Abstract
Japanese Angelica Root prepared from Angelica acutiloba var. acutiloba and A. acutiloba var. sugiyamae, known in Japan as “Toki” and “Hokkai Toki”, is an important crude drug used in Kampo medicine (traditional Japanese medicine). However, since these Angelica varieties have recently outcrossed with each other, it is unclear whether Japanese Angelica Root sold for use in Kampo medicine is a pure variety. Here, we describe DNA sequence polymorphisms that can be used to distinguish between A. acutiloba var. acutiloba and A. acutiloba var. sugiyamae. In our analyses, differences in the trnK region of chloroplast DNA distinguished among some A. acutiloba varieties and related species, but not between A. acutiloba var. acutiloba and A. acutiloba var. iwatensis. One geographical strain of A. acutiloba var. acutiloba and A. acutiloba var. sugiyamae showed identical sequences in three regions of chloroplast DNA, but differences in the internal transcribed spacer region of nuclear ribosomal DNA. One strain of A. acutiloba var. iwatensis and A. acutiloba var. sugiyamae had identical sequences in all of the chloroplast and nuclear ribosomal DNA regions examined. These findings show that A. acutiloba var. acutiloba has hybridized with A. acutiloba var. sugiyamae and that the “Hokkai Toki” variety resulted from outcrossing with A. acutiloba var. iwatensis. Molecular authentication based on analyses of chloroplast and nuclear ribosomal DNA sequences of A. acutiloba and related species is an efficient method to authenticate Japanese Angelica Root at the variety level. Therefore, these analyses can determine whether a product is derived from A. acutiloba var. acutiloba or A. acutiloba var. sugiyamae.
Highlights
The Japanese indigenous species of Angelica acutiloba Kitagawa var. acutiloba Kitagawa (Toki) or A. acutiloba Kitagawa var. sugiyamae Hikino (Hokkai Toki) are listed in the Japanese Pharmacopoeia, 16th Edition [1], and are precious crude drugs in Kampo medicine
Japanese Angelica Root prepared from Angelica acutiloba var. acutiloba and A. acutiloba var. sugiyamae, known in Japan as “Toki” and “Hokkai Toki”, is an important crude drug used in Kampo medicine
One strain of A. acutiloba var. iwatensis and A. acutiloba var. sugiyamae had identical sequences in all of the chloroplast and nuclear ribosomal DNA regions examined. These findings show that A. acutiloba var. acutiloba has hybridized with A. acutiloba var. sugiyamae and that the “Hokkai Toki” variety resulted from outcrossing with A. acutiloba var. iwatensis
Summary
The Japanese indigenous species of Angelica acutiloba Kitagawa var. acutiloba Kitagawa (Toki) or A. acutiloba Kitagawa var. sugiyamae Hikino (Hokkai Toki) are listed in the Japanese Pharmacopoeia, 16th Edition [1], and are precious crude drugs in Kampo medicine (traditional Japanese medicine). Iwatensis Hikino (Miyama Toki), which is genetically very close to “Yamato Toki”, and A. stenoloba (“Hosoba Toki” or “Tokachi Toki”) [6]. There are wild-growing species that that are closely related to A. acutiloba, including A. acutiloba Kitagawa var. These species readily outcross with A. acutiloba var. Identification of Angelica acutiloba and Related Species by Analysis of Inter- and Intra-Specific Sequence Variations in Chloroplast and Nuclear DNA Sequences ensis had two-nucleotide differences in the spacer region between the atpF and atpA genes in the plastid genome [11]. Acutiloba is called “Nisshiki Toki” and resembles the Japanese style of “Yamato Toki” This product has been imported into Japan. The DNA sequences were the atpF-atpA, rpl16-rpl, and trnK regions of chloroplast DNA (cpDNA), and the internal transcribed spacer (ITS) of nuclear ribosomal DNA, consisting of five regions; 18S, ITS1, 5.8S, ITS2, and 26S
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