基于ITS序列的竹亚科植物分子鉴定研究

Molecular Identification of Bambusoideae Plants Based on ITS Sequences

利用ITS序列对竹亚科13个属76个竹种进行DNA条形码分子鉴定.该研究提取慈竹属、刚竹属、箬竹属、唐竹属、矢竹属、大明竹属、赤竹属、巴山木竹属、倭竹属、短穗竹属、簕竹属、酸竹属及少穗竹属共13个属62个竹种DNA,对其内转录间隔区ITS序列进行PCR扩增和双向测序,所得序列与GenBank数据库下载的14条ITS序列共76个竹种序列用Clustal X软件比对,再利用MEGA6.06软件构建K2P距离法的NJ系统发育树.结果显示,种内遗传距离为0~3.907,平均遗传距离为0.370;种间遗传距离为0~4.394,平均遗传距离为2.287,种内遗传距离远小于种间遗传距离.ITS序列可用于竹类资源的物种鉴定研究.

Molecular identification was conducted of 76 species of 13 genera in Bambusoideae by nuclear internal transcribed spacer（ITS）sequence fragments.DNA was extracted from 62 bamboo species of 13 genera（Neosinocalamus,Phyllostachys,Indocalamus,Sinobambusa,Pseudosasa,Pleioblastu,Sas,Bashani,Shibataea,Brachystachyum,Bambusa,Acidosasa and Oligostachyum）,and PCR amplification and bidirectional sequencing of the 62 ITS sequences were carried out.Then the 62 ITS sequences obtained were aligned with 14 ITS sequences from GenBank database by Clustal X,and MEGA program（version 6.06）was employed to estimate the intraspecific and interspecific genetic distances.NJ（NeighborJoining）trees were constructed based on K2 P（Kimura-2-parameter）distance of the ITS sequences in order to evaluate the results.Results showed that the intraspecific genetic distance was much smaller than the interspecific genetic distance：the intraspecific genetic distance was 0-3.907,averaging 0.370;and the interspecific genetic distance was 0-4.394,averaging 2.287.In conclusion,ITS barcodes can be used to distinguish different species of Bambusoideae.