Identification and expression analysis of flavonoid O-methyltransferases gene family in Artemisia argyi / 药学学报

italic>Artemisia argyi (A. argyi) is a Chinese herbal medicine in China. The main active components are volatile oils, flavonoids, and other compounds, which have various pharmacological activities. Methoxylated flavonoids are the main active ingredients in A. argyi. Flavonoid O-methyltransferase (FOMT) is a key enzyme in the O-methylation of flavonoids. In order to further understand the function and characteristics of FOMT proteins, this paper carried out the whole genome mining and identification of FOMT genes in A. argyi and performed phylogenetic, chromosomal localization, gene sequence characterization, subcellular localization prediction, protein structure, gene structure analysis, and expression pattern analysis. The results showed that a total of 83 FOMT genes were identified in the genome of A. argyi. The phylogenetic tree shows that FOMT genes are divided into two subgroups, CCoAOMT (caffeoyl CoA O-methyltransferase) subfamily (32 genes) and COMT (caffeic acid O-methyltransferase) subfamily (51 genes). Gene sequence analysis showed that the number of amino acids encoded by FOMT was 70-734 aa, the molecular weight was 25 296.55-34 241.3 Da, and the isoelectric point was 4.51-9.99. Compared with 32 members of the CCoAOMT subfamily, nearly 1/3 of the 51 members of the COMT subfamily were hydrophobic proteins and 2/3 were hydrophilic proteins. Subcellular localization prediction showed that more than 80% of CCoAOMT subfamily members were located in the cytoplasm, and 96% of COMT subfamily members were located in the chloroplast. COMT subfamily members have more motifs than CCoAOMT subfamily members. The N-terminal motifs of COMT subfamily proteins are relatively variable, while the C-terminal motifs are relatively conserved. Expression pattern analysis showed that CCoAOMT subfamily members were mainly expressed in roots, while COMT members were mainly expressed in leaves. Some FOMTs showed the tissue expression specificity by real-time quantitative PCR analysis, especially in leaves. In this study, we identified and analyzed the FOMT gene family in A. argyi, and provided a theoretical basis for further research on the function of FOMTs and the biosynthesis of methylated flavonoids in A. argyi.

Analysis and evaluation of volatile oil content in leaves of different Artemisia argyi germplasm resources / 中国中药杂志

Volatile oil is the main effective component and an important quality indicator of Artemisia argyi leaves. In this study, 100 germplasm resources of A. argyi were collected from all the related habitats in China. The total volatile oils in A. argyi leaves were extracted by steam distillation and the content was determined by GC-MS. The result demonstrated that the content of total volatile oils was in the range of 0.53%-2.55%, with the average of 1.43%. A total of 39 chemical constituents were identified from the volatile oils, including 13 shared by the 100 germplasm resources. Clustering analysis of the 39 constituents showed that the 100 A. argyi samples were categorized into groups Ⅰ(9), Ⅱ(2), Ⅲ(66) and Ⅳ(23), and group Ⅲ had the most volatile medicinal components, with the highest content. Five principal components(PCs) were extracted from 13 shared constituents, which explained 73.454% of the total variance. PC1, PC2, and PC3 mainly reflected the pharmacological activity of volatile oils and the rest two the aroma information. The volatile oils identified in this study lay a foundation for variety breeding of and rational utilization of volatile oils in A. argyi leaves.

Diversity of Artemisia argyi germplasm resources based on agronomic and leaf phenotypic traits / 中国中药杂志

In this study, in order to evaluate the phenotypic diversity of Artemisia argyi germplasm resources and improve its efficiency of cultivation and breeding, 100 accessions of A. argyi germplasm resources from 58 regions in China were collected, 20 agronomic traits and leaf phenotypic traits were observed and described. The data were used for phenotypic diversity analysis, correlation analysis, principal component analysis and cluster analysis. The result showed that the genetic diversity index of 20 traits ranged from 0.82 to 4.37, among which the largest was the base depth and the smallest was the leaf width; the coefficient of variation of the 12 quantitative traits ranged from 10.55% to 41.47%. the highest coefficient of variation was the height of dead leaves, and the smallest was the content of chlorophyll, except for the angle of branches, all the quantitative characters tended to be normal distribution. The correlation analysis showed that 28 pairs of traits had significant correlation(P<0.01), and 13 pairs had significant correlation(P<0.05). According to principal component analysis, 20 traits were simplified into 9 principal components, and the cumulative contribution rate was 73.414%, nine traits including plant height, dead leaves heigh, stem diameter, symmetry of leave base, stipule, leaf tip shape, depth of the first pair of lobes, depth of the second pair of lobes and leaf yield were selected as key indexes for evaluating agronomic traits and leaf phenotypic traits of A.argyi germplasm resources. With cluster analysis, 100 accessions of A.argyi were classified into 3 groups, the groupⅠincluded the dwarf plants with thick stem and large leaf, the groupⅡincluded high plants with wide leaf and high yield, the group Ⅲ included dwarf plants with thin stem and flat bottom shape of leaf, which could provide the basis for cultivation identification and variety breeding of A.argyi germplasm resources.

Morphological comparison of glandular and non-glandular trichomes between Artemisia stolonifera and A. argyi / 中国中药杂志

The basic features of glandular and non-glandular trichomes on leaves of Artemisia argyi( germplasms from Qichun,Ningbo,Tangyin,and Anguo,respectively) and related species A. stolonifera were observed by scanning electron microscopy( SEM)and compared. There were significant differences in trichome characteristics of leaves at all parts of A. argyi and A. stolonifera,which were closely related to the difference in chemical components. The length of non-glandular trichomes and size of glandular trichomes on middle leaves were the stablest. A. argyi and A. stolonifera can be distinguished by the density of glandular trichome. Additionally,the four germplasms of A. argyi can be discriminated via the density and curvature of non-glandular trichome. The density of non-glandular trichomes was the highest in A. stolonifera. For A. argyi,the germplasm from Qichun had the highest density of non-glandular trichomes on the abaxial surfaces of upper leaves and that from Ningbo had the largest non-glandular trichome curvature. With regard to the germplasm from Anguo,the T-shaped non-glandular trichomes of long stalks on the adaxial surfaces of the middle leaves were lodging-susceptible,and those with slender heads were wave-like. Statistics results of A. argyi and A. stolonifera are as follows: largest glandular trichomes on the adaxial and abaxial surfaces and highest glandular trichome density on the abaxial surfaces of the lower leaves in A. argyi germplasm from Ningbo,highest density of non-glandular trichomes on the abaxial surfaces of upper leaves in A. stolonifera,and highest density of glandular trichomes and non-glandular trichomes on the adaxial surfaces of the upper leaves in A. argyi germplasm from Qichun. According to the observation result under fluorescence microscope( FM),flavonoids were closely related to the size and density of non-glandular trichomes and size of glandular trichomes. The fluorescence intensity was the strongest and fluorescence area was the largest for flavonoids in A. argyi germplasms from Qichun and Tangyin,while the fluorescence for flavonoids was the weakest in A. stolonifera. It was the first time to observe and analyze the trichome ultrastructure of A. argyi leaves at different positions by SEM and FM. This study clarifies the differences between A. stolonifera and four famous A. argyi germplasms,which provides new evidence for the microscopic identification of A. argyi and its related species and serves as a reference for the study of the relationship of A. argyi structure with its components and functions.

[Comparison of chemical components between Artemisia stolonifera and Artemisia argyi using UPLC-Q-TOF-MS].

The chemical component information of samples was analyzed by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS). The leaves and flosses of Artemisia argyi and A. stolonifera from different places, were distinguished by principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA). Then, compounds with significant differences(P<0.01) in abundance were screened out according to their VIP values and t-test. Eighteen components in all samples have been filtered and identified, including flavonoids and chlorogenic acids, the content of the 12 of which were measured by UPLC-UV, which are different in presence and content. Hispidulin in A. argyi is not detected in A. stolonifera. Schaftaside, isochlorogenic acid B, and isochlorogenic acid C are differential compounds of A. argyi and A. stolonifera leaves. Isochlorogenic acid A, isochlorogenic acid C and jaceosidin are differential compounds of A. argyi and A. stolonifera floss. There are significant differences in the contents of jaceosidin and schaftoside in the four famous A. argyi. In addition, the content of isochlorogenic acid A in wild A. stolonifera is higher than that in cultivated A. stolonifera. The results of the study successfully clarified the differences between A. stolonifera and A. argyi, and provided theoretical and data references for the further development and utilization of A. stolonifera.

[Textual research on traditional Chinese medicine Jiuniucao].

Jiuniucao in Qizhou, known as "Qiai", was precious and expensive in the Ming and Qing Dynasties. But the authentic plant of Jiuniucao is not mentioned in the Ben Cao Tu Jing and other medical books in the Song, Ming and Qing Dynasties. In history, mugwort leaf originates from many species of plants, Jiuniucao may be one of it. So this paper is to identify the original plant of Jiuniucao and clarify the historical origin of Jiuniucao and mugwort leaf. The textual research and geographical origin analysis of Jiuniucao in ancient literature was conducted. Then field investigation and sampling of Jiuniucao in Qizhou was proceeded, combing with feature comparison of Jiuniucao and Artemisia specimen in the Herbarium of Institute of Botany, Chinese Academy of Sciences. The study concluded that A. stolonifera is the authentic Jiuniucao for medical use. Jiuniucao was also an important original plant of mugwort leaf and it is worth further development and utilization.

Analysis of varieties and standards of Composite medicinal plants used in Dai medicine / 中国中药杂志

The study aims to analyze the varieties and standards of compositae medicinal plants used in Dai medicine. The results showed that there were 78 species (including varieties) compositae plants recorded in literatures, which belongs to 63 medicinal materials varieties. And 47 original plants (60.25%) were recorded in Chinese medicinal material standards. In those standards and literatures of Dai medicine, there are great differences in translated Chinese names, original plants, medicinal parts, and efficacy of medicinal plants. Therefore, the variety systematization and the quality standards of Dai medicine should be strengthened.