Identification, biological characteristics, and fungicide screening of pathogen of black spot in Aconitum carmichaelii / 中国中药杂志

In Hezhang county, Guizhou province, black spot tends to occur to Aconitum carmichaelii in the hot rainy summer, with the incidence up to 50%-70%, seriously impacting the yield and quality of the medicinal material. Thus, this study aims to clarify the pathogen and the occurrence characteristics. To be specific, the pathogen was isolated and identified according to Koch's postulates and the pathogenicity and biological characteristics were determined. In addition, the sensitivity of the pathogen to four microbial fungicides, four botanical fungicides, and five chemical fungicides was determined with the mycelium growth rate method for the purpose of screening out optimal fungicides. The pathogen was identified as Alternaria alternate, as evidenced by the similar colony morphology and microscopic characteristics and 99.55%-100% similarity in sequences of rDNA-ITS, LSU, 18S, and TEF of the two. The optimum growth conditions for A. alternata were 28 ℃, pH 8, and continuous darkness. Bacillus subtilis had strong inhibitory effect on the pathogen, and the inhibition rate was more than 90% when the concentration was 1 mg·L~(-1). In addition, difenoconazole and quinoline copper can also control the pathogen, with median effective concentration(EC_(50)) of 2.92 and 9.02 mg·L~(-1), respectively. This study lays a theoretical basis for the field control of black spot in A. carmichaelii.

Content of mineral elements in different Artemisia argyi germplasms and their relationship with quality properties / 中国中药杂志

To clarify the content characteristics of mineral elements in different Artemisia argyi germplasm resources and their relationship with the quality properties of Artemisiae Argyi Folium, this study measured the content of 10 mineral elements including nitrogen(N), phosphorus(P), potassium(K), calcium(Ca), magnesium(Mg), aluminum(Al), manganese(Mn), iron(Fe), copper(Cu), and zinc(Zn) in 100 Artemisia argyi germplasm samples. Besides, their relationship with the quality properties of Artemisiae Argyi Folium was explored by correlation analysis, path analysis, and cluster analysis. The results demonstrated that the variation coefficient of the 10 mineral elements in Artemisiae Argyi Folium ranged from 12.23% to 64.38%, and the genetic diversity index from 0.97 to 3.09. The genetic diversities of N, P, and Zn were obvious. As revealed by the correlation analysis, N, P, and K showed strong positive correlations with each other. Except that Mg and Al were negatively correlated, Ca, Mg, Al, Mn, Fe, Cu, and Zn were positively correlated. The correlation analysis of mineral elements with the quality properties of Artemisiae Argyi Folium proved the significant correlations of 17 pairs of characters. According to the path analysis, P, K, Ca, and Mn greatly affected the yield of Artemisiae Argyi Folium, P, K, and Mg the output rate of moxa, N, P, and K the content of total volatile oil, P and K the content of eucalyptol, and P, K, and Ca the content of eupatilin. The 100 germplasm samples were clustered into three groups. Specifically, in cluster Ⅰ, the enrichment capacity of P, K, and Mg elements was strong, and the comprehensive properties of mineral elements were better, implying good development potential. Ca, Mn, Fe, and Zn elements in cluster Ⅱ and N and Al in cluster Ⅲ displayed strong enrichment capacities. This study has provided new ideas for resource evaluation and variety breeding of A. argyi and also reference for fertilizer application.

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.

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.

Identification,biological characteristics and fungicide screening of pathogen of southern blight in Cynanchum stauntonii / 中国中药杂志

During the high-temperature and rainy season from June to October in 2017-2019,serious southern blight broke out in the Cynanchum stauntonii planting area in Tuanfeng county,Hubei province,which had a great impact on the yield and quality of medicinal materials. In this study,the pathogen of C. stauntonii was isolated,purified,and identified,and the pathogenicity was tested according to Koch's postulates. Meanwhile,the biological characteristics of the pathogen were analyzed. On this basis,the effective fungicides were screened in laboratory. Finally,the pathogen( BQ-1) was identified as Athelia rolfsii( Deuteromycotina,Basidiomycota,anamorph: Sclerotium rolfsii). The optimum growth conditions for BQ-1 were 25-30 ℃,p H 5-8,and alternating light and dark.The effective chemical fungicides were lime-sulphur-synthelic-solution( LSSS) and flusilazole,and the effective botanical fungicide was osthole. BQ-1 was highly homologous to the pathogen HS-1 of peanut southern blight,with the similarity of 18 S r DNA and TEF sequences at 99. 09%. The southern blight in C. stauntonii might be resulted from that in peanut. In the production of C. stauntonii,the following measures should be taken: avoiding rotation or neighboring with peanut,draining water from June to October to reduce humidity,and reasonably applying fungicides.

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.

Research strategies for endophytes in medicinal plants based on high-throughput sequencing and traditional culture and isolation methods / 中国中药杂志

The research on endophytes of medicinal plants mainly relies on the traditional culture and isolation methods. Because of their functions such as promoting host growth, improving stress resistance, promoting the accumulation of medicinal active ingredients or directly producing medicinal active ingredients, the endophytes of medicinal plants have gradually attracted wide attention. However, it was found that the strains isolated by traditional methods were not the true dominant endophytes of medicinal plants by comparing the results of traditional culture isolation with high-throughput sequencing. The blind and random nature of traditional methods leads to the lack of standards in terms of medium selection, culture time and interaction between species. On the contrary, high-throughput sequencing technology is an emerging molecular biology technology developed in recent decades. Due to its high resolution level and indepen-dent culture, it can be used for thorough analysis of the community structure and diversity of environmental microorganisms. Therefore, we proposed the strategy of using high-throughput sequencing technology to guide the traditional culture and isolation of endophytes from medicinal plants. Firstly, the endophytic structure and diversity of medicinal plants were completely clear by high-throughput sequencing technology, and the dominant endophytes of the host were unequivocal. Then according to the characteristics of each dominant endophytes design or query suitable medium for its growth to culture and isolation. Finally, the function of the isolates was studied. This method can prevent researchers from missing out on the important functional strains of the host, expand the research scope of endophytes of medicinal plants, and facilitate the in-depth excavation and utilization of endophytes of medicinal plants.

Effect of different drying methods on drying characteristics, appearance and active components of Belamcandae Rhizoma / 中国中药杂志

In order to explore the effect of different drying methods(drying-in-the-shade, sun-drying, and hot air drying) on appearance characteristics, internal structure and composition of Belamcandae Rhizoma, so as to provide a theoretical basis for screening out suitable drying methods for primary processing. In this study, the Belamcandae Rhizoma's dynamic changes of the moisture content ratio and drying rate with different drying time under different drying methods, as well as the effects of different drying methods on the appearance, drying rate, density, ash, extractives and the contents of six flavonoids(mangiferin, tectoridin, iridin, tectorigenin, irigenin, irisflorentin) were compared. The results showed that fresh Belamcandae Rhizoma consumed the longest time to reach the water balance point by traditional dry drying in the shade, whiche was about 311 h; that by sun drying was 19.3%, which was shorter than drying in the shade; both drying curves were smoother. The section color of the sun drying samples was the closest to that of fresh samples, but the interior is full of holes, with a low density and loose structure. Hot air drying(40, 60, 80 ℃) could save about 27% to 88% of the drying time, which was greatly shorter, with less pores, a larger density and compact structure. Compared with the traditional drying method, the drying rate of hot air drying was reduced by 13.7%. Ash was affected by temperature, the drying conditions under 40 ℃ and below were not significantly different from those of conventional drying. The ash content decreased by 7.73% to 18.5% compared with conventional drying at 60,80 ℃. After conventional drying and 40 ℃ hot air drying, the contents of tectoridin and iridin(glycosides) in the samples were significantly higher than those in 60,80 ℃ hot air drying, while the contents of tectorigenin, irigenin and irisflorentin(aglycones) dried at 60 ℃ were the best. Therefore, considering comprehensive appearance characteristics and content of medicinal ingredients, traditional Chinese medicinal materials after 60 ℃ hot air drying show a solid texture, tight internal structure, good appearance, appropriate reduction of toxic parasides and higher aglycone content.

Identification and biological characteristics of southern blight causing root rot on three medicine plants of Iridaceae in Dabie Mountains / 中国中药杂志

In order to identify the species and biological characteristics of the pathogen of southern blight from three kinds of Chinese medicine of Iridaceae(Belamcanda chinensis, Iris tectorum and I. japonica) in Dabie Mountains, the isolation, identification, pathogenicity and biological characteristics of the pathogens were studied according to Koch's postulates. In addition, 9 chemical fungicides, 3 botanical fungicides and 5 microbial fungicides were used to evaluate their inhibition to the isolates in vitro. The results showed that all the strains(SG-Q, YW-Q, and HDH-Q) isolated and purified from the diseased plants of B. chinensis, I. tectorum and I. japonica, respectively, were identified as Sclerotium rolfsii through morphological observation and sequence aligement of 18 S rDNA, rDNA-ITS and TEF. Field observations showed that the intensity of the disease incidence of three Iridaceae plants was B. chinensis>I. japonica> I. tectorum, and the pathogenicity of the strains was SG-Q>YW-Q>HDH-Q. For biological characteristics, SG-Q strain was suitable for growth under the 12 h light/12 h dark cycle, with the optimal growth temperature of 30 ℃ and pH of 5. Among the 9 tested chemical fungicides, 29% lime sulphure and 10% flusilazole had stronger inhibitory effect on mycelia growth of SG-Q. For 3 botanical fungicides, 1% osthol, 20% eugenol and 0.5% berberine could effectively inhibt the mycelial growth of SG-Q and cause the morphological variation of the pathogen. For 5 microbial fungicides, Trichoderma harzianum and Bacillus subtilis had better inhibition on the mycelium growth of SG-Q.

Content characteristics of mineral elements and its correlation with soil nutrients and effective components in Chrysanthemum morifolium from different producing areas in Macheng / 中国中药杂志

In this study, 24 copies of samples of Chrysanthemum morifolium and soil from two main production towns in Macheng city were collected, and the contents of 13 mineral elements, 5 effective components and 14 soil nutrient factors in Ch. morifolium were determined. The enrichment characteristics of available soil nutrients by mineral elements were analyzed and the dominant factors affecting the effective components of Ch. morifolium were screened. The results showed that the content of mineral elements and soil nutrients and effective components are very different, and variation of soil fertility was much greater than that of inorganic elements in chrysanthemum plants. In general, the level of element content in Ch. morifolium from different producing areas is K>N>P>Mg>Ca>Fe>Mn>Zn>Cu>Ni>Cr>Pb>Cd. The content of K, N and Mg is higher than that of common crops, and the content of Cu, Cd and Pb in Ch. morifolium from various producing areas does not exceed the relevant standards. The N, P and K enrichment capacity in soil was stronger than that of other elements, and the Ca enrichment ability was the worst. The content of AvCu in the soil was positively correlated with the contents of N, Mg, K, Fe and Cu elements in Ch. morifolium. The contents of chlorogenic acid, luteolin, 3,5-O-dicaffeoylquinic acid reached the pharmacopoeia standard. The percentage of chlorogenic acid and 3,5-O-dicaffeoylquinic acid in Ch. morifolium that from Huangtugang town in the active components were generally higher than that from Futianhe town, and the diffe-rences of luteolin contents in the two producing areas were relatively small. The correlation and regression analysis showed that the contents of Cu, Zn and Cr in Ch. morifolium were positively correlated with the active components, while the contents of Fe, Mn and Ni were negatively correlated with the contents of AvP, AvK, TK, AvMn and AvCu in soil. In general, Zn and Ca fertilizer should be added to the ecological planting of Ch. morifolium, K fertilizer should be added, and N and P fertilizer should be applied appropriately.

Analysis and evaluation of mineral elements of Chrysanthemum morifolium for medicinal and tea use of different germplasm resources / 中国中药杂志

In this study, 23 germplasm resources of Chrysanthemum morifolium used in medicine and tea were collected from Dabie Mountains and its surrounding producing areas, and the contents of 13 mineral elements were determined and compared. The thermal maps of correlation analysis, principal component analysis and cluster analysis were used for comprehensive evaluation. The results showed that the average content of each element in Ch. morifolium of different germplasm resources was: K>N>P>Mg>Ca>Fe>Mn>Zn>Cu>Ni>Cr>Pb>Cd, and the leaves were: K>N>Ca>Mg>P>Fe>Mn>Zn>Cr>Cu>Ni>Pb>Cd. There are rich contents of N, P, K, Ca, Mg and Fe in Ch. morifolium flowers and their leaves, among them, K element has the largest change range, while N, Ca, Fe, Mg and Zn elements have a larger change range. The absorption and accumulation of each element in the leaves of different germplasm resources varied greatly. The correlation analysis shows that there is a strong positive correlation between Ca element, Mg, Mn and Cd element.Principal component analysis in Ch. morifolium flowers characteristic elements for Mn, Cr, Cu, P, K, can be used as a Ch. morifolium resources to identify the characteristics of the elements, choose top five principal component(F1-F5) comprehensive evalua-tion of medicinal Ch. morifolium, scored in the top five varieties for Hangiu-Fuhuangju, Hangju-Xiaoyangju, Hangju-Sheyangju, Hangju-Dayanghua, Hangju-Subeiju,indicates that in terms of mineral elements, the five medicinal Ch. morifolium resources quality is better. The PCA score chart can divide 23 Ch. morifolium resources into 4 groups, and the cluster analysis heat map divides 23 Ch. morifolium resources into 5 groups. All the Ch. morifolium resources of the same type can be well clustered together, indicating that the difference in mineral element content of Ch. morifolium germplasm resources is closely related to genetic factors.

Analysis on Seed Quality Grading Standard of Belamcanda chinensis / 中国实验方剂学杂志

Objective:To study the quality inspection standard of seeds of <italic>Belamcanda chinensis</italic> and establish the quality grading standard of seeds of <italic>B. chinensis</italic>. Method:The purity of seeds of <italic>B. chinensis</italic> from different producing areas was analyzed，and the 1 000-grain weight，water content，viability and germination rate of seeds of <italic>B. chinensis</italic> with different diameters were determined after diameter classification. K-means was used for cluster analysis to preliminarily formulate the quality grading standard of seeds of <italic>B. chinensis</italic>. Result:There were obvious regional differences in the size of seeds of <italic>B. chinensis</italic>. The particle size of seeds of <italic>B. chinensis</italic> of Xiaochang，Dawu and Huanggang Academy of Agricultural Sciences in Hubei Province was larger and mostly between 4.5 mm to 5.5 mm. The seeds of<italic> B. chinensis </italic>from Tuanfeng，Qujing，Yunnan，and Anguo，Hebei province had smaller particle sizes than those from the other three producing areas，which were mostly between 3.5 mm to 5.0 mm. Arils in seeds of <italic>B. chinensis</italic> were removed by 10% NaClO to effectively break seed dormancy and significantly improve the seed germination rate. On this basis，the seed diameter was taken as the main grading index，and the seed moisture content，viability，germination rate and purity were taken as important reference indexes. The quality of seeds of <italic>B. chinensis</italic> was preliminarily divided into three grades，grade Ⅰ：seed diameter≥5.5 mm，moisture content≤10%，viability≥94%，germination rate≥60%，and cleanliness≥93%；grade Ⅱ：seed diameter≥4.5 mm，moisture content≤10%，viability≥90%，germination rate≥55%，and cleanliness≥85%；and grade Ⅲ：seed diameter≥3.5 mm，moisture content≤10%，viability≥84%，germination rate≥45%，cleanliness≥80%. Conclusion:In this study，the quality grading standard of seeds of <italic>B. chinensis</italic> was preliminarily established to provide reference for the quality evaluation of seeds of <italic>B. chinensis</italic> and the breeding of improved varieties. In addition，the maturity and the storage time of seeds of <italic>B. chinensis</italic> have a greater impact on the quality of seeds，so it is recommended to select fully mature（dark black） seeds and new seeds for production.

Analysis and Evaluation on Leaf Quality of Different Artemisia argyi Germplasm Resources / 中国实验方剂学杂志

Objective:To analyze the quality differences among different germplasm resources of <italic>Artemisia argyi </italic>and to screen out the specific germplasm by comprehensively evaluating 14 quality traits of 100 germplasm resources. Method:Germplasm resources of <italic>A. argyi </italic>were collected from all over the country. The output rate of moxa and the content of total volatile oil in <italic>A. argyi</italic> leaves were determined，and the contents of 12 flavonoids and phenolic acids in<italic> A. argyi </italic>leaves were detected by ultra performance liquid chromatography（UPLC）. The correlation analysis，principal component analysis and clustering analysis were used to comprehensively evaluate the quality of <italic>A. argyi</italic>. Result:There was rich genetic diversity of<italic> A. argyi</italic> germplasm resources，and the variation coefficients of 14 quality traits ranged from 25.67% to 127.34%，among which the coefficient of variation of chlorogenic acid，cryptochlorogenic acid，isoxiafotaside，isochlorogenic acid B and isochlorogenic acid A was more than 70%，with high variation. The output rate of moxa was negatively correlated with 9 quality traits，while the content of total volatile oil was positively correlated with 10 quality traits，and most of the flavonoids and phenolic acids had synergistic effects. 12 flavonoids and phenolic acids were analyzed by principal component analysis，and 4 principal components could be extracted. The highest contents of flavonoids and phenolic acids were found in S98（Hangzhou，Zhejiang province），S84（Longhui county，Shaoyang city，Hunan province），S66（Futian river town，Macheng city，Hubei province），S35 （Balihu town，Qichun county，Huanggang city，Hubei province），and S15 （Fudao town，Tangyin county，Anyang city，Henan province）. The systematic clustering analysis showed that the 100 germplasm could be divided into four groups when the euclidean distance was 8.0，with 90，3，3，3 and 4 accessions in group Ⅰ，Ⅱ，Ⅲ and Ⅳ，respectively. The germplasm resources in group Ⅱ contained the highest content of flavonoids and phenolic acids，the group Ⅲ contained the highest content of total volatile oil and the group Ⅳ contained the highest output rate of moxa. Conclusion:The leaf quality of different <italic>A. argyi </italic>germplasms is different. This study can provide the basis for the quality evaluation and variety breeding of <italic>A. argyi</italic> germplasm resources.

Fusarium Wilt Changes Microbial Community Structure in Rhizosphere Soil of Chrysanthemum morifolium / 中国实验方剂学杂志

Objective:To explore the differences in rhizosphere microbial community structure between <italic>Fusarium</italic> wilt-infected and healthy <italic>Chrysanthemum morifolium </italic>plants<italic>.</italic> Method:The rhizosphere soils of diseased and healthy<italic> C. morifolium </italic>plants were sampled and subjected to high-throughput 16S ribosomal DNA （rDNA） and internal transcribed spacer （ITS） sequencing， to identify the microbial community structure including bacteria and fungi. Result:<italic>Fusarium</italic> wilt reduced the bacterial abundance and diversity but had no significant effect on fungal alpha-diversity.The proportions of Acidobacteria， Gemmatimonadetes， and Nitrospirae in rhizosphere soil of healthy <italic>C.morifolium</italic> plants were higher than those of diseased plants， while the proportions of Proteobacteria and Bacteroidetes were lower（<italic>P</italic><0.05）. <italic>Fusarium</italic> fungi accounted for 27.49%， 14.53%， and 11.94% in diseased plants whereas 0.47%， 1.01%， and 0.67% in healthy plants.Pathogenic bacteria <italic>Pectobacterium</italic> and <italic>Dickeya</italic> were enriched in rhizosphere soil of diseased plants. The abundances of nitrifying， detoxifying， and photosynthetic bacteria in rhizosphere soil of healthy plants were higher than those of diseased plants. Conclusion:<italic>Fusarium</italic> wilt reduces the bacterial richness and diversity and triggers the enrichment of massive <italic>Fusarium</italic> fungi， <italic>Pectobacterium</italic>， and <italic>Dickeya</italic>. The proportion of beneficial bacteria in rhizosphere soil of healthy plants is significantly higher than that of diseased plants.

Analysis and Evaluation of Mineral Elements in Gastrodia elata with Different Specifications and Grades from Diverse Producing Areas / 中国实验方剂学杂志

Objective:To study the distribution characteristics of mineral elements in <italic>Gastrodia elata </italic>samples<italic> </italic>with different grades and specifications （variants） from diverse producing areas and their classification and identification evidences. Method:Fourteen mineral elements in 31 batches of <italic>Gastrodia elata</italic> samples of different grades and specifications （variants） from diverse producing areas were determined by atomic absorption spectrophotometry， Mo-Sb colorimetry， and curcumin colorimetry， and then subjected to correlation analysis （CA）， discriminant analysis （DA）， and principal component analysis （PCA）. Result:The content of K， N， and P in <italic>G. elata</italic> was the highest， enabling them to serve as the nutritional limiting factors affecting its growth. The <italic>G. elata</italic> samples could be identified by the variation trend of elements （K>N>P>Mg>Ca>Fe>B>Zn>Mn>Cu>Ni>Cr>Pb>Cd）. The comparison of <italic>G. elata</italic> samples from multiple producing areas showed that <italic>G. elata</italic> from Zhaotong has the highest P， Fe， and Cd content， that from Lijiang the highest K content， that form Luotian the highest Zn and Cr content， and that from Jinzhai the highest Cu and Pb content. The content of Mg， B， Pb， and Cr in <italic>G. elata</italic> f. <italic>elata</italic> was higher than that in <italic>G. elata </italic>f. <italic>glauca</italic>. It was found that the content of P， Cu， and Cd in commercially available <italic>G. elata </italic>products gradually increased with the decrease in the commercial grade， while that of Mg， Fe， B， and Ni mostly decreased. As revealed by CA， Fe was positively correlated with Mg， Cr， and B. The producing areas of <italic>G. elata</italic> samples could be effectively identified by DA with Cd， Pb， Cr， Cu， B， and Ni as the main variables， and the accuracy reached up to 85.71%. According to the PCA of mineral elements in <italic>G. elata</italic> f. <italic>glauca</italic> from Zhaotong， Yunnan Province， Fe， Cr， Mg， Cd， P， Mn， B， Pb， and Cu exerted a greater influence on <italic>G. elata</italic>. Conclusion:The determination of mineral elements in <italic>G. elata</italic> samples contributes to identifying their authenticity and origin due to the easy operation， accurate results， and good stability.

Effects of Combined Application Organic Fertilizer and Chemical Fertilizer on Agronomic Characters，Yield and Quality of Artemisiae Argyi Folium / 中国实验方剂学杂志

Objective:To explore the effect of different proportions of organic fertilizer and chemical fertilizer on the agronomic traits， yield and quality of Artemisiae Argyi Folium in Qichun county， and provide a theoretical basis for scientific fertilization of its planting. Method:Field plot experiment was carried out to set up 5 treatment methods with different proportions of organic fertilizers and chemical fertilizers［OM₀ （no combined application of biological organic fertilizer）， OM₁₇ （combined application of 17% biological organic fertilizer）， OM₃₃ （combined application of 33% biological organic fertilizer）， OM₆₇ （combined application of 67% biological organic fertilizer）， OM₁₀₀ （combined application of 100% biological organic fertilizer）］. The effects of different treatment methods on the agronomic characters， leaf yield， output rate of moxa， volatile oil content， flavonoid and phenolic acid contents and mineral element contents of Artemisiae Argyi Folium in Qichun county were determined. Result:With the increase of the proportion of organic fertilizer in application， the seedling number per unit area， plant height， stem diameter， leaf number， leaf width， leaf length， height of dead leaves and leaf yield of Artemisiae Argyi Folium were increased at first and then decreased. Among them， the yield of Artemisiae Argyi Folium in OM₃₃ treatment was 61.37% higher than that in OM₀ treatment. With the increase of the proportion of organic fertilizer， the output rate of moxa of Artemisiae Argyi Folium showed continuously increasing trend， contents of volatile oil and volatile components （eucalyptol， <italic>α</italic>-thujone， borneol， camphor and caryophyllene oxide） increased at first and then decreased， while the contents of <italic>α</italic>-caryophyllene and <italic>β</italic>-syringene decreased gradually， the contents of phenolic acids （neochlorogenic acid， chlorogenic acid， isochlorogenic acid B， A and C） increased at first and then decreased， while the contents of flavonoids （jaceosidin and eupatilin） increased continuously， and the contents of mineral elements （Ca， Cu and Zn） continued to increase， but the content of K decreased significantly at the high proportion of organic fertilizer. After treated with principal component analysis （PCA）， it was found that OM₁₇ treatment had the highest quality， while OM₁₀₀ and OM₀ treatment had low quality. Conclusion:Based on comprehensive analysis of agronomic traits， yield and quality indexes of Artemisiae Argyi Folium in Qichun county， it is suggested that 17%-33% proportion of organic fertilizer should be used in its production， in order to improve the quality and efficiency of Artemisiae Argyi Folium industry in Qichun county.

Active fractions of Camellia nitidissima inhibit non-small cell lung cancer via suppressing epidermal growth factor receptor / 中国中药杂志

The present study explored the effects and its underlying mechanisms of four active fractions of Camellia nitidissima(leaf polyphenols, leaf saponins, flower polyphenols, and flower saponins in C. nitidissima) in inhibiting the proliferation and migration of non-small cell lung cancer(NSCLC) by suppressing the epidermal growth factor receptor(EGFR). MTT assay was used to detect the effect of four active fractions on the proliferation of NCI-H1975 and HCC827 cells. Wound healing assay and Transwell assay were adopted to evaluate the effect of four active fractions on the migration of NSCLC. The effect of four active fractions on the enzyme activity of EGFR was detected. Molecular docking was carried out to explore the direct action capacity and action sites between representative components of the four active fractions and EGPR. Western blot assay was employed to investigate the effect of four active fractions on the protein expression in EGFR downstream signaling pathways. The results of the MTT assay indicated that the cell viability of NCI-H1975 and HCC827 cells was significantly inhibited by four active fractions at 50, 100, 150, and 200 μg·mL~(-1) in a dose-dependent manner. Wound healing assay and Transwell assay revealed that the migration of NCI-H1975 and HCC827 cells was significantly suppressed by four active fractions. In addition, the results of the protein activity assay showed that the enzyme activity of EGFR was significantly inhibited by four active fractions. The molecular docking results confirmed that various components in four active fractions possessed strong binding activity to EGFR enzymes. Western blot assay revealed that four active fractions down-regulated the protein expression of EGFR and its downstream signaling pathways. It is concluded that the four active fractions of C. nitidissima can inhibit NSCLC. The mechanism may be related to EGFR and its downstream signaling pathways. This study provides a new scientific basis for the clinical treatment of NSCLC with active fractions of C. nitidissima, which is of reference significance for further research on the anti-tumor mechanism of C. nitidissima.

Research progress and prospect of endophytes from medicinal plant Atractylodes lancea / 中国中药杂志

The endophytes of medicinal plants play an important role in promoting the quality formation of the host. Therefore, this paper made a review of endophytes of medicinal plant Atractylodes lancea. According to previous studies, A. lancea boasts endophytes, such as fungi, bacteria, and actinomycetes, among which the beneficial microorganisms help the growth and development of A. lancea. There is a close interaction between the volatile oil of A. lancea and endophytes. Different endophytes vary in regulating the composition and content of the volatile oil of A. lancea, which might contribute to the quality formation of A. lancea. However, the information of the endophytic flora of A. lancea obtained by traditional culture and isolation is not enough to reflect the real situation of the endophytes of A. lancea. Little is known about the endophytes of A. lancea from different chemical types and different habitats, which is not conducive to the study of the ecological relationship between A. lancea and endophytes and limits the development and utilization of the endophytes. Therefore, at the end of this paper, the authors put forward suggestions for future research on endophytes in A. lancea, including:(1)mining the core endophyte resources of A. lancea by combining high-throughput sequencing with traditional culture and isolation;(2)exploring the relationship between the diversity of endophytes and chemical types of A. lancea;(3)strengthening the application of endophytes in A. lancea cultivation, in order to facilitate the cultivation efficiency and quality of A. lancea.

Isochlorogenic acid (ICGA): natural medicine with potentials in pharmaceutical developments / 中国天然药物

Natural products have attracted a great deal of attention as significant resources in traditional Chinese medicine (TCM) and in chemical medicine, as well as in cosmetic ingredients, nutraceuticals and food products. Isochlorogenic acid (ICGA), which has medicinal value, has been discovered in various plants. As a widespread natural medicine, ICGA should be the subject of further research and development. However, there have been no systematic analyses of ICGA. According to our investigation, ICGA was initially isolated from green coffee extracts by Barnes et al. in 1950. To date, it has been discovered in a variety of tea, vegetables, medicinal diet and TCM materials. ICGA is used as a chemical marker for the quality control of these TCM materials. The metabolic process of ICGA has been studied in detail, conforming to be linear dynamics. Thus, the clear pharmacokinetics of ICGA offers a solid foundation for its research and development. ICGA has multiple biological and pharmacological effects, and studies have mainly focused on its antioxidant, anti-inflammatory, antimicrobial, hypoglycemic, neuroprotective, and cardiovascular protective effects, and hepatoprotective properties. The mechanisms underlying these effects are summarized in this review to provide scientific support and inspiration for the future research and development of ICGA and ICGA-rich natural products.

Isolation and Identification of Mycena of Pleurotus in Wumeng Mountain Area and Optimization of Its Production Conditions / 中国实验方剂学杂志

Objective:Isolate and identify Mycena, expand the resources of geminating fungus of Gastrodia elata and optimize the culture conditions of Mycena，in order to provide information and guidance for the production of geminating fungus of G. elata. Method:Juvenile tuber tissue mass transfer separation and purification technology was used for the separation and purification of strains，traditional morphology microscopy was used to isolate the colony mycelia spores morphological characteristics, such as identification，polymerase chain reaction（PCR） amplification rDNA （Ribosomal DNA） internal transcribed spacer（ITS） was used for sequencing analysis and further homology with NCBI database retrieval，MEGA6 software was used to establish Phylogenetic tree by the Maximum likelihood method （MaximumLikelihood，M-L）, so as to classify and identify isolated strains. At the same time，orthogonal test was used to optimize the optimal growth conditions of Mycena. Result:A total of 86 strains were isolated, which belong to 21 species in 12 genera. WMMFJ，SHXG，WMM-21 and MFJ8103 were identified as M. purpureofusca, and ZT01-6 and ZT01-8 were identified as M. cf. purpureofusca. The growth rate of Mycena in wheat bran medium was significantly higher than in PDA medium. The optimal medium composition for the growth of germinating bacteria was 100 g potato，150 g wheat bran and 20 g corn flour，100 g glucose. And 1，3，5-Trihydroxybenzene significantly promoted the growth of WMMFJ，and played a role in promoting the growth of WMM-21 and ZT01-6，and 2-Methoxyphenol promoted the growth of WMMFJ. Conclusion:Six strains of Mycena were isolated and identified，four of them are M. purpureofusca，and two of them are M. cf. Purpureofusca. The separation method improved the separation effect of germinating bacteria.