白木香良种‘热科2号’的DNA条形码分子鉴定

Molecular Identification of Aquilaria sinensis ‘Reke 2’Based on DNA Barcoding

奇楠沉香为极具收藏及药用价值的上品沉香,其鉴定目前以化学方法为主,但其种苗从苗期至采香之间则难以用传统分类和化学方法进行鉴定,DNA条形码的发展为这一产业难题的解决提供了新的思路。本研究在明确白木香良种‘热科2号’可产奇楠沉香的基础上,采用11对植物DNA条形码通用引物对‘热科2号’和普通白木香两个沉香品种进行分子鉴定探索,发现改良的CTAB法更适合白木香叶片样本的DNA提取,继而发现ITS、ITS2和源自叶绿体的8个DNA条形码在两个品种均无变异,而ITS1序列的第159个碱基在‘热科2号’中为T,在普通白木香中为A,多样本实验证明该变异位点可以作为‘热科2号’和普通白木香种苗鉴定的分子标记。本研究结果为白木香良种‘热科2号’及其幼苗的鉴定提供分子水平的技术支持,也为白木香良种的溯源提供新的思路。

‘Qi-Nan’is the top grade agarwood based on collectable and pharmaceutical value,its identification is primarily according to chemical methods,but they were hardly identified via traditional taxonomy and chemical analysis from seedling to agarwood harvest.DNA barcoding might provide the efficient method for identification of‘Qi-Nan’seedling.The modified CTAT method could be more suitable for DNA extraction from leaves of Aquilaria sinensis and then we developed 11 DNA barcoding from‘Reke 2’and normal A.sinensis after chemical analysis indicating that‘Reke 2’species could produce‘Qi-Nan’agarwood.The results showed that ITS,ITS2 and eight DNA barcoding were exactly same in‘Reke 2’and normal A.sinensis,only one variation site was identified in ITS1 and this site was located at 159 loci.The nucleotide of ITS 1at 159 loci in‘Reke 2’is thymine(T),whereas that in normal A.sinensis is adenine(A),and this variation site were validated in 20 plants of‘Reke 2’and 20 plants of normal A.sinensis.This analysis suggested that 159 loci of ITS1 could be identified‘Reke 2’and normal A.sinensis.This results provide technical support for excellent seedling of A.sinensis identification and would contribute to trace and identify superior seedling of A.sinensis.