Journal of Natural Medicines (2024) 78:792–798

https://doi.org/10.1007/s11418-024-01796-0

NATURAL RESOURCE LETTER

Identification of Angelica acutiloba, A. sinensis, and other Chinese

medicinal Apiaceae plants by DNA barcoding
Motoyasu Minami1 · Ryusaku Tanaka1 · Takako Mori1 · Taichi Fujii1 · Takashi Tsuchida2

Received: 15 January 2024 / Accepted: 25 February 2024 / Published online: 1 March 2024
© The Author(s) under exclusive licence to The Japanese Society of Pharmacognosy 2024

Abstract
Crude drug Angelicae acutilobae radix is one of the most important crude drugs in Japanese traditional medicine and is
used mainly for the treatment of gynecological disorders. In the listing in the Japanese Pharmacopoeia XVIII, Angelicae
acutilobae radix is defined as the root of Angelica acutiloba (Apiaceae), which has long been produced on an industrial scale
in Japan. With the aging of farmers and depopulation of production areas, the domestic supply has recently declined and
the majority of the supply is now imported from China. Due to having only slightly different morphological and chemical
characteristics for the Apiaceae roots used to produce dried roots for Chinese medicines, the plant species originating the
crude drug Apiaceae roots may be incorrectly identified. In particular, Angelicae sinensis radix, which is widely used in
China, and Angelicae acutilobae radix are difficult to accurately identify by morphology and chemical profiles. Thus, in order
to differentiate among Angelicae acutilobae radix and other radixes originated from Chinese medicinal Apiaceae plants,
we established DNA markers. Using DNA sequences for the chloroplast psbA–trnH intergenic spacer and nuclear internal
transcribed spacer regions, Angelicae acutilobae radix and other Chinese Apiaceae roots, including Angelicae sinensis radix,
can be definitively identified.

Keywords Angelica acutiloba · Angelica sinensis · DNA barcoding · Chloroplast psbA–trnH intergenic spacer region ·
Nuclear internal transcribed spacer region

Introduction produced for domestic consumption in Japan, but due to

aging of farmers and depopulation of production areas, the
Genus Angelica (Apiaceae) has over 90 species in the tem- domestic supply has recently decreased [6]. Consequently,
perate zone of the Northern hemisphere [1], and at least the domestic self-sufficiency rate of this drug has decreased
23 species grow in Japan [2]. Of these, Angelica acutiloba to 20.4% of domestic consumption demand (about 864 tons)
(Siebold et Zucc.) Kitag. is endemic to Japan and grows in 2018 [7]. In order to ensure a stable supply of this crude
in the mountainous north-central area of the main island drug in Japan, seeds of A. acutiloba from Japan are being
of Japan [2, 3]. The root of A. acutiloba is defined as the cultivated in China, mainly in Sichuan and Jilin Provinces
crude drug Angelicae acutilobae radix and is listed in the [8]. The crude drug processed in China is given the Japanese
Japanese Pharmacopoeia XVIII (JPXVIII) [4]. Angelicae common name “Nisshiki-toki” and is exported to Japan [8].
acutilobae radix, one of the most important crude drugs in Therefore, the majority of the supply of this crude drug used
Japanese traditional medicine, has been used mainly for in Japan is imported from China [7, 8].
treating gynecological disorders, as reviewed by Afendi et al. In China, genus Angelica has about 45 species, and
[5]. Angelicae acutilobae radix has long been industrially at least 22 species are used for medicinal purposes [1].
In particular, Angelicae sinensis radix originating from
* Motoyasu Minami the root of A. sinensis (Oliv.) Diels [9] has high impor-
[email protected] tance in traditional Chinese medicine and has been used
both for the treatment of various diseases and as a tonic
1
College of Bioscience and Biotechnology, Chubu University, medicine for thousands of years, as reviewed by Wei et al.
1200 Matsumoto‑cho, Kasugai, Aichi 487‑8501, Japan
[10]. Owing to the high similarity of morphological and
2
Central R&D Laboratory, Kobayashi Pharmaceutical Co., chemical characteristics of the dried roots of other Chinese
Ltd, 1‑30‑3 Toyokawa, Ibaraki, Osaka 567‑0057, Japan

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Journal of Natural Medicines (2024) 78:792–798 793

medicinal Apiaceae plants, several of these similar spe- obtained from China were identified and the usefulness of
cies are sometimes considered to be Angelicae sinensis this analytical method was discussed.
radix by error [11] or intentionally substituted in the mar-
ket for cost concerns [12]. This is especially the case for
Angelicae acutilobae radix [1] and the root of Levisticum Materials and methods
officinale W. D. J. Koch (Apiaceae) [11–13], which is the
main adulterant or substitute for Angelicae sinensis radix Materials
in China. As a result, Angelicae acutilobae radix, Angeli-
cae sinensis radix, and other Chinese medicinal Apiaceae A total of 111 Angelica specimens (9 Angelicae acutilobae
roots may be confused. Angelicae sinensis radix and other radix specimens obtained from Japanese medicinal markets,
Chinese medicinal Apiaceae roots are not defined as the and 63 Angelicae acutilobae radix and 39 Angelicae sinensis
crude drug Angelicae acutilobae radix in the JPXVIII [4]. radix specimens obtained from Japanese medicinal markets,
Notably, Angelicae sinensis radix is morphologically very and different production sites and medicinal markets in
similar, making correct identification of Angelicae acu- China) were examined for detecting inter- and intraspecific
tilobae radix based on morphology alone impossible. In DNA polymorphisms of psbA–trnH and ITSs sequences
addition, the major component of both radixes is ligusti- (Tables S1 to S3). Before importing Chinese samples
lide, and the chemical profiles of the other components obtained in China into Japan, all samples were taken in
are also similar, which are difficult to accurately identify quarantine at the General administration of customs of the
by chemical profiles, as reviewed by Zhou et al. [14]. People's Republic of China and were imported to Japan with
Therefore, when importing Angelicae acutilobae radix the permission of the Agriculture, Forestry and Fisheries
from the China, it is necessary to accurately distinguish it based on the Plant Protection Act of Japan (permission
from other Chinese medicinal Apiaceae roots, especially nos. shown in Tables S2 and S3). All specimens used in
Angelicae sinensis radix. this study are deposited at the College of Bioscience and
To address these problems, Mei et al. [11] established Biotechnology, Chubu University, Japan.
a random amplified polymorphic DNA (RAPD) and
inter-simple sequence repeat (ISSR) analysis method Screening for DNA sequence polymorphisms
for identifying A. sinensis and A. acutiloba cultivated in
China. However, the detection efficiency of RAPD and Total DNA was isolated from each specimen using a DNeasy
ISSR markers are greatly affected by gel electrophoresis Plant Mini Kit (QIAGEN, Hilden, Germany) and then
and PCR conditions, making it difficult to reproducibly purified using a GENECLEAN SPIN KIT (MP Biomedicals,
make judgments based on the presence or absence of PCR Santa Ana, CA, USA) according to the respective
products. Murata et al. [15] established a DNA marker manufacturer instructions. PCR of psbA–trnH and ITSs
(namely LOX), which was constructed from the putative regions was performed in 25-μl reaction mixtures containing
A. lipoxygenase gene, for identification of A. acutiloba and 0.5 units of Mighty Amp Polymerase ver.3 (Takara, Shiga,
A. sinensis. However, the LOX marker was only useful for Japan) and 0.32 μM of each universal primer; other
identifying A. acutiloba and A. sinensis. In another approach, reaction solutions were prepared according to manufacturer
combined analysis of rbcL and matK genes, along with the instructions. Primer pairs flanking the target sequences were
psbA–trnH genes intergenic spacer region (psbA–trnH) used for PCR reactions as follows: psbA–trnH was amplified
for maternally inherited chloroplast DNA, and the internal using forward primer psbA (5′-GTTA ​ TGC​ ATG ​ AAC​ GTA​ AT​
transcribed spacers (ITSs) of amphoteric inherited nuclear GCTC-3′) and reverse primer ­trnHGUG​ (5′-CGC​GCA​TGG​
DNA by direct sequencing can be used to accurately detect TGG​ATT​CAC​AATCC-3′) [16]; ITSs was amplified using
inter- and intraspecific DNA polymorphisms of Chinese forward primer ITS5 (5′-GGA​AGT​AAA​AGT​CGT​AAC​
Angelica plants [12]. Notably, since DNA polymorphisms of AAGG-3′) and a reverse primer ITS4 (5′-TCCT ​ CCG​ CTT​ AT​
psbA–trnH and ITSs regions in Chinese medicinal Angelica TGA​TAT​GC-3′) [12]. PCR amplification was performed
plants exhibited marked intra- and interspecific variation using a DNA Thermal Cycler (Mini Amp Plus Thermal
[12], combined analysis of both regions could be expected Cycler, Applied Biosystems, Beverly, MA, USA) under the
to distinguish A. acutiloba from other Chinese medicinal following conditions: initial denaturation step of 98 °C for
Apiaceae plants, including A. sinensis. The objectives of this 2 min, followed by 35 cycles of denaturation for 10 s at
study were to evaluate the usefulness of the identification 98 °C, annealing for 15 s at 60 °C, and extension for 30 s
of A. acutiloba, A. sinensis, and other Chinese medicinal at 68 °C. PCR products were separated electrophoretically
Apiaceae plants using DNA polymorphisms in psbA–trnH on a 1.5% (w/v) agarose gel containing ethidium bromide
and the ITSs sequences. Finally, the original plants of in 1X TBE buffer and photographed under UV light. The
Angelicae acutilobae and Angelicae sinensis radixes psbA–trnH and ITSs PCR products each showed a single
794 Journal of Natural Medicines (2024) 78:792–798

band on the electrophoresis profile. Prior to sequencing,

Total (bp)
the PCR products were purified using a NucleoMag NGS

262
Clean-up and Size Select (MACHEREY–NAGEL, Düren,

261
278
Germany) and subjected to direct sequencing using a

93 97 102 113 115 116 118 134 138 179 202 203 204 205 206 207 208 209 210 211 212 213 218 219 220 221 222 224 237
BigDye Terminator v3.1 Cycle Sequencing Kit (Applied

T
T
Biosystems) on a 3500 Genetic Analyzer System (Applied

A
A
T
Biosystems). The psbA–trnH and ITSs amplicons were
sequenced using the forward and reverse primers used for

–
–
the PCR. The complete psbA–trnH and ITSs sequences

–
–
obtained for the specimens were aligned using the MEGA11
software package [17] and haplotypes of both sequences for

–
–
each specimen were determined. For species identification

–
–
of the specimens used in this study, homology searches of
the NCBI (National Center for Biotechnology Information)

–
–
database using the blastn program were performed with

G
G
T
the obtained psbA–trnH and ITSs haplotype sequences.
In this study, only specimens for which both the complete

–
–
psbA–trnH and ITSs sequences could be determined were

A
used as plant species identification samples. To ensure

–
–
sufficiently robust accuracy, species identification was

–
–
only considered definitive for DNA sequences with more

Table 1  DNA polymorphisms of the psbA-trnH intergenic spacer sequences for Angelica specimens used in this study
than 99% for psbA–trnH and ITSs. If a specimen produced

–
–
different species identifications for psbA-trnH and ITSs

–
–
sequences, identification to the taxonomic rank that included
both identified plant species was assigned.

–
–
T

–
–

Accession number in parentheses after the haplotype name was deposited in DDBJ by the authors
Results
T

–
–
T

–
DNA polymorphisms of psbA–trnH sequences –
and homology search results
T

–
–
T

–
–

DNA polymorphisms of the psbA–trnH in the Angelica

Position of variable sites from the 5' end of the psbA-trnH intergenic spacer
Asterisks (*) indicate nucleotide identical to that in the top row of this table

specimens used in this study are summarized in

G

T
*

Table 1. Among the 110 Angelica specimen psbA–trnH

C

T
–

sequences, except for voucher specimen A82-18-1, DNA

polymorphisms were found at 29 sites in nucleotide
G
G
T

positions 93–237 from the 5′ end of the intergenic spacer

A
T
*

region, and the Angelica specimens were grouped into three

different haplotypes: AH-J1, AH-C1, and AH-C2. Sequences
G
T
*

of haplotypes AH-J1, AH-C1, and AH-C2 were deposited

A
C

in the DDBJ (DNA Data Bank of Japan) (accession nos.

*
Nucleotide ­positionb

LC773989 to LC773991; Table 1). Results of homology

A
T
*

searches in the blastn program for these three psbA–trnH

Dashes (–) denote alignment gaps
A

haplotypes identified the following species assignments

C
*

(Table 2): AH-J1 was identified as A. acutiloba (accession

A

C
C

nos. KX352468 and NC029391), AH-C1 was identified

AH-C2 (LC773991) A
AH-C1 (LC773990) A
AH-J1 (LC773989) G

as A. sinensis (accession nos. MK688989, MK688990,

MK688991, MH430891, MH778544, NC042826,
a
(accession no.)

BK059532, and KC887947), and AH-C2 was identified as A.

sinensis (accession no. MW820164). Intraspecific variation
Haplotype

was found in the psbA–trnH sequences of A. sinensis used

in this study.
b
a
Journal of Natural Medicines (2024) 78:792–798 795

Table 2  Homology search results in NCBI database for each haplotype of the psbA-trnH intergenic spacer in Angelica specimens used in this
study
Haplotype (accession no.)a Identified plant ­speciesb Accession no.c Identity (%)d

AH-J1 (LC773989) Angelica acutiloba KX352468, NC029391 278/278 (100)

AH-C1 (LC773990) Angelica sinensis MK688989, MK688990, MK688991, MH430891, 261/261 (100)
MH778544, NC042826, BK059532, KC887947
AH-C2 (LC773991) Angelica sinensis MW820164 262/262 (100)
a
Accession number in parentheses after the haplotype name was deposited in DDBJ by the authors
b
Plant species retrieved by homology search with blastn program
c
Accession numbers of plant species retrieved by homology search with blastn program
d
DNA sequence match ratio (%) between plant species retrieved by homology search with blastn program and each haplotype detected in this
study

DNA polymorphisms of ITSs sequences Identification of the origin plants of Angelicae

and homology search results sinensis radixes produced in China

DNA polymorphisms of ITSs in Angelica specimens used The original plant species of Angelicae sinensis radix
in this study are summarized in Table 3. Among the 111 specimens obtained from China are summarized in Table S3.
Angelica specimens, ITSs polymorphisms were found at Almost all Angelicae sinensis radix specimens, except for
104 sites in nucleotide positions 44–635 from the 5’ end voucher specimens A82-23-1, -2, and -3 from Sichuan
of the ITSs, and the Angelica specimens were grouped Province, were haplotype AH-C1 and ITS-C1. Therefore,
into four different haplotypes: ITS-J1, ITS-J2, ITS-C1, except for specimens A82-23-1, -2, and -3 from Sichuan
and ITS-C2. Sequences of haplotypes ITS-J1, ITS-J2, ITS- Province, almost all original plant species of Angelicae
C1, and ITS-C2 were deposited in DDBJ (accession nos. sinensis radix obtained from China were A. sinensis. On the
LC773993–LC773996; Table 3). Results of homology other hand, although the psbA–trnH sequences (haplotype
searches in the blastn program identified the following AH-C2) of specimens A82-23-1, -2, and -3 were identified
species assignments (Table 4): ITS-J1 was identified as A. as A. sinensis, those ITSs sequences (haplotype ITS-C2)
acutiloba (accession no. AY548227), ITS-J2 was identified were identified as P. aromaticum. Since specimens A82-23-
as A. acutiloba (accession no. AY548227), and ITS-C1 1, -2, and -3 showed different species identification based
was identified as A. sinensis (accession nos. JX022936, on psbA-trnH and ITSs haplotype sequences, those samples
GU289653, and GU289654). Intraspecific variation was were not identified to the species level. Therefore, those
found in the ITSs sequence of A. acutiloba used in this specimens were assigned to family Apiaceae.
study. The ITS-C2 haplotype sequence was identified as
Pleurospermum aromaticum W.W.Sm (Apiaceae) (accession
no. KP311496) by homology searches (Table 4). Discussion

Identification of the origin plants of Angelicae Angelicae sinensis radix is mainly produced in the southeast-
acutilobae radixes produced in China ern parts of Gansu Province and occurs in Yunnan, Sichuan,
Shaanxi, and Hubei Provinces in China [1, 10]. The main
All psbA–trnH haplotypes of Angelicae acutilobae radix production area of Angelicae acutilobae radix for export to
specimens from Japanese medicinal markets were haplotype Japan is Sichuan Province [8]. Thus, there is overlap in the
AH-J1 (Table S1). On the other hand, the ITSs haplotype production areas of Angelicae sinensis and Angelicae acuti-
was divided into two haplotypes (ITS-J1 and -J2) (Table S1). lobae radixes within Sichuan Province. Although this study
All Angelicae acutilobae radix specimens obtained from is somewhat limited by the small number of specimens, the
China were haplotype AH-J1 and ITS-J1 or -J2 (Table S2), Angelicae acutilobae radix obtained from Sichuan Province,
which is consistent with specimens in Japan (Table S1). which is the main area of production in China for export to
Therefore, all original plant species of Angelicae acutilobae Japan, were all identified as A. acutiloba. On the other hand,
radix specimens obtained from China were A. acutiloba almost all Angelicae sinensis radix specimens obtained from
(Table S2). Using the combined DNA sequences of AH-J1 Gansu, Qinghai, and Yunnan Provinces were identified as
and ITS-J1 or ITS-J2 haplotypes, A. acutiloba could be A. sinensis. However, three Angelicae sinensis radix speci-
correctly identified to the species level (Table 5). mens (voucher abbreviations A82-23-1, -2, and -3; Table S3)
 796

Table 3  DNA polymorphisms of the ITSs of Angelica specimens used in this study
Haplotype (accession no.)a Nucleotide ­positionb
44 54 67 70 75 76 78 83 86 87 89 90 95 96 97 99 104 106 110 117 131 133 135 137 139 140

ITS-J1 (LC773993) A T C C T T T G C G C G G C C C T C C T C A T C G G
ITS-J2 (LC773994) * * * * * * * * * * * * * G * * * * * * * * * * * *
ITS-C1 (LC773995) G C T A * A * A T * T C * * T T C T A – * C * T * *
ITS-C2 (LC773996) G * * A C A C A * * * * * * T T C T * – * C C T * *
Haplotype (accession no.)a Nucleotide ­positionb
148 149 157 162 166 188 196 206 210 212 223 227 229 230 232 246 251 377 393 414 417 426 427 437 441 443

ITS-J1 (LC773993) C T A C C C C C C T C C C C G C C C C G – A A A – A
ITS-J2 (LC773994) * * * * * * * * * * * * * * * * * * * * * * * * * *
ITS-C1 (LC773995) * * G * * A T * G C * * T * A * * T T A T – * T C T
ITS-C2 (LC773996) * * G A T A * * * C * * G * * * * T T A T – T T T *
Haplotype (accession no.)a Nucleotide ­positionb
445 448 452 453 454 457 458 460 470 471 426 427 437 441 443 445 448 452 453 454 457 458 460 470 471 483

ITS-J1 (LC773993) A T G C C T T – A C A A A – A A T G C C T T – A C A
ITS-J2 (LC773994) * * * * * * * * * * * * * * * * * * * * * * * * * *
ITS-C1 (LC773995) G C A * T * A G * T – * T C T G C A * T * A G * T G
ITS-C2 (LC773996) G A * T T * * G * T – T T T * G A * T T * * G * T G
Haplotype (accession no.)a Nucleotide position b Total (bp)
490 503 508 509 516 519 523 527 532 535 538 546 552 559 569 570 579 591 598 611 612 618 622 627 628 635

ITS-J1 (LC773993) C G A C C G C C C C C G G T G T T C C C A C C C G C 671

ITS-J2 (LC773994) * * * * * * * * * * * * * * * * * * * * * * * * * * 671
ITS-C1 (LC773995) T C G T * * * * T * T * T A * C G T A * G * T * A A 672
ITS-C2 (LC773996) T C G * * A * * T * * * * A A C G T T T G A * * * T 672

Asterisks (*) indicate nucleotide identical to that in the top row of this table
Dashes (–) denote alignment gaps
a
Accession number in parentheses after the haplotype name was deposited in DDBJ by the authors
b
Variable site positions are denoted from the 5' end of the ITSs
Journal of Natural Medicines (2024) 78:792–798
Journal of Natural Medicines (2024) 78:792–798 797

Table 4  Homology search Haplotype (accession no.) a Identified plant ­speciesb Accession no.c Identity (%)d
results by NCBI database of
obtained for each haplotype of ITS-J1 (LC773993) Angelica acutiloba AY548227 668/669 (99)
the ITSs for Angelica specimens
ITS-J2 (LC773994) Angelica acutiloba AY548227 669/669 (100)
used in this study
ITS-C1 (LC773995) Angelica sinensis JX022936, 672/672 (100)
GU289653,
GU289654
ITS-C2 (LC773996) Pleurospermum aromaticum KP311496 647/650 (99)
a
Accession number in parentheses after the haplotype name was deposited in DDBJ by the authors b Plant
species identified by homology search with blastn program
c
Accession number of plant species retrieved by homology search with blastn program
d
DNA sequence match ratio (%) between plant species retrieved by homology search with blastn program
and each haplotype sequenced in this study

Table 5  Combination of psbA-trnH and ITSs haplotypes for identifi- in order to establish a DNA barcoding method capable of
cation of Angelica acutiloba and A. sinensis identification to the species level in order to prevent con-
Scientific name Haplotype tamination with these Angelicae radixes.
Another concern in developing identification tools for
psbA-trnHa ITSsb
this study was whether Angelicae sinensis and Angelicae
A. acutiloba AH-J1 ITS-J1, ITS-J2 acutilobae radixes are contaminated with L. officinale
A. sinensis AH-C1 ITS-C1 roots. L. officinale roots contain ligustilide and ferulic
a
acid constituents similar to those in Angelicae sinensis
Haplotype information is shown in Table 1
b
and Angelicae acutilobae radixes [10]. L. officinale,
Haplotype information is shown in Table 3
which is native to southwest Asia and Europe, was
officially introduced to China from Europe in 1957 as
obtained from Sichuan Province, the main production area a substitute to satisfy the large demand for Angelicae
of Angelicae acutilobae radix, showed different species sinensis radix [12] and then was widely cultivated in Nei
identification based on psbA-trnH and ITSs haplotype Mongol, Shaanxi, Shandong, and Shanxi Provinces [13].
sequences: those psbA–trnH sequences (haplotype AH-C2) Later, pharmacological experiments determined a lower
were identified as A. sinensis and those ITSs sequences medicinal efficacy of L. officinale than for Angelicae
(haplotype ITS-C2) were identified as P. aromaticum. The sinensis radix, and thus, L. officinale was excluded from
psbA–trnH sequences of P. aromaticum were not registered the Chinese Pharmacopoeia (2005) and has been prohibited
in the NCBI database (accessed 25 December 2023). In as a substitute of Angelicae sinensis radix since 1983
addition, the psbA–trnH (accession nos. NC_071797 and [12]. However, intentional cultivation of L. officinale as
OP672456) sequences of Oreocomopsis aromatica (W. W. an alternative for Angelicae sinensis radix is prevalent
Smith) Pimenov et Kljuykov., which is the synonym of P. in Hebei, Shandong, and Henan Provinces [20]. The
aromaticum [18], registered in the NCBI database are com- psbA–trnH (accession nos. MT561024 and LC773992) and
pletely different from those of A. sinensis and A. acutiloba. ITSs (accession no. LC773997) sequences of L. officinale
Therefore, these three specimens (voucher abbreviations registered in the NCBI database are completely different
A82-23-1, -2, and -3) could not be identified to the species from those of A. sinensis and A. acutiloba. Therefore, both
level and were identified as Apiaceae at the family level. sequences can be used to distinguish the roots of L. officinale
Since A. sinensis and P. aromaticum are in different genera, from those of Angelicae sinensis and Angelicae acutilobae
it is unlikely that they retain the same psbA–trnH sequence. radixes.
Therefore, it is possible that these three specimens (voucher In this study, all Angelicae acutilobae radixes obtained
abbreviations A82-23-1, -2, and -3) are putative genus-level from the Chinese medicinal market were identified as having
hybrids of these two species, but the details are unknown. P. originated from the roots of Angelica acutiloba. However,
aromaticum is used for medicinal purposes and grows wild the origin plant species of some Angelicae sinensis radixes
in ditches of forests, open dwarf scrub, and alpine mead- obtained from the Chinese medicinal market are other
ows in high mountainous areas of southwestern Sichuan and Apiaceae plants. Therefore, though Angelica radixes
Xizang Provinces, and southwestern Yunnan Province [18, produced and distributed in China are difficult to accurately
19]. Since the habitats of P. aromaticum overlap with pro- identify to the species level by morphological characteristics
duction sites of Angelicae sinensis and Angelicae acutilobae and chemical component profile of their roots, the DNA
radixes, it is necessary to obtain the P. aromaticum plant barcoding method introduced in this study is a useful method
798 Journal of Natural Medicines (2024) 78:792–798

for identifying the plant species of not only Angelicae 9. State Pharmacopoeia Commission of the People’s Republic of
acutilobae radix but also the other Chinese medicinal China (2020) Angelicae sinensis Radix. In: State Pharmacopoeia
Commission of the People’s Republic of China (ed) The pharma-
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Springer, China, pp 139–185
Supplementary Information The online version contains supplemen- 10. Wei WL, Zeng R, Gu CM, Qu Y, Huang LF (2016) Angelica
tary material available at https://d​ oi.o​ rg/1​ 0.1​ 007/s​ 11418-0​ 24-0​ 1796-0. sinensis in China—a review of botanical profile, ethnopharmacol-
ogy, phytochemistry and chemical analysis. J Ethnopharmacol
Author contributions Drs. Minami and Tsuchida supervised the 190:116–141
research. Dr. Minami wrote the manuscript and analyzed the data. Dr. 11. Mei Z, Zhang C, Khan MA, Zhu Y, Tania M, Luo P, Fu J (2015)
Tsuchida contributed to the collection of Angelica specimens. Mr. Efficiency of improved RAPD and ISSR markers in assessing
Tanaka, Mrs. Mori, and Dr. Fujii performed DNA analysis and data genetic diversity and relationships in Angelica sinensis (Oliv.)
analysis. Diels varieties of China. Electron J Biotechnol 18:96–102
12. Yuan QJ, Zhang B, Jiang D, Zhang WJ, Lin TY, Wang NH, Chiou
Funding This research was supported by the Chubu University Grant SJ, Huang LQ (2015) Identification of species and materia medica
(K) (Grant Number 22212K). within Angelica L. (Umbelliferae) based on phylogeny inferred
from DNA barcodes. Mol Ecol Resour 15:358–371
Declarations 13. Pan Z, Watson MF (2005) Levisticum officinale W. D. J. Koch. In:
Wu ZY, Raven PH, Hong DY (eds) Flora of China, vol 14. Science
Conflict of interest Not applicable. Press, Beijing and Missouri Botanical Garden Press, St. Louis, p
172
14. Zhou SS, Xu J, Tsang CK, Yip KM, Yeung WP, Zhao ZZ, Zhu S,
Fushimi H, Chang HY, Chen HB (2018) Comprehensive quality
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