基于ITS 序列石斛材料的鉴定及系统进化分析

园艺学报 ›› 2012, Vol. 39 ›› Issue (8): 1539-.

基于ITS 序列石斛材料的鉴定及系统进化分析

栗丹, 李振坚, 毛萍, 严雪锋, 淳泽, 马欣荣

（1 中国科学院成都生物研究所，成都 610041；2 中国林业科学研究院林业研究所，国家林业局林木培育重点实验室，北京 100091）

出版日期:2012-08-25 发布日期:2012-08-25

Phylogenetic Analysis and Identification of Dendrobium Species Based on Ribosomal DNA Internal Transcribed Spacer（ITS）Sequence

LI Dan, LI Zhen-Jian, MAO Ping, YAN Xue-Feng, CHUN Ze, MA Xin-Rong

（1Chengdu Institute of Biology，Chinese Academy of Sciences，Chengdu 610041，China；2Research Institute of Forestry，Chinese Academy of Forestry，Key Laboratory of Tree Breeding and Cultivation，State Forestry Administration，Beijing 100091，China）

Online:2012-08-25 Published:2012-08-25

摘要/Abstract

摘要： 以核糖体DNA 内转录间隔区（Internal Transcribed Spacer，rDNA ITS 区）作为DNA 条形码对石斛种进行鉴定，并进行系统进化分析。收集获得43 个石斛样品，其中35 个为已知鲜样品，8 个为待确定种的干样品。从35 个鲜品中获得在GenBank 中未公布的海南石斛、华石斛以及秋石斛中的两个品种‘白花红心秋石斛’和‘紫红条纹秋石斛’ITS 序列。ITS 序列差异与形态特征的关系分析，结果显示，ITS 同源性的高低，与形态的相似性成正相关。以舌唇兰属为外类群，并从GenBank 中获得其他20 个石斛种的ITS 序列，对35 个已知样品和8 个待检测样品进行分析。结果显示，35 个已知样品分为5 支，其中大部分石斛种（24 个）聚在一支。竹叶石斛和苏瓣石斛聚在一支；华石斛、聚石斛和小黄花石斛聚在一支；短棒石斛单独为一支；竹枝石斛与‘白花红心秋石斛’和‘紫红条纹秋石斛’聚在一支；海南石斛和木石斛聚在一支。根据ITS 序列，大多数样品分组与传统分组相同，但按传统分组不在石斛组的重唇石斛、钩状石斛、鼓槌石斛和叉唇石斛分在了石斛组，并确定了檀香石斛分在石斛组。确定了8 个石斛干样品所属的种。

关键词: 石斛, ITS 序列, 系统进化, 物种鉴定

Abstract: Dendrobium is valued for its ornamental and medicinal purposes. However，the taxonomy of this genus is still in a state of confusion，and it is more difficult to authenticate after processing. Based on the emerging field of molecular systematics as a powerful classification tool，a phylogenetic analysis was conducted using sequences of the Internal Transcribed Spacer of nuclear ribosomal DNA（rDNA ITS） as DNA barcodes for species identification and phylogenetic analysis of Dendrobium plants. In this study， the ITS1-5.8S-ITS2 sequence database of the 43 Dendrobium samples was constructed. Among them，thirty-five fresh specimens were known and the other 8 processed dry samples were undetermined. The
results showed that four ITS sequences from Den. hainanense，Den. sinense and two Den. hybrida varieties of Den. Burana Charming and Den. Burana Stripe respectively were not promulgated in GenBank. Analyzing the relationship between ITS sequence discrepancies and the phenotypic characteristics revealed that the ITS homology was positively correlated to morphology traits. The phylogenetic trees were constructed based on the ITS sequences using Platanthera dilatata as an outgroup to root the tree. According to the analysis of phylogeny and cluster，the 35 Dendrobium known samples were divided into 5 clusters，and most of the samples（24 out of 35）were clustered together. Den. hancockii and Den. harveyanum were clustered together. Den. sinense Den. lindleyi and Den. jenkinsii were grouped to one branch. Den. capillipes was parted to a branch individually. Den. hainanense and Den. crumenatum were clustered together. The results displayed that the sections of most species divided by ITS were as the same as traditional classification. However，there were several species regrouped to different sections by ITS. And Den. salaccense was grouped to the same branch of Den. Burana Charming and Den. Burana Stripe， and the sections were not determined before. Moreover，the authentication of the eight unknown Dendrobium processed dry samples were also identified by rDNA ITS.

Key words: Dendrobium, ITS sequence, phylogenetic analysis, species identification

中图分类号:

S 682.31

栗丹, 李振坚, 毛萍, 严雪锋, 淳泽, 马欣荣. 基于ITS 序列石斛材料的鉴定及系统进化分析[J]. 园艺学报, 2012, 39(8): 1539-.

LI Dan, LI Zhen-Jian, MAO Ping, YAN Xue-Feng, CHUN Ze, MA Xin-Rong. Phylogenetic Analysis and Identification of Dendrobium Species Based on Ribosomal DNA Internal Transcribed Spacer（ITS）Sequence[J]. ACTA HORTICULTURAE SINICA, 2012, 39(8): 1539-.

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