Systematics of subtribe Gentianinae (Gentianaceae)

以自然生长环境中自然干燥的小蔓龙胆（Crawfurdia maculaticaulis）、长叶龙胆（Gentiana algida）、龙胆（Gentiana classicauris）、红花龙胆（Gentianantha）、云南龙胆（Gentianopsis grandis）、双蝴蝶（Tripterospermum distylum）、毛萼龙胆（T.hirticalyx）、日本龙胆（T.japonicum var. involubile，T. lanceolatum.在本研究的初始阶段，本研究旨在分析rbcL或matK基因，但实际上分析的是18 S-25 S核核糖体DNA的转录间隔区（ITS）。总DNA的提取、ITS区的扩增、PCR产物的克隆、ITS区的序列测定在东京都立大学牧野植物标本室完成。对于测序，使用本基金新购买的自动测序仪ALFexpress（Pharmacia）。用PAUP 3.0软件对获得的序列进行分析，构建最简约的系统发育树。结果，获得了32棵同样简约的树的共识树。该树的拓扑结构与目前对属间关系的认识基本一致，但龙胆属组Stenogyne与双蝴蝶属和Crawfurdia形成了一个单系分支，其自举概率为84%，这表明该属在目前的界定下不是一个单系群。在龙胆属狭蕊组、双蝴蝶属和蔓龙胆属中常见的三角形种子可能是它们的一种联会形态。内弯雄蕊可能与三棱种子同时出现，但在蔓苔属中消失。双蝴蝶属和蔓苔属属于一个单系分支，缠绕茎可能是它们的一个联会现象。浆果物种的单系性没有得到显着的支持。

Leaf samples immediately dried at the natural habitat of following species were used as samples to extract total DNA : Crawfurdia maculaticaulis, Gentiana algida, G.classicauris, G.rhodantha, G.yunnanense, Gentianopsis grandis, Tripterospermum distylum, T.hirticalyx, T.japonicum, T.japonicum var. involubile, T.lanceolatum. At the initial stage of this study rbcL or matK gene was intended to analyze but actually the inter transcribed spacer (ITS) regions of 18S-25S nuclear ribosomal DNA was analyzed. The extraction of total DNA,amplification of total ITS region, clcaning of the PCR products, sequencing of ITS region were carried out in the laboratory of Makino Herbarium, Tokyo Metropolitan University. For sequencing, the autosequensor ALFexpress (Pharmacia) , newly purchesed by this grant, was used. The sequences obtained were analyzed with PAUP 3.0s to construct the most parsimonious phylogenetic trees. As a result a consensus tree of the 32 equally parsimonious trees was obtained. The tree topology is generally conglueut with corrent understanding of the relationships between genera, excepting that Gentiana section Stenogyne joined to form a monophyletic clade with Tripterospermum and Crawfurdia with bootstrap probabilities 84%, which suggests that the genus Gentiana under present circumscription is not a monophyletic group. The triquetrous seeds commonly found in Gentiana section Stenogyne, Tripterospermum and Crawfurdiathe may be a synapomorphy of them. The recurved stamens might be gaind at the same time as triquetrous seeds but disappeared in Crawfurdia. Tripterospermum and Crawfurdia joined a monophyletic clade and the twining stem may be a synapomorphy of them. The monophyly of the species with berries was not significantly supported.