Efficacy of the combined method of chloroplast atpF-atpA and genome-wide SNP analysis using MIG-seq for identifying hybrids between A. acutiloba and A. acutiloba var. sugiyamae.

Angelicae acutilobae radix, which has traditionally been used to treat gynecological disorders, is listed in the Japanese Pharmacopoeia Eighteenth Edition as the root of Angelica acutiloba or A. acutiloba var. sugiyamae. There are constituent differences in the roots of these varieties, and the quality of the crude drug processed from the roots of A. acutiloba is considered to be superior to that processed from A. acutiloba var. sugiyamae based on traditional pharmacognostical evaluations. Although A. acutiloba and A. acutiloba var. sugiyamae have different botanical characteristics, interbreeding of these varieties easily produces hybrids. The presence of hybrids within cultivated A. acutiloba populations or the same lot can result in variability of this crude drug. In order to identify their hybrids, we developed a combined method for the analysis of DNA polymorphisms in the atpF-atpA intergenic region of chloroplast DNA using direct sequencing to identify the maternal lineage and genomic components using genome-wide SNP analysis by MIG-seq. Mixtures of hybrids found in cultivated A. acutiloba plants and processed Angelicae acutilobae radices in Japan and China could be identified by the combined method. This combined method is expected to be useful for accurately identifying hybrids in a wide variety of plant-derived medicines globally, both in cultivated fresh medicinal plant populations and in dried herbal materials.