TYPE Review

PUBLISHED 11 August 2022

DOI 10.3389/fpls.2022.973197

The genus Chrysanthemum:

OPEN ACCESS Phylogeny, biodiversity,
phytometabolites, and
EDITED BY
Wanchai De-Eknamkul,
Chulalongkorn University,
Thailand

REVIEWED BY
chemodiversity
Xiwen Li,
China Academy of Chinese Medical Sciences,
China
Da-Cheng Hao 1,2*†, Yanjun Song 3†, Peigen Xiao 3,4, Yi Zhong 3,
Wenpan Dong, Peiling Wu 3 and Lijia Xu 3,4*
Beijing Forestry University,
China 1
School of Environment and Chemical Engineering, Biotechnology Institute, Dalian Jiaotong
University, Dalian, China, 2 Institute of Molecular Plant Science, University of Edinburgh, Edinburgh,
*CORRESPONDENCE
United Kingdom, 3 Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences
Da-Cheng Hao
and Peking Union Medical College, Beijing, China, 4 Key Laboratory of Bioactive Substances and
[email protected]
Resources Utilization of Chinese Herbal Medicine, Ministry of Education, Beijing, China
Lijia Xu
[email protected]
†
These authors have contributed equally to The ecologically and economically important genus Chrysanthemum contains
this work
around 40 species and many hybrids and cultivars. The dried capitulum of
SPECIALTY SECTION
Chrysanthemum morifolium (CM) Ramat. Tzvel, i.e., Flos Chrysanthemi, is
This article was submitted to
Plant Metabolism and Chemodiversity, frequently used in traditional Chinese medicine (TCM) and folk medicine for at
a section of the journal least 2,200years. It has also been a popular tea beverage for about 2,000years
Frontiers in Plant Science
since Han Dynasty in China. However, the origin of different cultivars of CM and
RECEIVED 19 June 2022
the phylogenetic relationship between Chrysanthemum and related Asteraceae
ACCEPTED 18 July 2022
PUBLISHED 11 August 2022 genera are still elusive, and there is a lack of comprehensive review about the
CITATION association between biodiversity and chemodiversity of Chrysanthemum. This
Hao D-C, Song Y, Xiao P, Zhong Y, article aims to provide a synthetic summary of the phylogeny, biodiversity,
Wu P and Xu L (2022) The genus
phytometabolites and chemodiversity of Chrysanthemum and related
Chrysanthemum: Phylogeny, biodiversity,
phytometabolites, and chemodiversity. taxonomic groups, focusing on CM and its wild relatives. Based on extensive
Front. Plant Sci. 13:973197. literature review and in light of the medicinal value of chrysanthemum, we give
doi: 10.3389/fpls.2022.973197
some suggestions for its relationship with some genera/species and future
COPYRIGHT
applications. Mining chemodiversity from biodiversity of Chrysanthemum
© 2022 Hao, Song, Xiao, Zhong, Wu and
Xu. This is an open-access article containing subtribe Artemisiinae, as well as mining therapeutic efficacy and
distributed under the terms of the Creative other utilities from chemodiversity/biodiversity, is closely related with sustainable
Commons Attribution License (CC BY). The
use, distribution or reproduction in other
conservation and utilization of Artemisiinae resources. There were eight main
forums is permitted, provided the original cultivars of Flos Chrysanthemi, i.e., Hangju, Boju, Gongju, Chuju, Huaiju, Jiju,
author(s) and the copyright owner(s) are Chuanju and Qiju, which differ in geographical origins and processing methods.
credited and that the original publication in
this journal is cited, in accordance with Different CM cultivars originated from various hybridizations between multiple
accepted academic practice. No use, wild species. They mainly contained volatile oils, triterpenes, flavonoids, phenolic
distribution or reproduction is permitted
which does not comply with these terms.
acids, polysaccharides, amino acids and other phytometabolites, which have
the activities of antimicrobial, anti-viral, antioxidant, anti-aging, anticancer, anti-
inflammatory, and closely related taxonomic groups could also be useful as food,
medicine and tea. Despite some progresses, the genetic/chemical relationships
among varieties, species and relevant genera have yet to be clarified; therefore,
the roles of pharmacophylogeny and omics technology are highlighted.

KEYWORDS

Chrysanthemum, Chrysanthemum morifolium, phylogenetic relationship,

phytochemistry, chemodiversity, pharmacophylogeny

Frontiers in Plant Science 01 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

Introduction Due to its ornamental, medical and nutritional value, CM

appreciation had created an unique chrysanthemum culture in East
Anthemideae is a taxonomically controversial Asteraceae Asia. The varieties of CM have increased from 300 in Qing Dynasty
(Compositae) tribe, in which phylogenetic relationships are still not to over 1,000 nowadays (Zhang, 2008; Mao, 2020). The CM’s medical
settled (Criado Ruiz et al., 2021). In Anthemideae, Artemisiinae and nutritional values were recognized around 2,300 years ago
and Matricariinae are the two most widely distributed subtribes (Chen, 2005; Zhang, 2008); in the Spring and Autumn Period CM
(Watson et al., 2002; Shen et al., 2021). Artemisiinae (Figure 1) firstly appeared in “Li Sao,” written by the great poet Yuan Qu, as an
includes 1/3 species of tribe Anthemideae, consisting of more than edible vegetable. Later it was recorded as a medicine in “Shen Nong’s
600 species of 18–19 genera, which are mainly in the northern Classic of the Materia Medica” in Han Dynasty. Presently, CM is one
hemisphere, especially in Central Asia and East Asia. of the most commonly consumed food, herbal medicine and tea
Chrysanthemum, a small Artemisiinae genus, is native to East Asia beverage in China and adjacent countries (Lai et al., 2007; Jiang et al.,
and northeastern Europe. It contains around 40 species and many 2013). CM is extensively used by healthcare providers to treat
hybrids and cultivars, most of which originate from East Asia, and conditions such as dizziness, photophobia with lacrimation,
China is the center of diversity (Liu et al., 2012). Innumerable conjunctivitis, headache with fever, red eyes, swollen poison and
horticultural varieties and cultivars of Chrysanthemum exist. The boils, among others (Chinese Pharmacopoeia Commission, 2020).
Chrysanthemum plants are perennial herbs. Their leaves are Modern studies found that Chrysanthemum and related
undivided or palmately/pinnately divided once or twice. The genera contain significant amounts of volatile oils, flavonoids and
capitulum is heteromorphic, solitary at the top of the stem, a few or hydroxycinnamoyl-quinic acids (Beninger et al., 2004; Jiang et al.,
more of which are arranged into corymbose or compound 2004; Chen et al., 2007; Clifford et al., 2007; Lai et al., 2007; Wang
corymbose inflorescences at the top of stem and branch. The et al., 2008a,b; Duan et al., 2022), and have extensive biological
marginal flowers are female, tongue shaped, 1-layer (multi-layer in activities (Figure 1), e.g., anti-inflammation, antioxidation,
cultivated varieties), and the central disk flowers are bisexual and vasodilation, protecting cardiovascular system, anticancer,
tubular. The involucre is shallowly discoid, rarely campanulate. The inhibiting aldose reductase and anti-mutagenic (Zhang et al.,
involucral bracts are 4–5 layers, with white, brown, black brown or 2000; Rajic et al., 2001; Peng et al., 2006; Shao et al., 2020). Given
brownish black edges. The receptacle is protuberant, hemispherical the long use history and important role of Chrysanthemum taxa,
or conical, without stipules. The tongue shaped flowers are yellow, and in order to better preserve and utilize Chrysanthemum
white or red, and the tongue can be as short as 1.5 mm, and as long resources, a comprehensive review is necessary to synthesize
as 2.5 cm or longer. All tubular flowers are yellow, with five teeth at information from existing studies of their phylogeny, biodiversity,
the top. The style branches are linear, apically truncate. The anther chemodiversity and chemotaxonomy. In this study, the up-to-date
base is obtuse, and apical appendages are lanceolate ovate or oblong. information of Chrysanthemum were collected from major
All achenes are isomorphic, nearly cylindrical and narrowed to the databases including NCBI PubMed, Google Scholar, Web of
lower part, with 5–8 longitudinal veins and no coronal crown hair. Science, SciFinder, Wiley online, Elsevier ScienceDirect and
As the most remarkable taxon of Chrysanthemum, China National Knowledge Infrastructure (CNKI), using the
Chrysanthemum morifolium Ramat. Tzvel (CM, known as “Ju Hua” keywords such as “Chrysanthemum,” “juhua,” “phylogenetic,”
in Chinese) originated in China as a medicinal, food and ornamental “biosynthesis,” the respective phytometabolite name, the
plant. Its history began about 3,000 years ago, since Xia and Shang respective genus/species name, and the like. Additionally, materia
Dynasties (Zhang and Dai, 2009). CM was introduced to Japan as a medica books and patents have made their contribution to the
famous spice during Tang Dynasty (AD 710 to 784), and later summary of botany, traditional uses and cultural significance of
entered Europe and then America in the 17th century (Li, 1993). C. morifolium. The phylogenetic kinship, origin of genuine
varieties, and the chemical ingredients and their bioactive effects
of Chrysanthemum taxa were elaborated from the perspective of
Abbreviations: Acn, Acacetin; AFLP, Amplified fragment length polymorphism; pharmacophylogeny, so as to facilitate a holistic understanding of
AG, Acetylglucoside; ANS, Anthocyanidin synthase; Api, Apigenin; CDDP,
Conserved DNA-derived polymorphism; CHI, Chalcone isomerase; CHS, Chalcone
Chrysanthemum and related genera and promote the sustainable
synthase; CIF, Chrysanthemi Indici Flos; CM, Chrysanthemum× morifolium; cp, conservation and utilization of relevant natural resources.
chloroplast; CYP, Cytochrome p450; DCQA, Dicaffeoylquinic acid; DFR,
Dihydroflavanol 4-reductase; Dio, Diosmetin; EST, Expressed sequence tag; F3′H,
Flavonoid 3′-hydroxylase; F3′5′H, Flavonoid 3’5’-hydroxylase; FISH, Fluorescence
in situ hybridization; GT, Glycosyltransferase; IGS, Intergenic spacer; ISSR, Inter- Biodiversity and phylogenetic
simple sequence repeat; Lut, Luteolin; MG, Malonylglucoside; MW, Molecular relationship
weight; nr ITS, Nuclear ribosomal internal transcribed spacer; PCA, Principal
component analysis; QTP, Qinghai-Tibetan Plateau; RAPD, Random amplified
polymorphic DNA; RFLP, Restriction fragment length polymorphism; SCAR, Intergeneric relationship within
Sequence-characterized amplified region; SNP, Single nucleotide polymorphism; Asteraceae tribe Anthemideae
SSR, Simple sequence repeat; TCM, Traditional Chinese medicine; TPS, Terpene
synthase; TS, Terpenoid synthase; UGT, UDP-glucuronosyl and -glucosyltransferase;
UPGMA, Unweighted pair group method with arithmetic means; WGD, Whole The family Asteraceae, also called Compositae, consists of
genome duplication. over 32,000 known species of flowering plants in over 1,900 genera

Frontiers in Plant Science 02 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

FIGURE 1
Phylogenetic relationship of Artemisiinae species, including Chrysanthemum and its wild relatives. The distribution of therapeutic efficacy of
ethnomedicinal species on the phylogenetic tree of Chinese taxa is shown. In the inner circle, Artemisia species are in green area, and non-
Artemisia species are in red area. From the inside to the outside of the outer circle are poisoning, circulatory diseases, gastrointestinal diseases,
nervous system diseases, eye diseases, other/general diseases, hepatobiliary diseases, musculoskeletal diseases, oral diseases, ear/nose/throat
diseases, pediatric diseases, reproductive system diseases, respiratory diseases, skin diseases, and urinary diseases, indicated by stars of different
colors, square or circle.

within the order Asterales (Mandel et al., 2019). There are more intergeneric crossing between cultivated C. × grandiflorum and
than 200 genera and more than 2,000 Asteraceae species in China, Ajania pacifica was conducted, as well as self-crosses and
which are distributed all over the country. Anthemideae is a tribe backcrosses of hybrid F1. The genetic performance and cytological
of the subfamily Asteroideae, plants of which are distributed behavior of traits in different hybrid combinations and generations
worldwide with concentrations in central Asia, Mediterranean were probed to elucidate the relationship between parental
Basin, and southern Africa. Most species of plant known as genomes and their roles in different generations of hybrids. The
chamomile belong to genera of this tribe. There are about 1,800 East Asian Anthemideae taxa are mainly diploid and tetraploid,
species classified in 111 Anthemideae genera (Oberprieler et al., and many monotypic genera native to East and Central Asia, e.g.,
2007). This tribe is divided into 14 subtribes, and the genus Opisthopappus Shih and Crossostephium Less., etc., are diploid. It
Chrysanthemum belongs to the subtribe Artemisiinae (Figure 1). is speculated that the ancestor of the tribe should be diploidy.
Artemisiinae has 19 genera, 15 of which are distributed in East Within the genus Ajania Polj., the chromosome ploidy differs
Asia, and there are eight endemic genera. This subtribe is the most between species and intraspecific populations.
important Anthemideae subtribe in East Asia, including more East Asian Anthemideae can be divided into three groups
than 80% of the Anthemideae genera. Based on the extensive according to the outer wall decoration of pollen (Zhao, 2007). The
collection of East Asian Anthemideae resources, the systematic first group has obvious thorn-like decoration on the surface, the
evolution was studied by means of pollen morphology, molecular thorns are larger, longer than 1.5 μm, the base is expanded, and
systematics, molecular markers and so on (Zhao, 2007). The there are hole-like perforations from middle of the thorn to the

Frontiers in Plant Science 03 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

thorn base. This group includes genera Glebionis and obvious differentiation between genera and species. In the cluster
Argyranthemum of the subtribe Chrysanthemidae, Hippolytia, diagram, Elachanthemum and Neopallasia are at the base of the
Opisthopappus, Pyrethrum and Tanacetum of subtribe Tanacetinae, whole subtribe, while Brachanthemum is at the base of
Leucanthemum and Nipponanthemum of subtribe Chrysanthemum-Ajania branch. The population of Ajania is
Leucanthemidae, Matricaria of subtribe Matricariinae, Achillea of always at the base of Chrysanthemum population, and the latter
Achilleinae, Chrysanthemum, Ajania and Brachanthemum of may have evolved from different species or populations of Ajania
subtribe Artemisiinae. The second group has degenerate small in parallel. The close relatedness between Opisthopappus and
spines or inconspicuous spines, basically less than 0.5 μm. This Chrysanthemum-Ajania was also confirmed by AFLP results
group includes genera Artemisia, Seriphidium, Crossostephium, (Zhao et al., 2010). Based on the results of cytology, pollen
Elachanthemum, Neopallasia, Ajaniopsis, Filifolium and morphology, molecular sequence analysis and AFLP, it is believed
Kaschgaria. The third group has shorter surface thorn-like that the East Asian Anthemideae could originate in the Eurasian
ornamentation, which is, however, very obvious, and its length is continent, and the ancestral species of Eurasian continent spread
between the first and second groups, 1.0–1.5 μm; it is conical, the eastward and gradually evolved throughout Central Asia and East
base does not descend, and the apex is pointed. This group Asia. Some genera distributed in Central Asia, e.g., Kaschgaria,
includes genus Stilpnotepis, A. salicifolia and A. variifolia of Ajania. may be the relatively primitive groups of Artemisiinae.
Based on the outer wall decoration of pollen, Phaeostigma is The geography and ecology play a key role in driving species
proposed to be a separate genus from Ajania. diversification of Anthemideae and in shaping genotype/
To better understand the intergeneric relationships and phenotype of species. The multi-phased orogenesis of Qinghai-
taxonomic position of small Asian genera of Anthemideae, the Tibetan Plateau (QTP) and global climate changes over late-
sequences of nuclear ribosomal internal transcribed spacer (nr Miocene has profoundly influenced the environments and
ITS) and chloroplast (cp) trnL-F intergenic spacer (IGS) were evolution of Anthemideae plants in QTP and adjacent regions
used to infer the phylogenetic relationship of 48 Anthemideae (Shen et al., 2021). The DNA sequences of seven low-copy nuclear
taxa (Zhao et al., 2010). The trnL-F sequence was of poor genes and nrITS were combined to reconstruct a time-calibrated
resolving power, but some deletions and insertions were useful in phylogeny of subtribe Artemisiinae (Figure 2). In the
interspecific and intergeneric circumscriptions. Both ITS and monophyletic Chrysanthemum group, Chrysanthemum and
ITS/IGS phylogenies suggest two major clades. The clade A is Ajania were well resolved, agreeing with capitulum morphology
subtribe Artemisiinae, consisting of two groups; one group and suggesting the low resolving power of cp and ITS markers
includes Chrysanthemum, Arctanthemum, Ajania, Opisthopappus (Zhao et al., 2010; Hong et al., 2015; Tyagi et al., 2020; Masuda
and Elachanthemum, while the other includes Artemisia, et al., 2022). Within Chrysanthemum, the C. indicum complex and
Crossostephium, Neopallasia and Sphaeromeria. Within the C. zawadskii complex diverged, which temporally coincide with
former group, Chrysanthemum, Arctanthemum and Ajania were the late Cenozoic uplift of Northern QTP and associated climatic
closely related to each other, and the delimitation of genera heterogeneity between eastern and central Asia. The origin of
Chrysanthemum and Ajania was ambiguous. The successful Chrysanthemum group might be in Central Asia, then
hybridization between C. × grandiflorum and A. pacifica and the Chrysanthemum migrated eastward, in contrast to the in situ
normal meiotic chromosome behavior of the hybrid also indicate diversification of Ajania (Figure 3). The C. indicum complex and
the close relatedness between two genera (Zhao, 2007). C. zawadskii complex have distinct distributions in East Asia, i.e.,
Phaeostigma was not in Chrysanthemum group, which was the former in more southern and the latter in more northern
confirmed by the 6-bp insertion in trnL-F. Brachanthemum was regions. The distribution patterns are related with the niche
excluded, and Elachanthemum was included in this group. differentiation of different lineages and environmental
Opisthopappus of subtribe Tanacetinae was proposed to be in the heterogenization.
subtribe Artemisiinae, Chrysanthemum group (Hong et al., 2015; Through distant hybridization and aggregating germplasms,
Figure 2). The molecular phylogeny facilitates the inference of the wild Anthemideae genetic resources with excellent
evolution of pollen and capitulum characters. Disentangling old resistance, e.g., cold resistance, drought resistance, waterlogging
hybridization events may be compromised by the set of resistance, etc., can be used in modern chrysanthemum breeding
evolutionary processes accumulated subsequently (Criado Ruiz (Sun et al., 2010a; Hong, 2014) to realize the innovation of
et al., 2021), particularly in areas with past climatic instability. The germplasm resources. While breeding new varieties, some basic
high-throughput sequencing data will facilitate understanding data for exploring the genetic relationship between
the role and impact of reticulate evolution in the Chrysanthemum and related genera can be obtained (Wu, 2014),
phylogenetic puzzle. and the phylogenetic positions of some ambiguous taxa can also
In an amplified fragment length polymorphism (AFLP) be clarified. With conventional hybridization methods or
analysis of Artemisiinae, 1705 polymorphic bands were obtained embryo rescue conditions, distant hybrids were obtained
(Zhao, 2007). According to the similarity coefficient, both within between Chrysanthemum and genera such as Ajania,
the whole subtribe, its related groups, and within Chrysanthemum Crossostephium (subtribe Artemisiinae), Opisthopappus,
and Ajania, the high genetic diversity was revealed, indicating Tanacetum, Pyrethrum (subtribe Tanacetinae), Argyranthemum,

Frontiers in Plant Science 04 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

FIGURE 2
Phylogenetic relationship of Chrysanthemum taxa. The number after the taxon name represents the ploidy level. This schematic is compiled
mainly based on Shen et al., 2021, as the phylogenetic topology inferred from low-copy nuclear genes and nrITS could be more convincing than
those inferred from cp sequences (Liu et al., 2012; Lu et al., 2022a). It should be noted that it is challenging to distinguish taxa from each other
within C. indicum complex and C. zawadskii complex, respectively.

Glebionis (subtribe Chrysanthemidae), Leucanthemella (subtribe crossed with Ajania myriantha (2x = 18), Crossostephium Chinense
Leucanthemidae; Zhao et al., 2008; Sun et al., 2010a; Shao, 2018). (2x = 18), Opisthopappus taihangensis (2x = 18) and Tanacetum
Chrysanthemum, Ajania and Opisthopappus are closely related vulgare (2x = 18), respectively, to explore the genetic relationship
(Figures 1, 2), and their hybridization is easier to be successful. between Chrysanthemum and its Anthemideae relatives (Tang,
Brachanthemum and Hippolytia are close to the clade of 2009). The F1 progeny of C. indicum×Crossostephium Chinense
Chrysanthemum group, and their hybridization with grew healthily and bloomed luxuriantly; the F1 progeny of
Chrysanthemum (e.g., CM, C. chanetii and C. indicum×O. taihangensis grew well, but there was no flower bud
C. naktongense×C. argyrophyllum) can be successful (Hong, differentiation. The F1 progeny of C. indicum×T. vulgare did not
2014; Xie, 2016). The hybrid without emasculation between grow normally in the field. It is speculated that the genetic
Hippolytia and Nipponanthemum was obtained (Hong et al., relationship between Chrysanthemum and related genera from
2015); on the ITS tree, both genera are closer to Brachanthemum close to far is Ajania (Wu, 2007), Crossostephium, Opisthopappus
than to Chrysanthemum. Chrysanthemum sensu lato includes and Tanacetum. In genomic two-color fluorescence in situ
around 38 genera, of which 27 are distributed in China (Shao, hybridization (FISH), the chromosomes of Crossostephium and
2018). There are abundant wild resources and excellent stress Tanacetum were not observed in F1 progenies of CM cultivar
resistance in Chrysanthemum sensu lato, which is of great “Tianzhuiyulu (falling jade dew)” × (C. indicum×Crossosstephium
significance to improve cultivated chrysanthemum breeding. Chinense) and “Tianzhuiyulu” × (C. nakingense×T. vulgare),
The chromosomal ploidy changes are abundant in indicating that the chromosomes of two related genera may have
Chrysanthemum (Figure 2), from diploid to decaploid, and there been excluded. However, in F1 progeny of
are also ploidy differences in different populations within the “Tianzhuiyulu” × (C. crassum×Crossostephium Chinense) the
species. C. crassum (2n = 10x =90), C. indicum (4x = 36) and genomic chromosomes of three parents were observed, which
C. nakingense (2x = 18), as the female parent, respectively, were realized the innovation of CM germplasm and enable full use of

Frontiers in Plant Science 05 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

A B

FIGURE 3
Geographical distribution of representative taxa of Asteraceae subtribe Artemisiinae. (A) Genus Ajania is produced in the vast areas of China except
the southeast, Mongolia, North Korea, northern Afghanistan and Russia; Opisthopappus is endemic to Taihang Mountains, China; Elachanthemum
is distributed in northern China, northwestern China and Mongolia; Hippolytia is produced in central Asia and the Himalayas. (B) C. indicum is
produced in Northeast China, North China, Central China, South China, Southwest China, India, Japan, North Korea and Russia; C. vestitum is
produced in western Henan, western Hubei and western Anhui, China; C. zawadskii is produced in Heilongjiang, Jilin, Liaoning, Hebei, Shanxi,
Inner Mongolia, Shaanxi, Gansu and Anhui of China, Mongolia, Russia and Europe; C. lavandulifolium is produced in Jilin, Liaoning, Hebei,
Shandong, Shanxi, Shaanxi, Gansu, Qinghai, Xinjiang (eastern), Jiangxi, Jiangsu, Zhejiang, Sichuan, Hubei and Yunnan of China, Korea and Japan.
The geographic distribution data are retrieved from Global Biodiversity Information Facility (https://www.gbif.org/).

the excellent germplasm of Chrysanthemum related genera to Ajania may be relatively recent, presumably temporally similar to
improve CM varieties. the divergence of C. zawadskii. Hybridization and gene flow
between taxa occurred frequently during the evolutionary history
of Chrysanthemum and Ajania, causing considerable incongruency
Intrageneric relationship of of cp and ITS trees (Li et al., 2014; Lu et al., 2022a). The gene flow
Chrysanthemum and origin of CM had occurred between C. indicum, C. nankingense and
C. lavandulifolium (Yang et al., 2006; Liu et al., 2012), and between
The genus Chrysanthemum, associated with polyploidy and Cheiracanthium mongolicum and C. chanetii (Luo et al., 2017),
hybridization, undergoes rapid speciation and has about 40 which could be involved in the origin of polyploids of C. indicum
species, most of which are distributed in East Asia (Liu et al., and C. zawadskii complex, respectively.
2012). Many taxa are narrowly distributed and grow in specific The ITS and cp rpl16 sequences were combined to resolve the
habitats. The Chrysanthemum taxa vary greatly in morphology interspecific relationship of Chrysanthemum (Wu, 2007).
and ploidy (Figure 2), and the interspecific relationships are not C. indicum and C. nankingense (once regarded as a variety of the
fully understood. The genome size values of 15 species fall into former), C. dichrum and C. lavandulifolium, C. japonicum and
three groups (Luo et al., 2017), which positively correlate with C. yoshinaganthum, C. crassum and C. rhombifolium are closely
three ploidy levels (2×, 4×, 6×), and there was a genome related. The meiotic behavior of several Chrysanthemum taxa and
downsizing after polyploidization in Chrysanthemum. Two major their hybrids suggested that three diploid species are closely
phylogenetic clades were inferred based on ITS and trnL-F related to each other (Cui, 2004), among which C. dichrum and
sequences, i.e., C. indicum complex and C. zawadskii + Ajania C. lavandulifolium are the closest (Figure 2), followed by
group. The genome size and 1Cx values (DNA content of one C. lavandulifolium and C. nankingense, and C. nankingense and
non-replicated monoploid genome with chromosome number x) C. dichrum are the furthest. C. japonense and C. vestitum,
in the former group were significantly lower than those in the C. lavandulifolium and C. boreale, C. zawadskii and C. weyrichii
latter group, even though both have same ploidy level, and they are also closely related. However, they are still independent species
were also correlated with latitude and morphology. The 1Cx values and should not be combined into the single species. In neighbor-
of diploid Chrysanthemum species were similar to those of joining analysis of DNA sequences, C. japonicum, C. arcticum,
C. indicum (Mt. Tianzhu) and C. indicum (Fujian) indicate that C. japonense, C. vestitum and their respective subspecies/variety
some populations of tetraploid C. indicum originated by got together in pairs, showing a close kinship. The small genetic
autoploidy (Li et al., 2013). The genome size of C. chanetii is distance between C. zawadskii and C. zawadskii var. latilobum
significantly larger than that of other tetraploids, while C. vestitum shows their close genetic relationship. However, in the
is smaller than other hexaploids, but the relationship between morphological cluster analysis, these species and their variety/
genome size and evolutionary time cannot be determined due to subspecies were not well aggregated, indicating the inconsistency
limited sampling. The divergence between Chrysanthemum and between the phenotypic evolution and genetic evolution of

Frontiers in Plant Science 06 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

Chrysanthemum, with large differentiation of morphological traits According to pollen morphology and DNA data, A. purpurea
and small genetic differentiation of DNA sequences. is a member of Phaeostigma. The species differentiation in
In the clustering analysis based on 24 morphological Chrysanthemum could be correlated with geographic and
characters and one biological character, Chinese C. indicum, environmental factors. The C. zawadskii complex is distributed
C. lavandulifolium, C. nankingense and C. dichrum clustered in northern China and C. indicum complex in southern China
together (Wu, 2007), and most Chrysanthemum taxa originating (Figure 3). Many polyploid species, e.g., C. argyrophyllum,
in Japan clustered together, indicating that there is an obvious could originate from divergent ancestors via
geographical isolation between Chinese and Japanese allopolyploidization. The geographic/ecological conditions,
Chrysanthemum plants. The Japanese wild species evolutionarily hybridization and polyploidy play important roles in the
appeared later than Chinese ones. In PCR-RFLP, three CM divergence and speciation of Chrysanthemum. The comparative
varieties “Yinxing,” “Jinlingwanxia” and “Shenma” had the same transcriptome analysis of CM and diploid C. boreale revealed
enzyme digestion pattern as that of wild Chrysanthemum in whole-genome duplication (WGD) and gene selection patterns
China, indicating their close genetic relationship. These CM in these taxa (Won et al., 2017); the transcriptomes of
cultivars have close genetic relationship with C. indicum, C. rhombifolium and C. dichrum were also preliminarily
C. vestitum and C. nankingense, but they have the closest characterized (Liu et al., 2021a; Zhang et al., 2021a), but the
relationship with C. chanetii, and are far from C. ornatum and phylotranscriptomic analysis covering more Chrysanthemum
C. japonense, although wild species native to Japan have also species has not been reported.
played a positive role in enriching modern CM varieties. In order to clarify the origin of CM, the investigation and
According to the morphological characteristics and molecular collection of Chinese medicinal Chrysanthemum varieties with
phylogeny, the wild Chrysanthemum species were classified into less germplasm erosion, ancient ornamental big CM (Daju in
four groups, the indicum group, makinoi group, zawadskii group, Chinese) varieties, Chrysanthemum sensu stricto and related
and Ajania group (Nakano et al., 2021). genera were highlighted (Zhou, 2009); these plants were used as
The multiple differentiation and hybridization/ the materials of various experiments for the origin of CM by
polyploidization cycles characterize the reticulate evolution of using different technical methods. Natural hybridization and
C. indicum complex (Yang et al., 2006) and C. zawadskii complex cultivation play an important role in the origin of CM. In the
(Lu et al., 2022a), causing the difficulties in systematic distant hybridization of four years, 47 combinations were carried
classification. The DNA sequences of single-copy nuclear CDS out, and 3,225 seeds, 675 hybrid seedlings and 30 flowering
(chrysanthemyl diphosphate synthase) gene and seven cpDNA variant seedlings were obtained. C. vestitum and C. indicum,
loci were used to infer the phylogenetic relationship of 32 C. vestitum and C. zawadskii, C. vestitum and C. lavandulifolium
Chrysanthemum taxa and 11 species of the allied genera (Liu et al., were easy to cross successfully, and seeds were obtained. In the
2012). It was found that the affinity between Chrysanthemum and same distribution area of C. vestitum and C. indicum, the
Ajania is very close, and the resolving power of CDS and cpDNA flowering period of the former is slightly earlier than that of
markers were not enough. When eight nuclear sequences were latter, and they meet in flowering period (Wang, 2020). The F1
used (Shen et al., 2021), Chrysanthemum and Ajania were well generation had great variation, and the inflorescence, flower
separated with only a few exceptions (Figure 2), which agrees with color and leaf type were close to modern CM. The F2 progeny
traditional taxonomy mainly based on capitulum morphology. In showed greater variation after natural pollination, which was
Chrysanthemum, two Chinese endemic species with white ray very similar to ornamental small CM (Xiaoju). Hybridization is
flowers, C. rhombifolium and C. vestitum, are sister to C. indicum the main approach of origin of CM. C. vestitum, C. indicum,
complex with yellow/white ray flowers (Figure 2). The C. indicum C. zawadskii, C. lavandulifolium and C. dichrum are closely
complex has a and b branches; species of ‘b’ have island related and play a role in the origin of CM to varying degrees.
distributions, and species of ‘a’ are mostly distributed in mainland There are more frequent natural crosses between C. indicum and
China. The C. zawadskii complex (Figure 2) has white to purple C. vestitum, possibly due to the weak reproductive isolation
ray flowers with continuous color variation, and Opisthopappus between tetraploid and hexaploid (Wang, 2020). The
falls into this clade. A Hyb-Seq based phylogenetic study found phylogenetic analysis of nuclear LFY sequences suggested that
that C. zawadskii, C. naktongense, C. chanetii and C. maximowiczii different cultivars had different ancestors (Ma et al., 2016).
intermingled in a clade with highly complicated phylogenetic C. indicum, C. zawadskii and C. nankingense might be the direct
relationships (Lu et al., 2022a), and the leaf morphology and ancestors of most CM cultivars examined. C vestitum and
phylogenetic analyses were difficult to distinguish them from each C. lavandulifolium might be the ancestor of some cultivars.
other, arguing that they have not yet differentiated into Another line of support is from simple sequence repeat (SSR)
independent species and supporting the taxonomic treatments of markers mined from expressed sequence tags (ESTs; Fan et al.,
C. zawadskii complex. 2019) of CM. Transferability of EST-SSR primers among
When nuclear CDS was used to infer the phylogenetic Chrysanthemum species was identified in C. indicum (96–100%),
relationship, Phaeostigma is more closely related to C. nankingense (100%), C. chanetii (96%), C. zawadskii (96%),
Chrysanthemum + Ajania than other genera (Liu et al., 2012). and C. indicum var. aromaticum (92%).

Frontiers in Plant Science 07 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

The cluster analysis of ISSR (inter-simple sequence Intraspecific relationship of CM and

repeat)-PCR showed that the genetic relationship between origin of cultivars
C. vestitum and Daju/medicinal CM is the closest (Zhou, 2009),
followed by C. vestitum of Yichang, Hubei Province (C. vestitum Several genus names have been used for CM, such as
var. vestitum, Wang, 2020), C. zawadskii and C. nankingense. Matricaria, Anthemis, Pyrethrum, Chrysanthemum and
The main evolution mode of CM could be: Wild Dendranthema, reflecting its ambiguous phylogenetic position.
Chrysanthemum → medicinal CM (economic original The species names of grandiflora vs. morifolium were contentious.
chrysanthemum) → ornamental CM. Based on the research The International Code of Botanical Nomenclature changed the
results on the origin of CM for half a century, the macro- defining species of genus to C. indicum (Figure 1), and restored
morphology is combined with cytology, palynology, isozyme, the florist’s chrysanthemum to the genus Chrysanthemum (Poe
DNA molecular markers, which are further combined with and Swofford, 1999). CM is a perennial herb with 50–140 cm
artificial hybridization, field resource investigation, height, has erect and slightly purplish red stems (FRPS, 1983); the
introduction and domestication, and historical analysis, and it base of the plant is woody, with much branching covered by
is concluded that the cultivated CM (original CM beginning in densely white pubescent. The leaves of CM have a shape of ovate
the era of poet Tao Yuanming, Jin Dynasty) originated in to lanceolate, with a color of dark green above and greenish
China; it is a hybrid cultigen complex, and is mainly produced beneath covered by densely white short hair on both sides. The
by natural interspecific hybridization of some wild species in flower heads are on the branches separately or aggregated with
the middle reaches of Yangtze River (Hubei, Anhui and each other, with 2.5–15 cm in diameter; the involucral bracts of
Henan). After long-term artificial repeated selection of some CM have multiple layers and ligulate flowers have diverse colors.
special variation types, it is carefully cultivated. The main The tubular flowers are yellow, blossoming in September to
parents are C. vestitum and C. indicum. Later, C. zawadskii, October in the northern hemisphere.
C. lavandulifolium, C. indicum var. aromaticum, C. nankingense CM has been cultivated for more than 1,600 years and is one
and C. dichrum participate in the evolution of CM through of the top ten traditional famous flowers in China. It has rich
pollen to varying degrees; the germplasm introgression variation, strong adaptability, wide distribution and complex
enhanced the diversification of CM genes, so today’s ever- genetic background, which has brought great difficulties to the
changing CM is formed. The recurrent hybridizations between investigation and protection of germplasm resources,
several wild progenitor species contributed to the evolutionary identification and classification, genetic diversity analysis, and the
novelty of CM (Song et al., 2018), and cultivation/selection also like. The genetic diversity of 53 CM germplasm samples with
play a significant role in the formation of CM. different flower diameter, petal type and flower color type was
Many wild Chrysanthemum species may be involved in the studied using conserved DNA-derived polymorphism (CDDP)
complex formation process of CM. The ISSR-based UPGMA markers (Li, 2014) to provide a basis for the classification and
(unweighted pair group method with arithmetic means) identification of chrysanthemum varieties. In the cluster analysis
clustering suggested that the Chrysanthemum species evolved of CDDP amplification results, 53 chrysanthemum materials were
from low ploidy to high ploidy (Liu, 2010); it can generally divided into six groups. Group 1 includes 30 Daju varieties and
be inferred that the flat petal is the basic petal shape of two Xiaoju varieties, group 2 contains 16 Xiaoju varieties, and
ornamental CM. C. nankingense and CM had the closest group 3 includes two Xiaoju varieties. C. nakingense, C. indicum
genetic relationship, C. chanetii, C. japonicum, C. japonicum and C. vestitum were in groups 4, 5 and 6, respectively. The CM
var. wakasaense were also closely related to CM, and varieties were basically clustered according to flower diameter,
C. indicum var. aromaticum was farthest related to other which has no direct correlation with petal type and flower color.
samples. The random amplified polymorphic DNA (RAPD) The phylogenetic analysis based on large sets of single nucleotide
analysis showed that the genetic diversity of wild species was polymorphisms (SNPs) revealed that the Xiaoju types and potted/
more abundant than that of CM. The cultivated Xiaoju, rather ground chrysanthemum, instead of Daju, are more closely related
than Daju, was genetically closer to the wild Chrysanthemum to the wild progenitor species (Chong et al., 2016).
species, and the petal shape and flower diameter can be used The CM varieties are classified in terms of the parameters such
in the classification of CM. AFLP markers were used to detect as flowering stage, flower diameter, flower color, appearance of
the relationships among 12 wild accessions and 62 petals, cultivation form, leaf shape, among others. Both medicinal
groundcover cultivars (Chen et al., 2013). The genetic CM, i.e., Flos Chrysanthemi in TCM, and ornamental CM can
variation is abundant in chrysanthemums. The 74 accessions be classified according to these external characteristics. Before
were classified by UPGMA; the genetic relationship was the Song Dynasty, the artificially cultivated CM was for food/medicine
most relevant factor in AFLP-marker clustering, and and viewing, and the wild relatives were also used as medicine.
petal type was also informative. AFLP technology could With the increased use of medical varieties, the cultivation
be very efficient for discriminating Chrysanthemum flourished in Qing Dynasty, when CM’s genuine production areas
species and related genera and reconstructing their were formed (Figure 4). The changes of CM producing areas in
genetic relatedness. ancient times are shown in Supplementary Figure S1. With the

Frontiers in Plant Science 08 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

passage of time, changes of cultivation sites and processing and sesquiterpenoids (Wang et al., 2008b). Gongju is introduced
methods, eight famous CM categories are gradually formed for over 100 years mainly as tea beverage. Gongju grows in an
(Figure 4); Chinese Pharmacopoeia contains five Daodi medicinal environment of high altitude and high humidity during harvest,
materials (geoherbs) of CM: Hangju, Boju, Gongju, Chuju, Huaiju and the main processing method was drying by charcoal fire. In a
(Chinese Pharmacopoeia Commission, 2020). There are also local ISSR analysis (Lyu et al., 2008), Gongju was closer to Hangju,
varieties such as Qiju, Jinsi Huangju, Wuyuan Huangju and so on agreeing with morphological and metabolomic investigations
(Liu, 2020). Gongju and Hangju could be closely related (Wang (Wang et al., 1999, 2001; Nie et al., 2019).
et al., 1999, 2001), Huaiju, Jiju, Boju and Qiju are more closely Huaiju is considered as the ancestor of medicinal CM because
related, while Chuju is an independent variety. Interestingly, of its ancient records in medicinal uses. In a RAPD analysis (Dai
among 26 antifungal terpenoids of 13 CM cultivars, et al., 2017), Huaiju was not clustered with Hangju, Gongju and
longifolenaldehyde and β-selinene were identified only in Chuju C. indicum (Chrysanthemi Indici Flos (CIF) in TCM; Song et al.,
(Xue et al., 2019). At present, Hang Baiju (Hangzhou white CM) 2020), indicating the rich genetic diversity in chrysanthemum
and Gongju in the south of Yangtze River are mainly used for tea, germplasm resources. Huaiju samples were clustered into a large
while Huaiju, Boju, Chuju, Qiju and Jiju in the north of Yangtze group, indicating its high genetic purity. In a ISSR analysis (Lyu
River are mainly used for medicinal purposes. et al., 2008), C. indicum, C. nankingense and a hybrid between
Hangju, including Hang Huangju (Hangzhou yellow CM) and C. indicum and Gongju were grouped together. The 19 medicinal
Hang Baiju, has been cultivated for about 400 years. Huangju was CM accessions were divided into two groups according to the
mainly used as medicine while Baiju was consumed as tea. In the origin. Most accessions originating in the north of Yangtze River
1950s and 1960s in Futianhe Town, Macheng, Hubei Province, had relatively close genetic relationships, while most accessions
Hangju (Big Baiju) was introduced from Tongxiang, Zhejiang cultivated in the south also had relatively close relationships. The
Province (Chang et al., 2019). The large-scale planting began in resolving power of ISSR is generally higher than that of RAPD (Xu
the 1980s. The local CM is produced in Futianhe Town as the core et al., 2006; Luo et al., 2013), facilitating the clarification of inter-
production area, so they are also called “Macheng Fubaiju” or cultivar relationship of CM. Medicinal CM germplasm resources
“Fubaiju.” The cultivars of Fubaiju mainly include Baiju and Jinju are indeed distinct at the molecular level; the differences among
(golden CM), which can also be divided into two major varieties: medicinal CM cultivation types are related to environmental
“local early-flowering” and “local late-flowering” (Xiong, 2014); factors (Huang et al., 2020), but to a greater extent are determined
they were mainly used for tea. Surprisingly, the ISSR cluster by their genetic factors. The quantitative analysis of morphological
analysis showed that the genetic distance between Fubaiju and variation (Shao et al., 2011) and isozyme analysis (Ding et al.,
Hang Baiju was relatively far (Xiong, 2014), and it was closer to 2008) also help much in elucidating the inter-cultivar relationship,
heterophyllous chrysanthemum and Chuju in Chuzhou, variety identification and conservation at different phenotypic
Anhui Province. levels.
Boju has been cultivated for at least 240 years, but its
processing method of sulfur-fumigation arouses controversy.
Twenty-one medicinal CM cultivars were heteroploidy (Wang Phytometabolites and
et al., 2012), mostly hexaploid, and some were tetraploid chemodiversity of Chrysanthemum
(Huangyaoju) and pentaploid (Dahuangju). The karyotype of Da
Boju, Xiao Boju, Chuju, “Xenogeneic Dabaiju” and “Little Baiju” Plant pharmacophylogeny is an interdisciplinary subject that
is 2A, and the other 16 types are 2B. The chromosomes of 21 CM integrates plant systematics, phytochemistry, TCM, pharmacology
cultivars showed polymorphism, and the chromosome length, and other multidisciplinary knowledge systems (Hao and Xiao,
centromere position and satellite varied among cultivars. In a ISSR 2017, 2020), and plays an important role in guiding the conservation,
analysis (Lyu et al., 2008), both Da Boju and Xiao Boju were closer development and utilization of medicinal plant resources. The core
to different cultivars of Hangju. A cultivar-specific sequence- idea of plant pharmacophylogeny is that species in adjacent
characterized amplified region (SCAR) marker was developed to phylogenetic groups have relatively close genetic characteristics; the
detect Da Boju (Cai et al., 2022), which is morphologically similar similar gene sequences lead to relatively close biosynthetic pathways
to ornamental Xiaoju, and could be artificially evolved of various phytometabolites in closely related taxa, manifesting a
from Xiaoju. high degree of similarity in the chemical composition, i.e., (1)
Chuju, Hangju and Boju have deep notch correctitude-leaves specific secondary metabolites are more likely to be distributed in
(Li et al., 2010); unlike other cultivars, Chuju and Boju have no multiple species that are genetically close and (2) natural products in
auricle. In a UPGMA analysis based on the DNA fingerprint closely related taxa have higher similarity in molecular skeleton
patterns of SSR markers (Feng et al., 2016), Chuju was closer to composition and the relationship of derivation (Liu et al., 2021b).
Boju, followed by Gongju and Huaiju. In a ISSR analysis (Lyu The similarity of the above two tiers of chemicals is manifested in the
et al., 2008), Chuju was closer to Jiju than to other medicinal overall similarity of biological activity or therapeutic effect in clinical
CM. The uniqueness of Chuju was also validated by the quality application. In practice, the concept of plant pharmacophylogeny has
evaluation and identification studies based on monoterpenoids effectively guided the in-depth development and utilization of

Frontiers in Plant Science 09 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

FIGURE 4
Geographical distribution, chemical and morphological characteristics of representative CM cultivars. The medicinal quality (Q) markers of each
cultivar are shown: Boju, Huaiju, Gongju and Hangju: chlorogenic acid, 3,5-DCQA, and kaempferol-3-O-rutinoside (Lu et al., 2022b); Chuju:
chlorogenic acid, Lut-7-O-G, quercetin, 3,5-DCQA (Yang et al., 2018b); Qiju: chlorogenic acid, 3,5-DCQA, and Lut-7-O-G (Peng et al., 2019); Jiju:
chlorogenic acid, luteolin and 3,5-DCQA (Kang et al., 2022). The Q-marker of Chuanju of Zhongjiang, Sichuan Province is not reported.
Morphology of medicinal CM: Boju: Inverted conical or cylindrical shape, sometimes slightly flattened and fan-shaped, 1.5–3cm in diameter,
discrete (Chinese Pharmacopoeia Commission, 2020). Involucral bracts dish-shaped; involucral bracts 3–4 layers, ovate or elliptic, herbaceous,
yellow-green or brown-green, pubescent outside, margin membranous. Chuju: Irregular spherical or oblate spherical, diameter 1.5–2.5cm.
Ligulate flowers are white, irregularly twisted, involute, with shriveled edges, sometimes with light brown glandular dots; tubular flowers are mostly
hidden. Gongju: Oblate spherical or irregular spherical, 1.5–2.5cm in diameter. Ligulate flowers white or off-white, obliquely ascending, upper part
reflexed, margin slightly involute and shriveled, usually without glandular dots; tubular flowers few, exposed. Hangju: It is dish-shaped or oblate
spherical, with a diameter of 2.5–4cm, and is often connected in several pieces. Ligulate flowers white or yellow, spreading or slightly folded,
adhering to each other, usually without glandular dots; tubular flowers numerous, exposed. Huaiju: Irregular spherical or oblate spherical,
diameter 1.5–2.5cm. Ligulate flowers are the most, white or yellow, irregularly twisted, involute, with shriveled edges, and sometimes glandular
dots can be seen; most of the tubular flowers are hidden.

medicinal plant resources (Hao and Xiao, 2017, 2020), especially in displayed lower emission rates. The differences were principally
the development of new medicinal plant species, so as to avoid the determined by seven monoterpenes (camphor, endo-borneol,
blindness in traditional research methods and guarantee the targeted bornyl acetate, sabinene 1,8-cineole, filifolone, β-myrcene;
research. Fully understanding the phytometabolites and Figure 5) and five sesquiterpenes (germacrene D, β-ylangene,
chemodiversity of Chrysanthemum and related taxonomic groups is (E)-β-farnesene, β-copaene, β-caryophyllene). At least 183
an indispensable basis for pharmacophylogenetic research. monoterpenoids have been reported in Chrysanthemum (Jiang
et al., 2021b), e.g., the abundant borneol, camphor, β-pinene,
α-thujone, and verbenone, etc. (Xie et al., 2012). CDS catalyzes the
Volatile oils condensation of two molecules of dimethylallyl diphosphate to
chrysanthemyl diphosphate, an irregular monoterpene (Liu et al.,
Volatile oils were one of the most important bioactive 2012). Irregular monoterpenes are mainly found in plants of the
components in Chrysanthemum species, which are mainly tribe Anthemideae. The monoterpene glycosides were identified
composed of hydrocarbons, terpenoids, alcohols, esters, ketones, from flowers of edible C. “Kamiohno” (Kurimoto et al., 2021).
aldehydes, among others (Ryu et al., 2019; Wang et al., 2021a). The In C. indicum, 17 monoterpenes and 27 sesquiterpenes were
herbivory-induced emission of volatile terpenes in identified (Zhou et al., 2021). The recombinant CiTPS1 and 2
Chrysanthemum is an indirect defense against pest by attracting produced α-pinene, CiTPS3 was responsible for the production of
natural enemies (Xu et al., 2021). The candidate terpene synthase sesquiterpenoids β-farnesene, petasitene, and α-bisabolene, while
(TPS) genes were identified by comparing the transcriptomes of CiTPS4 contributed to the production of four monoterpenoids
healthy and pest-infested CM plants. Totally 46 terpenoids were and three sesquiterpenoids. These functionally redundant genes
identified in flower heads of 44 Chrysanthemum species/varieties were derived from WGD and segmental duplication of
and 43 were identified in the emission (Zhang et al., 2021b). The Chrysanthemum (Song et al., 2018; Nakano et al., 2021). Among
CM flowers had less terpenoids than their wild relatives, and the expanded C. nankingense gene families, the genes involved in

Frontiers in Plant Science 10 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

the biosynthesis of monoterpenoids, sesquiterpenoids, monomer, but the content of ketones in chamomile was
triterpenoids are highly enriched (Song et al., 2018). The terpenoid relatively low; ethyl 2-methylbutanoate and ketone 6-methyl-5-
synthase (TS) and cytochrome p450 (CYP) families are among the hepten-2-one were more abundant in “Xiaokuixiang.” The
most highly enriched functional categories, and 219 TS genes (7 terpenoids 6-methyl-5-hepten-2-one and 1-menthol monomer
squalene synthases, 158 TPSs, and 54 triterpene cyclases) and 708 were salient in Hangju and Huangju, but very few esters were
CYP genes were identified in C. nankingense genome, suggesting detected in both, agreeing with their close phylogenetic relationship.
a significant expansion of TPS genes, explicitly in the TPS-a/−b
subfamilies. Unlike Chrysanthemum, much less TS and CYP
genes were identified in the other three Asteraceae genomes, i.e., Triterpenes and other terpenoids
artichoke, sunflower, and lettuce.
At least 207 sesquiterpenoids have been identified from More than fifty triterpenes were identified from
Chrysanthemum (Chen et al., 2019; Jiang et al., 2021a,b), Chrysanthemum, including tetracyclic and pentacyclic
including 26 germacrane-type, 26 eudesmane-type, 64 triterpenes (Figure 5). The former includes stigmastanes,
guaianolide-type, 4 bisabolane-type, and 15 other-type lanostanes, dammaranes, cycloartanes (more than others),
sesquiterpenoids (Figure 5), with anti-inflammatory, antibacterial, tirucallanes. The latter includes lupanes, taraxeranes (more than
anti-tumor, insecticidal, and anti-viral activities, etc. CM, others), oleananes, and ursanes. Squalene, camphor, DL-α-
C. indicum, C. lavandulifolium, C. zawadskii, and C. ornatum are tocopherol were among the top 10 volatile compounds of 12
rich in sesquiterpenes, especially the former two. γ-irradiated mutant cultivars of CM (Ryu et al., 2019), which are
Angeloylcumambrin B, cumambrin A and handelin were reported useful for classification and identification of chrysanthemum
from more than one species. Germacrane-type sesquiterpenes are mutant cultivars. Though, the triterpene biosynthesis genes of
monocyclic, composing of a 10-member carbon ring, a methyl Chrysanthemum and related taxonomic groups have not been
group at C-4 and C-10, and an isopropyl group at C-7. Eudesmane- reported, and the bioactivity of Chrysanthemum triterpenes is
type sesquiterpenes are bicyclic, consisting of two six-member less studied. One diterpene (C20) was found in roots of 12 CM
carbon rings with methyl groups at C-4 and C-10, and an cultivars (Zhang et al., 2020).
isopropyl group at C-7. Guaianolide-type sesquiterpenes have
ternary rings consisting of a five-member ring, a seven-member
ring, a five-member γ-lactone ring, and methyl groups at C-4 and Flavonoids
C-10. They are more abundant than other types of sesquiterpene.
The cyclization of farnesyl diphosphate to germacrene A could More than 78 CM flavonoids (Figure 6) are identified, and
be the first committed step in sesquiterpene biosynthesis (Jiang flavones are the most, followed by flavonols and flavonones. They
et al., 2021b), and oxidations of (+)-germacrene A determine fall into 12 categories (Jo et al., 2020; Mao, 2020), i.e., apigenin
where additional cyclization occurs to generate guaianolides or (Api) and derivatives (mainly glycosides), diosmetin (Dio) and
eudesmanes. In GC/MS analysis of CM, β-humulene derivatives, acacetin (Acn) and derivatives, kaempferol and
(β-caryophyllene) showed the highest contents, accounting for derivatives, quercetin and derivatives, luteolin (Lut) and
16.3% of the total 58 detected volatiles (Sun et al., 2010b); ledene derivatives, hesperetin and derivatives, eriodictyol and their
oxide-(I) was also abundant, amounting to 9.0% of total volatiles. glycosides, isorhamnetin, baicalin, eupatilin, and anthocyanins.
The constituent and yield of CM essential oils varied a lot Among them, apigenin and derivatives are the most, mainly
depending on the harvest time, origin, processing methods, etc. Api-7-O-6-AG (acetylglucoside), Api-7-O-G and the like, which
(Xue et al., 2007; Yang et al., 2007; Wu et al., 2015). The flowers is followed by luteolin and its derivatives, e.g., Lut-7-O-G
were blue-green in the early bloom stage and contained the most (glucoside), Lut-7-O-MG (malonylglucoside) and so on. These are
abundant volatile oil (Xue et al., 2007). There is correlation also abundant in C. indicum and C. nankingense (Zou et al., 2022).
between floral volatile components and antioxidant activity of In C. pacificum, luteolin conjugates were mostly enriched in
different CM cultivars (Yang et al., 2017a), therefore it is important flowers (Farag et al., 2015), and non-flowering aboveground parts
to quantify contents of volatile oils in the respective cultivar. were rich in quercetin and methoxylated flavone conjugates. Root
Chuju, Gongju and Qiju had 2.0–4.0 ml/kg volatile oil, Huangyaoju sample had the lowest contents of all flavones and dicaffeoylquinic
(a kind of Gongju), Boju and Huaiju had more (4.5 ml/kg), and acids (DCQAs).
Jiju had the highest volatile oil content of >10.0 ml/kg, whereas Among 477 metabolites identified in C. mongolicum and
two kinds of Hangju had the lowest content of <1.0 ml/kg (Xu C. rhombifolium (Duan et al., 2022), 69 are listed in the Chinese
et al., 2005). Pharmacopoeia (2015) and YaTCM database (Li et al., 2018). The
Based on the volatile chemical profile of flower tea, Hangju, flavonoids, e.g., linarin (11.64%), chrysanthin (5.03%), farnesin
Huangju, chamomile (Matricaria chamomilla), and a new (3.03%) and genkwanin (2.75%), are abundant in C. mongolicum.
CM “Xiaokuixiang” were well distinguished from each other The contents of genkwanin, trigonelline, Dio, narcissoside,
(Wang et al., 2021a). The main volatile components in chamomile 3,4-dihydroxyphenylacetic acid, linarin, N′,N′-p-coumarin,
were the ester ethyl 2-methylbutanoate and alcohol 1,8-cineole C-hexosyl-tricetin O-pentoside, chrysoeriol, Acn and

Frontiers in Plant Science 11 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

FIGURE 5
The molecular structure of representative terpenoid components of Chrysanthemum. A, Monoterpene: camphor, endo-borneol, bornyl acetate,
sabinene,1,8-cineole, filifolone, β-myrcene. B, Sesquiterpene: germacrane-type: zawadskinolide F (anti-inflammatory, C. zawadskii),
chrysanthediol A (anti-viral, CM); eudesmane-type: chrysanthemumin A (anti-viral, C. indicum), chrysanthemumin D (anti-viral, C. indicum);
guaianolide-type: chrysanthemulide A (anti-tumor, anti-inflammatory, C. indicum), cumambrin A (treating osteoporosis, C. ornatum, C. indicum,
C. zawadskii); bisabolene-type: jinsidajuol A (CM), tunefulin (C. indicum); others: handelin (anti-aging, C. ornatum, C. indicum),
8,8′-ditigloylchrysanolide D (anti-tumor, C. indicum). C, Triterpene: tetracyclic: stigmastanes: (24R)-saringosterol, (24S)-saringosterol (CM);
lanostane: (24S)-lanost-9(11)-ene-3β,24,25-triol (CM); dammaranes: dammarenediol II (CM), 3-epicabraleadiol (anti-viral, CM); cycloartanes:
cycloartenol (various bioactivities, CM), (24S)-25-methoxycycloartane-3β,24-diol (anti-inflammatory, CM); tirucallanes: 4,5α-epoxyhelianol
(antitubercular, CM), helianol (anti-inflammatory, CM). pentacyclic: lupanes: lupeol (anti-inflammatory, C. indicum, CM), 3-epilupeol
(antitubercular, CM); taraxeranes: heliantriol C (anti-tumor, CM), arnidiol (anti-tumor, CM); oleananes: maniladiol (antitubercular, CM), coflodiol
(anti-tumor, CM); ursanes: α-amyrin, brein (CM).

kaempferol-3-O-gentiobioside were significantly different (colorless co-pigments modifying flower color). Higher
between C. mongolicum and C. rhombifolium (Duan et al., 2022), accumulation of taxifolin could easily result in higher
which could be biomarkers and inspire drug/food development. accumulation of cyanidin and/or quercetin derivatives by
Lut-7-G and quercitrin were the top two abundant compounds mutagenesis, causing changes in flower color. The anthocyanins
of CM, accounting for 85.7% of total flavonoids (Sun et al., 2010b). cyanidin 3-G and cyanidin 3-(3′-malonoyl) glucoside were
More Acn 7-O-galactoside resulted in lighter colored flowers and identified in 23 CM cultivars (Park et al., 2015), among which
less Acn and kaempferol was associated with yellow flowers (Chen “Magic,” “Angel” and “Relance” had high amounts of anthocyanins
et al., 2021). The principal component analysis (PCA) of metabolic and showed a wide range of red and purple colors in their petals.
profile data separated 10 CM cultivars according to flower color Dihydroflavanol 4-reductase (DFR) and anthocyanidin synthase
rather than mutation rates (Sawada et al., 2019). Hot-H2O (ANS) convert dihydrokaempferol to pelargonidin, a pink
extraction of CM tea showed that most flavonoids and CQAs anthocyanidin. In CM, the B-ring of dihydrokaempferol is further
dissolved out at 30 min (Chen et al., 2021), with 20.977 and hydroxylated to cyanidin by flavonoid 3′-hydroxylase (F3′H;
8.958 mg/g gross weight (Learn how to make good CM tea). Mekapogu et al., 2020). Due to the absence of flavonoid
The contents of taxifolin (dihydroquercetin) were clearly 3′5’-hydroxylase (F3′5′H), there is no delphinidin-based
associated with the variation in mutation rates among 10 CM anthocyanin accumulated in CM petals. The accumulation of
cultivars (Sawada et al., 2019). Taxifolin is a precursor of both cyanidin and pelargonidin imparts pink to red-purple, orange to
cyanidin derivatives (pigments) and quercetin derivatives red, respectively. To date, the anthocyanin biosynthesis genes of

Frontiers in Plant Science 12 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

FIGURE 6
The molecular structure of representative flavonoid components of Chrysanthemum. Flavones: Lut-7-O-G (anti-inflammatory, antioxidant,
relieving asthma, xanthine oxidase inhibitor, CM, C. indicum), linarin (antioxidant, anti-inflammatory, preventing acute lung injury, promoting
osteogenic differentiation, inhibiting acetylcholinesterase activity, CM, C. indicum, C. zawadskii), Api-7-O-(6″-O-acetyl)-β-D-glucopyranoside
(antioxidant, CM). Flavonols: kaempferol (antioxidant, anti-inflammatory, anticancer, CM, C. indicum), isorhamnetin 3-O-β-D-G (anti-
inflammatory, CM, C. indicum), quercetin 7-O-β-D-G (various bioactivities, CM, C. indicum). C, flavonones: naringenin (treating depression, CM),
eriodictyol-7-O-β-D-glucuronide (immunoregulation, C. zawadskii, C. indicum), 5,7,3″,5″-tetrahydroxyflavanone-7-O-β-D-glucopyranoside (C.
indicum). D, anthocyanins: cyanidin-3-O-(6″-O-malonyl)glucoside (C. grandiflorum), cyanidin-3-O-(3″,6″-di-O-dimalonyl-β-glucopyranoside)(C.
grandiflorum).

wild Chrysanthemum taxa have not been reported; it is unknown obvious after the end of flowering. The content of linarin in CIF
whether F3′5′H is present in these taxa and how the variation of was significantly different among different habitats (Wei et al.,
biosynthesis genes is involved in the contents of anthocyanins as 2021), where climatic factors, especially average annual
well as spectrum of flower color. precipitation, annual average sunshine hours, and annual average
The total flavonoid content of Chrysanthemum sensu lato temperature, pointedly impacted on the linarin content. Some
varied between 4 and 10% (Shao, 2018), and its change was not abundant flavonoids with content differences among CM cultivars

Frontiers in Plant Science 13 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

can be used as indicators for quantitative identification the spatial and temporal regulation of flavonoid biosynthesis.
(Mao, 2020), e.g., Lut-7-O-G, Api-7-O-G, Lut-7-O-β-G, Api-7-O- Numerous flavonoid UDP-glucuronosyl and -glucosyltransferase
6-AG. The components with high content, large dispersion among (UGT) gene copies were also identified in C. nankingense genome,
cultivars, and large contribution to classification can be used as which is consistent with the phytochemical findings.
evaluation indicators for the quality of CM and cultivar
identification, e.g., Api-7-O-6-AG, 3,5-DCQA, 4,5-DCQA,
3,4-DCQA, 3 -CQA. The content of some phytometabolites is Phenolic acids
relatively low, and their differences between CM cultivars are
large, e.g., Api, Dio, 5-CQA, 4-CQA, Acn; some components were Common phenolic acids of CM have long been known, e.g.,
not detected in some CM cultivars, e.g., Api, Acn. These are not chlorogenic acid, 4-CQA, 3,4-DCQA, 3,5-DCQA, 3,5-DC-epi-QA,
suitable as indicators for Chrysanthemum quality evaluation. The 1,3-DC-epi-QA and ethyl caffeate (Xie et al., 2012; Yang et al.,
CM cultivar “XiaoYangJu” (a kind of Hangju) had the highest total 2018a; Lu et al., 2022b). A rare 8-oxa-bicyclo[3.2.1]oct-3-en-2-one
flavone, total flavonol and total CQA, which is very useful in ring was reported (Yang et al., 2017b), which is formed through a
investigating the biosynthesis pathway of active ingredients and [5 + 2] cycloaddition of CQA with a D-glucose derivative
breeding cultivars with the highest specialized metabolite yield. (Figure 7). C. indicum is rich in cryptochlorogenic acid and
Whether the flavonoid/anthocyanin biosynthesis is 3-O-p-coumaroyl quinic acid (Zou et al., 2022).
conserved across Chrysanthemum species is actually unknown CQA and flavonoids showed a certain correlation with the
(Mekapogu et al., 2020). In general, the biosynthesis of flavonoids stress tolerance of CM plants (Huang, 2010). Importantly, the
begins with the condensation of 4-coumaroyl-CoA and medicinal quality evaluation indexes of Chrysanthemum can
malonyl-CoA by chalcone synthase (CHS) and chalcone be identified from these two categories (Mao, 2020; Chen et al.,
isomerase (CHI) to yield naringenin (Hao et al., 2015). In CM, 2021; Kang et al., 2022). Among the common phytometabolites
the flavonoid 3-hydroxylase and F3′H are responsible for the shared by different medicinal cultivars, compounds with high
synthesis of dihydrokaempferol and dihydroquercetin as the content and great differences among cultivars, e.g., 4,5-DCQA,
essential precursors for the corresponding flavonols. Naringenin flavonoids Acn and Lut, are suitable for screening the index
could also be transformed into flavones via flavone synthase components of quality evaluation, which also have high
(FS) 1. Flavonols and flavones, via glycosyltransferase (GT), contribution to CM classification. The content of 4,5- DCQA is
methyltransferase, hydroxyl transferase, and acyltransferase, are high in most cultivars, so it is suitable to be used as a quantitative
subject to diverse structural modifications to generate various index component for Chrysanthemum quality evaluation. The
flavonoids. In CM, the common feature of flavonol modification content of 3-CQA in all samples was high (Mao, 2020), and
was 3-O-glycosylation (Chen et al., 2021), while flavones were the difference between cultivars was small. It is suitable to
involved in 7-O-glycosylation and 7-O-6′-acylation; the be used as a qualitative index component for Chrysanthemum
metabolic intermediates are then shunted into seven quality evaluation. The low-abundance phytometabolites of
sub-pathways. There were significant positive correlations Chrysanthemum cannot represent its chemical characteristics,
between flavonoids and corresponding flavonoid glycosides, and e.g., 1-CQA, anthocyanins and carotenoids, which are not
there were meaningfully positive correlations between Api and suitable for the quality evaluation. The anti-hepatotoxic
Aca, Api and Lut, Lut and Dio, indicating coordinated 1,5-DCQA was at high levels in C. pacificum flowers and aerial
sub-pathways in CM, and possibly in its wild relatives. However, parts reaching 3,145 and 1,390 μg/g, respectively (Farag et al.,
few data of other Artemisiinae species are available. The content 2015), suggesting an alternative natural resource of
of total flavonoids in different tissues of O. taihangensis was medicinal compounds.
higher than that in corresponding tissues of C. indicum (Liu, The correlation analysis between the pharmacologically active
2013), but biosynthetic pathways have not been well investigated CM ingredients showed that there was a very significant positive
in these species. Seven C. indicum samples from different habitats correlation between chlorogenic acid, isochlorogenic acid B,
were distinct in their flavonoid profile (Zou et al., 2022), which isochlorogenic acid A and isochlorogenic acid C (Liu, 2020), and
was also significantly different from C. nankingense. So far, a chlorogenic acid, isochlorogenic acid B/C and tilianin was
comprehensive metabolomic similarity analysis of as many significantly positively correlated. There was a very significant
Chrysanthemum taxa as possible is still lacking, and the role of positive correlation between isochlorogenic acid C and tilianin,
genetic vs. climatic factors in metabolic differences of while cynaroside and Lut-7-O-β-D-glucuronic acid had a very
populations/species is elusive. significant positive correlation. There was a significant positive
The enrichment of flavonoid biosynthesis genes in the association between Dio-7-O-G and linarin, and a very significant
C. nankingense genome was revealed by whole genome sequencing positive correlation between the former and tilianin. These
(Song et al., 2018). Two key genes, CHS and CHI, are significantly interesting results imply some commonly shared biosynthesis
expanded in C. nankingense genome (17 CHS genes and eight CHI pathway of phenolics and flavonoids, as well as coordinated
genes), which showed the differential expression among tissues, regulation of metabolic networks in Chrysanthemum. The
and there were significantly high expressions in flowers, suggesting phylogenetically related taxa not only have analogous chemical

Frontiers in Plant Science 14 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

constituents, their biosynthesis pathways and regulatory and they were not detected in leaves. In different harvesting
components, as well as metabolic networks and signal periods, the contents of five phenolics in different tissues of
transduction networks, might also be similar. O. taihangensis were higher than those in corresponding tissues of
The anti-inflammatory components of CM could C. indicum. Interestingly, O. taihangensis is often locally used as
be 3,5-DCQA, 4,5-DCQA, Lut-7-O-β-G, 1,3-DCQA, 3-CQA, and C. indicum in folk medicine, and the people in the producing area
the like (Peng et al., 2019; Mao, 2020). Some of them were induced report that its aroma is strong, and its tea and medicinal value is
by jasmonic acid against western flower thrips (Chen et al., 2019). far superior to that of C. indicum.
These compounds can be used as indicator components for quality
evaluation of CM, which are also useful in studying the chemical
relatedness between CM cultivars (Chen et al., 2021), as well as Oligosaccharide, polysaccharide
CM and wild relatives. According to the evaluation results of CM
grades, 50 kinds of CM on the market, including Gongju, Hangju, The sugar components of CM contribute to its dual value of
Taiju (made from CM bud), etc., were divided into three grades food/medicine; the expressions of most glycolysis-, GT-related
(Mao, 2020), which was positively correlated with the content of genes were induced by ultraviolet-B radiation (Yang et al., 2018b).
index components. The lower the grade, the lower the index The health-promoting raffinose and 1-kestose were abundant in
component content. This grading provided a reference for the C. dichrum leaves, flower buds, and blooming flowers (Liu et al.,
establishment of TCM quality grade evaluation system. 2021a). The immunomodulatory and anti-inflammatory JFP1-1-2,
Interestingly, in the abundance of CQA, flavonoids and a homogenous polysaccharide of non-medicinal parts of CM, is
carotenoids, Boju was largely equivalent to Gongju and many composed of mannose, glucose and galactose in a molar ratio of
cultivars of Hangju (Chen et al., 2021), although the former is 4.53:3.06:1.00 (Tao, 2017); another bioactive polysaccharide
phylogenetically distinct from the latter two (Lyu et al., 2008). JFP1-2-2 is composed of mannose, galactose, glucose, xylose and
Taiju and full bloom CM of the same cultivar were relatively galacturonic acid in 3.0:2.3:1.2:1.0:1.0.
close in the PCA diagram based on index components (Mao, The content of polysaccharide prepared by membrane
2020), indicating that there is a great similarity between their separation method of six CM cultivars of Kaifeng, Henan Province
chemical components. Among 13 cultivars, Gongju was the most varied between 47.8 and 64.3% (Zhao, 2015), while those obtained
unique. Gongju, Boju and the characteristic cultivar #1 were quite by alcohol precipitation were 18.9 and 25.3%. The 295 g boiling
different from others. The clustering analysis based on contents of water-extracted crude polysaccharide CMJA (yield 7.8%) and 96 g
13 components largely agreed with PCA results. Most Hangju alkali-extracted polysaccharide JHB (2.5%) were obtained from
samples were clustered into one group, and Huaiju, Chuju and 3.8 kg of dried Huaiju (Zheng, 2015). CMJA contained eight
some Hangju samples were clustered together, indicating a certain homogeneous polysaccharide fractions: CMJA0S1 (110 mg, yield
similarity between them. The characteristic cultivars had certain 0.4%), CMJA0S2 (2.2%), CMJA1a S2 (2.3%), CMJA1a S3 (0.4%),
similarities with Boju, suggesting the kinship between them; CMJA1b S2 (0.5%), CMJA1b S3 (2.0%), CMJA2S2 (2.0%) and
alternatively, these might be partially explained by similar CMJA2S3 (2.2%). JHB contained a polysaccharide JHB0S2
cultivation conditions, and/or similar terroir factors (Suo et al., (532 mg, yield 1.9%). CMJA0S1 is an arabinogalactan with
2011; Hao and Liu, 2021). molecular weight (MW) of 4.7 × 104 Da. The MW of CMJA0S2 is
The cluster analysis based on chlorogenic acid, Lut, 6.5 × 103 Da; its main chain is a galactomannan glucan composed
3,5-DCQA, etc., showed that different CM cultivars were basically of 1,4-β-Glcp, 1,4-β-Galp, 1,4-β-Manp, with the branched chains
clustered by cultivar (Xiong, 2014), and the subgroup composed composed of 1,6-β-Galp, T-α-Glcp, T-α-Araf, 1,5-α-Araf, etc.
of Fubaiju cultivars “local early-flowering” and “local late- CMJA1a S2, CMJA1a S3, CMJA1b S2, CMJA1b S3 and CMJA2S2
flowering,” derived from Hang Baiju, clustered with Hangju and are a series of RG-I type pectin polysaccharides with different
Gongju. The chemical cluster of Hangju and Gongju was also MWs, and their galacturonic acid content and branched-chain
confirmed by other independent studies (Liu et al., 2019), which glycosyl composition varied. 1,2-α-Rhap and 1,4-α-Gal Ap
is basically consistent with the traditional classification and constitute the main chain, and at the O-4 position of rhamnose,
molecular marker-based classification of CM (Lyu et al., 2008; there are branched chains composed of arabinose and galactose.
Mao, 2020). CMJA2S3 is a HG-type pectin polysaccharide of 1.1 × 104 Da and
The content of chlorogenic acid varied greatly among composed of galacturonic acid. JHB0S2 is a xyloglucan of
Chrysanthemum sensu lato plants, the highest was 0.975 g/100 g 1.6 × 104 Da, with 1,4-β-Glcp constituting the main chain and a
(Shao, 2018), and the lowest was only 0.086 g/100 g. After branch at the O-6 position, connecting xylose residues.
flowering, the content of chlorogenic acid in the plant will increase
to a certain extent. The contents of chlorogenic acid, rutinoside,
quercetin, Lut, Api in the leaves of O. taihangensis were higher Mineral elements
than those in flowers, stems and C. indicum (Liu, 2013). The
contents of chlorogenic acid, rutinoside and quercetin were higher Mineral elements absorbed by Chrysanthemum plants play
in the stems and leaves, Lut and Api were less abundant in stems, an important role in regulating the processes of primary and

Frontiers in Plant Science 15 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

FIGURE 7
The molecular structure of representative phenolic components of Chrysanthemum. chlorogenic acid (=3-CQA), 5-CQA, 3,5-DCQA, 4,5-DCQA,
isochlorogenic acid B, 3-O-p-coumaroyl quinic acid, cryptochlorogenic acid, chrysanthemorimic acid A/B/C.

secondary metabolism (Liu, 2020). They are the constituent cultivar clustered together well, indicating that the differences in
factors of active ingredients in traditional medicine, and have a the mineral element content of CM germplasm resources are
great impact on the formation and accumulation of bioactive closely related to genetic factors.
metabolites in TCM plants. CM contained mineral elements There was a significant positive correlation between CM
such as calcium, magnesium, phosphorus, sulfur, potassium, and isochlorogenic acid A/tilianin and N/P element contents (Liu,
indispensable trace elements like copper, iron, zinc, cobalt, 2020; Liu et al., 2021c), isochlorogenic acid B and N element were
manganese, strontium, selenium. The total beneficial mineral significantly positively correlated, whereas chlorogenic acid,
elements (K, Ca, Mg, Fe, Na) contained in six CM cultivars of isochlorogenic acid A, C and cymaroside were significantly
Kaifeng were between 32,008.7 and 40,183.1 mg/kg (Zhao, 2015), negatively correlated with heavy metal Pb. Cynaroside had a very
while the hazardous As, Pb, Hg and Cd were between 0.01–0.04 significant negative relationship with Mn; Dio-7-O-G had a very
and 0.63–1.53 mg/kg. The CM flowers and leaves are rich in N, significant positive correlation with Mn and Mg, and had a
P, K, Ca, Mg, and Fe (Yan et al., 2021), among which K element significant positive correlation with Ca and Pb. There was a
had the largest variation, and N, Ca, Fe, Mg, and Zn had a large significant positive correlation between linarin and Cd/Cr heavy
variation range. The absorption and accumulation of various metals. These imply the potential roles of phenolics and flavonoids
elements in leaves of different germplasm resources are distinct. in Chrysanthemum stress/defense response, as well as possible
There is a strong positive correlation between Ca and Mg/Mn/ approaches of regulating phytometabolite yields.
Cd. The PCA found that CM cultivars were separated based on
Mn, Cr, Cu, P, K. From the perspective of mineral elements,
Hangju-Fuhuangju, Hangju-Xiaoyangju Late-ripening, Hangju- Nutrition constituents
Sheyangju Late-ripening, Hangju-Dayanghua, Hangju-Subeiju
were of good quality. The PCA score map divided the 23 CM Seventeen amino acids were identified in CM, and contents
samples into four groups, and the cluster analysis heat map of Lys, Phe and Leu were relatively high. The contents of seven
divided them into five categories. The samples of the same essential amino acids, i.e., Thr, Val, Met, Ile, Leu, Phe and Lys, in

Frontiers in Plant Science 16 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

the manufactured goods were 2.3–6 times those in the fresh be applied in more Chrysanthemum species to obtain a holistic
flowers (Wang et al., 2016). The total amino acids of six CM and comparative view of their nutritional value.
cultivars of Kaifeng varied between 10.5 and 11.9% (Zhao, 2015),
and the essential amino acids were between 2.4 and 3.0%.
Asparagine and glutamine were abundant in C. dichrum leaves, Other constituents
flower buds, and blooming flowers (Liu et al., 2021a).
Among 28 nucleobases, nucleosides, nucleotides and amino Coumarin, umbelliferone, lignan, tonghaosu, pyrethrin,
acids of CM of nine geographical origins, eight crucial quality jasmolin, cinerin, etc., were also found in CM (Yang et al., 2019;
markers, i.e., 2′-deoxyadenosine, guanosine, adenosine 3′,5′- Zhou et al., 2019b; Mao, 2020; Figure 8). Pyrethrins, a natural
cyclic phosphate (cAMP), guanosine 3′,5′-cyclic monophosphate insecticide, are biosynthesized by the Anthemideae plants such
(cGMP), arginine, proline, glutamate and tryptophan, were as Tanacetum cinerariifolium (Matsuda, 2012) and CM (Mao,
identified (Chang et al., 2021), which could be used to discriminate 2020). Their monoterpenoid acid and rethrolone-type oxylipin
geographical origin of CM. alcohol moieties are biosynthesized via the 2-C-methyl-D:
The crude fat content of six CM cultivars of Kaifeng was -erythritol 4-phosphate (MEP) and oxylipin pathways,
between 3.6 and 7.1% (Zhao, 2015). Twenty chemical constituents respectively (Lybrand et al., 2020). The organic acids, e.g., azelaic
were detected after methylation of fatty acids, including five acid, 4-guanidinobutyric acid, are abundant in C. indicum (Zou
saturated fatty acids; unsaturated fatty acids accounted for 69.9% et al., 2022). The anti-diabetic polyacetylene glycosides from
of the total fatty acids. The contents of fatty acids, e.g., capric acid flowers of edible C. “Kamiohno” are also reported (Kurimoto
(C10.0), pentadecanoic acid (C15.0), α-linolenic acid (C18.3 N3), et al., 2021).
eicosanoic acid (C20.0), docosanoic acid (C22.0) and lignoceric
acid (C24.0), flavonoids and CQAs were increased in CM by UV-B
radiation (Yang et al., 2018b). The contents of moisture, protein, Quality control methods and standards
fat, ash and carbohydrate in C. mongolicum tea were 8.2, 13.7, 4.1,
6.1 and 67.7%, respectively (Xiao, 2019); all essential amino acids Five kinds of medicinal CM are listed in the Chinese
were detected, and the content of amino acids was 11.3 g/100 g. Pharmacopoeia (2020), including Boju, Hangju, Chuju,
Twenty-two kinds of fatty acids were detected, with abundant Huaiju, Gongju. The diversity of species, origins, cultivation,
linoleic acid (22.8%) and α-linolenic acid (7.4%). processing and harvesting methods directly affects the
The carotenoids, e.g., zeaxanthin, carotene, cryptoxanthin, medicinal components and effects. However, the traditional
lutein, etc., were identified in different CM cultivars (Park et al., 2015; HPLC method of measuring the contents of chlorogenic acid,
Zhou et al., 2019b; Mao, 2020; Chen et al., 2021). The average total Lut-7-O-G, 3,5-DCQA is not enough to evaluate the quality of
content of carotenoid was highest in the 10% open, followed by 70 medicinal CM comprehensively. The concept of quality
and 100% bloom stages at 0.50, 0.43, and 0.36 mg/g FW, respectively. marker (Q-marker) provides a new idea for quality control of
The CM cultivars with higher carotenoid contents displayed yellow TCM. Q-marker is based on the perspective of biological
or green petal colors (Park et al., 2015). The expression of carotenoid activity, and various methods are utilized to find the chemical
biosynthetic genes in the petals of different CM cultivars at components that best reflect biological effects of TCM (Liu
mid-development showed no differences (Kishimoto and Ohmiya, et al., 2018). Multiple methods, especially multi-ingredients
2006). The antioxidant vitamin C contents of six CM cultivars of quantification, chromatographic fingerprint and/or their
Kaifeng varied between 21.7 and 62.7 mg/100 g (Zhao, 2015). CM is combination (Nie et al., 2019; Peng et al., 2019), have been
beneficial to human health and is expected to be widely developed applied extensively in the quality control of CM. In recent
and applied in plant food, tea making, health care products, and such years, quality verification methods based on the activity-
like. The nutrient composition measurement results were graded integrated fingerprints have received wide attention. For
(Zhao, 2015), the nutritional index values of each CM cultivar were instance, an antioxidant activity based method combining the
accumulated to obtain the comprehensive nutritional evaluation online HPLC-DPPH, ESI-MS, and NIR spectra analysis
index, so as to compare various CM cultivars and select cultivars that identified 16 antioxidants of five CM cultivars (Zhang et al.,
are prioritized for human consumption. 2022). Phenolic acids play a more important role in antioxidant
When compared with C. indicum, quinones, flavonoids (e.g., activity, and chlorogenic acid, Lut-7-O-G, 3,5-DCQA,
naringenin 7-O-G, quercetin 4’-O-G), steroids, lipids and 4,5-DCQA were observed as the main contributors to the
carbohydrates were more abundant in C. nankingense (Zou et al., overall antioxidant capacity. The chromatographic fingerprint
2022), which is usually eaten as a vegetable in Nanjing, China, and and in vitro antioxidant activity assay were combined to show
its phenylpropanoid, terpene, indole derivatives and alkaloid that chlorogenic acid, 3,5-DCQA, 1, 4,5-O-DCQA and
contents were relatively low. Among identified 477 metabolites, 72 kaempferol-3-O-rutinoside could be Q-markers of Hang-
showed significant differences between C. mongolicum and baiju, Gongju, Huaiju, Taiju and Boju (Lu et al., 2022b). In the
C. rhombifolium (Duan et al., 2022), mainly flavonoids, organic future, the strategy based on activity-integrated fingerprints
acids and nucleotides. The metabolomic techniques should has great potential in quality control of traditional medicine,

Frontiers in Plant Science 17 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

food and other fields, as it can fully reflect the pharmacological may be the principal material basis for these effects. For example,
information of active ingredients. total flavonoids of CM cultivar Bianliang ziyu prevented
hepatotoxicity by inhibiting oxidative stress and apoptosis via the
activation of Nrf2 signaling (Tian et al., 2019); caffeoylquinic acids,
Pharmacological properties chlorogenic acid, gallocatechin, Lut-7-OG, Acn-7-O-rutinoside,
and anthocyanins of Chrysanthemum showed strong antioxidant
The plants of Chrysanthemum and related taxonomic groups activities (Supplementary Table S1.). Acn-7-O-rutinoside, Lut-7-
are traditionally used as ethnomedicine (Figure 1; Jia and Zhang, OG, and chlorogenic acid also showed the anti-inflammatory
2016). Chrysanthemum plants have a wide range of pharmacological activity (Zhang et al., 2019). Flavonoids (including linarin, Dio-7-G,
activities (Figure 9; Supplementary Table S1). In recent years, the tilianin, etc.) and phenolic acids (including isochlorogenic acid C,
main studied taxa include CM (Khan et al., 2020), C. indicum (Tian isochlorogenic acid A, 1,3-DCQA, etc.) of Chrysanthemum
et al., 2020), C. boreale (Kim et al., 2022) and C. zawadskii (Kim improved the inflammatory bowel disease of zebrafish by regulating
et al., 2019), among others. The most reported activity of the expressions of IL-1β, IL-8 and MMP9 (Li et al., 2022).
Chrysanthemum is its anti-inflammatory and immunomodulatory Eriodictyol-7-O-β-d-glucuronide and 5,7-dihydroxy-4-chromene
effects (Xue et al., 2021; Kim et al., 2022), which are consistent with of C. zawadskii var. latilobum had antiallergic effects in FcεRI-
traditional efficacy; it also shows great potential for improving mediated human basophilic KU812F cells (Lee and Shim, 2020).
chronic metabolic diseases, neurodegenerative diseases, etc. As the The activities of Chrysanthemum flavonoids in metabolic regulation
main active ingredients, flavonoids, phenolic acids and terpenoids are also salient. For instance, Lut and luteoloside improved blood

FIGURE 8
Examples of other phytometabolites of Chrysanthemum. Coumarin, umbelliferone, azelaic acid (C. indicum Zou et al., 2022), 4-guanidinobutyric
acid, chrysanthelignanoside A (neuroprotection, CM), dendranlignan A (anti-inflammatory, CM), tonghaosu, pyrethrin II, jasmolin II, cinerin I,
kamiohnoyneosides A and B (anti-diabetic, CM).

Frontiers in Plant Science 18 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

FIGURE 9
Illustration of pharmacological mechanisms exerted by Chrysanthemum extracts and compounds. Various bioactive compounds produced in
Chrysanthemum plants are shown in left panel; their diverse pharmacological activities and possible mechanisms of action are exemplified in the
central part; two representative signaling pathways regulated by Chrysanthemum components are shown on the right part. ARE: Antioxidant
response element; NF-κB: Nuclear factor kappa-B; Nrf2: NF-E2-related factor 2; TLR4: Toll-like receptor 4.

lipids and hepatic steatosis in hyperlipidemia rats by regulating discovered sesquiterpenes are subject to the in vitro activity
antioxidant levels and lipid metabolism (Sun et al., 2021); screening (Jiang et al., 2021a), whose efficacies and corresponding
naringenin and naringenin-7-O-G inhibited the intracellular lipid mechanisms of action warrant further explorations.
accumulation by the activation of PPARγ and phosphorylation of The activities of Chrysanthemum polysaccharides have also
the PI3K/Akt pathway (Nishina et al., 2019); Lut, Acn, and attracted attention (Wang et al., 2021b), e.g., antioxidant, anti-
buddleoside controlled the postprandial glucose concentration by inflammatory and anti-viral (Supplementary Table S1), and other
inhibiting the activity of α-amylase (Li et al., 2019). Other activities compounds and extracts exhibited diversified bioactivities. The
of Chrysanthemum flavonoids are also intriguing. Linarin and antimicrobial, anti-inflammatory, anti-hypertension and antioxidant
scutellarein displayed anticancer activity (Jung et al., 2019; Li et al., effects (Supplementary Table S1) of CM have been utilized by
2020); buddleoside reduced blood pressure in spontaneously ancient doctors and herbalists to help maintain body balance and
hypertensive rats by inhibiting the vascular TLR4/MyD88 pathway relieve much of the sufferings. The other tantalizing effects of CM
and improving vascular endothelial function (Wang et al., 2021c); components, e.g., anti-aging and anti-diabetic, are being revealed by
Acn-7-O-β-D-rutinoside prevented dexamethasone-evoked contemporary polypharmacology investigations, which facilitate
muscle atrophy via the Akt/mTOR pathway and decreasing the drug repurposing and benefit more people. The phylogenetically
mitochondrial respiration (Lee et al., 2021). related Chrysanthemum species are more likely to possess similar
When compared with flavonoids and phenolics, the chemical profiles, despite the impact of environmental factors. The
antioxidant activity and metabolic regulation of Chrysanthemum most typical examples are CM and wild relatives. The medicinal
terpenoids are less reported (Supplementary Table S1.), but their compounds and therapeutic efficacy of CM are closer to C. indicum
anti-inflammatory, anticancer and other properties are than to C. zawadskii, and are more dissimilar to those of C. boreale.
remarkable. For example, chrysanthemulide A (sesquiterpenoid) Significant or subtle differences in chemical composition between
of C. indicum induced apoptosis of osteosarcoma by upregulating closely related species could be useful in tailoring personalized
death receptor 5 via JNK-mediated autophagosome accumulation therapeutic protocol for each patient.
(Zhuo et al., 2019); it also showed the anti-inflammatory activity
via suppressing the LPS-induced NF-κB pathway and down-
regulating MAPK activation (Xue et al., 2018); cumambrin A Conclusions and prospects
(sesquiterpenoid) suppressed the osteoclast formation, bone
resorption, and RANKL-induced signaling pathways in the This article summarized the phylogeny, biodiversity,
treatment of osteoporosis (Zhou et al., 2019a). The recently phytochemistry, and chemodiversity of Chrysanthemum, especially

Frontiers in Plant Science 19 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

the cultivated hybrid CM. As a unique Chinese culture, the formation of CM. The assembly and annotation of more high-
chrysanthemum culture has witnessed the development of history quality Chrysanthemum genomes will help to elucidate the genus
and had a profound influence on human health. The evolution and its contributions to gene abundance/function.
ethnomedicinal experiences, phytochemistry and bioactivity The genome editing enables the induction of mutations in a
studies are the prerequisite of product research and development; targeted genomic region and has recently played a substantial role
to date more than 120 CM compounds have been isolated and in Asteraceae functional genomics and biotechnology (Bernard
identified, and these monomeric compounds and crude extracts et al., 2019; Park et al., 2019). As precise mutations can
are screened for pharmacological activities in vivo and in vitro. The be generated in the targeted sequence, it is considered to be more
experimental studies validated the traditional medicinal uses of effective than conventional mutation breeding. This breakthrough
CM, but the responsible chemicals have not been thoroughly technology has been adopted for various Asteraceae crops such as
determined. Thus, there is a need of bioassay-guided identification chicory and lettuce (Bernard et al., 2019; Park et al., 2019), and
of the bioactive components. What’s more, the relationship could be used in Chrysanthemum for desired phenotypes. Unlike
between traditional uses and recent pharmacological findings is transgenic technology, genome editing does not require the
not always clear, and it is imperative to investigate the biochemical transgene to be integrated with the genome, and the integrated
and physiological mechanisms of components of CM and transgene could be segregated in the progeny.
evolutionarily related taxa, especially their cardiovascular At present, there are very few reports on the use of
protection, anti-tumor and antioxidant activities. Efforts should metabolomics in Chrysanthemum taxa. The metabolomics
also be made to determine the modes of action, bioavailability, technology can be used to comprehensively characterize the
pharmacokinetics and physiological pathways of specific functional chemical components of Chrysanthemum species and varieties,
compounds in Chrysanthemum and related taxonomic groups. obtain quantitative information of each phytometabolite, and
Moreover, clinical studies, e.g., randomized controlled trial, should quickly screen the characteristic chemical, so as to provide
be encouraged to identify any side effects and possible interactions effective data for the quality evaluation and control of
between Chrysanthemum herbal medicine and other natural Chrysanthemum samples, and contribute new ideas and
medicines/synthetic drugs. Further safety verification and clinical references for the elucidation of medicinal material basis of
trials should be carried out to expand the application scope of CM Chrysanthemum. The evaluation and control provide effective
and better integrate it into medicinal practice. With multiple data, inspire new ideas and contribute references for the
chemical ingredients, CM may exert its beneficial effects by gently elucidation of the medicinal material basis of chrysanthemum.
interacting with different cell signaling pathways and networks, The metabolic research is conducive to the realization of the
which achieve the same therapeutic efficacy as that of mono- integrity and traceability of chrysanthemum quality control. The
ingredient agents, whereas CM doses are much lower than those of metabolic makeup of fresh flowers and processed products may
single compounds. In order to better mine the medicinal potential vary greatly; how to achieve the transferability of chrysanthemum
and edible value of different Chrysanthemum cultivars and Daodi quality in multiple links such as harvesting, crude processing, fine
medicinal materials, the intricate link between phylogenetic processing, and production of patent medicines still needs to
relationship, chemical profile, ethnomedicinal knowledge and be studied from multiple hierarchies. However, the relative
pharmacological activities should be scrutinized within the quantification using the peak area as an index cannot reflect the
pharmacophylogenetic framework (Hao and Xiao, 2017, 2020). exact content of compounds in chrysanthemum, therefore it is
The emerging pharmacophylogenomics (Hao and Xiao, 2017, necessary to establish the quantitative analysis method for the
2020) calls for much more genomic data of Chrysanthemum and differential components of different species/varieties of
related taxonomic groups. The rapid development of reduced- Chrysanthemum. The compounds detected by UPLC-Q-TOF-MS
representation genome sequencing enables the cost-effective in plant metabolomics are more comprehensive (Hao et al.,
sequencing of Asteraceae plant (Mérot, 2020). It is of low cost and 2021). To find the quality control indicators of chrysanthemum
strong sequencing performance. In the future, it can be used in from numerous compounds, it is obligatory to combine content
Chrysanthemum phylogeny, plant identification and genome determination and pharmacokinetics/pharmacodynamics for
assisted breeding. The cp genome with maternal genetic comprehensive evaluation, which warrant deeper studies in the
characteristics has the advantages of multi-copy and conservative pharmacologically active constituents of chrysanthemum.
structure. The combination of cp genome and high-throughput The current highly intensive industrial systems of agricultural
sequencing technology has become an effective means of plant and horticultural production are counter-sustainable, as the
genetic resources research, which facilitates the study of energy consumption is particularly intensive for cultivation, and
Chrysanthemum classification and genetic resources (Tyagi et al., the excessive use of nitrogen fertilizer leads to the worrying
2020; Masuda et al., 2022). The accumulation of whole genome emission of greenhouse gas, N2O (Hao et al., 2021; Wang et al.,
sequencing data of Chrysanthemum (Song et al., 2018; Nakano et al., 2022). The genetic modification (GM) is transforming the
2021; van Lieshout et al., 2022) and relevant taxonomic groups will prospects of sustainable crop protection, the specialized metabolite
enable the comprehensive phylogenomic analyses to reveal the (e.g., pyrethrins) biosynthesis genes can be integrated to the target
hybridization/polyploidization events leading to the speciation and Chrysanthemum plants to enhance the stress response and disease

Frontiers in Plant Science 20 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

resistance, so as to realize the full potential of GM and offer LX: resources and supervision. D-CH, YS, and PW: data curation
greener crop protection. and visualization. D-CH and YS: writing—original draft
This article applies the systematic and holistic characteristics preparation. All authors contributed to the article and approved
of plant pharmacophylogeny to the regular arrangement of the submitted version.
biodiversity and chemodiversity explorations on Chrysanthemum.
On the basis of scientific and technological innovation, and with
the integration of holistic view of traditional medicine, we have Funding
deepened our understanding of the following points: The
phylogenetically close taxa are more likely to have similar This work is supported by the Scientific Research Funds
metabolic profile, and the similar metabolic makeup could result Project of Liaoning Education Department (JDL2019012), China
in analogous pharmacokinetic behavior and clinical efficacy (Hao Scholarship Council (202108210156), CAMS Innovation Fund for
and Xiao, 2020). The studies of pharmacophylogeny are conducive Medical Sciences (CIFMS 2021-I2M-1-032), and Hainan
to the sustainable conservation and rational utilization of Academician Innovation Platform Scientific Research Project and
Chrysanthemum resources, as well as the inheritance and National Science & Technology Fundamental Resources
innovation of traditional medicine. The medicinal properties of Investigation Program of China (2018FY100700).
plants can be predicted by virtue of phylogenetic methods (Hao
et al., 2022a,b), which has been utilized to explore the regularity
of bioactivities of Asteraceae and Ranunculales plants against Conflict of interest
diseases of multiple human organs. The mapping of different types
of Chrysanthemum (and related taxonomic groups) compounds The authors declare that the research was conducted in the
onto the species phylogenetic tree could also be beneficial to absence of any commercial or financial relationships that could
mining novel sources of economically important chemicals. be construed as a potential conflict of interest.
Disentangling recent speciation events and distribution of
compounds/therapeutic effects usually requires a reliable
phylogenetic framework, therefore research in phylogeny/ Publisher’s note
evolution of Chrysanthemum and related taxonomic groups
cannot be overemphasized. Besides the intensive studies of CM, All claims expressed in this article are solely those of the
the evaluation of other Chrysanthemum species should also keep authors and do not necessarily represent those of their affiliated
pace with the times and develop scientifically. The ability to organizations, or those of the publisher, the editors and the
correlate the chrysanthemum metabolome with the medicinal reviewers. Any product that may be evaluated in this article, or
efficacy in the context of pharmacophylogeny will help to develop claim that may be made by its manufacturer, is not guaranteed or
chrysanthemum cultivars with better curative effect and more endorsed by the publisher.
commercial value in the near future.

Supplementary material
Author contributions
The Supplementary material for this article can be found
D-CH and PX: conceptualization and writing—review and online at: https://www.frontiersin.org/articles/10.3389/fpls.2022.
editing. D-CH, YS, and YZ: methodology and analysis. PX and 973197/full#supplementary-material

References
Beninger, C. W., Abou-Zaid, M. M., Kistner, A. L., Hallett, R. H., Iqbal, M. J., Chang, X., Zhang, Z., Yan, H., Su, S., Wei, D., Guo, S., et al. (2021). Discovery of
Grodzinski, B., et al. (2004). A flavanone and two phenolic acids from quality markers of nucleobases, nucleosides, nucleotides and amino acids for
Chrysanthemum morifolium with phytotoxic and insect growth regulating Chrysanthemi Flos from different geographical origins using UPLC-MS/MS
activity. J. Chem. Ecol. 30, 589–606. doi: 10.1023/B:JOEC.0000018631. combined with multivariate statistical analysis. Front. Chem. 9:689254. doi: 10.3389/
67394.e5 fchem.2021.689254
Bernard, G., Gagneul, D., Alves Dos Santos, H., Etienne, A., Hilbert, J. L., and Chen, J. Y. (2005). Contributions of Chinese Chrysanthemum to the world in the
Rambaud, C. (2019). Efficient genome editing using CRISPR/Cas9 technology in past and future. J. Chin. Landscape Architect. 9, 73–75.
chicory. Int. J. Mol. Sci. 20:1155. doi: 10.3390/ijms20051155
Chen, G., Kim, H. K., Klinkhamer, P. G., and Escobar-Bravo, R. (2019). Site-
Cai, Y., Gao, Y., Zhang, Z., Liu, H., Wang, Y., Ma, Y., et al. (2022). Development dependent induction of jasmonic acid-associated chemical defenses against
and application of a cultivar-specific sequence-characterized amplified region western flower thrips in Chrysanthemum. Planta 251:8. doi: 10.1007/
(SCAR) marker for the detection of Chrysanthemum morifolium Ramat. 'Daboju'. s00425-019-03292-2
Plants. 11:604. doi: 10.3390/plants11050604
Chen, T., Li, L. P., Lu, X. Y., Jiang, H. D., and Zeng, S. (2007). Absorption and
Chang, X. W., Wei, D. D., Chen, D. J., Yan, H., Sun, X. D., Zhu, W. B., et al. (2019). excretion of luteolin and apigenin in rats after oral administration of
Historical origin and development of medicinal and tea Chrysanthemum morifolium Chrysanthemum morifolium extract. J. Agri. Food Chem. 55, 273–277. doi:
resource. Modern Chin. Med. 21, 116–123. doi: 10.13313/j.issn.1673-4890.20181224005 10.1021/jf062088r

Frontiers in Plant Science 21 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

Chen, S., Liu, J., Dong, G., Zhang, X., Liu, Y., Sun, W., et al. (2021). Flavonoids and Hong, G. (2014). Preliminary study on intergeneric hybridization of
caffeoylquinic acids in Chrysanthemum morifolium Ramat flowers: A potentially Chrysanthemum and its related genera (VIII). MSc thesis. [Beijing]: Beijing Forestry
rich source of bioactive compounds. Food Chem. 344:128733. doi: 10.1016/j. University, 1–56.
foodchem.2020.128733
Hong, G., Wu, X. B., Liu, Y. C., Xie, F., Liu, Z. H., Liu, W. C., et al. (2015).
Chen, X. J., Sun, M., Liang, J. G., Xue, H., and Zhang, Q. X. (2013). Genetic Intergeneric hybridization between Hippolytia kaschgarica (Krascheninnikov)
diversity of species of Chrysanthemum and related genera and groundcover Poljakov and Nipponanthemum nipponicum (Franch. ex Maxim.) Kitam. Genet.
cultivars assessed by amplified fragment length polymorphic markers. HortScience Resour. Crop. Evol. 62, 255–263. doi: 10.1007/s10722-014-0150-1
48, 539–546. doi: 10.21273/HORTSCI.48.5.539 Huang, Y. (2010). Preliminary study on cross breeding of medicinal Chrysanthemum.
Chinese Pharmacopoeia Commission. (2015). Pharmacopoeia of the People’s MSc thesis. [Nanjing(Jiangsu)]: Nanjing Agricultural University, 1–87.
Republic of China. Beijing: China Medical Science Press, 1, 310–311. Huang, Z., Liu, Z. Y., Wang, S. L., Xue, Y. Q., Xue, J. Q., and Zhang, X. X. (2020).
Chinese Pharmacopoeia Commission. (2020). Pharmacopoeia of the People’s Resources investigation of Chinese medicinal Chrysanthemum morifolium
Republic of China. Βeijing: China Medical Science Press, 1, 323–324. varieties and its industry status analysis. J. Chin. Medi. Materia. 43, 1325–1329.
doi: 10.13863/j.issn1001-4454.2020.06.007
Chong, X., Zhang, F., Wu, Y., Yang, X., Zhao, N., Wang, H., et al. (2016). A SNP-
enabled assessment of genetic diversity, evolutionary relationships and the Jia, M.R., and Zhang, Y. (2016). Dictionary of Chinese Ethnic Medicine. Beijing:
identification of candidate genes in Chrysanthemum. Genome Biol. Evol. 8, China Medical Science Press.
3661–3671. doi: 10.1093/gbe/evw270 Jiang, S., Wang, M., Jiang, Z., Zafar, S., Xie, Q., Yang, Y., et al. (2021a). Chemistry
Clifford, M. N., Wu, W., Kirkpatrick, J., and Kuhnert, N. (2007). Profiling the and pharmacological activity of sesquiterpenoids from the Chrysanthemum genus.
chlorogenic acids and other caffeic acid derivatives of herbal Chrysanthemum by Molecules 26:3038. doi: 10.3390/molecules26103038
LC-MSn. J. Agri. Food Chem. 55, 929–936. doi: 10.1021/jf062314x Jiang, H., Xia, Q., Xu, W., and Zheng, M. (2004). Chrysanthemum morifolium
Criado Ruiz, D., Villa Machío, I., Herrero Nieto, A., and Nieto Feliner, G. (2021). attenuated the reduction of contraction of isolated rat heart and cardiomyocytes
Hybridization and cryptic speciation in the Iberian endemic plant genus induced by ischemia/reperfusion. Pharmazie 59, 565–567.
Phalacrocarpum (Asteraceae-Anthemideae). Mol. Phylogenet. Evol. 156:107024. doi: Jiang, B. P., Xu, L. J., Wang, Q. L., and Xiao, P. G. (2013). Chrysanthemum
10.1016/j.ympev.2020.107024 morifolium--- Ethnopharmacological use, phytochemistry and pharmacological
Cui, N. X. (2004). Meiosis behavior and genetic relationship of several activity. Modern Chin. Med. 15, 523–530. doi: 10.13313/j.issn.1673-4890.2013.06.016
Chrysanthemum plants and their hybrids. MSc thesis. [Nanjing(Jiangsu)]: Nanjing Jiang, Y. F., Zhang, W. B., Chen, X. L., Wang, W., Köllner, T. G., Chen, S., et al.
Agricultural University, 1–61. (2021b). Diversity and biosynthesis of volatile terpenoid secondary metabolites in
Dai, Z., Li, S. X., Chen, S. Q., and Zhu, Y. (2017). Quality evaluation of Huaiju the Chrysanthemum genus. Criti. Rev. Plant Sci. 40, 422–445. doi:
germplasm resources. Chin. J. Expt. Trad. Med. Formulae. 23, 48–54. doi: 10.13422/j. 10.1080/07352689.2021.1969504
cnki.syfjx.2017060048 Jo, Y. D., Ryu, J., Kim, Y. S., Kang, K. Y., Hong, M. J., Choi, H. I., et al. (2020).
Ding, L., Chen, F. D., Teng, N. J., Fang, W. M., and Zhao, J. Y. (2008). Isozyme Dramatic increase in content of diverse flavonoids accompanied with down-
analysis of wild and different purpose Chrysanthemum. J. Nanjing Agri. Univ. 2008, regulation of F-box genes in a Chrysanthemum (Chrysanthemum × morifolium
37–42. (Ramat.) Hemsl.) mutant cultivar producing dark-purple ray florets. Genes. 11:865.
doi: 10.3390/genes11080865
Duan, X. X., Zhang, W. J., Li, J. J., Xu, H., Hu, J., Zhao, L., et al. (2022). Comparative
metabolomics analysis revealed biomarkers and distinct flavonoid biosynthesis Jung, C. H., Han, A. R., Chung, H. J., Ha, I. H., and Um, H. D. (2019). Linarin
regulation in Chrysanthemum mongolicum and C. rhombifolium. Phytochem. Anal. inhibits radiation-induced cancer invasion by downregulating MMP-9 expression
33, 373–385. doi: 10.1002/pca.3095 via the suppression of NF-κB activation in human non-small-cell lung cancer A549.
Nat. Prod. Res. 33, 3582–3586. doi: 10.1080/14786419.2018.1484460
Fan, M., Gao, Y. K., Gao, Y. H., Wu, Z., Liu, H., and Zhang, Q. X. (2019).
Kang, S. H., Nie, J., Chen, K. L., and Deng, H. Y. (2022). Comparison of
Characterization and development of EST-SSR markers from transcriptome
morphological characteristics and quality evaluation of Chrysanthemum morifolium.
sequences of chrysanthemum (Chrysanthemum× morifolium Ramat.). Hort Science
J. Chin. Medi. Mate. 2022, 49–57. doi: 10.13863/j.issn1001-4454.2022.01.009
54, 772–778. doi: 10.21273/HORTSCI13694-18
Khan, I. A., Xu, W., Wang, D., Yun, A., Khan, A., Zongshuai, Z., et al. (2020).
Farag, N. F., Farag, M. A., Abdelrahman, E. H., Azzam, S. M., and
Antioxidant potential of chrysanthemum morifolium flower extract on lipid and
El-Kashoury, E. S. A. (2015). Metabolites profiling of Chrysanthemum pacificum
protein oxidation in goat meat patties during refrigerated storage. J. Food Sci. 85,
Nakai parts using UPLC-PDA-MS coupled to chemometrics. Nat. Prod. Res. 29,
618–627. doi: 10.1111/1750-3841.15036
1342–1349. doi: 10.1080/14786419.2015.1025396
Kim, K. Y., Oh, T. W., Yang, H. J., Kim, Y. W., Ma, J. Y., and Park, K. I. (2019).
Feng, S. G., He, R. F., Lu, J., Jiang, M., Shen, X., Jiang, Y., et al. (2016).
Ethanol extract of Chrysanthemum zawadskii Herbich induces autophagy and
Development of SSR markers and assessment of genetic diversity in medicinal
apoptosis in mouse colon cancer cells through the regulation of reactive
Chrysanthemum morifolium cultivars. Front. Genet. 7:113. doi: 10.3389/
oxygen species. BMC Complement. Altern. Med. 19:274. doi: 10.1186/
fgene.2016.00113 s12906-019-2688-0
Flora Reipublicae Popularis Sinicae. (1983). Beijing: Science Press, 76, 35–39. Kim, D. Y., Won, K. J., Hwang, D. I., Kim, N. Y., Kim, B., and Lee, H. M. (2022).
Hao, D. C., Chen, S. L., Osbourn, A., Kontogianni, V. G., Liu, L. W., and 1-Iodohexadecane alleviates 2,4-dinitrochlorobenzene-induced atopic dermatitis in
Jordán, M. J. (2015). Temporal transcriptome changes induced by methyl jasmonate mice: Possible involvements of the skin barrier and mast cell SNARE proteins.
in Salvia sclarea. Gene 558, 41–53. doi: 10.1016/j.gene.2014.12.043 Molecules 27:1560. doi: 10.3390/molecules27051560
Hao, D. C., Li, P., Xiao, P. G., and He, C. N. (2021). Dissection of full-length Kishimoto, S., and Ohmiya, A. (2006). Regulation of carotenoid biosynthesis in
transcriptome and metabolome of Dichocarpum (Ranunculaceae): implications in petals and leaves of chrysanthemum (Chrysanthemum morifolium). Physiologia
evolution of specialized metabolism of Ranunculales medicinal plants. PeerJ. Plant. 128, 436–447. doi: 10.1111/j.1399-3054.2006.00761.x
9:e12428. doi: 10.7717/peerj.12428
Kurimoto, S. I., Fujita, H., Kawaguchi, S., Sasaki, Y. F., Nakamura, T., and
Hao, D. C., and Liu, C. X. (2021). Back to beginning: Searching for Rosetta Stone Kubota, T. (2021). Kamiohnoyneosides A and B, two new polyacetylene glycosides
of enhancing herbal medicine quality. Chin. Herb. Med. 13, 299–300. doi: 10.1016/j. from flowers of edible Chrysanthemum "Kamiohno". J. Nat. Med. 75, 167–172. doi:
chmed.2022.03.005 10.1007/s11418-020-01443-4
Hao, D. C., Lyu, H. Y., Wang, F., and Xiao, P. G. (2022a). Evaluating potentials of Lai, J. P., Lim, Y. H., Su, J., Shen, H. M., and Ong, C. N. (2007). Identification and
species rich taxonomic groups in cosmetics and dermatology: Clustering and characterization of major flavonoids and caffeoylquinic acids in three Compositae
dispersion of skin efficacy of Asteraceae and Ranunculales plants on the species plants by LC/DAD-APCI/MS. J. Chromatography B. 848, 215–225. doi: 10.1016/j.
phylogenetic tree. Curr. Pharm. Biotech. 23:926. doi: 10.2174/1389201023666220324 jchromb.2006.10.028
123926
Lee, H., Kim, Y. I., Nirmala, F. S., Jeong, H. Y., Seo, H. D., Ha, T. Y., et al. (2021).
Hao, D. C., and Xiao, P. G. (2017). An Introduction of Plant Pharmacophylogeny. Chrysanthemum zawadskil Herbich attenuates dexamethasone-induced muscle
Beijing: Chemical Industry Press. atrophy through the regulation of proteostasis and mitochondrial function. Biomed.
Pharmacother. 136:111226. doi: 10.1016/j.biopha.2021.111226
Hao, D. C., and Xiao, P. G. (2020). Pharmaceutical resource discovery from
traditional medicinal plants: Pharmacophylogeny and pharmacophylogenomics. Lee, M., and Shim, S. Y. (2020). Inhibitory effects of eriodictyol-7-O-β-d-
Chin. Herb. Med. 12, 104–117. doi: 10.1016/j.chmed.2020.03.002 glucuronide and 5,7-dihydroxy-4-chromene isolated from Chrysanthemum
zawadskii var. latilobum in FcεRI-mediated human basophilic KU812F cell
Hao, D. C., Zhang, Y. L., He, C. N., and Xiao, P. G. (2022b). Distribution of
activation. Molecules 25, 994. doi: 10.3390/molecules25040994
therapeutic efficacy of Ranunculales plants used by ethnic minorities on the
phylogenetic tree of Chinese species. Evid. Based Complement. Altern. Med. 2022, Li, H. (1993). Chinese Chrysanthemum. Nanjing: Jiangsu Scientific and
9027727. doi: 10.1155/2022/9027727 Technological Publishing Co.

Frontiers in Plant Science 22 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

Li, T. (2014). Construction of CDDP fingerprints and genetic diversity analysis of Lybrand, D. B., Xu, H., Last, R. L., and Pichersky, E. (2020). How plants synthesize
Chrysanthemum. MSc thesis. [Taian(Shandong)]: Shandong Agricultural pyrethrins: Safe and biodegradable insecticides. Trends Plant Sci. 25, 1240–1251.
University, 1–64. doi: 10.1016/j.tplants.2020.06.012
Li, P., Huang, Z., She, Y., Qin, S., Gao, W., Cao, Y., et al. (2019). An assessment of Lyu, L., Qin, M. J., He, D. X., and Gu, Y. H. (2008). Analyses of ISSR
the interaction for three Chrysanthemum indicum flavonoids and α-amylase by molecular marker and genetic relationship of different provenances of
surface plasmon resonance. Food Sci. Nutri. 8, 620–628. doi: 10.1002/fsn3.1349 Dendranthema morifolium, D. indicum and D. nankingese. J. Plant Resour.
Environ. 17, 7–12.
Li, Y., Liu, X. J., Su, S. L., Yan, H., Guo, S., Qian, D. W., et al. (2022). Evaluation of
anti-inflammatory and antioxidant effects of Chrysanthemum stem and leaf extract Ma, Y. P., Chen, M. M., Wei, J. X., Zhao, L., Liu, P. L., Dai, S. L., et al. (2016). Origin
of Chrysanthemum cultivars—Evidence from nuclear low-copy LFY gene
on zebrafish inflammatory bowel disease model. Molecules 27, 2114. doi: 10.3390/
sequences. Biochem. Syst. Ecol. 65, 129–136. doi: 10.1016/j.bse.2016.02.010
molecules27072114
Mandel, J. R., Dikow, R. B., Siniscalchi, C. M., Thapa, R., Watson, L. E., and
Li, B., Ma, C., Zhao, X., Hu, Z., Du, T., Xu, X., et al. (2018). YaTCM: Yet another
Funk, V. A. (2019). A fully resolved backbone phylogeny reveals numerous
traditional Chinese medicine database for drug discovery. Comput. Struct. dispersals and explosive diversifications throughout the history of Asteraceae. Proc.
Biotechnol. J. 16, 600–610. doi: 10.1016/j.csbj.2018.11.002 Natl. Acad. Sci. U. S. A. 116, 14083–14088. doi: 10.1073/pnas.1903871116
Li, J., Wan, Q., Abbott, R. J., and Rao, G. Y. (2013). Geographical distribution of Mao, C. Y. (2020). Study on quality evaluation standard of the flower of
cytotypes in the Chrysanthemum indicum complex as evidenced by ploidy level and Chrysanthemum morifolium Ramat based on the correlation of ingredients and
genome-size variation. J. Syst. Evol. 51, 196–204. doi: 10.1111/j.1759-6831. efficacy. [PhD dissertation. [Beijing]: China Academy of Chinese Medical Sciences,
2012.00241.x 1–246.
Li, J., Wan, Q., Guo, Y. P., Abbott, R. J., and Rao, G. Y. (2014). Should I stay or Masuda, Y., Nakano, M., and Kusaba, M. (2022). The complete sequence of the
should I go: biogeographic and evolutionary history of a polyploid complex chloroplast genome of Chrysanthemum rupestre, a diploid disciform capitula
(Chrysanthemum indicum complex) in response to Pleistocene climate change in species of Chrysanthemum. Mitochondrial DNA B Resour. 7, 603–605. doi:
China. New Phytol. 201, 1031–1044. doi: 10.1111/nph.12585 10.1080/23802359.2022.2057252
Li, Y., Wang, J., Zhong, S., Li, J., and Du, W. (2020). Scutellarein inhibits the Matsuda, K. (2012). Pyrethrin biosynthesis and its regulation in Chrysanthemum
development of colon cancer via CDC4-mediated RAGE ubiquitination. Int. J. Mol. cinerariaefolium. Top. Curr. Chem. 314, 73–81. doi: 10.1007/128_2011_271
Med. 45, 1059–1072. doi: 10.3892/ijmm.2020.4496 Mekapogu, M., Vasamsetti, B. M. K., Kwon, O. K., Ahn, M. S., Lim, S. H., and
Li, D. L., Zhu, H. W., Ren, Q. J., and Xu, Z. L. (2010). Comparative analysis on the Jung, J. A. (2020). Anthocyanins in floral colors: Biosynthesis and regulation in
configuration of vegetative organs of medicinal Chrysanthemum from different Chrysanthemum flowers. Int. J. Mol. Sci. 21, 6537. doi: 10.3390/ijms21186537
original locations and species. Zhong Yao Cai 33, 1845–1849. doi: 10.13863/j. Mérot, C. (2020). Making the most of population genomic data to understand the
issn1001-4454.2010.12.005 importance of chromosomal inversions for adaptation and speciation. Mol. Ecol. 29,
Liu, R. (2010). A study on the relationship between some wild species of 2513–2516. doi: 10.1111/mec.15500
Chrysanthemum and cultivated chrysanthemum. MSc thesis. [Baoding(Hebei)]: Nakano, M., Hirakawa, H., Fukai, E., Toyoda, A., Kajitani, R., Minakuchi, Y., et al.
Hebei Agricultural University, 1–58. (2021). A chromosome-level genome sequence of Chrysanthemum seticuspe, a
model species for hexaploid cultivated chrysanthemum. Commun. Biol. 4, 1167. doi:
Liu, Q. J. (2013). Accumulation and antioxidant activity of phenolic substances in
10.1038/s42003-021-02704-y
Opisthopappus taihangensis and related Chrysanthemum indicum. PhD dissertation.
[Zhengzhou(Henan)]: Henan Agricultural University, 1–92. Nie, J., Xiao, L., Zheng, L., Du, Z., Liu, D., Zhou, J., et al. (2019). An integration of
UPLC-DAD/ESI-Q-TOF MS, GC-MS, and PCA analysis for quality evaluation and
Liu, Y. (2020). Comparative study on agronomic characters, chemical identification of cultivars of Chrysanthemi Flos (Juhua). Phytomedicine 59:152803.
components and pharmacological effects of different chrysanthemum germplasm doi: 10.1016/j.phymed.2018.12.026
resources. MSc thesis. [Wuhan(Hubei)]: Hubei University of Chinese Medicine,
1–124. Nishina, A., Sato, D., Yamamoto, J., Kobayashi-Hattori, K., Hirai, Y., and
Kimura, H. (2019). Antidiabetic-like effects of naringenin-7-O-glucoside
Liu, H., Chen, X., Chen, H., Lu, J., Chen, D., Luo, C., et al. (2021a). Transcriptome from edible Chrysanthemum 'Kotobuki' and naringenin by activation of the
and metabolome analyses of the flowers and leaves of Chrysanthemum dichrum. PI3K/Akt pathway and PPARγ. Chem. Biodivers. 16:e1800434. doi: 10.1002/
Front. Genet. 12:716163. doi: 10.3389/fgene.2021.716163 cbdv.201800434
Liu, Y., Dai, M., Bao, W. Z., Huang, B. S., Guo, L. P., and Liu, D. H. Oberprieler, C., Himmelreich, S., and Vogt, R. (2007). A new subtribal
(2021c). Characteristics of mineral elements in chrysanthemums from different classification of the tribe Anthemideae (Compositae). Willdenowia 37, 89–114. doi:
origins in Macheng and their correlation with soil nutrients and effective 10.3372/wi.37.37104
components. China J. Chin. Mate. Med. 46, 281–289. doi: 10.19540/j.cnki. Park, C. H., Chae, S. C., Park, S. Y., Kim, J. K., Kim, Y. J., Chung, S. O., et al. (2015).
cjcmm.20200717.101 Anthocyanin and carotenoid contents in different cultivars of Chrysanthemum
Liu, Y., Gong, W. L., Bao, W. Z., Guo, L. P., Xu, Y., Liu, Y. M., et al. (2019). (Dendranthema grandiflorum Ramat.) flower. Molecules 20, 11090–11102. doi:
Establishment of Chrysanthemum morifolium HPLC fingerprint of Hubei and 10.3390/molecules200611090
comparison of active components among varieties. China J. Chin. Mate. Med. 44, Park, J., Choi, S., Park, S., Yoon, J., Park, A. Y., and Choe, S. (2019). DNA-free
3711–3717. doi: 10.19540/j.cnki.cjcmm.20190701.106 genome editing via ribonucleoprotein (RNP) delivery of CRISPR/Cas in lettuce.
Liu, C. X., Guo, D. A., and Liu, L. (2018). Quality transitivity and traceability Methods Mol. Biol. 1917, 337–354. doi: 10.1007/978-1-4939-8991-1_25
system of herbal medicine products based on quality markers. Phytomedicine 44, Peng, A., Lin, L., Zhao, M., and Sun, B. (2019). Classification of edible
247–257. doi: 10.1016/j.phymed.2018.03.006 chrysanthemums based on phenolic profiles and mechanisms underlying the
Liu, H. B., Ma, P., Xu, L. J., and Xiao, P. G. (2021b). Informatics and big data: A protective effects of characteristic phenolics on oxidatively damaged erythrocyte.
Food Res. Int. 123, 64–74. doi: 10.1016/j.foodres.2019.04.046
new stage of pharmacophylogeny in medicinal plants. Mod. Chin. Med. 23,
1506–1511. doi: 10.13313/j.issn.1673-4890.20210901002 Peng, Y. R., Shi, L., and Luo, Y. H. (2006). Protective effect of the total flavones
from Chrysanthemum on isoprenaline-induced myocardial ischemiain rats.
Liu, P. L., Wan, Q., Guo, Y. P., Yang, J., and Rao, G. Y. (2012). Phylogeny of the
Lishizhen Med. Materia Med. Res. 2006, 1131–1132.
genus Chrysanthemum L.: evidence from single-copy nuclear gene and chloroplast
DNA sequences. PLoS One 7:e48970. doi: 10.1371/journal.pone.0048970 Poe, S., and Swofford, D. L. (1999). Taxon sampling revisited. Nature 398,
299–300. doi: 10.1038/18592
Lu, W. X., Hu, X. Y., Wang, Z. Z., and Rao, G. Y. (2022a). Hyb-Seq provides new
insights into the phylogeny and evolution of the Chrysanthemum zawadskii species Rajic, A., Akihisa, T., Ukiya, M., Yasukawa, K., Sandeman, R. M., Chandler, D. S.,
complex in China. Cladistics. doi: 10.1111/cla.12514 et al. (2001). Inhibition of trypsin and chymotrypsin by anti-inflammatory
triterpenoids from Compositae flowers. Planta Med 67, 599–604. doi:
Lu, Y. F., Li, D. X., Zhang, R., Zhao, L. L., Qiu, Z., Du, Y., et al. (2022b). Chemical 10.1055/s-2001-17350
antioxidant quality markers of Chrysanthemum morifolium using a spectrum-effect
approach. Front. Pharmacol. 13:809482. doi: 10.3389/fphar.2022.809482 Ryu, J., Nam, B., Kim, B. R., Kim, S. H., Jo, Y. D., Ahn, J. W., et al. (2019).
Comparative analysis of phytochemical composition of gamma-irradiated mutant
Luo, C., Chen, D. L., Cheng, X., Zhao, H. E., and Huang, C. L. (2017). Genome cultivars of Chrysanthemum morifolium. Molecules 24:3003. doi: 10.3390/
size estimations in Chrysanthemum and correlations with molecular phylogenies. molecules24163003
Genet. Resour. Crop. Evol. 64, 1451–1463. doi: 10.1007/s10722-016-0448-2
Sawada, Y., Sato, M., Okamoto, M., Masuda, J., Yamaki, S., Tamari, M., et al.
Luo, X. Y., Wang, C., Dai, S. L., Li, B. Q., Liu, Q. Q., Zhu, J., et al. (2013). Genetic (2019). Metabolome-based discrimination of chrysanthemum cultivars for the
diversity of large-flowered Chrysanthemum cultivars revealed by ISSR analysis. Sci. efficient generation of flower color variations in mutation breeding. Metabolomics
Agri. Sin. 46, 2394–2402. 15, 118. doi: 10.1007/s11306-019-1573-7

Frontiers in Plant Science 23 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

Shao, B. J. (2018). Preliminary study on germplasm resources innovation of forage Wang, D. Q., Liu, S. J., and Liang, Y. M. (2001). Research on medicinal taxa of
species within Chrysanthemum in broad sense. MSc thesis. [Beijing]: Beijing Chinese chrysanthemum. J. Anhui College TCM. 20, 45–48.
Forestry University, 1–114.
Wang, Y. J., Su, J., Yu, J. J., Yan, M. Q., Shi, M. L., Huang, Q. D., et al. (2021c).
Shao, Q. S., Guo, Q. S., Li, Y. C., and Mao, P. F. (2011). Quantitative analysis of Buddleoside-rich Chrysanthemum indicum L. extract has a beneficial effect on
morphological variation of medicinal Chrysanthemum germplasm resources. China metabolic hypertensive rats by inhibiting the enteric-origin LPS/TLR4 pathway.
J. Chin. Mate. Med. 36, 1261–1265. Fronti. Pharmacol. 12:755140. doi: 10.3389/fphar.2021.755140
Shao, Y. H., Sun, Y. D., Li, D., and Chen, Y. P. (2020). Chrysanthemum indicum L.: Wang, Y. S., Wang, M. X., Han, S., Zhang, J., Shen, X., Zhou, J., et al. (2016).
A comprehensive review of its botany, phytochemistry and pharmacology. Am. J. Differences in amino acid contents in fresh flowers and manufactured goods among
Chin. Med. 48, 871–897. doi: 10.1142/S0192415X20500421 four Chrysanthemum cultivars in Tongxiang City. Zhejiang Province. J. Anhui Agri.
Univ. 43, 1024–1028. doi: 10.13610/j.cnki.1672-352x.20161205.011
Shen, C. Z., Zhang, C. J., Chen, J., and Guo, Y. P. (2021). Clarifying recent adaptive
diversification of the Chrysanthemum-group on the basis of an updated multilocus Wang, Y. J., Yang, X. W., and Guo, Q. S. (2008a). Studies on chemical constituents
phylogeny of subtribe Artemisiinae (Asteraceae: Anthemideae). Front. Plant Sci. in Huangjuhua (flowers of Chrysanthemum morifolium). China J. Chin. Materia
12:648026. doi: 10.3389/fpls.2021.648026 Med. 33, 526–530.
Song, C., Liu, Y. F., Song, A., Dong, G., Zhao, H. B., Sun, W., et al. (2018). The Wang, Z., Yuan, Y., Hong, B., Zhao, X., and Gu, Z. (2021a). Characteristic volatile
Chrysanthemum nankingense genome provides insights into the evolution and fingerprints of four Chrysanthemum teas determined by HS-GC-IMS. Molecules
diversification of Chrysanthemum flowers and medicinal traits. Mol. Plant 11, 26:7113. doi: 10.3390/molecules26237113
1482–1491. doi: 10.1016/j.molp.2018.10.003
Watson, L. E., Bates, P. L., Evans, T. M., Unwin, M. M., and Estes, J. R. (2002).
Song, Y. J., Xu, L. J., Miao, J. H., and Xiao, P. G. (2020). Research progress in Molecular phylogeny of Subtribe Artemisiinae (Asteraceae), including Artemisia
Chrysanthemum indicum. Modern Chin. Med. 22, 1751–1756. doi: 10.13313/j. and its allied and segregate genera. BMC Evol. Biol. 2:17. doi: 10.1186/1471-2148-2-17
issn.1673-4890.20190722006
Wei, M., Zhang, Y. J., Wang, T., Guo, Q. S., Zou, Q. J., Chen, F. R., et al. (2021).
Sun, C. Q., Chen, F. D., Fang, W. M., Liu, Z. L., and Teng, N. J. (2010a). Advances Correlations between content of linarin in Chrysanthemum indicum and climatic
in research on distant hybridization of Chrysanthemum. Sci. Agri. Sin. 43, factors in habitats. China J. Chin. Materia Med. 46, 2167–2172. doi: 10.19540/j.cnki.
2508–2517. doi: 10.3864/j.issn.0578-1752.2010.12.015 cjcmm.20210320.104
Sun, Q. L., Hua, S., Ye, J. H., Zheng, X. Q., and Liang, Y. R. (2010b). Flavonoids Won, S. Y., Kwon, S. J., Lee, T. H., Jung, J. A., Kim, J. S., Kang, S. H., et al. (2017).
and volatiles in Chrysanthemum morifolium Ramat flower from Tongxiang County Comparative transcriptome analysis reveals whole-genome duplications and gene
in China. Afr. J. Biotechnol. 9, 3817–3821. selection patterns in cultivated and wild Chrysanthemum species. Plant Mol. Biol.
95, 451–461. doi: 10.1007/s11103-017-0663-z
Sun, J., Wang, Z., Chen, L., and Sun, G. (2021). Hypolipidemic effects and
preliminary mechanism of Chrysanthemum flavonoids, its main components Wu, G. S. (2007). Study on the genetic relationship between some Chrysanthemum
luteolin and luteoloside in hyperlipidemia rats. Antioxidants. 10:1309. doi: 10.3390/ and Ajania plants. MSc thesis. [Nanjing (Jiangsu)]: Nanjing Agricultural University,
antiox10081309 1–101.
Suo, F. M., Chen, S. L., Yu, H., Xie, C. X., and Sun, C. Z. (2011). Study on the origin Wu, X. B. (2014). Preliminary study on distant hybridization of Chrysanthemum
suitability of four famous Chrysanthemums in China. Moderni. Trad. Chin. Med. sensu lato (VII). MSc thesis. [Beijing]: Beijing Forestry University, 1–62.
Mater. Med.-World Sci. Technol. 13, 332–339. Wu, X., Sun, Y., Shen, X., and Wang, Z. (2015). Study on combined effects of
Tang, F. P. (2009). Study on distant hybridization between Chrysanthemum and chemical components for different flowers blossoming degree of yellow medicinal
four related genera. PhD dissertation. [Nanjing(Jiangsu)]: Nanjing Agricultural Chrysanthemum morifolium from Zhejiang. China J. Chin. Mate. Med. 40, 3174–3178.
University, 1–121. Xiao, Y. (2019). Research on nutritional and functional properties of
Tao, J. H. (2017). Study on the effect mechanism of polysaccharides from non- Chrysanthesum mongolicum tea and its application in whey tea beverage. MSc
medicinal parts of chrysanthemum on inflammatory bowel disease. PhD thesis. [Hohhot (Inner Mongolia)]: Inner Mongolia Agricultural University, 1–62.
dissertation. [Nanjing(Jiangsu)]: Nanjing University of Chinese Medicine, Xie, F. (2016). Preliminary study on distant hybridization of Chrysanthemum
1–174. (IX): Germplasm utilization of Hippolytia and other genera. MSc thesis. [Beijing]:
Tian, Z., Jia, H., Jin, Y., Wang, M., Kou, J., Wang, C., et al. (2019). Chrysanthemum Beijing Forestry University, 1–82.
extract attenuates hepatotoxicity via inhibiting oxidative stress in vivo and in vitro. Xie, Y., Qu, J., Wang, Q., Wang, Y., Yoshikawa, M., and Yuan, D. (2012).
Food Nutr. Res. 63:1667. doi: 10.29219/fnr.v63.1667 Comparative evaluation of cultivars of Chrysanthemum morifolium flowers by
Tian, D., Yang, Y., Yu, M., Han, Z. Z., Wei, M., Zhang, H. W., et al. (2020). Anti- HPLC-DAD-ESI/MS analysis and antiallergic assay. J. Agri. Food Chem. 60,
inflammatory chemical constituents of Flos Chrysanthemi Indici determined by 12574–12583. doi: 10.1021/jf304080v
UPLC-MS/MS integrated with network pharmacology. Food Funct. 11, 6340–6351. Xiong, Y. X. (2014). Research on the germplasm resources of Fubaiju and the
doi: 10.1039/D0FO01000F selection of excellent lines. MSc thesis. [Wuhan(Hubei)]: Hubei University of
Tyagi, S., Jung, J. A., Kim, J. S., and Won, S. Y. (2020). A comparative analysis of Chinese Medicine, 1–125.
the complete chloroplast genomes of three Chrysanthemum boreale strains. PeerJ. Xu, W. B., Guo, Q. S., Li, Y. N., and Wang, T. (2005). Comparative study on
8:e9448. doi: 10.7717/peerj.9448 internal quality of various Chrysanthemum morifolium. China J. Chin. Mate. Med.
van Lieshout, N., van Kaauwen, M., Kodde, L., Arens, P., Smulders, M. J. M., 30, 1645–1648.
Visser, R. G. F., et al. (2022). De novo whole-genome assembly of Chrysanthemum Xu, W. B., Guo, Q. S., and Wang, C. L. (2006). RAPD analysis of genetic diversity
makinoi, a key wild chrysanthemum. G3. 12, jkab358. doi: 10.1093/g3journal/ of medicinal Chrysanthemum morifolium. China J. Chin. Mate. Medi. 2006, 18–21.
jkab358
Xu, M., Jiang, Y., Chen, S., Chen, F., and Chen, F. (2021). Herbivory-induced
Wang, L. Z. (2020). Preliminary identification of natural hybrids between emission of volatile terpenes in Chrysanthemum morifolium functions as an indirect
Chrysanthemum indicum and Chrysanthemum vestitum. MSc thesis. defense against Spodoptera litura larvae by attracting natural enemies. J. Agri. Food
[Taian(Shandong)]: Shandong Agricultural University, 1–56. Chem. 69, 9743–9753. doi: 10.1021/acs.jafc.1c02637
Wang, T. Y., Guo, Q. S., and Wang, T. (2012). Karyotype analysis of 21 medicinal Xue, H., Jiang, Y., Zhao, H., Köllner, T. G., Chen, S., Chen, F. D., et al. (2019).
Chrysanthemum cultivation types. J. Nanjing Agri. Univ. 35, 13–18. Characterization of composition and antifungal properties of leaf secondary
Wang, Y. J., Guo, Q. S., Yang, X. W., Xu, W. B., and Tao, H. Y. (2008b). metabolites from thirteen cultivars of Chrysanthemum morifolium Ramat. Molecules
Characterization of chemical components of essential oil from flowers of 24:4202. doi: 10.3390/molecules24234202
Chrysanthemum morifolium produced in Anhui province. China J. Chin. Materia Xue, G. M., Li, X. Q., Chen, C., Chen, K., Wang, X. B., Gu, Y. C., et al. (2018).
Med. 33, 2207–2211. Highly oxidized guaianolide sesquiterpenoids with potential anti-inflammatory
Wang, L., Hao, D. C., Fan, S. S., Xie, H. T., Bao, X. L., Jia, Z. J., et al. (2022). N2O activity from Chrysanthemum indicum. J. Nat. Prod. 81, 378–386. doi: 10.1021/acs.
emission and nitrification/denitrification bacterial communities in upland black soil jnatprod.7b00867
under combined effects of early and immediate moisture. Agriculture 12:330. doi: Xue, L. M., Qin, X. M., and Guo, J. G. (2007). Comparative study on main
10.3390/agriculture12030330 components of Chrysanthemum morifolium introduced in Ruicheng in Shanxi
Wang, J., Liang, Q., Zhao, Q., Tang, Q., Ahmed, A. F., Zhang, Y., et al. (2021b). The Province. Chin. Trad. Herb. Drug. 2007, 751–754.
effect of microbial composition and proteomic on improvement of functional Xue, G. M., Xue, J. F., Zhao, C. G., Zhao, Z. Z., Zhi, Y. L., Du, K., et al. (2021).
constipation by Chrysanthemum morifolium polysaccharide. Food Chem. Toxicol. 1,10-seco guaianolide-type sesquiterpenoids from Chrysanthemum indicum. J. Asian
153:112305. doi: 10.1016/j.fct.2021.112305 Nat. Prod. Res. 23, 877–883. doi: 10.1080/10286020.2020.1787388
Wang, D. Q., Liu, S. J., and Liang, Y. M. (1999). A study on producing areas of Yan, H. Y., Liu, Y., Xu, Y., Fang, Y., Guo, L. P., and Liu, D. H. (2021). Analysis and
Chinese flos Dendranthematis. China J. Chin. Mate. Medi. 24:573. evaluation of mineral elements of Chrysanthemum morifolium for medicinal and tea

Frontiers in Plant Science 24 frontiersin.org

Hao et al. 10.3389/fpls.2022.973197

use of different germplasm resources. China J. Chin. Mate. Med. 46, 272–280. doi: Zhang, W., Xu, H., Duan, X., Hu, J., Li, J., Zhao, L., et al. (2021a). Characterizing
10.19540/j.cnki.cjcmm.20201023.101 the leaf transcriptome of Chrysanthemum rhombifolium (Ling et C. Shih), a
drought resistant, endemic plant from China. Front. Genet. 12:625985. doi: 10.3389/
Yang, L., Aobulikasimu, N., Cheng, P., Wang, J. H., and Li, H. (2017a). Analysis of
fgene.2021.625985
floral volatile components and antioxidant activity of different varieties of
Chrysanthemum morifolium. Molecules 22:1790. doi: 10.3390/molecules22101790 Zhang, Z., Zhang, Y., Wang, L., Cui, T., Wang, Y., Chen, J., et al. (2022). On-line
screening of natural antioxidants and the antioxidant activity prediction for the
Yang, C. F., Dong, C. M., Xing, B., Xia, W., and Tian, Y. J. (2018a). Study on quality
extracts from flowers of Chrysanthemum morifolium ramat. J. Ethnopharmacol.
of Chrysanthemum morifolium from different habitats. Mod. Chin. Med. 20,
294:115336. doi: 10.1016/j.jep.2022.115336
716–720. doi: 10.13313/j.issn.1673-4890.20170519008
Zhao, H. B. (2007). Phylogeny of tribe Anthemideae (Asteraceae) from East Asia
Yang, P. F., Feng, Z. M., Yang, Y. N., Jiang, J. S., and Zhang, P. C. (2017b).
and intergeneric cross between Dendranthema×grandiflorum (Ramat.) Kitam. and
Neuroprotective caffeoylquinic acid derivatives from the flowers of Chrysanthemum
Ajania pcifica (Nakai) K. Bremer & Humphries. PhD dissertation. [Nanjing
morifolium. J. Nat. Prod. 80, 1028–1033. doi: 10.1021/acs.jnatprod.6b01026
(Jiangsu)]: Nanjing Agricultural University, 1–181.
Yang, W., Glover, B. J., Rao, G. Y., and Yang, J. (2006). Molecular evidence for
Zhao, S. H. (2015). Analysis of nutrient composition of six kinds of
multiple polyploidization and lineage recombination in the Chrysanthemum
chrysanthemum. MSc thesis. [Kaifeng(Henan)]: Henan University, 1–81
indicum polyploid complex (Asteraceae). New Phytol. 171, 875–886. doi: 10.1111/j.
1469-8137.2006.01779.x Zhao, H. B., Chen, F. D., Chen, S. M., Wu, G. S., and Guo, W. M. (2010). Molecular
phylogeny of Chrysanthemum, Ajania and its allies (Anthemideae, Asteraceae) as
Yang, X. W., Han, M. H., Tao, H. Y., Wang, Z. A., Yang, Z., and Xiao, S. Y. (2007).
inferred from nuclear ribosomal ITS and chloroplast trnL-F IGS sequences. Plant
GC-MS analysis of essential oil from anthodiums of Chrysanthemum morifolium
Syst. Evol. 284, 153–169. doi: 10.1007/s00606-009-0242-0
processed by microwave-airflow and steam calefaction. China J. Chin. Mate. Medi.
32, 227–230. doi: 10.1021/acs.jnatprod.6b01026 Zhao, H. B., Chen, F. D., Guo, W. M., Tang, F. P., and Fang, W. M. (2008).
Yang, Y. J., Jiang, Z. F., Guo, J., Yang, X., Xu, N., Chen, Z., et al. (2018b). Preliminary study on hybrid compatibility between Chrysanthemum and some
Transcriptomic analyses of Chrysanthemum morifolium Ramat under UV-B genera of Anthemideae. J. Nanjing Agri. Univ. 2008, 139–143.
radiation treatment reveal variations in the metabolisms associated with bioactive Zheng, C. P. (2015). Isolation, purification, structure identification and bioactivity
components. Indus. Crop Prod. 124, 475–486. doi: 10.1016/j.indcrop.2018.08.011 of chrysanthemum polysaccharide. MSc thesis. [Nanchang(Jiangxi)]: Nanchang
Yang, P. F., Yang, Y. N., Feng, Z. M., Jiang, J. S., and Zhang, P. C. (2019). Six new University, 1–107
compounds from the flowers of Chrysanthemum morifolium and their biological Zhou, J. (2009). Studies on the problem of origin of Chinese garden
activities. Bioorg. Chem. 82, 139–144. doi: 10.1016/j.bioorg.2018.10.007 chrysanthemum. PhD dissertation. [Beijing]: Beijing Forestry University, 1–148.
Zhang, R. D. (2008). The Chrysanthemum culture research in ancient China. PhD Zhou, L., Liu, Q., Hong, G., Song, F., Zhao, J., Yuan, J., et al. (2019a). Cumambrin
dissertation. [Nanjing(Jiangsu)]: Nanjing Normal University, 16–25. A prevents OVX-induced osteoporosis via the inhibition of osteoclastogenesis, bone
Zhang, L. J., and Dai, S. L. (2009). Research advance on germplasm resources of resorption, and RANKL signaling pathways. FASEB J. 33, 6726–6735. doi: 10.1096/
Chrysanthemum morifolium. Chin. Bull. Bot. 44, 526–535. doi: 10.3969/j. fj.201800883RRR
issn.1674-3466.2009.05.002 Zhou, H. P., Ren, M. X., Guan, J. Q., Liu, Y. L., Xiong, Y. X., Zhong, Q. F., et al.
Zhang, E. X., Fang, L., Zhang, J., Yu, L. J., and Xiao, X. (2000). The antioxidant (2019b). Research progress on chemical constituents and pharmacological effects of
activity research of Chrysanthemum extract. Food Sci. 21, 6–9. Chrysanthemum morifolium and predictive analysis on quality markers. Chin. Trad.
Herb. Drug. 50, 4785–4795.
Zhang, N., He, Z., He, S., and Jing, P. (2019). Insights into the importance of
dietary chrysanthemum flower (Chrysanthemum morifolium cv. Hangju)- Zhou, Z., Xian, J., Wei, W., Xu, C., Yang, J., Zhan, R., et al. (2021). Volatile
wolfberry (Lycium barbarum fruit) combination in antioxidant and anti- metabolic profiling and functional characterization of four terpene synthases reveal
inflammatory properties. Food Res. Int. 116, 810–818. doi: 10.1016/j. terpenoid diversity in different tissues of Chrysanthemum indicum L. Phytochemistry
foodres.2018.09.015 185:112687. doi: 10.1016/j.phytochem.2021.112687
Zhang, W., Jiang, Y., Chen, S., Chen, F. D., and Chen, F. (2021b). Concentration- Zhuo, F. F., Zhang, C., Zhang, H., Xia, Y., Xue, G. M., Yang, L., et al. (2019).
dependent emission of floral scent terpenoids from diverse cultivars of Chrysanthemulide A induces apoptosis through DR5 upregulation via JNK-
Chrysanthemum morifolium and their wild relatives. Plant Sci. 309:110959. doi: mediated autophagosome accumulation in human osteosarcoma cells. J. Cellu.
10.1016/j.plantsci.2021.110959 Physiol. 234, 13191–13208. doi: 10.1002/jcp.27991
Zhang, K., Jiang, Y., Zhao, H., Köllner, T. G., Chen, S., Chen, F. D., et al. (2020). Zou, Q., Guo, Q., Wang, T., Chen, J., Yang, F., and Yang, C. (2022). Comparison
Diverse terpenoids and their associated antifungal properties from roots of different of metabolome characteristics and screening of chemical markers in Chrysanthemum
cultivars of Chrysanthemum morifolium Ramat. Molecules 25:2083. doi: 10.3390/ indicum from different habitats. Physiol. Mol. Biol. Plants 28, 65–76. doi: 10.1007/
molecules25092083 s12298-022-01137-z

Frontiers in Plant Science 25 frontiersin.org