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以南岭黄檀(Dalbergia balansae)为研究对象,采用改良CTAB法对南岭黄檀边材进行总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,构建南岭黄檀及其近缘树种的分子系统发育树。结果表明:用改良CTAB法可以成功提取南岭黄檀边材总DNA,并在模板稀释倍数为1×10-2~1×10-3倍时可成功PCR扩增出r DNA-ITS序列,并基于r DNA-ITS序列测定和系统发育树构建的分子生物学方法,利用ITS序列对南岭黄檀进行分子鉴定,为南岭黄檀的种类鉴定和种间的分类地位提供分子生物学根据。
Taking Dalbergia balansae from Nanling as the research object, the total DNA of Dalbergia balansae from Nanling was extracted by modified CTAB method. The PCR amplification and sequencing of r DNA-ITS sequence were carried out by using primers and MEGA software Developmental analysis, the construction of Nanling Dalbergia and its related species of molecular phylogenetic tree. The results showed that the total DNA of Dalbergia glutinosa sapwood could be successfully extracted by modified CTAB method and the r DNA-ITS sequence was successfully amplified by PCR at a dilution of 1 × 10-2 ~ 1 × 10-3. Based on the rDNA-ITS sequence analysis and molecular phylogenetic tree construction, the ITS sequence was used to identify the species of Dalbergia hupehensis and provide the molecular biological basis for the species identification and interspecific taxonomic classification of Dalbergia hupensis .