基于单核苷酸多态性位点的土沉香物种鉴定

Identification of Aquilaria sinensis by Single Nucleotide Polymorphism

对土沉香Aquilaria sinensis,云南沉香A. yunnanensis和马来沉香A. malaccensis 3种沉香属Aquilaria植物的ITS序列进行PCR扩增和测序,并通过DNAMAN软件筛选出单核苷酸多态性(SNP)位点,土沉香序列第236位上的碱基为C,而云南沉香和马来沉香为T,设计出土沉香特异性引物AS-F,AS-R和探针AS-P。实时荧光定量PCR实验表明,该引物探针能特异性扩增土沉香,且检测灵敏度可以达到0.01ng·μL^(-1),是一种快速、灵敏的鉴定方法。

Leaves were collected and timbers of Aquilaria sinensis, A. yunnanensis and A. malaccensis were quarantined from March to July of 2017 from different provinces in China, and were amplified and sequenced. Screening of single nucleotide polymorphisms (SNP) by DNAMAN software, specific primers and probes were designed for. The real-time PCR test showed that the primer and probe could specifically amplify A. sinensis, and the sensitivity reached 0.01 ng·μL-1.