Systematics of subtribe Gentianinae (Gentianaceae)

以在自然栖息地立即干燥的叶片样品为样本提取总DNA，这些物种包括：马尾草、龙胆、古典龙胆、红花龙胆、云南龙胆、龙胆、雷公藤、粗毛龙胆、日本龙胆、日本龙胆。对开、轮叶杉木属。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18S-25S核糖体DNA的转录间隔区(ITS)。总DNA的提取、总ITS区的扩增、聚合酶链式反应产物的克隆、ITS区序列的测定均在东京市立大学Makino标本馆实验室完成。对于测序，使用了由该赠款新购买的自动测序仪ALFExpress(Pharmacia)。用PAUP3.0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，得到了32棵同样节俭的树的共识树。除了龙胆组与雷公藤属和爬山龙胆属组成一个单系支系，自举概率为84%，表明龙胆属不是一个单系群外，树的拓扑结构与属间关系的认识基本一致。龙胆组Stenogyne、雷公藤属Tripterosperum和CrawFurdiathe中常见的三棱种子可能是它们的同形异体。下弯的雄蕊可能与三棱种子同时增加，但在Crawfudia中消失。Tripterosperum和CrawFurdia加入一个单系分支，缠绕的茎可能是它们的同形同形。有浆果的物种的单调性没有得到显著支持。

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.