›› 2011, Vol. 23 ›› Issue (6): 0-1112.

• 园艺科学 •    

天台山4种鹅耳枥属植物ITS序列的克隆与分析


陈贝贝,李温平,周晶,汪安乐

  

  1. 台州学院 生命科学学院, 浙江 临海 317000
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-11-25 发布日期:2011-11-25

Cloning and analysis of ITS sequences from four Carpinus plants in Mount Tiantai

CHEN Beibei;LI Wenping;ZHOU Jing;WANG Anle   

  1. College of Life Science, Taizhou University, Linhai 317000, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2011-11-25 Published:2011-11-25

摘要:

通过克隆和分析雷公鹅耳枥(Carpinus viminea)、短尾鹅耳枥(C. londoniana)、多脉鹅耳枥(C. polyneura)和天台鹅耳枥(C. tientaiensis)的ITS序列,为鹅耳枥属植物的分子鉴定和遗传多样性研究提供依据。利用PCR法从4种鹅耳枥叶片DNA中克隆到各自的ITS,以ClustalX,EXCEL和MEGA等软件分析和比较序列特征,并从NCBI数据库下载另外7种鹅耳枥的ITS序列用于进化分析。序列已上传至NCBI,登录号为JF796531JF796534。结果表明,4种鹅耳枥属植物ITS的全长为600~602 bp,具26个变异位点,部分位点有明显的物种特征,可用作DNA指纹特异性识别。进化分析的结果表明,11种植物的遗传距离为0.0048~0.0681,表现为种间关系。4种采自天台山的鹅耳枥分属3个不同进化枝,多脉鹅耳枥与阿里山鹅耳枥和云南鹅耳枥处于同一分枝,天台鹅耳枥与欧洲鹅耳枥和湖北鹅耳枥归为一类;而短尾鹅耳枥、雷公鹅耳枥则与美洲鹅耳枥聚为一组。

关键词: 鹅耳枥属, ITS序列, 序列分析, 分子鉴定

Abstract:

To provide evidence for molecular identification and genetic diversity studies, sequencing and analysis of ITS from Carpinus tientaiensis, C. londoniana, C. polyneura and C. viminea were performed. ITS sequences were amplified from leaf DNA of four Carpinus plants by PCR. Sequence were compared and analyzed using ClustalX, EXCEL and MEGA softwares. Seven Carpinus ITS sequences were downloaded from NCBI database for evolution analysis. Sequence analysis results revealed that these four ITSs were 600-602 bp in length with 26 variable sites. Some sites showed prominent species characteristics and could be used for specific DNA fingerprint identification. Evolution analysis results indicated that the genetic distance among 11 Carpinus plants were 0.0048-0.0681, confirming their interspecific relationship. The four Carpinus plants collected from Mount Tiantai belonged to 3 different clades, one for C. polyneura, C. kawakamii and C. monbeigiana, another for C. tientaiensis, C. betulus and C. hupeana, and the other for C. londoniana C. viminea and C. caroliniana.


Key words: Carpinus, ITS sequence, sequence analysis, molecular identification