Rapid identification of Atractylodes macrocephala Koidz., Atractylodes lancea (Thunb.) DC. and hybrids with MLPA-qPCR based on nuclear genome

Atractylodes macrocephala and A. lancea are traditional medicinal herbs with a long history in the treatment of diseases. Although A. macrocephala and A. lancea have similar chemical components, their efficacies are different. Confusion of A. macrocephala and A. lancea is one of the hidden dangers of drug safety. Besides, the hybrids of A. macrocephala and A. lancea were bred and sold for bigger rhizomes, grow faster and cheaper prices. which is harmful to correct breeding and market regulation of medicinal materials. In order to correctly identify A. macrocephala, A. lancea and their hybrids, a molecular method combining multiplex ligation-dependent probe amplification and Real-time Quantitative PCR had been established in this study. Highly species-specific MLPA probes were designed based on ITS2 region within nuclear genome without cross-reaction, and three species could be distinguished according to the melting curve within 3 h. This study suggests that MLPA-qPCR is an accurate, feasible and high-throughput method for the identification of A. macrocephala, A. lancea and hybrids, and provides a reference for identification of species and even hybrids.