1.Genetic diversity analysis and DNA fingerprinting of Artemisia argyi germplasm resources based on EST-SSR molecular markers.
Yu-Yang MA ; Chang-Jie CHEN ; Ming-Xing WANG ; Yan FANG ; Yu-Huan MIAO ; Da-Hui LIU
China Journal of Chinese Materia Medica 2025;50(9):2356-2364
This study investigates the genetic diversity and evolutionary relationships of different Artemisia argyi germplasm resources to provide a basis for germplasm identification, variety selection, and resource protection. A total of 192 germplasm resources of A. argyi were studied, and EST-based simple sequence repeat(EST-SSR) primers were designed based on transcriptomic data of A. argyi. Polymerase chain reaction(PCR) amplification was performed on these resources, followed by fluorescence capillary electrophoresis to detect genetic diversity and construct DNA fingerprints. From 197 pairs of primers designed, 28 pairs with polymorphic and clear bands were selected. A total of 278 alleles were detected, with an average of 9.900 0 alleles per primer pair and an average effective number of alleles of 1.407 2. The Shannon's diversity index(I) for the A. argyi germplasm resources ranged from 0.148 1 to 0.418 0, with an average of 0.255 7. The polymorphism information content(PIC) ranged from 0.454 5 to 0.878 0, with an average of 0.766 9, showing high polymorphism. Cluster analysis divided the A. argyi germplasm resources into three major groups: Group Ⅰ contained 136 germplasm samples, Group Ⅱ contained 45, and Group Ⅲ contained 11. Principal component analysis also divided the resources into three groups, which was generally consistent with the clustering results. Mantel test results showed that the genetic variation in A. argyi populations was to some extent influenced by geographic distance, but the effect was minimal. Structure analysis showed that 190 germplasm materials had Q≥ 0.6, indicating that these germplasm materials had a relatively homogeneous genetic origin. Furthermore, 8 core primer pairs were selected from the 28 designed primers, which could distinguish various germplasm types. Using these 8 core primers, DNA fingerprints for the 192 A. argyi germplasm resources were successfully constructed. EST-SSR molecular markers can be used to study the genetic diversity and phylogenetic relationships of A. argyi, providing theoretical support for the identification and molecular-assisted breeding of A. argyi germplasm resources.
Artemisia/classification*
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Microsatellite Repeats
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Genetic Variation
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Expressed Sequence Tags
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DNA Fingerprinting
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Phylogeny
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Polymorphism, Genetic
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DNA, Plant/genetics*
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Genetic Markers
2.Development of DUS testing guidelines of Artemisia argyi.
Chang-Jie CHEN ; Yu-Huan MIAO ; Xiu-Fu WAN ; Lan-Ping GUO ; Da-Hui LIU
China Journal of Chinese Materia Medica 2024;49(21):5835-5842
Artemisia argyi is a perennial herbaceous herb of the Artemisia family, with leaves for medical use. However, the germplasm of A. argyi is seriously unclear and mixed during production, and it is urgent to protect new varieties of A. argyi. The distinctness, uniformity, and stability(DUS) testing of the new varieties of plants is the basis for the protection of new varieties of plants, and the development of the DUS testing guidelines is the technical basis for DUS testing. To develop the DUS testing guidelines of A. argyi, A. argyi of 100 germplasm was used as the research objects, and their agronomic and medicinal quality characters were observed and measured during six growth stages, and each character was graded and described. A total of 53 test characters were determined, including 19 characters that must be tested; there were four plant characters, two rhizome characters, five stem characters, three branching characters, 29 leaf characters, three floral characters, five medicinal quality characters, and two other characters. It also involved 16 quality characters, 22 quantitative characters, and 15 pseudo-quantitative characters. Seven grouping characters were determined from 53 characters, including "emergence period" "plant-plant type" "branching-primary branching site" "stem-color" "middle leaf-number of leaf splits" "budding period", and "plant-height". By searching for standard characters, 16 standard varieties were ultimately determined. The preparation of this guideline was of great significance for the review and protection of new A. argyi varieties, the protection of breeders' rights, and the promotion of the development of A. argyi industry.
Artemisia/chemistry*
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Plant Leaves/chemistry*
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Quality Control
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Plants, Medicinal/classification*
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Guidelines as Topic
3.Mini-barcode combined with ITS2 for identification of bulk Artemisiae Scopariae Herba.
Xin-Yi LI ; Hua GUO ; Ming-Xue MA ; Liu-Wei XU ; Yu-Hua HUANG ; Yun ZHANG ; Cui-Ping YANG ; Feng HE ; Xiao-Xuan TIAN
China Journal of Chinese Materia Medica 2024;49(24):6685-6691
Artemisiae Scoporiae Herba is derived from Artemisia scoparia or A. capillaris. The accurate identification of the herbs, particularly when dealing with bulk samples, is critical for ensuring the quality and efficacy of the medicinal product. This study aimed to establish a comprehensive molecular approach by combining multiple markers for the precise identification of Artemisiae Scoporiae Herba. The ITS2 from A. scoparia, A. capillaris, and other common Artemisia species were retrieved from GenBank. MEGA was used to build a phylogenetic tree with these sequences, and the effectiveness of ITS2 in species identification was assessed. The analysis revealed that while ITS2 could distinguish Artemisiae Scoporiae Herba from other closely related species of Artemisia, it was insufficient to differentiate between A. scoparia and A. capillaris. To address this limitation, the chloroplast genome of A. capillaris was assembled and compared with the published chloroplast genomes of A. scoparia and A. capillaris, on the basis of which a DNA mini-barcode was developed. The rpoA-rps11 region was selected as the target for the development of mini-barcode due to its potential for distinguishing between these two species. Specific primers were designed to differentiate A. scoparia from A. capillaris. The ITS2 sequences and the newly developed mini-barcode were used together for Sanger sequencing to identify individual samples of Artemisiae Scoporiae Herba, while DNA metabarcoding was employed for the identification of bulk samples. The identification results of representative individual samples and bulk samples from different regions consistently confirmed A. capillaris. This study established a method that combined ITS2 and mini-barcode to identify bulk samples of Artemisiae Scoporiae Herba from different regions. This approach overcomes the limitations of morphological and chemical methods, enhancing species identification accuracy and supporting a stable supply of medicinal materials.
Artemisia/classification*
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DNA Barcoding, Taxonomic/methods*
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Phylogeny
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DNA, Plant/genetics*
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DNA, Ribosomal Spacer/genetics*
4.Development of Analytical Method and Validation using HPLC/PDA for Discrimination between Artemisiae Argyi Folium and Artemisiae Iwayomogii Herba
Duc Dat LE ; Duc Hung NGUYEN ; Bing Tian ZHAO ; Byung Sun MIN ; Mi Hee WOO
Natural Product Sciences 2019;25(3):275-283
In this study, we described the new developed method to simultaneously discriminate two herbal drugs of Artemisiae Argyi Folium and Artemisiae Iwayomogii Herba using eight marker compounds (1 – 8) on an HPLC-PDA system. The developed method was applied to quantify the major components of two herbal drugs. The pattern analysis successfully discriminated and evaluated different components between Artemisiae Argyi Folium and Artemisiae Iwayomogii Herba. Results were used for classification of different species from collected samples.
Artemisia
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Classification
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Discrimination (Psychology)
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Methods
5.Screening and taxonomic identification of endophytic fungi with antitumor and antioxidant activities from Artemisia lactiflora.
Yi-Xin QIAN ; Ji-Chuan KANG ; Bang-Xing LEI ; Lu WANG ; Ying HUANG
China Journal of Chinese Materia Medica 2014;39(3):438-441
Artemisia lactiflora is an important medicinal plant in China. The antitumor and antioxidant activities of the extracts of 54 endophytic fungi from the plant were screened via MTT assay and DPPH scavenging radical assay, respectively. The bioactive strains were identified based on similarity of 5.8S gene and internal transcribed spacer (ITS) sequences. The results showed that extracts from ten (18.5%) isolates exhibited antitumor activity, and which from two (3.7%) isolates exhibited antioxidant activity. The Alternaria sp. GYBH47 strain was simultaneously having antagonistic activity against HL-60 leukemia, MCF-7 breast and COLO205 colon cell lines, and Phomopsis sp. GYBH42 strain having cytotoxic and antioxidant activities. The results indicated that endophytic fungi from Artemisia lactiflora are potential resources to find valuable bioactive components.
Antineoplastic Agents
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chemistry
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pharmacology
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Artemisia
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microbiology
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Biphenyl Compounds
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metabolism
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Cell Line, Tumor
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Endophytes
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chemistry
;
classification
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physiology
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Free Radical Scavengers
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chemistry
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pharmacology
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Fungi
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classification
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physiology
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Humans
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Picrates
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metabolism
6.Genetic structure and genetic diversity of Artemisia annua varieties (strains) populations based on SCoT markers.
Da-xia CHEN ; Guang-lin CUI ; Xue ZHANG ; Long-yun LI
China Journal of Chinese Materia Medica 2014;39(17):3254-3258
To reveal the genetic diversity and genetic structure in Artemisia annua varieties (strains) populations, we detected the genetic polymorphism within and among eight varieties (strains) populations (192 individuals) by the approach of Start Codon Targeted Polymorphism (SCoT). The associated genetic parameters were calculated by POPGENE1.31 and the relationship was constructed based on UPGMA method. The results showed that, using 20 screened primers, a total of 145 bands were produced, of which 122 were polymorphic loci. At species level, there was a high level of genetic diversity among eight varieties (strains) populations (PPB = 84.1% ,H = 0.217 3 and H(sp) = 0.341 9). However, at the variety (strains) population level, genetic diversity was lower, the average of genetic parameters was PPB = 41.9%, H = 0.121 5, H(pop) = 0.186 8. The Nei's genetic differentiation coefficient was 0.441 0, indicate that most of the genetic variation in this species existed within the variety populations. The gene flow (N(m) = 0.633 9) was less among populations, indicating that the degree of genetic differentiation was higher. Genetic similarity coefficient were changed from 0.755 1 to 0.985 7. By clustering analysis, eight varieties (strains) were clustered into two major categories and it was also showed the same or similar genetic background varieties (strains) have a tendency to gather in the same group. Results suggest that, in variety breeding, breeders should strengthen the exchange of bred germplasm and increase mutual penetration of excellent genes, which would broaden the genetic base of A. annua.
Artemisia annua
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classification
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genetics
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Codon, Initiator
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genetics
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Genetic Markers
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genetics
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Genetic Structures
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Genetic Variation
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Genetics, Population
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methods
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Phylogeny
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Polymorphism, Genetic
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Species Specificity
7.Diversity and cluster analysis on agronomic traits of Artemisia annua germplasm resources in Yun-Gui plateau.
Meiquan YANG ; Dahui LIU ; Aijuan SHAO ; Hang JIN ; Xiao WANG ; Zhenghua FAN ; Luqi HUANG
China Journal of Chinese Materia Medica 2010;35(23):3097-3102
OBJECTIVETo evaluate the diversity of germplasm resources of Artemisia annua and provide the basis for improving utilization of germplasm resources, the agronomic traits of germplasm resources of A. annua were studied in Yun-Gui plateau.
METHODThe agronomic traits of 67 A. annua germplasm resources were measured by the visual observation and measurement methods. And the germplasm resources were clustered using flexible-beta method to analysis their genetic background.
RESULTThe result showed that 67 germplasm resources had a relatively wide variation on the 22 agronomic traits. Among 22 agronomic traits, the dry weight of branch had the greatest coefficient of variation, which was 53. 63, and the next were the dry weight of leaf, total plant weight, the length of pinnules and the length of leaflet, which were 42.74, 41.61, 39.54 and 39.22 respectively. The smallest coefficient of variation was the leaf corlor. Based the result of cluster analysis, these 67 germplasm resources were classed into 5 groups, and each group had its respective character. The first group showed early-maturing resources, dwarf stalk, slender rod, long bipinnata, high leaf-stem ratio and moderate leaf weight The third group showed late-maturing resources, tall and thick stalk, much-branch, bushy accessory pinna, high leaf weight and yield. The fifth group showed very late-maturing resources, strong lateral shoot, high leaf yield.
CONCLUSIONThere were significant genetic difference and diversity in the germplasm resources of A. annua. The result of cluster analysis showed that the resources of group 1, group 3 and group 5 were suitable as breeding material of A. annua.
Artemisia annua ; classification ; genetics ; growth & development ; Biodiversity ; Biomass ; China ; Cluster Analysis
8.Cloning and bioinformatics analysis of P450 cDNA in Artemisia annua.
Jian-Qiang KONG ; Wei WANG ; Xiao-Dong ZHENG ; Ping ZHU ; Ke-Di CHENG
China Journal of Chinese Materia Medica 2007;32(21):2227-2231
OBJECTIVECloning and bioinformatics analysis of P450 cDNA in Artemisia annua.
METHODA P450 cDNA gene was cloned from A. annua by RT-PCR. The bioinformatics analysis of the P450 gene was performed.
RESULTThe complete ORF of this P450 cDNA is 1 464 bp and encodes 488 aa. The sequence was reported to GenBank and coded as DQ667171. Bioinformatics analysis of the P450 cDNA showed it encoded an A-type P450 protein with 54. 992 kDa, it's isoelectric point was 8.83 and the possibility of export to mitochondria was 0.893 2.
CONCLUSIONThe comparable analysis of the P450 with CYP71AV1 revealed that the two proteins probably performed the same function because of the similar character.
Amino Acid Sequence ; Artemisia annua ; enzymology ; genetics ; Base Sequence ; Cloning, Molecular ; Cytochrome P-450 Enzyme System ; classification ; genetics ; DNA, Complementary ; chemistry ; genetics ; Molecular Sequence Data ; Open Reading Frames ; genetics ; Phylogeny ; Plant Proteins ; genetics ; Plants, Medicinal ; enzymology ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid
9.Studies on chemical constituents in bud of Artemisia scoparia (II).
Sheng LIN ; Yong-qing XIAO ; Qi-wei ZHANG ; Ning-ning ZHANG
China Journal of Chinese Materia Medica 2004;29(2):152-154
OBJECTIVETo investigate the chemical constituents in flower bud of Artemisia scoparia.
METHODThe constituents were isolated and purified by means of various chromatographic methods, and spectroscopic methods were used to identify their structures.
RESULTFour flavones were isolated and their structures were identified as hyperin (V), eupafolin (VI), pedalitin (VII), 5,7,2',4'-tetrahydroxy-6,5'-dimethoxyflavone (VIII).
CONCLUSIONCompounds V, VI, VII, VIII were obtained from this plant for the first time.
Artemisia ; chemistry ; classification ; Flavones ; chemistry ; isolation & purification ; Flowers ; chemistry ; Plants, Medicinal ; chemistry ; Quercetin ; analogs & derivatives ; chemistry ; isolation & purification
10.Studies on chemical constituents in buds of Artemisia scoparia.
Qi-wei ZHANG ; Yong-xi ZHANG ; Ying ZHANG ; Yong-qing XIAO ; Zhi-min WANG
China Journal of Chinese Materia Medica 2002;27(3):202-204
OBJECTIVETo investigate the chemical constituents in flower buds of Artemisia scoparia.
METHODThe constituents were separated and purified by means of various chromatographic techniques, and their structures were determined on the basis of physical and chemical properties and spectral data.
RESULTFour flavones were isolated and their structures were identified as cirsilineol (I), cirsimaritin(II), arcapillin(III) and cirsiliol(IV) respectively.
CONCLUSIONCompounds I, III and IV were isolated from this plant for the first time.
Artemisia ; chemistry ; classification ; Flavones ; Flavonoids ; chemistry ; isolation & purification ; Flowers ; chemistry ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry

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