报春花属藏报春组、毛茛叶报春组的界定和系统发育关系:核糖体DNA ITS序列证据

报春花属(Primula L.)藏报春组(sect. Auganthus Pax ex Balf. f.)和毛茛叶报春组(sect. Ranunculoides Chen et C. M. Hu)的界定一直是没有解决的问题.应用核糖体DNA ITS序列数据探讨其系统发育关系.取样包括藏报春组和毛茛叶报春组的全部5个种以及其他一些相关组的代表种.ITS系统树表明,陕西羽叶报春(P.filchnerae Knuth)应与藏报春(P. sinensis Sabine ex Lindl.)和野藏报春(P. rupestris Balf. f. et Farrer)一起置于藏报春组;毛茛叶报春组只含两种:毛茛叶报春(P. cicutariifolia Pax)和安徽羽叶报春(P. merrilliana Schltr.).这两组并不具密切亲缘关系,它们在报春花属中与其他组的关系还需进一步研究.研究也表明ITS序列可以为报春花属的系统发育重建提供大量可靠资料.     

报春花属藏报春组、毛莨叶报春组的界定和系统发育关系：核糖体DNAITS序列证据

报春花属（PrimulaL.)藏报春组（sect.Auganthus Pax ex Balf.f.)和毛莨叶报春组（sect.Ranunculoides Chen et C.M.Hu)的界定一直是没有解决的问题,应用核糖体DNAITS序列数据探讨其系统发育关系。取样包括藏报春组和毛莨叶报春组的全部5个种以及其他一些相关组的代表种,ITS系统树表明,陕西羽叶报春（P.filchnerae Knuth)应与藏报春（P.sinensis Sabine ex Lindl.)和野藏报春耕（P.rupestris Balf.f.et Farrer)一起置于藏报春组;毛莨叶报春组只含两种;毛莨叶报春（P.cicutariifoliaPax)和安徽羽叶报春（P.merrilliana Schltr.)。这两组并不具密切亲缘关系,它们在报春花属中与其他组的关系还需进一步研究。研究也表明ITS序列可以为报春花属的系统发育重建提供大量可靠资料。     

中国四川报春花属一新种

报道了中国四川报春花属(Primula L.)一新种——小繸瓣报春(P.hydrocotylifolia G.Hao,C.M.Hu&Y.Xu)。小繸瓣报春植株无毛、无粉,叶片多少呈圆形,具纤细的叶柄,开花期叶丛基部无鳞片,球形蒴果藏于宿存花萼,成熟时裂成不规则碎片,显然属于脆蒴报春组(sect.Petiolares Pax)革叶报春亚组(subsect.Chartacea W.W.Sm.&Forrest),并与该亚组的川西繸瓣报春(P.veitchiana Petitm.)最为接近,但后者植物体的各部分均远大于此新种,花序多花,花亦较大,二者之间无过渡类型。     

安徽羽叶报春和毛茛叶报春的微形态特征

在扫描电子显微镜和光学显微镜下,观察了安徽羽叶报春(Primula merrilliana Schltr.)与毛茛叶报春(Primula cicutariifolia Pax)的种子、花粉形态及表面纹饰与叶表皮形态结构.结果表明:2种类的种子都呈不规则七面体或多面体,多具五边形纹饰;安徽羽叶报春的种皮雕纹浅,网眼具许多小的乳头状突起;毛茛叶报春的种皮雕纹明显,网眼粗糙,具许多大的乳头状突起.2种类花粉均为球形,具散孔(孔数达10个以上),具网状雕纹.2种类的叶表皮细胞呈凹凸镶嵌状不规则排列,而在中脉处呈长方形或略带微波状长方形,上表皮细胞稍大;叶两面被腺毛,且顶端具水孔.气孔的分布属于偏叶下表面生长,在边缘处尤密,而在中脉处近无;相对而言,安徽羽叶报春的气孔略比毛茛叶报春的气孔小,且密度也低;电子显微镜下2种类的气孔外拱盖内缘光滑,保卫细胞外缘隆起明显,中间凹陷,其上着生的乳头状突起安徽羽叶报春不如毛茛叶报春明显.     

应用ITS序列分析探讨偏花报春的系统位置

对于偏花报春Primula secundiflora Franch. 的系统位置,主要有两种意见,一种认为偏花报春具有典型的钟状花冠,应置于钟花报春组Sect．Sikkimensis;而另一种意见则依据其他特征将其置于灯台报春组sect．Proliferae。通过对偏花报春、灯台报春组4种植物和钟花报春组3种植物核糖体DNA中的内转录间隔区(ITS)的序列测定及分析,并结合形态学及染色体特征的比较论证,认为偏花报春应置于灯台报春组。     

安徽羽叶报春与毛茛叶报春的遗传多样性研究与新资源评估

采用RAPD方法对分布于安徽的珍稀濒危植物安徽羽叶报春(Primula merrilliana Schltr.)和毛茛叶报春(P．cicutariifolia Pax)的10个居群进行了DNA基因组多态性分析,从50条10碱基随机引物中筛选出14条能扩增出稳定性和重复性好的DNA片段的引物,共扩增出85条带,其中多态性带67条,占总数的78．8％。安徽羽叶报春与毛茛叶报春的多态位点分别达到71．6％与66．7％,因而推测主要致濒因素可能不是通常认为的低水平的遗传多样性。采用N—J法对遗传距离构建的树状图进行聚类分析,从分子角度证明了安徽羽叶报春与毛茛叶报春是2个自然的物种,同时也说明了它们的遗传多样性与生境多样性的密切相关性。由于这2个物种具有地处黄山风景区的地理优势和花朵多、花期长、花姿优美等观赏性特点以及遗传优势,如加以合理的开发与利用,可以作为观赏花卉新资源。     

小报春与岩生报春种间杂交亲和性研究

以报春花属报春花组的小报春(Primula forbesii)和指叶报春组的岩生报春(Primula saxatilis)为亲本,对种间杂交的结实性及花粉管行为进行了观察。结果表明:小报春与岩生报春种间杂交表现为不亲和,正反交组合正常结实率为0,但正交、反交组合花粉在柱头表面的萌发和花粉管伸长过程有明显差异,小报春花粉授粉4h后可以在岩生报春柱头表面萌发,但花粉管伸长的速度明显比对照组[岩生报春(P)×岩生报春(T)、岩生报春(T)×岩生报春(P)]慢,并且花粉管生长弯曲,授粉192h后花粉管仍未到达子房;岩生报春的花粉可以在小报春柱头上正常萌发,授粉48h后花粉管到达子房;4个对照授粉组合均分别于24h(短花柱为母本)、48h(长花柱为母本)时完成受精过程。研究表明,岩生报春×小报春杂交存在受精前障碍,小报春×岩生报春杂交亲和性较好,并可通过幼胚拯救的方法获得组间杂种后代;花柱长度可能是影响种间杂交结实能力的因素之一。     

七种灯台报春组植物的细胞学研究

对报春花科（Primulaceae）报春花属（Primula）灯台报春组植物Section Proliferoe的7种报春进行了细胞学研究,其中腾冲灯台报春和川东灯台报春的核型分析为首次报道,结合已报道的灯台报春组其它植物的细胞学资料进行统计分析,结果表明灯台报春组植物在染色体基数、染色体形态、着丝点位置及染色体对称性上都具有很高的一致性,在灯台报春组有核型记录的种类中,其核型都属于Stebbins的2A型或2B型,核型差异很小,染色体基数均为x=11,推测其可能与报春花属中具有相同染色体基数的组亲缘关系更近。结合已发表的灯台报春组植物的细胞学资料及它们的形态特征,对其系统演化关系进行了比较分析,以期对该组的系统学及演化关系提供一些证据。     

鄂报春(Primula obconica Hance) 一新亚种

报春花属植物广布于我国各地,惟福建、江苏未曾有记录.本文作者最近在福建首次采得此属标本,其体态极小,高不及6 cm, 花葶短于叶丛, 甚为特殊,但无疑与鄂报春Primula obconica 有密切的亲缘关系;其外形与小型鄂报春P. obconica subsp. parva尤为相似, 但花更小,白色,花同型(花柱更长),可以区别,其地理分布和生殖也与鄂报春其他亚种存在明显的隔离,因此作一新亚种发表.     

云南报春花属（报春花科）——新变种―长萼铁梗报春

报道了云南报春花属一新变种,即长萼铁梗报春Primula sinolisteri Balf. f. var. longicalyx D. W. Xue & C. Q. Zhang。新变种与原变种的不同之处在于：花冠喉部具环状附属物,外面无毛,花萼与冠筒近等长,长7-10 mm。     

The monophyly and rapid evolution of Gentiana sect. Chondrophyllae Bunge s.l. (Gentianaceae): evidence from the nucleotide sequences of the internal transcribed spacers of nuclear ribosomal DNA

The nucleotide sequences of the internal transcribed spacers (ITS) of nuclear ribosomal DNA of 24 representative species of sect. Chondrophyllae s.l. have been determined and analysed phylogenetically, together with some species of other sections of the genus Gentiana. The ITS sequences strongly support the monophyly of the sect. Chondrophyllae s.l. as a whole complex including various different dysploid cytotypes. Species, such as G. boryi and G. pyrenaica , that had been split into distinct genera by some cytotaxonomists have been proven to be closely related. However, the ITS sequences do not provide sufficient information to make a robust estimation of the phylogenetic relationships among the closely related species and dysploid cytotypes of the complex, beyond recognizing their monophyly and rapid evolution.     

Circumscription and phylogeny of Saxifraga sect. Ciliatae: Evidence from nrDNA ITS sequences

The largest section of the genus Saxifraga (Saxifragaceae), sect. Ciliatae, consists of 175 morphologically diverse species. This section is mainly distributed in the Qinghai-Tibetan Plateau and adjacent regions of southwest China and more than 80% of the total number of species are endemic to this region. It remains unknown whether this section is monophyletic and up to now no study has been conducted on the infra-sectional phylogeny. In this study, ITS sequences of the nuclear ribosomal DNA were firstly determined for 33 species mainly from this section and related sections. We further downloaded the corresponding sequences of the same DNA region for the other 22 species of Saxifraga and Mitella from GenBank. All sequences were together used to construct the phy-logenetic trees. The main implications of the phylogenetic analyses include: (1) sect. Ciliatae, as traditionally defined, constitutes as a monophyletic clade and its sister group is a well supported clade that includes species from 8 sections such as sect. Porphyrion, sect. Saxifraga and sect. Mesogyne; (2) three morphological subsections, i.e., subsect. Gemmiparae, subsect. Hirculoideae and subsect. Rosulares were tentatively recovered despite the relatively low statistic bootstrap support for the last one; however, subsect. Flagellares and subsect. Hemi-sphaericae were not recognized as separate entities, and nested within subsect. Gemmiparae; (3) subsect. Hircu-loideae and subsect. Rosulares clustered together as sister subclades while subsect. Gemmiparae diverged early. In addition, our results suggest that the paired variation of ITS sequences in sect. Ciliatae is relatively low between the sampled species in spite of their diverse morphology. It is suggested that such a scenario may mirror rapid speciation in this section that probably trigged by the uplifts of the Qinghai-Tibetan Plateau and the extensive selection pressure under the alpine environments.     

DNA barcoding in closely related species: A case study of Primula L.sect.Proliferae Pax (Primulaceae) in China

DNA barcoding is a method of identifying species by analyzing one or a few short standardized DNA sequences. There are particular challenges in barcoding plants, especially for distinguishing closely related species. Hence, there is an urgent need to evaluate the performance of candidate loci for distinguishing between species, especially closely related species, to complement the rbcL + matK combination suggested as the core barcode for land plants. We sampled 48 individuals representing 12 species in Primula sect. Proliferae Pax in China to evaluate the performance of eight leading candidate barcode loci (matK, rbcL, rpoB, rpoCl, trnH-psbA, psbK-psbI, atpFatpH, and internal transcribed spacer (ITS)). The core combination rbcL + matK gave only 50% species resolution in sect. Proliferae. In terms of intraspecies and interspecies divergence, degree of monophyly, and sequence similarity, ITS, trnH-psbA, and psbK-psbI showed good performance as single-locus barcodes. Internal transcribed spacer displayed the highest genetic divergence and best discriminatory power, both alone and in combination with rbcL +matK (83.3% species resolution). We recommend evaluating the use of ITS for barcoding in other species. Low or single copy nuclear regions would provide more sophisticated barcoding tools in the long term, even though further research is required to find suitable loci.     

DNA barcoding in closely related species: A case study of Primula L. sect. Proliferae Pax (Primulaceae) in China

DNA barcoding is a method of identifying species by analyzing one or a few short standardized DNA sequences. There are particular challenges in barcoding plants, especially for distinguishing closely related species. Hence, there is an urgent need to evaluate the performance of candidate loci for distinguishing between species, especially closely related species, to complement the rbcL + matK combination suggested as the core barcode for land plants. We sampled 48 individuals representing 12 species in Primula sect. Proliferae Pax in China to evaluate the performance of eight leading candidate barcode loci (matK, rbcL, rpoB, rpoC1, trnH-psbA, psbK-psbI, atpF-atpH, and internal transcribed spacer (ITS)). The core combination rbcL+matK gave only 50% species resolution in sect. Proliferae. In terms of intraspecies and interspecies divergence, degree of monophyly, and sequence similarity, ITS, trnH-psbA, and psbK-psbI showed good performance as single-locus barcodes. Internal transcribed spacer displayed the highest genetic divergence and best discriminatory power, both alone and in combination with rbcL+matK (83.3% species resolution). We recommend evaluating the use of ITS for barcoding in other species. Low or single copy nuclear regions would provide more sophisticated barcoding tools in the long term, even though further research is required to find suitable loci.     

Classification of Primula sect. Auricula (Primulaceae) based on two molecular data sets (ITS, AFLPs), morphology and geographical distribution

On the basis of the study of c . 1100 herbarium specimens, field observations and molecular (ITS, AFLPs) evidence, Primula sect . Auricula is classified into two subsections, 25 species and six subspecies. Primula auricula L. ssp. widmerae (Pax) L. B. Zhang stat. nov. , P. auricula Linn. ssp. tatriaca L. B. Zhang ssp. nov. , and P. latifolia Lapeyr. ssp. cynoglossifolia (Widmer) L. B. Zhang stat. nov. are newly described or combined; P. auricula s.l. is divided into two species: P. auricula Linn. and P. balbisii Lehm., representing the northern and the southern populations of P. auricula s.l. resolved in the molecular study, respectively; P. cottia Widmer and P. balbisii Lehm. are recovered from synonymy; P. grignensis D. M. Moser and P. pedemontana E. Thomas ex. Gaudin ssp. iberica Losa et P. Monts. are synonymized with P. hirsuta All. and P. pedemontana , respectively; P. clusiana is postulated to be an allopolyploid species. Phylogenetic relationships in the section are discussed based on morphological and molecular variation and geographical distribution.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 146 , 1–26.     

两栖蓼的分子系统学研究

两栖蓼是一种水陆两栖植物,植株在不同生态环境下外部形态差异较大,同时两栖蓼的系统位置存在争议,被归入春蓼组(sect.Persicaria)或提升为两栖蓼组(sect.Amphibium)。该文选取两栖蓼及春蓼组植物12种,以及刺蓼组、头状蓼组、神血宁组、拳参组、萹蓄组和外类群掌叶大黄共23种植物进行研究。植物总DNA的提取采用改进的CTAB法,所测序列以及从Genbank数据库下载的序列,以掌叶大黄为外类群,采用最大简约法和贝叶斯法对核糖体ITS序列和叶绿体trn L-F序列进行了系统发育分析。ITS序列对位排列的长度为735 bp,包括489个可变位点,272个位点是信息位点。简约法得到9个简约树,步长为1 084,CI指数为0.680,RI指数为0.614。trn L-F序列对位排列的长度为1 121 bp,包括427个可变位点,239个位点是信息位点。简约法寻找到9个简约树,步长为551(CI=0.911,RI=0.910)。贝叶斯法和简约法得到的树基本一致。分子序列分析结果显示,trn L-F序列树类似于ITS序列树。ITS序列构建的发育树上,两栖蓼与刺蓼组植物、春蓼组其他植物形成3个并列的分支;在trn L-F序列树上,两栖蓼则与其他春蓼组植物形成两个并列的分支。由此可见,两栖蓼与春蓼组其他植物的亲缘关系较远,成一独立的分支。两个分子证据支持将两栖蓼提升为两栖蓼组的处理意见。此外,两栖蓼的花粉具散沟,与典型的春蓼组的具散孔花粉不一致。再加上两栖蓼水陆两栖的特性,因此支持把两栖蓼提升为两栖蓼组的观点。两栖蓼组的界定为多年生草本,水陆两栖,根状茎横生,生于水中茎漂浮,叶长圆形或椭圆形,生于陆地茎直立,叶披针形或长圆状披针形,托叶鞘为筒状、薄膜质,总状花序穗状,瘦果近圆形,花粉具散沟。     

用ITS序列确定小麦B基因组的可能供体间的关系

对小麦B基因组的可能供体山羊草属Aegilops sect．Sitopsis的5个种的核糖体DNA的内部转 录区(ITS)进行了PCR扩增和克隆,井测定ITSl和ITS2的序列,用ITSl+ITS2的序列重建了 Aegilops sect．Sitopsis中5个种的系统发育关系。结果表明,斯卑尔脱山羊草Ae．speltoides是sect． Sitopsis中特殊的一个种,它与该组其余4种间的平均遗传距离是后者彼此间平均遗传距离的3倍,Ae． speltoides与同组其余4个种的分离要比后者相互间的分离早得多;在拟斯卑尔脱组Sect．Sitopsis的5 个种中,长柱山羊草Ae．longissima与沙融山羊草Ae．sharonensis的关系最近。ITS序列可以进一步用来作为确定B基因组起源的分子标记。