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。     

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.     

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.     

What is the phylogenetic placement of Dipteronia dyerana Henry? An example of plant species placement based on nucleotide sequences

Abstract

DNA sequence data have been widely used to evaluate species delimitations and examine infraspecific relationships. However, species placements inferred from different nucleotide sequences are frequently in conflict. As an example of plant species placement based on nucleotide sequences, the phylogenetic placement of Dipteronia dyerana Henry (Aceraceae) was analyzed in the present study. The study species included eight Acer species (from different sections of Acer), two Dipteronia species, and two outgroup taxa. Phylogenetic trees based on five datasets (ITS, trnL‐F, trnD‐trnT, psbM‐trnD, and rpl16 regions) as well as their combined datasets were generated by using maximum parsimony (MP) and maximum likelihood (ML) analyses. Further analyses were conducted to compare the strict consensus trees based on single regions and the combination of different regions. The results revealed a significant discrepancy among the phylogenetic placements of D. dyerana, inferred from various sequences. Phylogenetic trees using MP analysis based on trnD‐trnT, rpl16, and the four chloroplast combined sequences supported the genus Dipteronia as a monophyletic group, while in the other trees D. dyerana was positioned either in parallel with D. sinensis and Acer species or within the genus Acer. In ML analysis, only rpl16 and the four chloroplast combined sequence datasets supported the genus Dipteronia as a monophyletic group. We concluded that, although significant genetic differentiation occurred between D. dyerana and D. sinensis, D. dyerana was more advanced than D. sinensis. However, whether Dipteronia is monophyletic remains to be further investigated, e.g., by using more closely related taxa and more sequences. Furthermore, in addition to internal transcribed spacer sequences, more chloroplast gene sequences should be used for phylogenetic analyses of species.     

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.     

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。     

nrDNA ITS和cpDNA rpl16基因测序法对六种鼠尾草的检测

为从鼠尾草属植物中鉴别丹参品种,采用基因测序方法,用核糖体核酸内转录间隔区基因(nrDNA ITS),编码核蛋白体大亚基多肽L16的基因(rpl16)及叶绿体DNA上包含trnL以及trnL和trnF间隔区的区域基因(trnL-trnF)的序列,检测六种鼠尾草属新鲜植物.由于nrDNA ITS和rpl16突变率较高,可以做为6种鼠尾草的基源鉴定标记,依此设计了两对特异引物,从6种鼠尾草中鉴定出丹参(Salvia miltiorrhiza)和云南鼠尾草(S.yunnanensis).但trnL-trnF突变率太低,未能用于鉴别.商品干燥中药材因加工和储藏的方式致使DNA降解严重,基因测序法难于应用.     

五种金花茶组植物的耐寒性比较研究

金花茶组植物是世界珍稀、濒危的观赏植物,具有极高的观赏价值和药用价值.为了比较金花茶组植物的耐寒性,以五种金花茶组植物为材料,采用人工模拟低温环境的方法对其2年生叶片进行低温胁迫处理,应用电导法研究五种金花茶种质在20℃(常温对照)、8℃、-2℃、-7℃、-12℃、-17℃、-22℃和-27℃低温下相对电导率的变化,配合Logistic方程,测定其低温半致死温度(LT50),以及叶片中游离脯氨酸、可溶性糖和丙二醛的含量.结果表明:五种金花茶的低温半致死温度(LT50)范围为-14.58~-12.74℃,其中金花茶为-14.58℃、龙州金花茶为-14.27℃、柠檬黄金花茶为-13.44℃、直脉金花茶为-13.09℃、东兴金花茶-12.74℃.低温半致死温度能反映金花茶种质的耐寒性,金花茶和龙州金花茶耐寒性强,其次为柠檬黄金花茶和直脉金花茶,东兴金花茶耐寒性较弱.在降温过程中,五种金花茶叶片相对电导率随温度降低呈S型上升,与温度呈负相关;脯氨酸、可溶性糖和丙二醛含量均呈现先上升后下降的趋势.同一低温条件下,半致死温度低的金花茶脯氨酸和可溶性糖的含量更高,而丙二醛含量更低.该研究结果为金花茶组植物耐寒种质选育提供了科学依据,为人工种植金花茶提供了技术支持.     

十四种金花茶组植物叶总黄酮含量比较

金花茶叶含有丰富的黄酮类化合物.以70%乙醇为提取溶剂,对14种金花茶叶中的黄酮进行超声波提取,并以芦丁为对照品,采用紫外分光光度法于273 nm处测定吸光度,得出各种金花茶叶总黄酮的含量并进行了比较.结果表明:不同种金花茶叶黄酮的含量差异较大,平果金花茶中新叶的总黄酮含量(22.011%)最高,而东兴金花茶中一年叶龄...     

四种金花茶的核型比较

<正> 金花茶是山茶属的珍稀植物,目前在我国广西南部已发现20个种和变种,但做过染色体核型研究的仅有5个种。本文首次报道凹脉金花茶(Comellia impressinervis)的核型,并结合显脉金花茶(C. euphlebia),金花茶(C. chrysantha),小金花茶(C. microcarpa)的核型作了分析比较,意在为研究金花茶的来源、分类和杂交育种提供     

基于ITS和matK序列的獐牙菜亚族 (龙胆科龙胆族)分子系统学

为探讨獐牙菜亚族（subtribe Swertiinae）各属之间和一些属内的系统关系,本研究选取了该亚族14属68种1变种,采用最大简约法（maximum parsimony）和贝叶斯法（Bayesian inference）对样品核基因ITS和叶绿体基因matK的两个片段进行独立和联合分析。结果显示：Bartonia位于亚族的最基部;喉毛花属（Comastoma）、肋柱花属（Lomatogonium）和假龙胆属（Gentianella）都非单系,处于同一个较为进化的分支中;獐牙菜属折皱组（Swertia sect. Rugosa）和獐牙菜组（S. sect. Swertia）亲缘关系最近,宽丝组（S. sect. Platynema）和藏獐牙菜组（S. sect. Kingdon Wardia）亲缘关系最近;口药花属（Jaeschkea）与獐牙菜属多枝组（S. sect. Ophelia）的大籽獐牙菜（Smacrosperma）亲缘关系最近。同时讨论了獐牙菜亚族形态分类与分子数据不一致的原因。     

Phylogeny of korean rubus (rosaceae) based on its (nrDNA) and trnL/F intergenic region (cpdna)

We examined the phylogeny of the genusRubus in Korea using an internal transcribed spacer (ITS) of the nuclear ribosomal DNA and a trnL-trnF (trnL/F) intergenic region of the chloroplast DNA. In all, 21 ingroup species (1.2 kb for each species) were analyzed using parsimony, resulting in 672 aligned sequences from ITS, and 502 bases with trnL/F. Individual and combined analysis of ITS and trnL/F data proved that the genusRubus is a monophyletic group. This phylogeny also substantiated a previous sectional classification scheme rather than a subgerius classification scheme. However, our results did not support the earlier sectional classification by Focke (sect.Corchorifolii), but did support the sectional classification of Nakai: sect.Crataegifolii (R. crataegifolius, R. takesimensis andR. trifidus), and sect.Villosii (R. corchorifolius). Most of these species, which are found in Korea and belong to subg.Idaeobatus, appeared in two different groups in all data sets. This suggests that this subgenus is a polyphyletic group that has gone through at least two independent evolutionary processes. The taxa, when mapped onto the combined tree, showed that the occurrence of their morphological characters of simple and compound leaves was concurrent in KoreanRubus. ITS sequence data were consistent overall with the geographical distribution of each species. Furthermore, the trnL/F sequence data provided phylogenetic information within closely related species.     

山茶科系统发育诠释——Ⅲ.关于金花茶组及山茶属演化若干问题

作者通过比较认为山茶属金花茶组的模式和古茶组的模式不是同一分类单位,因而取消金花茶组是不恰当的。金花茶组是一个自然的集合体。作者详细地分析了山茶属内演化的四个阶段:第一阶段表现为苞被不分化、大型、宿存、子房5室、心皮部分分离;第二阶段演化出苞被宿存和苞被脱落两个类群,前者较为原始的代表是离蕊茶织和短蕊茶组,较为进化的代表是管蕊茶组;后者较为原始的代表是半宿萼茶组、瘤果茶组、糙果茶组,较为进化的代表是油茶组、短柱茶组、红山茶组;第三阶段是苞被分化为小苞片和萼片的类群,金花茶组和长柄茶组是较为原始而茶组、超长柄茶组则是较为进化的代表;第四阶段的连蕊茶组和毛蕊茶组苞萼小型化且宿存,的代表,雌、雄蕊均高度连合,子房室不完全发育 它们只能由第三阶段具有多数小苞片的原始类群发展出来,认为把山茶属划分为四个亚属的系统是合理的。 作者还认为,花的颜色以及其他相似性状的集合是划分山茶属次级分类单位的重要依据;分类系统的自然性和实用性相结合是分类学家始终应该追随的目标,混淆不同差异的做法是不可取的。最后,作者认为山茶属没有真正的顶生花。     

Systematic position of Gyrocheilos and some odd species of Didymocarpus (Gesneriaceae) inferred from molecular data,with reference to pollen and other morphological characters

A phylogenetic analysis was carried out to clarify the systematic position of Gyrocheilos and Didymocarpus, particularly the species placed in Didymocarpus sect.Heteroboea by Wang et al. Based on sequencing the internal transcribed spacer and the chloroplast spacer trnL-F, parsimony and Bayesian inference analyses were carried out using separate nuclear and chloroplast datasets, as well as a combined dataset. Our results showed that the two sections of Didymocarpus in China andGyrocheilos did not form separate monophyletic subclades, but turned up in three different places in the phylogenetic trees. In the frame of the present study, the pollen morphology of the species included in the analysis was studied. It proved inconsistent with the delimitation between Didymocarpus and Gyrocheilos. Furthermore, pollen and other morphological characters indicate that Gyrocheilos and some taxa of Didymocarpus should be placed within Didymocarpus.     

Molecular taxonomy of Camellia (Theaceae) inferred from nrITS sequences

Camellia, comprising more than 200 species, is the type genus of the family Theaceae. Currently, the interspecies relationship of the economically important genus is still a matter of great debate and controversy. In an attempt to help settle this dispute using molecular phylogeny, we analyzed ITS sequences of 112 species of Camellia. The maximum parsimony and Bayesian trees grouped these species into eight major clades and four isolates. The current study supported the monophyly of sections Thea and Furfuracea, a merged section of Theopsis and Eriandra and the formation of section Oleifera by H, -t. Chang (Flora of Reipulicae Popularis Sinicae. Tomus 49 (3), Science Press, China). The study suggested the polyphyletic nature of the sections Camellia, Paracamellia, Pseudocamellia, and Tuberculata and the paraphyletic nature of the section Chrysantha but did not support the sectional status of the three small sections, Archecamellia, Piquetia, and Sterocarpus. We also discuss the results in terms of morphology, geographic distribution and the results from an earlier molecular phylogeny analysis.     

Tackling speciose genera: species composition and phylogenetic position of Senecio sect. Jacobaea (Asteraceae) based onplastid and nrDNA sequences

The molecular phylogeny of Senecio sect. Jacobaea (Asteraceae; Senecioneae) was studied to clarify species composition and interspecific relationships of Senecio sect. Jacobaea. This information is necessary for studies seeking explanations of the evolutionary success of Senecio, in terms of high species numbers and the evolution of chemical defense mechanisms. Parsimony analyses with 60 species of the tribe Senecioneae, representing 23 genera and 11 sections of Senecio, based on DNA sequence data of the plastid genome (the trnT-L intergenic spacer, the trnL intron, and two parts of the trnK intron, flanking both sides of the matK gene) and nuclear genome (ITS1, 5.8S, and ITS2 gene and spacers) show that sect. Jacobaea is a strongly supported monophyletic group. Fifteen species have been identified as members of section Jacobaea, including three species that have been consistently ascribed to this section in taxonomic literature and 12 species that were either placed in other sections of Senecio or not exclusively ascribed to sect. Jacobaea. This section was traditionally circumscribed as a group of European, biennial, or perennial herbs with pinnately incised leaves, but the results of this study show that one annual species, a species from northeastern Asia, and a species growing in the Himalayas are members of sect. Jacobaea as well. Furthermore, not all species in the section have pinnately incised leaves. The genera Emilia, Packera, and Pseudogynoxys form the sister clade of sect. Jacobaea, but this relationship lacks strong bootstrap support and thus remains provisional.     

Systematic positions of Lamiophiomis and Paraphlomis (Lamiaceae) based on nuclear and chloroplast sequences

Genera Lamiophlomis and Paraphlomis were originally separated from genus Phlomis s.l. on the basis of particular morphological characteristics. However, their relationship was highly contentious, as evidenced by the literature. In the present paper, the systematic positions of Lamiophlomis, Paraphlomis, and their related genera were assessed based on nuclear internal transcribed spacer (ITS) and chloroplast rpl16 and trnL-F sequence data using maximum parsimony (MP) and Bayesian methods. In total, 24 species representing six genera of the ingroup and outgroup were sampled. Analyses of both separate and combined sequence data were conducted to resolve the systematic relationships of these genera. The results reveal that Lamiophlomis is nested within Phlomis sect. Phlomoides and its genetic status is not supported. With the inclusion of Lamiophlomis rotata in sect. Phlomoides, sections Phlomis and Phlomoides of Phlomis were resolved as monophyletic. Paraphlomis was supported as an inde-pendent genus. However, the resolution of its monophyly conflicted between MP and Bayesian analyses, suggesting the need for expended sampling and further evidence.     

山茶属金花茶组金花茶系植物叶表皮形态学研究

在光学显微镜下观察山茶属金花茶组金花茶系25个分类群的叶表皮形态。结果表明:25个分类群的叶上表皮形态结构相似,气孔仅分布于下表皮。下表皮的细胞垂周壁式样和气孔形状在一些种间存在着明显的差异。分布于不同地貌类型的金花茶种类的气孔密度存在显著差异,分布在石灰岩山地的种类的气孔密度在整体上明显高于分布在非石灰岩山地的种类。     

基于ITS序列探讨山茶属金花茶组的系统发育关系

测定了分布于我国的 2 2个山茶属金花茶组的种或变种的nrDNAITS区序列 ,它们的序列长度在 476～ 496之间。GC含量都超过了 70 % ,应用Kimura2 模型计算了序列间的分化程度 ,构建了最大简约树、邻接树和最大似然树 ,分析结果表明 :( 1 )淡黄金花茶、毛籽金花茶、陇瑞金花茶、弄岗金花茶、大样金花茶和凹脉金花茶的关系较近 ;( 2 )小瓣金花茶、小花金花茶、薄叶金花茶、多瓣金花茶、夏石金花茶和龙州金花茶的关系较近。ITS区序列分析结果与AFLP分析结果相近     

三种金花茶花粉形态的扫描电镜研究

金花茶分类系统尚未完善,金花茶又极具观赏价值,潜在价值极高,该研究应用扫描电镜观察三种金花茶花粉形态,即平果金花茶、直脉金花茶、金花茶,并运用数理统计和图片分析的方法对其花粉形态进行了探讨.结果表明:三种金花茶花粉均属于大花粉粒,花粉平均极轴在50μm以上,花粉极面观为钝凹三角形,萌发孔为三孔沟,即N3P4C5类型.平果金花茶与直脉金花茶花粉外壁纹饰为拟网状纹饰,平果金花茶网眼较密,网脊稍不平,直脉金花茶网脊稍隆起;金花茶为蠕虫状纹饰,短条状突起.花粉形态对其分类有重要意义,通过花粉外壁纹饰的类型可知金花茶与其二者亲缘关系较远,花粉形态各指标具有种间变异性,种内具有保守性,而平果金花茶和直脉金花茶花粉外壁纹饰相同但具体分类还需进一步研究.该研究结果为金花茶种质资源分类和利用提供了参考,在一定程度上促进了金花茶资源的开发和利用.