基于rbcL基因序列的欧洲菟丝子分子检测

Molecular detection of Cuscuta europaea based on rbcL sequence

菟丝子属(Cuscuta L.)被列为我国禁止进境的植物检疫性有害生物,目前进出境口岸使用常规形态学鉴定方法对菟丝子属的检测很难鉴定到种。本文通过扩增寄生于大豆上的6种菟丝子的1,5-二磷酸核酮糖羧化酶/加氧酶大亚基基因(rbcL)序列,通过Clustal X比对分析该6种菟丝子基因序列的差异,设计了针对欧洲菟丝子的特异引物,实现了对欧洲菟丝子(Cuscuta europaea)快速、准确的PCR分子检测,灵敏度达到单粒种子,满足各口岸对欧洲菟丝子的检测需求。

All species of the genus Cuscuta are plant quarantine pests that are forbidden to enter China. Identification of the genus Cuscuta is mainly based on morphological method at port presently, but it is difficult to identify samples to species. The sequence of the ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit （rbcL） of six dodders that were sheltered in soybeans were amplified. The rbcL sequences were aligned and analyzed by Clustal X. The special primers for Cuscuta europaea were subsequently designed, and quick and accurate PCR method was established to distinguish the C. europaea from the other five dodders. The detection sensitivity was high enough to detect one seed. This method can meet the detection requirements for C. europaea on port.