铁刀木DNA提取及其rDNA-ITS序列的分析  

DNA extraction and ribosomal DNA-ITS sequence analysis from wood tissue of Cassia siamea Lam.

作　　者：李奇威[1] 谭贵良 孙瑾[3] 苏越骁 王业胜[1] 周林[1] 朱爽[1] 

机构地区：[1]广东药学院生命科学与生物制药学院/广东省生物技术候选药物研究重点实验室,广东广州510006 [2]中山市质量计量监督检测所,广东中山528403 [3]华南农业大学林学院,广东广州510120

出　　处：《广东药学院学报》2015年第6期733-736,758,共5页Academic Journal of Guangdong College of Pharmacy

基　　金：中山市科技计划项目(2013A3FC0249);国家自然科学基金(21207021)

摘　　要：目的以核糖体转录间隔区（r DNA-ITS）序列为分子标记,对红木木材铁刀木进行遗传分析,并结合Gen Bank数据库中已有的序列阐明黄檀属、紫檀属、决明属之间及属内的分子系统发育关系。方法通过改良CTAB法对铁刀木边材进行总DNA提取,利用引物对r DNA-ITS序列进行PCR扩增和序列测定,用Mega软件进行系统发育分析,构建红木木材的分子系统发育树。结果用改良CTAB法可以成功提取铁刀木边材总DNA,并在模板稀释倍数为1×102~1×103时可成功PCR扩增出r DNA-ITS序列,片段长度大约为730 bp。结论基于通过r DNA-ITS序列进行序列测定和系统发育树构建的分子生物学方法,可利用ITS序列对红木木材铁刀木进行准确的分子鉴定,为红木的种类鉴定和种间分类提供分子生物学根据。Objective In order to clarify the phylogenetic relationships among the different species of genus Dalbergia,genus Pterocarpus,and genus Cassia by genetic analysis for Cassia siamea Lam. with the r DNAITS sequence as molecular marker. Methods Total DNA was extracted with modified CTAB method and r DNA-ITS regions were amplified with universally conserved primers. The r DNA-ITS sequences were characterized by phylogentic analysis using Mega software. Results Total DNA could be extracted from sapwood by modified CTAB method and r DNA-ITS region could be amplified successfully with template concentration in the dilution range of 1× 102- 1 × 103. Conclusion Based on molecular biology methods of r DNA-ITS sequence analysis,accurate molecular identification could be performed on Cassia siamea and provide molecular evidence for the taxonomy and identification of different species in rosewood.

关 键 词：铁刀木 红木 DNA提取 ITS序列 分子鉴定 

分 类 号：S567.19[农业科学—中草药栽培]