Integrated phenotypic, transcriptomics and metabolomics: growth status and metabolite accumulation pattern of medicinal materials at different harvest periods of Astragalus Membranaceus Mongholicus.

BACKGROUND: Astragalus membranaceus var. mongholicus (Astragalus), acknowledged as a pivotal "One Root of Medicine and Food", boasts dual applications in both culinary and medicinal domains. The growth and metabolite accumulation of medicinal roots during the harvest period is intricately regulated by a transcriptional regulatory network. One key challenge is to accurately pinpoint the harvest date during the transition from conventional yield content of medicinal materials to high and to identify the core regulators governing such a critical transition. To solve this problem, we performed a correlation analysis of phenotypic, transcriptome, and metabolome dynamics during the harvesting of Astragalus roots. RESULTS: First, our analysis identified stage-specific expression patterns for a significant proportion of the Astragalus root genes and unraveled the chronology of events that happen at the early and later stages of root harvest. Then, the results showed that different root developmental stages can be depicted by co-expressed genes of Astragalus. Moreover, we identified the key components and transcriptional regulation processes that determine root development during harvest. Furthermore, through correlating phenotypes, transcriptomes, and metabolomes at different harvesting periods, period D (Nov.6) was identified as the critical period of yield and flavonoid content increase, which is consistent with morphological and metabolic changes. In particular, we identified a flavonoid biosynthesis metabolite, isoliquiritigenin, as a core regulator of the synthesis of associated secondary metabolites in Astragalus. Further analyses and experiments showed that HMGCR, 4CL, CHS, and SQLE, along with its associated differentially expressed genes, induced conversion of metabolism processes, including the biosynthesis of isoflavones and triterpenoid saponins substances, thus leading to the transition to higher medicinal materials yield and active ingredient content. CONCLUSIONS: The findings of this work will clarify the differences in the biosynthetic mechanism of astragaloside IV and calycosin 7-O-ß-D-glucopyranoside accumulation between the four harvesting periods, which will guide the harvesting and production of Astragalus.

Ecological niche modeling of Astragalus membranaceus var. mongholicus medicinal plants in Inner Mongolia, China.

Radix Astragali is commonly used in traditional Chinese medicine, and its quality is closely related to ecological factors, such as climate and soil, in the production area. To provide high-quality Radix Astragali to Chinese and foreign markets, we used maximum entropy model and statistical analysis method, combined with data on ecological factors, Astragalus membranaceus var. mongholicus geographical distribution, and index component content to predict the ecological suitability distribution of A. membranaceus var. mongholicus and establish the relationship between astragaloside IV and calycosin-7-glucoside in this species and ecological factors. Subsequently, we could determine the suitability regionalization of high-quality A. membranaceus var. mongholicus in Inner Mongolia, China. The results showed that the standard deviation of seasonal changes in temperature (40.6%), precipitation in October (15.7%), vegetation type (14.3%), soil type (9.2%), and mean sunshine duration in the growing season (9.1%) were the top five most influential factors out of the 17 main ecological factors affecting the distribution of A. membranaceus var. mongholicus. The standard deviation of seasonal changes in temperature, precipitation in October, precipitation in April, soil pH, and mean sunshine duration in the growing season were found to be the key ecological factors affecting the accumulation of astragaloside IV and calycosin-7-glucoside in A. membranaceus var. mongholicus. The regions with the highest-quality A. membranaceus var. mongholicus were distributed in Baotou (Guyang County), Hohhot (Wuchuan County), and central Wulanchabu (Chahar Right Middle Banner, Chahar Right Back Banner, and Shangdu County) and its surroundings in Inner Mongolia. Baotou, Hohhot, and their surrounding areas were the main traditional production areas of A. membranaceus var. mongholicus, and central Wulanchabu was a potentially suitable distribution area of this species. The main production areas were consistent with the actual production base of A. membranaceus var. mongholicus. This study therefore provides a scientific basis to guide the cultivation of A. membranaceus var. mongholicus.

Transcriptome sequencing and characterization of Astragalus membranaceus var. mongholicus root reveals key genes involved in flavonoids biosynthesis.

BACKGROUND: Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao is a traditional medicinal herb of Leguminosae since it contains bioactive compounds such as flavonoids, which have significant pharmacological effects on immunity and antioxidant. However, the scanty genomic and transcriptome resources of Astragalus membranaceus have hindered further exploration of its biosynthesis and accumulation mechanism. OBJECTIVE: This project aim to further improve our understanding of the relationship between transcriptional behavior and flavonoids content of A. mongholicus. METHODS: The accumulation of flavonoids and related gene expression in five different developmental stages (A: vegetative, B: florescence, C: fruiting, D: fruit ripening and E: defoliating stages) of A. mongholicus root were studied by combining UV spectrophotometry and transcriptomic techniques. The de novo assembly, annotation and functional evaluation of the contigs were performed with bioinformatics tools. RESULTS: After screening and assembling the raw data, there were a total of 158,123 unigenes with an average length of 644.89 bp were finally obtained, which has 8362 unigenes could be jointly annotated by NR, SwissProt, eggNOG, GO, KEGG and Pfam databases. KEGG enrichment analysis was performed on differentially expressed genes(DEGs)in the four groups (A vs. B, B vs. C, C vs. D, D vs. E). The results showed that many DEGs in each group were significantly enriched to flavonoids biosynthesis related pathways. Among them, a number of 86 were involved in the biosynthesis of isoflavonoid (12), flavonoid (5) and phenylpropanoid (69). Further analysis of these DEGs revealed that the expression levels of key genes such as PAL, 4CL, CCR, COMT, DFR, etc. were all down-regulated at the fruiting stage, and then raised at the fruit ripening stage. This expression pattern was similar to the accumulation trend of total flavonoids content. CONCLUSIONS: In summary, this comprehensive transcriptome dataset allowed the identification of genes associated with flavonoids metabolic pathways. The results laid a foundation for the biosynthesis and regulation of flavonoids. It also provided a scientific basis for the most suitable harvest time and resource utilization of A. mongholicus.

Discovery of chemical markers for identifying species, growth mode and production area of Astragali Radix by using ultra-high-performance liquid chromatography coupled to triple quadrupole mass spectrometry.

BACKGROUND: Astragali Radix (AR) is a well-known Chinese herbal medicine. The quality of AR can be affected by many factors such as species, growth mode and production area, but there are still no chemical markers to distinguish it. PURPOSE: To explore chemical markers for improving the quality assessment of AR and discover chemical markers for identifying species, growth mode and production area of AR. METHODS: A highly sensitive, efficient and accurate method based on ultra-high performance liquid chromatography coupled to triple quadrupole mass spectrometry (UHPLC-QQQ-MS/MS) for simultaneous quantitative determination of 14 major chemical components (five flavonoids and nine triterpene saponins) in 94 batches of AR from China, Republic of Korea and Germany was developed for the first time. To explore chemical markers and assess changes in the contents of 14 compounds in the 94 batches of AR samples from different regions, hierarchical clustering analysis (HCA) and principal component analysis (PCA) were performed. RESULTS: Astragaloside III was not only an important chemical marker for distinguishing two species of AR, i.e.: Astragalus mongholicus and A. membranaceus, but also a potential chemical marker for the classification of cultivated and semi-wild AR. In addition, in the batches of cultivated AR, the content of isoastragaloside II and cyclocephaloside II were greater in batches from the region of Shaanxi Province than that of other Provinces in China, but the content of calycosin-7-O-ß-D-glucoside and astragaloside IV, which are the quality control markers of AR required by the Chinese Pharmacopoeia, were higher than that of other Provinces in China. In addition, the content of calycosin-7-O-ß-D-glucoside, ononin, calycosin and astragaloside I could be used to identify samples of AR collected from China, Republic of Korea and Germany. CONCLUSION: This UHPLC-QQQ-MS/MS method could be applied to the quantitative evaluation of AR and could be an important and meaningful reference to develop chemical markers for quality control of AR.

Enhanced Production of Two Bioactive Isoflavone Aglycones in Astragalus membranaceus Hairy Root Cultures by Combining Deglycosylation and Elicitation of Immobilized Edible Aspergillus niger.

A cocultivation system of Astragalus membranaceus hairy root cultures (AMHRCs) and immobilized food-grade fungi was established for the enhanced production of calycosin (CA) and formononetin (FO). The highest accumulations of CA (730.88 ± 63.72 µg/g DW) and FO (1119.42 ± 95.85 µg/g DW) were achieved in 34 day-old AMHRCs cocultured with immobilized A. niger (IAN) for 54 h, which were 7.72- and 18.78-fold higher than CA and FO in nontreated control, respectively. IAN deglycosylation could promote the formation of CA and FO by conversion of their glycoside precursors. IAN elicitation could intensify the generation of endogenous signal molecules involved in plant defense response, which contributed to the significantly up-regulated expression of genes in CA and FO biosynthetic pathway. Overall, the coupled culture of IAN and AMHRCs offered a promising and effective in vitro approach to enhance the production of two health-promoting isoflavone aglycones for possible nutraceutical and pharmaceutical uses.

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In order to explore the ecological factors affecting the growth of Astragalu smembranaceus var. mongholicus, we investigated the resource distribution, habitat characteristics and growth conditions of wild and cultivated A. membranaceus var. mongholicus by fixed-plot observation, survey method, and literature. These data were analyzed by traditional Chinese medicine GIS-2 (TCMGS-2) to obtain the most suitable areas of A. membranaceus var. mongholicus within Inner Mongolia. The results showed that the production areas of cultivated A. membranaceus var. mongholicus were mainly located in Wuchuan County, Guyang County and other 15 counties, which were cha-racterized by the altitude higher than 1000 m, with soil type of sand, gravel and calcareous clay. The wild A. membranaceus var. mongholicus was distributed mainly in the eastern Inner Mongolia and germinated in sunny place, which preferred to the cold dry climate and sandy loam soil or gra-vel but avoided damp heavy clay soils. There are 43 counties of 94460.30 km2 for wild A. membranaceus var. mongholicus, and 32 counties of 76013.93 km2 for cultivated one within Inner Mongolia, with a similarity coefficient of ecological factors greater than 95%.

Triterpene and Flavonoid Biosynthesis and Metabolic Profiling of Hairy Roots, Adventitious Roots, and Seedling Roots of Astragalus membranaceus.

Astragalus membranaceus is an important traditional Chinese herb with various medical applications. Astragalosides (ASTs), calycosin, and calycosin-7-O-ß-d-glucoside (CG) are the primary metabolic components in A. membranaceus roots. The dried roots of A. membranaceus have various medicinal properties. The present study aimed to investigate the expression levels of genes related to the biosynthetic pathways of ASTs, calycosin, and CG to investigate the differences between seedling roots (SRs), adventitious roots (ARs), and hairy roots (HRs) using quantitative real-time polymerase chain reaction (qRT-PCR). qRT-PCR study revealed that the transcription level of genes involved in the AST biosynthetic pathway was lowest in ARs and showed similar patterns in HRs and SRs. Moreover, most genes involved in the synthesis of calycosin and CG exhibited the highest expression levels in SRs. High-performance liquid chromatography (HPLC) analysis indicated that the expression level of the genes correlated with the content of ASTs, calycosin, and CG in the three different types of roots. ASTs were the most abundant in SRs. CG accumulation was greater than calycosin accumulation in ARs and HRs, whereas the opposite was true in SRs. Additionally, 40 metabolites were identified using gas chromatography-time-of-flight mass spectrometry (GC-TOF-MS). Principal component analysis (PCA) documented the differences among SRs, ARs, and HRs. PCA comparatively differentiated among the three samples. The results of PCA showed that HRs were distinct from ARs and SRs on the basis of the dominant amounts of sugars and clusters derived from closely similar biochemical pathways. Also, ARs had a higher concentration of phenylalanine, a precursor for the phenylpropanoid biosynthetic pathway, as well as CG. TCA cycle intermediates levels including succinic acid and citric acid indicated a higher amount in SRs than in the others.

[Effect of Water Extracts from Rhizosphere Soil of Cultivated Astragalus membranaceus var. mongholicus on It's Seed Germination and Physiological Characteristics].

OBJECTIVE: To explore the relationship between continuous cropping obstacle and autotoxicity of Astragalus membranaceus var. mongholicus. METHODS: Distilled water(CK), water extracts of rhizosphere soil(50, 100, 200 and 400 mg/mL) were applied to test their effect on early growth and physiological characteristics of Astragalus membranaceus var. mongholicus. RESULTS: The water extracts from rhizospher soil of cultivated Astragalus membranaceus var. mongholicus significantly increased seedling emergence rate, root length and vigor index of Astragalus membranaceus var. mongholicus seedling when at the concentration of 100 mg/mL or below, however,there was no significant effect at 200 mg/mL or higher. The water extracts from rhizosphere soil of cultivated Astragalus membranaceus var. mongholicus significantly reduced the SOD activity in Astragalus membranaceus var. mongholicus seedling at 400 mg/mL and POD activity at 200 mg/mL and 400 mg/mL,while significantly increased the MDA content. CONCLUSION: Water extracts from Astragalus membranaceus var. mongholicus rhizosphere soil significantly affected Astragalus membranaceus var. mongholicus germination and seedling growth in a concentration-dependent manner, generally, low concentrations increased the SOD and POD activity which improved seed germination and seedling growth, while high concentrations caused cell membrane damage of the seedling.

Transcriptional Profiling and Molecular Characterization of Astragalosides, Calycosin, and Calycosin-7-O-ß-D-glucoside Biosynthesis in the Hairy Roots of Astragalus membranaceus in Response to Methyl Jasmonate.

We used the next-generation Illumina/Solexa HiSeq2000 platform on RNA analysis to investigate the transcriptome of Astragalus membranaceus hairy roots in response to 100 µM methyl jasmonate (MeJA). In total, 77,758,230 clean reads were assembled into 48,636 transcripts (average length of 1398 bp), which were clustered into 23,658 loci (genes). Of these, 19,940 genes were annotated by BLASTx searches. In addition, DESeq analysis showed that 2127 genes were up-regulated, while 1247 genes were down-regulated by MeJA. Seventeen novel astragaloside (AST) biosynthetic genes and seven novel calycosin and calycosin-7-O-ß-D-glucoside (CG) biosynthetic genes were isolated. The accumulation of ASTs, calycosin, and CG increased significantly in MeJA-treated hairy roots compared with control hairy roots. Our findings will provide a valuable resource for molecular characterization of AST, calycosin, and CG biosynthetic pathways and may lead to new approaches to maximize their production and biomass productivity in the hairy roots of A. membranaceus.

[Investigation on Main Cultivation of Astragalus membranaceus var. mongholicus and Commercial Specifications Grades].

OBJECTIVE: To investigate the main cultivation areas of Astragalus membranaceus var. mongholicus, and to clarify its present cultivation status and commercial specification grades. METHODS: Astragalus membranaceus var. mongholicus cultivated in Hunyuan and Ying County of Shanxi, Longxi, Weiyuan and Min County of Gansu and Wuchuan, Guyang and Chifeng of Inner Mongolia were investigated, including their ecological environment, cultivation history and present situation, harvesting and processing as well as commercial specification grades. RESULTS: The cultivation methods of Astragalus membranaceus var. mongholicus changed greatly in the main cultivation areas. In Gansu and Inner Mongolia, the current major cultivation method was 2 - 3 years of seedling transplantation method, while the 3 - 5 years of seed-directly-sowed method in Inner Mongolia had been almost abandoned, and the 6 - 8 years of mountain direct seeding method was still in application in Hunyuan county of Shanxi province. On the other hand, the harvest method of Astragalus membranaceus var. mongholicus had been gradually changed from manual harvesting in ancient times to the current mechanical harvesting although the latter method did more damage to environment of cultivation areas and the resource of Astragalus membranaceus var. mongholicus in regions Hunyuan county of Shanxi. Through investigating the processing methods of Astragalus membranaceus var. mongholicus in different cultivation areas, it was found that processing method varied from area to area, and no unified specifications had been formulated on processing methods in market. In addition, the current processing method was too complicated in processing the slices of prepared Astragali Radix and most were self processing by farmers themselves, creating market disorders. Moreover, the Astragali Radix products were basically without grading classification but sold as bulk cargo. CONCLUSION: It is urgent to unify the specifications and grades of Astragali Radix product, and standardize its processing methods.

Efficient production of isoflavonoids by Astragalus membranaceus hairy root cultures and evaluation of antioxidant activities of extracts.

In this study, Astragalus membranaceus hairy root cultures (AMHRCs) were established as an attractive alternative source for the efficient production of isoflavonoids (IF). A. membranaceus hairy root line II was screened as the most efficient line and was confirmed by PCR amplification of rolB, rolC and aux1 genes. Culture parameters of AMHRCs were systematically optimized, and five main IF constituents were quali-quantitatively determined by LC-MS/MS. Under optimal conditions, the total IF accumulation of 34 day old AMHRCs was 234.77 µg/g dry weight (DW). This yield was significantly higher compared to that of 3 year old field grown roots (187.38 µg/g DW). Additionally, in vitro antioxidant assays demonstrated that AMHRC extracts exhibited antioxidant activities with lower IC50 values (1.40 and 1.73 mg/mL) as compared to those of field grown roots (1.96 and 2.17 mg/mL). Overall, AMHRCs may offer a promising and continuous product platform for naturally derived, high quality and valuable nutraceuticals.

[Study on method and its optimization of improving seed germination of Astragalus membranaceus as gansu traditional medicinal herb].

OBJECTIVE: To break the hard testa and improve seed germination situation of Astragalus membranaceus var. mongholicus, in order to solve the problems of low success rate of seed germination and seedling. METHODS: Longxi Astragalus membranaceus var. mongholicus seed was treated by soaking seed with 75% alcohol and concentrated sulfuric acid, warm-water incubating, grinding and comprehensive treating with warm-water incubating, grinding and sand culture. Its seed germination situation was evaluated by germination potential, germination rate and germination index. RESULTS: Different processing methods significantly improved seed germination with different effect. Comprehensive treatment with warm-water incubating, grinding and sand culture was the best one on Astragalus membranaceus var. mongholicus seed germination. Its germination potential, germination rate and germination index was 66.04%, 87.70% and 1.34,respectively. CONCLUSION: Comprehensive treatment with warm-water incubating, grinding and sand culture is an economic and effective processing method, which is suitable for actual production.

[Autotoxicity of aqueous extracts from plant of cultivated Astragalus membranaceus var. mongholicus].

OBJECTIVE: To exploring the relationship between continuous cropping obstacle and autotoxicity of Astragalus membranaceus var. mongholicus, autotoxic effect of plant aqueous extract were determined. METHODS: Distilled water (CK), aqueous extract of plant, including root, stem and leaf (12.5, 25, 50 and 100 mg/mL respectively)were applied to testing their effect on early growth of Astragalus membranaceus var. mongholicus. Specifically, seed germination rate, germination index, emergence rate, elongation of radical and embryo, and seedling vigor index were determined. RESULTS: The aqueous extract of root, stem, and leaf at 25 mg/mL significantly inhibited the seed germination and seedling growth of Astragalus membranaceus var. mongholicus, and this inhibitory effect generally increased with the increase of the concentration of aqueous extracts. To the comprehensive allelopathic effect, the extracts from Astragalus membranaceus var. mongholicus stem were more inhibitory than those from leaf and root. The germination index and seedling vigor index were more sensitive to extract than other determined parameters. CONCLUSION: Aqueous extracts from Astragalus membranaceus var. mongholicus plant gave inhibitory effects on Astragalus. membranaceus var. mongholicus germination and seedling growth, and this inhibitory effect generally increased with the increases of aqueous extract concentration at a certain ranges. In conclusion, there is an autotoxicity in continuous cropping of Astragalus membranaceus var. mongholicus.

[Status and analysis of Astragali Radix resource in China].

To explore the status of the resources of Astragali Radix, a survey on its germplasm resources was carried out. Some conclusions can be drawn for Astragali Radix: the major source is the cultivated Astragalus mongolicus. The new major cultivation areas for A. mongolicus and A. membranaceus are Shandong and Gansu province. The semi-wildly planting model in Shanxi province maintains the genuine trait of Astragali Radix, but its yield is limited, and now a combination model has been developed. The major problems for Astragali Radix are the selection of planting sites, the rot root and difficulty in collecting and processing. Several developmental proposals for Astragali Radix were put forward including rational distribution of planting areas, establishment of standard system, development and standardization of producing technologies.

[The FTIR spectral characteristics and comparison study of Astragalus menbranceus soil].

To study the genuine soil of Astragalus menbranceus grows, FTIR spectrometry was used, which is accurate, simple and efficient and has high resolution. The genuine soils include six areas in Hunyuan of Shanxi province, three areas in Yingxian of Shanxi province, Fansi of Shanxi province, and Guyang of Inner Mongolia. Different growth years of two to five for each area were also studied. The results show that there are significant differences between Astragalus menbranceus soil FTIR spectrometry and general soil's, between soil of Astragalus menbranceus growth and radix codonopsitis growth, between different soil of Astragalus menbranceus growth, providing useful information for the area chose of Chinese herb cultural and transplantation.

[Regulation of Vitreoscilla hemoglobin on biosynthesis of astragaloside IV].

In the present study, the regulation of Vitreoscilla hemoglobin (VHb) on astragaloside IV biosynthesis was investigated. An intermediate expression vector consisting of the CaMV35S promoter fused to the vgb and nopaline synthase terminator was transferred into Astragalus membranaceus via Agrobacterium rhizogenes. The transgenic hairy roots were confirmed by PCR amplification and Southern blot hybridization. The expression of vgb in transgenic hairy roots was confirmed by RT-PCR. After 15 days cultivation, the dry weight and growth rate of transgenic hairy roots were higher than that of the non-transgenic hairy root. ELSD-HPLC analysis showed that astragaloside IV content of transgenic hairy roots was 5 to 6 times of non-transgenic hairy root control and 10 to 12 times of Radix Astragali from Shanxi Province. These results suggested that the expression of vgb promoted the growth of transgenic hairy roots, and increased the content of astragaloside IV.

[Effects of inoculating AM fungi on physiological characters and nutritional components of Astragalus membranaceus under different N application levels].

A pot culture with unsterilized soil as growth substrate showed that AM fungi had significant effects on the growth of Astragalus membranaceus (Fabaceae) under different N application levels. Inoculation with AM fungi promoted the AM infection of A. membranaceus roots, but high N application level suppressed the infection. Inoculating AM fungi increased the growth rate, soluble protein and sugar contents, and SOD, POD and CAT activities of A. membranaceus. Under 50 and 100 mg x kg(-1) of N application, new bands of POD isozyme occurred in inoculated plants, and the contents of flavonoid, N, and P in A. membranaceus also had definite increase. The best inoculation effect was observed under the N application level of 50-100 mg N x kg(-1) soil.

[Effects of nitrogen, phosphorus and potassium application on growth and active ingredient of Astragalus membranaceus].

OBJECTIVE: To study the effects of fertilizers with the different proportional of nitrogen, phosphorus and potassium on growth and active ingredient of Astragalus membranaceus. METHOD: Field experiment was conducted based on the D-saturation optimal design with three factors of nitrogen, phosphorus and potassium. The effects on growth and active ingredient of A. membranaceus were analyzed. RESULT: Fertilization promoted the seedling growth and provided abundant supply of nutrition for growth of root, yield and accumulation of active ingredient at the later growth stage, and increased the accumulation of dry matter of stem-leaf and root system. The effect of nitrogen, phosphorus and potassium application on the total dry matter accumulation of A. membranaceus was as following: nitrogen > potassium > phosphorus; the effect on the stem-leaf dry matter accumulation was as following: nitrogen > phosphorus > potassium; the effect on the root dry matter accumulation was as following: nitrogen > potassium > phosphorus. Nitrogen, phosphorus and potassium fertilizer increased the root yield of A. membranaceus. Obviously, the effect on the root yield was as following: nitrogen > potassium > phosphorus. The application of different proportional with nitrogen, phosphorus and potassium increased the content of polysaccharide and astragaloside, but had no distinct effect on the content of total flavonoids. The effect on the content of polysaccharide was as following: potassium > phosphorus > nitrogen, but the effect on the conten,t of astragaloside was as following: nitrogen > potassium > phosphorus. CONCLUSION: Nitrogen and potassium fertilizer application had more important effect on growth, yield and the contents of polysaccharide and astragaloside in A. membranaceus. During medicinal plants cultivation process, it should pay attention to the application of nitrogen fertilizer and potassium fertilizer and make balance application of nitrogen, phosphorus and potassium fertilizer.

[Study on difference of biological characteristics and resistance to powdery mildew of different Astragalus populations].

OBJECTIVE: To study difference among populations which belong to Astragalus membranaceus and A. membranaceus var. mongholicus on morphology, habit, characteristics of physiology and resistance to powdery mildew, and classify them in order to provide theoretical basis for breeding and improving varieties. METHOD: Morphology, habits, isozyme and soluble protein electrophoretograms were compared among the populations. They were categorized by cluster analysis based on those electrophoretograms. Different ability of resistance to powdery mildew was also studied through comparing disease indices among six populations. RESULT: The results showed A. membranaceus var. mongholicus was distinctly different from A. membranaceus. There was a special type in colonies of A. membranaceus, which showed hairy upper epidermis of leaflets and later florescence. CONCLUSION: Astragalus for medicine could be categorized in three types A. membranaceus var. mongholicus, A. membranaceus early florescence type and A. membranaceus late florescence type. Among them A. membranaceus var. mongholicus is most resistant to powdery mildew, while A. membranaceus is easily infected, and the early florescence type is even more easily infected.

[Quantitative analysis of calycosin glycoside and formononetin in Radix astragali from different sources].

OBJECTIVE: To establish the quantitative methods for calycosin glycoside and formononetin in Radix Astragali, and the samples from different sources were analyzed, in order to supply the basis for the quality control of Radix Astragali. METHOD: The content of calycosin glycoside and formononetin in 59 samples of Radix Astragali from eight with different provinces was analyzed by HPLC-DAD. RESULT: The contents of calycosin glycoside and formononetin in Radix Astragali from different sources, with different cultivating method or in different ages differed markedly, and the results showed that the quality of samples from Shannxi, Innermongolia and Shanxi were better than other sources, and the semi-wild samples were better than other cultiving samples, moreover the shorter age, the better quality. CONCLUSION: This simple, accurate and reproducible method could use to determine the contents of isoflavanoids in Radix Astragali.