Germplasm resources and secondary metabolism regulation in Reishi mushroom (Ganoderma lucidum) / 中草药·英文版

Ganoderma lucidum is a valuable medical macrofungus with a myriad of diverse secondary metabolites, in which triterpenoids are the major constituents. This paper introduced the germplasm resources of genus Ganoderma from textual research, its distribution and identification at the molecular level. Also we overviewed G. lucidum in the components, the biological activities and biosynthetic pathways of ganoderic acid, aiming to provide scientific evidence for the development and utilization of G. lucidum germplasm resources and the biosynthesis of ganoderic acid.

Evaluation of genetic diversity of ginseng fruit color germplasm resources: based on SSR analysis / 中国中药杂志

Illumina Xten was employed for shallow sequencing of Panax ginseng(ginseng) samples, MISA for screening of SSR loci, and Primer 3 for primer design. Polymorphic primers were screened from 180 primers. From the successfully amplified polymorphic primers, 15 primers which featured clear peak shape, good polymorphism, and ease of statistics were selected and used to evaluate the genetic diversity and germplasm resources of 36 ginseng accessions with different fruit colors from Jilin province. The results showed that red-fruit ginseng population had high genetic diversity with the average number of alleles(N_a) of 1.031 and haploid genetic diversity(h) of 0.172. The neighbor-joining cluster analysis demonstrated that the germplasms of red-fruit and yellow-fruit ginseng populations were obviously intermixed, and pick-fruit ginseng germplasms clustered into a single clade. The results of STRUCTURE analysis showed high proportion of single genotype in pick-fruit ginseng germplasm and abundant genotypes in red-fruit and yellow-fruit ginseng germplasms with obvious germplasm mixing. AMOVA revealed that genetic variation occurred mainly within populations(62.00%, P<0.001), and rarely among populations(39%, P<0.001), but homogenization was obvious among different populations. In summary, pink-fruit ginseng population may contain rare genotypes, which is the basis for breeding of high-quality high-yield, and multi-resistance varieties, genetic improvement of varieties, and sustainable development and utilization of ginseng germplasm resources.

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.

Identification and activity evaluation of Astragalus Radix from different germplasm resources based on specific oligosaccharide fragments / 中草药·英文版

Objective: Based on trifluoroacetic acid (TFA) hydrolysis, polyacrylamide gel electrophoresis (PAGE) and high performance thin layer chromatography (HPTLC) analysis, the carbohydrate responsible for immunomodulatory activity are used as quality indicators for Astragalus Radix (AR). Methods: In this study, 24 batches of AR from different germplasm resources were selected as the research object, and AR polysaccharides were extracted. PAGE and HPTLC methods were used to analyze the partial acid hydrolyzate of AR polysaccharides and obtain a series of saccharide fingerprints. The data were analyzed by principal component analysis to obtain the difference between AR from different germplasm resources. Results: The results showed that trisaccharide and tetrasaccharide could be used as differential fragments to distinguish AR of different cultivation methods; Disaccharides and trisaccharides can be used as differential fragments to distinguish different species of AR. The immunological activity analysis of the specific oligosaccharide fragment of AR showed that the specific oligosaccharide fragment of AR could promote the secretion of TNF-α, IL-1β, IL-6, and NO in THP-1 cells in a concentration-dependent manner. Conclusion: Both PAGE and HPTLC methods can be used to evaluate AR from different germplasm resources. This study laid the foundation for the quality evaluation of AR medicinal herbs.

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.

Fruit variation and geographical distribution of citron / 中国中药杂志

The ripe dried fruit of citron(Citrus medica) is one of the important sources of Chinese herb Citri Fructus. At the same time, it is also grown for edible and ornamental uses. There are many species and abundant genetic variation. To clarify the intraspecific variation and resource distribution of citron, this study investigated the variation in 11 citron fruits, basically covering the main species in China, including Xiaoguo citron(C. medica var. ethrog), Goucheng(C. medica var. yunnanensis), Muli citron(C.medica var. muliensis), Dehong citron(C.medica×Citrus spp.), Fuzhou citron(C.medica×C.grandis?), Mawu(C.medica×C.grandis?), Cangyuan citron, Binchuan citron, Sweet citron, Big citron, and Small citron. The natural communities of citron were proved to be mainly distributed in the southwestern and western Yunnan and southeastern Tibet of China, with Yunnan, Sichuan, Guangxi, Chongqing, Hubei, and Zhejiang identified as the main production areas. Citron has also been widely grown in India, the Mediterranean region, and the Caribbean coast countries. The field investigation revealed the large-scale intraspecific variation of citron fruits. Most of the fruits are oval-like or sphere-like in shape. The fruits are green when raw and yellow when ripe, with oil cell dots on the skin, stripe-likes running from top to bottom, and bulge at the top. Usually, in the smaller citron fruits, the pulp and juice vesicles are better developed and the central columella is tighter. By contrast, the juice vesicles and central columella in larger fruits became more vacant, with carpels visible, and the apex segregation and development of the carpels is one of the reasons for variation. These variations should be given top priority in the future variety selection and breeding, and the quality differences of different citron species and their mechanisms should be further studied. In particular, variety selection and classification management according to their medicinal or edible purposes will provide scientific and technological supports for the orderly, safe, and effective production of citron products consumed as food and medicine.

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 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.

Guarantee of Quality and Clinical Efficacy of Chinese Materia Medica Preparation： Control of Quality Transfer Process of Chinese Materia Medica / 中国实验方剂学杂志

Traditional Chinese medicine （TCM） is the treasure of the Chinese nation. As an important raw material for clinical treatment of diseases， Chinese materia medica plays an extremely important role. However， in the process of transformation from traditional wild collection of animals and plants to modern artificial cultivation and industrial production of preparations， whether the quality of Chinese materia medica is fully transferred will directly affect the quality and clinical efficacy of Chinese materia medica preparation. From the field to the sickbed， process control of quality transfer of Chinese materia medica is the key to guarantee quality and curative effect. In this paper， the whole process that affects the quality of Chinese materia medica preparations such as seed and seedling， planting and breeding， harvesting and processing， processing of decoction pieces and preparation production was analyzed. Paying attention to the whole process of quality control of Chinese materia medica is of great significance to improve the quality of Chinese materia medica preparations and promote the rapid development of TCM. Based on this， the author intended to analyze the key control links in the quality transfer process of Chinese materia medica （breeding， planting areas and field management， timely harvesting and intensive primary processing， appropriate processing， optimization of preparation technology， standardization of packaging and informationization of storage and transportation）， in order to provide reference for the design and development of Chinese materia medica preparations guided by clinical value.

Genetic Diversity Analysis of Pogostemon cablin Based on AFLP Markers / 中国实验方剂学杂志

Objective:This paper aims to study the genetic diversity of <italic>Pogostemon cablin</italic> by amplified fragment length polymorphism （AFLP） markers. Method:The 12 pairs of primers were used for AFLP analysis of 212 samples from 14 varieties，and biological analysis software such as POPGENE 32，Arlequinver 3.5，MEGA 7 and NTSYSpc 2.10e were used for polymorphism parameter calculation，principal coordinate analysis and cluster analysis. Result:A total of 2 238 loci were amplified by 12 pairs of primers. 2 226 of them were polymorphic loci， accounting for 99.38%. At the inter-population level，the values of effective alleles（<italic>Ne</italic>），Nei's gene diversity index（<italic>H</italic>），Shannon polymorphic information index（<italic>I</italic>） were 1.365 6±0.066 3， 0.220 7±0.036 4， and 0.343 7±0.050 2，respectively；and 1.118 5±0.038 7，0.071 3±0.023 0，0.109 4±0.035 0，respectively at the intra-population level. Analysis of molecular variance（AMOVA）showed that 71.57% of the total variation of <italic>P. cablin</italic> was of inter-population nature， and 28.43% was of intra-population nature. The 14 populations could be divided into four groups by cluster analysis. Conclusion:The results of AFLP molecular markers showed that abundant genetic diversity was present at inter-population level of <italic>P. cablin</italic>，however，relatively low at intra-population level； the genetic differentiation at the inter-population level was significant，which could provide a reference for the subsequent study of good germplasm selection of <italic>P. cablin</italic>.

Analysis and evaluation on main active ingredients of Asarum heterotropoides var. mandshuricum from different habitats / 中草药

Objective: Active ingredients of 47 Asarum samples from different habitats with different phenotypes were analyzed in this paper in order to reveal the relationship between the formation of active ingredients and genetics and geography, and provide some scientific basis for the breeding and untilizing of Asarum. Methods: Ethanol extract and volatile oil were extracted according to the methods of Chinese Pharmacopoeia. Asarinin was detected by HPLC. Volatile oil compositon was determined by GC-MS. The between-groups linkage clustering method was carried out. Results: Volatile oil content of 47 samples was 0.81%-3.32%, and five samples’ volatile oil content exceeded 3.0 %. Ethanol extract content of samples was 9.87%-29.40%, and ethanol extract content of 30 resources accounting for 63.8 percent of all samples exceeded 20%. Asarinin content of 26 resources was more than 0.25%. Totally 48-77 compounds from the volatile oil could be determined and relative content of each compound was different. Forty-seven samples from different habitats with different phenotypes were divided to ten groups by between-groups linkage clustering method according to the active ingredients. All the accessions could be classified clearly. The cluster of accessions was associated with resource region. Asarum heterotropoides var. mandshuricum and Asarum sieboldii var. seoulense from the same origin were grouped together. Conclusion: There were much more differences in active ingredients content among Asarum resources which were in different regions cultivated with same way and environment with different phenotypes. We could select the perfect fundamental material according to the phenotypes and introduce germplasm from different regions for breeding and utilizing of Asarum. Most of samples clustered together associated with their origins instead of phenotype or specie.

Comprehensive Evaluation of Differences of Pinelliae Rhizoma Germplasm Resources Based on AHP and Grey Correlation Method / 中国实验方剂学杂志

Objective::To establish the evaluation method of Pinelliae Rhizoma (PR) resources by the analytic hierarchy process (AHP) and grey correlation degree (GCD) method, so as to explore the differences in quality components, production efficiency and appearance traits of different germplasm resources cultivated in the same environment. Method::The quality component index, efficiency index, and appearance traits index of 15 germplasm resources were measured, including moisture, total ash, extractives, total acid content, harvest index, yield, drying rate, commodity rate, decay rate and deformability of tubers. Based on AHP and grey correlation method, each indicator data was processed in a comprehensive way, its comprehensive correlation value was calculated, and different PR germplasm resources were comprehensively evaluated. Result::Based on three factors-quality composition, production efficiency and appearance traits, the comprehensive correlation value of A6 was the highest, reaching 0.749 4, which was followed by A14, A15, A7, and their comprehensive correlation values were 0.736 6, 0.726 2, 0.718 2, respectively. Therefore, the source of A6 could be used as an excellent provenance introduced to the cultivation of PR, and the provenance of A14, A15, and A7 could be used as a useful supplement. Conclusion::The method of AHP and GCD-based multi-index comprehensive evaluation is simple and comprehensive to evaluate the diversity of different PR germplasm resources, and could provide a reference for the development and utilization of resources and the screening of high-quality provenances.

Chinese herbal medicine resources: Where we stand / 中草药·英文版

In recent years, the development of Chinese herbal medicine (CHM) has been challenged by shortages of CHM resources and drug safety concerns related to end products. There have been significant efforts by Chinese scholars to tackle these challenges, which are revealed by analyzing the research trend of CHM resources via surveying Chinese Traditional and Herbal Drugs (Zhong Cao Yao), a representative journal in CHM. Our study focused on 781 articles in CHM resources from 2013 to 2018 and included four subject areas: germplasm resources, quality analysis and evaluation, cultivation, and bioengineering of CHM. Discussion and prospective for future investigations were also presented, including: construct the core germplasm of medicinal plants and expand germplasms; combine molecular research with field experiments and promote the deeper study of cultivation of CHM plants; improve the quality evaluation method of CHM and strengthen the identification of Chinese patented medicines; promote the sustainable development of CHM resources by utilizing bioengineering and synthetic biology. This study helps international scholars understand the status quo of CHM research and provides theoretical support for the healthy, modern, and international development of CHM, and it will facilitate the sustainable development of the traditional Chinese medicine industry.

Genetic diversity analysis of germplasm resouces of Paris genus in Shaanxi Province by SCoT molecular makers / 中草药

Objective: To study the genetic diversity and genetic structure of Paris genus in Shaanxi Province. Methods: Start Codon Targeted Polymorphism (SCoT) molecular markers were performed on 48 samples from six taxas of Paris genus in Shaanxi. Genetic distance was calculated by POPGENE 3.2 software, and cluster analysis was generated by NTSYS 2.10 software based on unweighted mean distance method (UPGMA) method. Results: A total of 152 bands were produced by 12 primers, among which 135 bands were polymorphic bands, and the percentage of polymorphic bands was 88.82%. The average value of Nei’s genetic diversity index (H) was 0.267 4, Shannon’s information index (I) was 0.404 1, genetic differentiation coefficient (Gst) was 0.517 9, and the gene flow (Nm) was 0.465 4. Six taxas were ranked by genetic diversity: Paris polyphylla > Paris polyphylla var. latifolia > Paris fargesii var. petiolata > Paris polyphylla var. stenophylla > Paris polyphylla var. yunnanensis > Paris verticillata. Cluster analysis distinguished others from Paris verticillata; When the genetic distance was certain, six taxas of the genus were completely separated. Conclusion: SCoT molecular markers can obtain polymorphic and clear band amplification map, which indicates that the technology can be used for molecular identification and genetic relationship of major taxas in Paris genus in Shaanxi Province, providing a scientific basis for screening alternative resource types close to the Pharmacopoeia collection and guiding the rational use of local species.

Industrialization condition and sustainable development strategies of Bletillae Rhizoma / 中草药

In recent years, the industry of Bletillae Rhizoma has developed rapidly and widely in medicine, health care, beauty, etc., but there are still problems that restrict its sustainable development. Based on field investigating in development status of Bletillae Rhizoma market and industry, consulting relevant literature and our research results, and main constraints were discovered as follows: wild resources shortage, uneven varieties, unclear mechanism, low comprehensive utilization, unclear quality standards, difficulty of quality control, weak industrial level, slow cluster development and so on. Therefore, we can strengthen the protection of germplasm resources and selection of superior varieties, carry out the mechanism of its effective ingredients, improve quality evaluation system and expand sales market using brand strategy to promote the healthy and sustainable development of Bletillae Rhizoma.

Genetic diversity analysis and germplasm identification of Paris species by CDDP markers / 中草药

Objective: To explore genetic diversity and phylogenic relationship among different types of Paris species and provide an effective method for the rapid identification of germplasm resources. Methods: CDDP marker was used to evaluate the genetic diversity and phylogenic relationship among 13 types of Paris species. And the coding of Paris genus plants was carried out based on method of CODE128 barcode. Results: Our results indicated that 73 polymorphic bands were amplified by 11 primers among 80 bands, and the ratio of polymorphic band was 91.25%. The observed number of alleles (Na), effective number of alleles (Ne), Nei’s gene diversity (H), and Shannon’s information index (I) was 1.912 5, 1.589 6, 0.342 3, and 0.507 0, respectively. The results of UPGMA analysis showed that there were great differences among 13 types of Paris species on genetic diversity. Based on CDDP markers, 13 barcode molecular identity cards were constructed for Paris species based on method of CODE128 barcode. Conclusion: There was abundant polymorphism among Paris species, and CDDP markers were effective to analyze the genetic diversity of Paris species, and the established barcode molecular identity for Paris species was sensitive and fast, which can be used for scientific research and industrial production of Paris genus.

AFLP analysis of genetic diversity of Aconitum brachypodum / 中草药

Objective To analyze the genetic diversity of Aconitum brachypodum, an endangered medicinal plants. Methods Three pair primers were used to evaluate the relevant genetic parameters and clustering pattern for four natural populations containing 105 individuals by using POPGENE32, MEGA4, NTSYS and other biometric softwares. Results For A. brachypodum, the Nei’s genetic diversity index (He), Shannon’s information index (I), percentage of polymorphic loci (PPB) were 0.322 9 ± 0.179 8, 0.472 0 ± 0.251 7, and 80.57%, respectively; and the population level were 0.115 4 ± 0.044, 0.168 0 ± 0.065 3, and 12%, respectively. The genetic differentiation index (Gst) was 0.864 2. The clustering results showed that, Luquan and Dongchuan populations are grouped together, Lijiang and Huize populations are clustered together. Conclusion A.brachypodum possesses the relatively high level of genetic diversity at the species level, however, low within populations. This study should provide an initial insights for A.brachypodum germplasm conservation

Establishment of fingerprint of Astragali Radix polysaccharides based on endo-1,3-β-glucanasehydrolysis and identification of Astragali Radix of different germplasm resources / 中草药

Objective Enzymatic hydrolysis of Astragali Radix polysaccharides from different germplasm resources Astragalusmembranaceus var. mongholicus (MG) (cultured and natural) or Astragalusmembranaceus (MJ) (cultured and natural) was carried out by the best enzymolysis conditions of endo-1,3-β-glucanase. Saccharide fingerprints were obtained for the identification and evaluation of the germplasm resources of Astragali Radix by Fluorophore-assisted Carbohydrate Electrophoresis (FACE). Methods The data were analyzed by principal component analysis and t test using SMICA software to distinguishdifferential sugar segments among different germplasm resources of Astragali Radix. Results Pentasaccharide and hexasaccharide of endo-1,3-β-glucanasehydrolyzate could be used as differentiated saccharide fragments between natural MG and MJ.Trisaccharide, tetrasaccharide, and pentasaccharide could be used as differentiated saccharide fragments to distinguish the cultured MG and MJ.The pentasaccharide and hexasaccharide can be used as differential fragments to distinguish MJ (culturedandnatural). Conclusion Thepolysaccharide products degraded by endo-1,3-β-glucanase can well distinguish Astragali Radix species (MG and growth mode (cultured and natural Astragali Radix). This study laid the foundation for the quality evaluation of Astragali Radix and screening of active oligosaccharides.

Genetic diversity analysis of Tetrastigma hemsleyanum germplasm resources by ISSR / 中草药

Objective In this paper, the genetic diversity of 64 samples of Tetrastigma hemsleyanum germplasm resources in Chinese was analyzed. Methods ISSR-PCR was firstly used to amplify, and then POPGENE 32 software and NTSYS software was used to analyze the genetic diversity and phylogenetic relationship of 64 samples of T. hemsleyanum germplasm resources, and phylogenetic tree was constructed according to the UPGMA method. Results Ten primers with clear and reproducible bands were screened from 30 primers and used for genomic DNA amplification of 64 sample materials. A total of 83 polymorphic locis were amplified, whose polymorphic percentages were 71.43%-100% and average polymorphism percentage was 94.31%. The amplification polymorphic locis of primer S17 were the most (11) and the amplification polymorphic locis of primer P6 were the least (5), the average amplified polymorphic locis of 10 primers were 8.3. Genetic diversity analysis showed that the average number of alleles (Na) of 64 samples was 1.943 1, the average effective allele number (Ne) was 1.381 08, the average Nei’s gene diversity index (H) was 0.242 98, and the average Shannon diversity index (I) was 0.385 83. The variation range of the genetic similarity coefficient of the 64 samples was 0.431 8-0.988 6. A total of 64 samples were divided to six groups by UPGMA clustering method according to the similarity coefficient matrix when the genetic similarity coefficient was 0.715 5, which showed the abundant genetic diversity and relative gene stability of T. hemsleyanum germplasm resources. In addition, amplification figures by primers ISSR20, UBC857, and S17 were screened based on amplification result of 10 ISSR primers, and DNA fingerprinting was constructed, which can be used to identify 64 samples of T. hemsleyanum tested. Conclusion There are abundant genetic diversity and relative gene stability in T. hemsleyanum germplasm esources in China. ISSR analysis can reveal the genetic relationship among T. hemsleyanum germplasm resources in China, and provide certain reference for the evaluation, identification and new variety breeding of T. hemsleyanum germplasm resources in China.

Genetic diversity analysis of Tetrastigma hemsleyanum germplasm resources based on fluorescently labeled SSR markers / 中草药

Objective To understand the genetic diversity and genetic relationship of 64 samples of Tetrastigma hemsleyanum germplasm resources. Methods Fluorescently labeled SSR were used for PCR amplification, POPGENE32 software was used to analyze genetic diversity and genetic relationship of 64 samples of T. hemsleyanum germplasm resources, UPGMA method was used to construct their genetic tree map, NTSYS software was used to construct their two-dimensional principal component analysis map and three-dimensional scatter map. Result Eight pairs of primers with clear and reproducible bands were screened from 14 pairs of primers and used for genomic DNA amplification of 64 sample materials. The observed number of alleles (Na) ranged from 3 to 13, and the mean value was 7.875 0; The effective number of alleles (Ne) ranged from 1.424 9 to 6.087 4, and the mean value was 3.605 2; The Shannon information index (I) ranged from 0.689 5 to 2.082 4, and the mean value was 1.424 0; The observed heterozyghosity (Ho) ranged from 0.206 3 to 0.734 4, and the mean value was 0.524 7; The expected heterozygosity (He) ranged from 0.300 6 to 0.842 4, and the mean value was 0.658 4; The Nei’s gene diversity (H) ranged from 0.298 2 to 0.835 7, and the mean value was 0.653 2; The polymorphism information content (PIC) ranged from 0.288 0 to 0.817 5, and the mean value was 0.614 5; The genetic similarity coefficient ranged from 0.115 4 to 0.954 5; The genetic distance ranged from 0.000 0 to 3.218 1. It was indicated that the genetic relationship of 64 T. hemsleyanum germplasm resources was far, and the degree of genetic differentiation was high. At the genetic distance of 1.018 9, 64 T. hemsleyanum germplasm resources could be divided into five groups. Conclusion There was no necessary connection between geographical difference and genetic difference of germplasm. Themsleyanum germplasm resources are rich in genetic diversity. The results of fluorescently labeled SSR analysis can provide some references for the utilization and variety breeding of T. hemsleyanum germplasm resources.