menispermeae range from 527 to 601 bp. The aligned length is 667 bp, which provides 281 phylogenetically informative sites when gaps are treated as missing. The results of phylogenetic analyses show that: ① trib. Menispermeae is a monophyletic group strongly supported by a bootstrap value of 100%; ② Pachygone valida, whose systematic position was uncertain in the previous classification, should be placed in the Cocculus. ③Sinomenium and Menispermum are two close genera of the tribe. Their sequcences are very similar to each other, with ITS1 having 41 to 73 bp longer than that of the other genera in trib. Menispermeae. ④ Stephania and Cyclea are also closely related. The former forms two major clades, which are approximately consistent with the two traditional subgenera: subgen. Stephania and subgen. Tuberiphania. The species of Cyclea are mutually little diverged in complete ITS sequences, and they com-prise a sister clade to the genus Stephania.     

Pollination biology of Cymbidium goeringii (Orchidaceae) in China

The genus Cymbidium comprises three subgenera with ca. 50 species. Interactions between pollinators and plants have been studied in the two subgenera Cymbidium and Cyperorchis, but only a few studies in the subgenus Jensoa have been reported. Here we report on the reproductive characteristics of C. goeringii (in sub-genus Jensoa) in three populations in the southwest of Hunan Province, China, during the winter from December 2005 to March 2006. Floral phenological and morphological features, behavior of visitors, the breeding system, and fruit sets under natural conditions were studied. The flowers of C. goeringii were strongly fragrant, but did not present any rewards to the visitors. The flowering period of C. goeringii in the studied populations lasted about 40 days, and most flowers (about 60%) opened within 30 days. Flowers opened immediately when temperature increased obviously and reached to about 16 ℃. The flowering time of the pollinated flowers, unpollinated flow-ers and the flowers with pollinarium removed were similar. Two bees of Apidae, Apis cerana cerana (honeybee) and Anthophora melanognatha, were observed visiting flowers of C. goeringii, but only the honeybees performed as pollinators. The honeybees mostly visited the orchid flowers at 10:00–17:00 on sunny days with temperature above 10 ℃. A total of only thirteen visits were observed during 20 days of observation, indicating pollinators were rare. Honeybees directly landed on the upper surface of the labellum and inserted their heads into the flowers between column and labellum, while the hind legs trod on the surface of the curved downward mid-lobe of label-lum. When a honeybee landed on the labellum of a flower, the labellum moved up and down slightly. After the honeybee entered the flower further, its head might touch the foot of the column. At this time, the body of the honeybee was parallel with the upper surface of the labellum. Then the honeybee used its front legs to scratch on the callus ridges on the upper surface of the labellum, and its hind legs hooked the edges of the side lobe of label-lum, trying to exit forcibly from the flower. During exiting process, sometimes the honeybee’s body was arched owing to tension. In this case, the surface of the scutellum came into close contact with the viscidium and then pollinaria, together with the anther cap, were removed. When this honeybee visited next flower, the pollinaria would be adhered to the stigma when it arched its body during the exiting process. Three plant species flowered synchronously with C. goeringii in the studied areas, but their flowers were different with C. goeringii in color or shape. Because C. goeringii is rewardless to pollinators, the flowers probably attract visitors by olfactory stimu-lus. Breeding system experiments showed no spontaneous autogamy and pollination success relying on pollinators in C. goeringii. Artificial pollination resulted in 90% fruit set by induced autogamy, 100% by pollinating within clone, and 100% by xenogamous pollination, respectively. These results indicate that C. goeringii is highly self-compatible and the fruit production is pollen limited. Because pollinators are essential for fruit set of C. goeringii in natural habitats, protection of wild honeybee populations or apiculture is likely a simple but effective strategy to maintain the orchid populations.     

Phylogeny of Rubus (rosaceae) based on nuclear ribosomal DNA internal transcribed spacer region sequences

We used nuclear ribosomal DNA internal transcribed spacer region (ITS 1 - 5.8S - ITS 2; ITS) sequences to generate the first phylogeny of Rubus based on a large, molecular data set. We sampled 57 taxa including 20 species of subgenus Rubus (blackberries), one to seven species from each of the remaining 11 subgenera, and the monotypic and closely related Dalibarda. In Rubus, ITS sequences are most informative among subgenera, and variability is low between closely related species. Parsimony analysis indicates that Rubus plus Dalibarda form a strongly supported clade, and D. repens may nest within Rubus. Of the subgenera with more than one species sampled, only subgenus Orobatus appears monophyletic. Three large clades are strongly supported: one contains all sampled species of nine of the 12 subgenera; another includes extreme Southern Hemisphere species of subgenera Comaropsis, Dalibarda, and Lampobatus; and a third clade consists of subgenus Rubus plus R. alpinus of subgenus Lampobatus. Rubus ursinus appears to be a hybrid between a close relative of R. macraei (subgenus Idaeobatus, raspberries) and an unidentified subgenus Rubus species. ITS sequences are generally consistent with biogeography and ploidy, but traditionally important morphological characters, such as stem armature and leaf type, appear to have limited phylogenetic value in Rubus.