应用分子生物学方法鉴定马来沉香  

Identification of Aquilaria malaccensis with mollecular biological method

作　　者：张明哲 徐莉莉 陈曦 陈吴健 林晓佳 吴志毅 武扬 吴蓉 

机构地区：[1]浙江省检验检疫科学技术研究院,浙江杭州310016 [2]浙江出入境检验检疫局 [3]杭州出入境检验检疫局

出　　处：《植物检疫》2015年第3期72-76,共5页Plant Quarantine

基　　金：浙江省科技厅重大专项(2011C12023);国家质检总局科技计划项目(2011IK249);浙江省科技厅公益性项目(2011C22065)

摘　　要：DNA条形码技术是利用基因组中一段短的标准序列进行物种鉴定的新技术。本研究对截获的一批非法入境木材样品的ITS1及TrnL-TrnF序列进行PCR扩增和测序。将测序结果与Gen Bank中的同源序列进行比对分析,发现未知样品与马来沉香（Aquilaria malaccensis）的序列相似性最高,达到99%~99.7%,遗传距离最小,为0.00~0.01。用邻接（NJ）法构建ITS1和TrnL-TrnF序列的系统进化树,未知样品均与马来沉香聚为一枝,bootstrap values分别为77%和100%。上述结果表明,未知样品为马来沉香,可利用ITS1和TrnL-TrnF从分子水平区分马来沉香与其他混伪种。DNA barcoding is a technique of species identification by using DNA sequences from a small fragment of the genome. In the study, we used this technique to identify a batch of smuggled wood and selected ITS1 and TrnL-TrnF, were amplified and sequenced. Alignment analysis with the homologous sequences in GenBank revealed that the unknown samples had the highest sequences similarity, which was 99%-99.7%, with the sequences of Aquilaria malaccensis. The genetic distance that was computed by MEGA5.05 between unknown samples and Aquilaria malaccensis was 0.00-0.01, which was the minimum. Moreover, analyses of phylogenetic trees by NJ methods based on ITS1 and TrnL-TrnF regions indicated that the unknown samples in the tests clustered together with the Aquilaria malaccensis species from the GenBank, with bootstrap values of 77% and 100%, respectively. Consequently, the unknown samples is Aquilaria malaccensis and it is feasible to identify Aquilaria malaccensis wood on a species level based on DNA barcoding.

关 键 词：马来沉香 DNA条形码 TRNL-TRNF ITS1 物种鉴定 

分 类 号：S41[农业科学—植物保护]