Comprehensive evaluation of two Astragalus species (A. campylosema and A. hirsutus) based on biological, toxicological properties and chemical profiling.

Astragalus L. (Fabaceae) is an important genus with numerous species having various traditional medicinal uses making them of interest for scientific investigations to ascertain their therapeutic benefits. In the present study, the quantitative polyphenolic profiles of methanolic extracts from different parts (leaves, flowers, and roots) of two endemic Astragalus species growing in Turkey, i.e. A. campylosema Boiss. and A. hirsutus Vahl were determined, along with their antioxidant and enzyme inhibitory properties. A. campylosema and A. hirsutus extracts showed varying total phenolic (25.80-40.60 and18.59-29.46 mg GAE/g, respectively) and total flavonoid (11.21-105.91 and 16.06-131.91 mg RE/g, respectively) contents. HPLC-MS/MS revealed rutin to be the predominant phenolic compound in all the extracts of A. campylosema and leaf extract of A. hirsutus (133.53-752.42 µg g-1), while hyperoside was the major one in the flower and root extracts of A. hirsutus (2014.07 and 123.13 µg g-1, respectively). In DPPH and ABTS assays, radical scavenging capacity was demonstrated by all extracts of A. campylosema (47.13-48.10 and 87.03-115.36 mg TE/g, respectively) and A. hirsutus (17.82-38.67 and 47.84-57.29 mg TE/g, respectively). Reducing activity was also displayed by the extracts in CUPRAC and FRAP assays (A. campylosema: 83.06-135.20 and 59.15-90.19 mg TE/g, respectively; A. hirsutus: 53.02-83.42 and 31.25-43.25 mg TE/g, respectively). All extracts were also found to act as metal chelators (12.32-21.45 mg EDTAE/g) and exhibited total antioxidant capacity ranging from 1.16 to 1.60 mmol TE/g, in phosphomolybdenum assay. Acetyl- and butyryl-cholinesterase inhibitory effects were observed by all the extracts of the two species (1.56-4.99 mg GALAE/g). Anti-hyperpigmentation potential by inhibiting tyrosinase (54.55-67.35 mg KAE/g) was reported as well. Carbohydrate hydrolyzing enzymes, amylase and glucosidase were also inhibited (0.22-1.03 mmol ACAE/g). Overall, A. campylosema extracts showed relatively better antioxidant and enzyme inhibitory potentials compared to A. hirsutus extracts. Strikingly, A. hirsutus extracts was found to have higher AGE inhibition activity than A. campylosema. Although the cytotoxic effect of three different organs obtained from A. campylosema and A. hirsutus increased depending on the dose (from 10 to 200 µg/mL), it was found that both plant extracts did not show a genotoxic effect at the highest concentration of 200 µg/mL. Indeed, data amassed from this current scientific work showed the two selected Astragalus species to be rich in bioactive polyphenols that could be responsible for the various pharmacological activities and hence demands to be further explored for their possible applications as natural health promoting agents.

Chloroplast phylogenomics and character evolution of eastern Asian Astragalus (Leguminosae): Tackling the phylogenetic structure of the largest genus of flowering plants in Asia.

Astragalus, as the largest genus of the flowering plants, is well-known for its high species richness and morphological diversity. Previous studies suggested that many of the subgenera of Astragalus are not monophyletic and the phylogenetic relationships within the genus are still poorly known. In this study, we sampled 117 accessions of Astragalus and its close relatives, covering 55 sections of the genus plus 30 outgroup taxa to recover the main clades of eastern Asian Astragalus based on sequences of the whole chloroplast genome and 65 chloroplast CDSs. Astragalus is supported to be monophyletic and it is sister to the Oxytropis + Coluteoid clade. Within Astragalus, we recovered ten clades, and the ten clades differ substantially from Bunge's subgenera. The former segregate genus Astracantha is also monophyletic, but embedded within Astragalus s. str., supporting the merge of the spiny former genus Astracantha with Astragalus. We detected the atpF intron losses in the chloroplast genome of the Oxytropis + Coluteoid clade, i.e., the sister clade to Astragalus. Furthermore, we estimated the ancestral states of the trichome morphology and habit via the Bayesian Binary Method. The medifixed hair type is inferred to have developed at least five times and the annual habit originated at least six times. In addition, Astragalus is estimated to have originated in the mid Miocene (stem age, 16.09 Ma, 95% HPD: 12.46-20.50 Ma). The divergence times of the medifixed hair groups ranged from 4.03 to 0.87 Ma, mostly 2-1 Ma, which are correlated with the estimated phased uplifts of the Qinghai-Tibetan Plateau (QTP). We hypothesize that the uplifts of the QTP, which contributed to aridification in eastern Asia and the adjacent regions, may have accelerated the rapid speciation of Astragalus, especially the xerophilous groups (i.e. the medifixed hair groups).

Multiple auto- and allopolyploidisations marked the Pleistocene history of the widespread Eurasian steppe plant Astragalus onobrychis (Fabaceae).

The Eurasian steppes occupy a significant portion of the worldwide land surface and their biota have been affected by specific past range dynamics driven by ice ages-related climatic fluctuations. The dynamic alterations in conditions during the Pleistocene often triggered reticulate evolution and whole genome duplication events. Employing genomic, genetic and cytogenetic tools as well as morphometry we investigate the intricate evolution of Astragalus onobrychis, a widespread Eurasian steppe plant with diploid, tetraploid and octoploid cytotypes. To analyse the heteroploid RADseq dataset we employ both genotype-based and genotype-free methods that result in highly consistent results, and complement our inference with information from the plastid ycf1 region. We uncover a complex and reticulate evolutionary history, including at least one auto-tetraploidization event and two allo-octoploidization events; one of them involved also genetic contributions from other species, most likely A. goktschaicus. The present genetic structure points to the existence of four main clades within A. onobrychis, which only partly correspond to different ploidies. Time-calibrated diffusion models suggest that diversification within A. onobrychis was associated with ice age-related climatic fluctuations during the last million years. We finally argue for the usefulness of uniparentally inherited plastid markers, even in the genomic era, especially when investigating heteroploid systems.

Genetic analyses of Astragalus sect. Humillimi (Fabaceae) resolve taxonomy and enable effective conservation.

PREMISE OF THE STUDY: Astragalus sect. Humillimi is distributed across the southwestern United States and contains two endangered taxa, A. cremnophylax var. cremnophylax and A. humillimus. The former was originally described from the South Rim of the Grand Canyon. Analysis of individuals discovered on the North Rim of the Grand Canyon yielded some evidence that the population represented a distinct species. To enable effective conservation, we clarify the group's taxonomy and characterize the genetic diversity of A. cremnophylax and A. humillimus. METHODS: We used AFLPs to genotype most species in sect. Humillimi, focusing on the two endangered forms. We examined patterns of genetic diversity using complementary analytical approaches. KEY RESULTS: Our results demonstrate that North Rim populations group with A. c. var. cremnophylax. We found low levels of genetic diversity at certain localities and strong differentiation among populations. Astragalus humillimus, which has suffered recent and severe population declines, exhibits weak differentiation among and low diversity within populations. CONCLUSIONS: Our results clarify the taxonomy of sect. Humillimi and define the boundaries of A. c. var. cremnophylax, which is shown to inhabit both rims of the Grand Canyon. This clarification, and detailed analysis of genetic variation within both endangered taxa, may advance ongoing efforts to conserve these taxa. Our results suggest that range-wide genetic analysis of A. humillimus may inform recovery strategies for this taxon.

Screening for swainsonine among South American Astragalus species.

Swainsonine is a toxic alkaloid found in several plant genera worldwide. The objective of this study was to screen several South American Astragalus species for the toxin swainsonine. Swainsonine was detected in 16 of 30 Astragalus species using liquid and gas chromatography coupled with mass spectrometry. Information in regard to the swainsonine content of these species may provide important information on the risk of grazing these toxic species.

[New records of medicinal plants in Hubei].

In this paper, we make a report on new records of medicinal plants in Hubei, which include one newly recorded genera and seven newly recorded species and a newly recorded variety. The newly recorded genera is Anoectochilus and its corresponding species is Anoectochilus roxburghii; These newly recorded species are Euphorbia micractina, Astragalus wulingensis, Blumea megacephala, Potentilla saundersiana, Blumea formosana, Lycoris houdyshelii and Colocasia gigantea ; The newly recorded variety is Neottia puberula var. maculata. Among these species, Anoectochilus roxburghii and N. puberula var. maculata are considered as the second-class protection in our country, A. roxburghii is regarded as Endangered（EN）and Astragalus wulingensis is regarded as Critically Endangered (CN) by IUCN. The report of these newly recorded plants borden the distribution and enrich the plant diversity of Hubei.

A swainsonine survey of North American Astragalus and Oxytropis taxa implicated as locoweeds.

Swainsonine, an indolizidine alkaloid with significant physiological activity, is an α-mannosidase and mannosidase II inhibitor that causes lysosomal storage disease and alters glycoprotein processing. Swainsonine is found in a number of plant species worldwide, and causes severe toxicosis in livestock grazing these plants, leading to a chronic wasting disease characterized by weight loss, depression, altered behavior, decreased libido, infertility, and death. Swainsonine has been detected in 19 Astragalus and 2 Oxytropis species in North America by thin layer chromatography, gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry and a jack bean α-mannosidase inhibition assay. In addition, 5 species in North America are presumed to contain swainsonine based upon reports from field cases. Many of these plant species have not been analyzed for swainsonine using modern instrumentation such as gas or liquid chromatography coupled with mass spectrometry. To provide clarification, 22 Astragalus species representing 93 taxa and 4 Oxytropis species representing 18 taxa were screened for swainsonine using both liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry. Swainsonine was detected in 48 Astragalus taxa representing 13 species and 5 Oxytropis taxa representing 4 species. Forty of the fifty-three swainsonine-positive taxa had not been determined to contain swainsonine previously using liquid or gas chromatography coupled with mass spectrometry. The list of swainsonine-containing taxa reported here will serve as a reference for risk assessment and diagnostic purposes.

Molecular Phylogeny and Morphological Analysis Support a New Species and New Synonymy in Iranian Astragalus (Leguminosae).

As a result of a taxonomic and phylogenetic revision of Astragalus section Hymenostegis we identified a new species of Astragalus from northwestern Iran, namely A. remotispicatus spec. nov., which is described and illustrated here. It is morphologically similar to A. karl-heinzii in possessing a lax inflorescence. Phylogenetic inference of the nuclear ribosomal DNA internal transcribed spacer (ITS) region support A. remotispicatus as a clearly distinct species within the lax-inflorescence group of this section. Also the placement of A. sciureus var. subsessilis was found to be wrong and this taxon should be treated as a synonym within A. kohrudicus.

Cycloartane-Type Saponins from Astragalus tmoleus var. tmoleus.

Five known cycloartane-type glycosides were isolated from the roots of A. tmoleus Boiss. var. tmoleus. The identification of these compounds was mainly achieved by 1D and 2D NMR spectroscopic techniques and FABMS. The results of our studies confirm that triterpene saponins with the cycloartane-type skeleton might be chemotaxonomically significant for the genus Astragalus.

Preserving a Comprehensive Vegetation Knowledge Base--An Evaluation of Four Historical Soviet Vegetation Maps of the Western Pamirs (Tajikistan).

We edited, redrew, and evaluated four unpublished historical vegetation maps of the Western Pamirs (Tajikistan) by the Soviet geobotanist Okmir E. Agakhanjanz. These maps cover an area of 5,188 km2 and date from 1958 to 1960. The purpose of this article is to make the historic vegetation data available to the scientific community and thus preserve a hitherto non available and up to now neglected or forgotten data source with great potential for studies on vegetation and ecosystem response to global change. The original hand-drawn maps were scanned, georeferenced, and digitized and the corresponding land cover class was assigned to each polygon. The partly differing legends were harmonized and plant names updated. Furthermore, a digital elevation model and generalized additive models were used to calculate response curves of the land cover classes and to explore vegetation-topography relationships quantitatively. In total, 2,216 polygons belonging to 13 major land cover classes were included that are characterized by 252 different plant species. As such, the presented maps provide excellent comparison data for studies on vegetation and ecosystem change in an area that is deemed to be an important water tower in Central Asia.

[Study on identification of Astragali Radix and Hedysari Radix by PCR amplification of specific alleles].

To explore the new method of discriminating Astragali Radix and Hedysari Radix by using PCR amplification of specific alleles, 30 samples of the different Astragali Radix materials and 28 samples of Hedysari Radix were collected. The total DNA of all samples were extracted, trnL-trnF sequence from Astragali Radix and Hedysari Radix was amplified by PCR and sequenced unidirectionally. These sequences were aligned by using Clustul W. Primer was designed and the PCR reaction systems including annealing temperature, dNTP, etc were optimized. All samples were amplified by PCR with specific primer, DNA from Astragali Radix would be amplified 136 bp, whereas PCR products from all of Hedysari Radix were 323 bp. This method can detect 10% of intentional Hedysari Radix DNA into Astragali Radix. PCR amplification of alleles can be used to identify Astragali Radix and Hedysari Radix successfully and is an efficient molecular marker for authentication of Astragali Radix and Hedysari Radix.

Geographic isolation affects inter- and intra-specific seed variability in the Astragalus tragacantha complex, as assessed by morpho-colorimetric analysis.

The effect of geographical isolation on the inter- and intra-specific seed variability of the Astragalus tragacantha complex was investigated by using a computer-aided imaging system. In particular, seed morphometric and colorimetric features of Astragalus balearicus, A. tegulensis, A. terraccianoi, A. thermensis and A. tragacantha were measured and the data were used to discriminate among regions of provenance, taxa and populations. Discriminant analysis of the data supports the accepted taxonomy of the group and the hypothesis that interspecific differentiation resulted from geographical isolation. The results also showed a positive correlation between the wideness of species' distribution and the levels of seed intra-specific variability.

Chemical and genetic assessment of variability in commercial Radix Astragali (Astragalus spp.) by ion trap LC-MS and nuclear ribosomal DNA barcoding sequence analyses.

Radix Astragali (Huangqi) has been demonstrated to have a wide range of immunopotentiating effects and has been used as an adjuvant medicine during cancer therapy. Identity issues in the collection of Radix Astragali exist because many sympatric species of Astragalus occur in the northern regions of China. In order to assess the quality, purity, and uniformity of commercial Radix Astragali, 44 samples were purchased from herbal stores in Hong Kong and New York City. The main constituents, including four isoflavonoids and three saponins, were quantitatively determined by liquid chromatography mass spectrometry (LC-MS). There was significant sample-to-sample variability in the amounts of the saponins and isoflavonoids measured. Furthermore, DNA barcoding utilizing the variable nuclear ITS spacer regions of the 44 purchased Radix Astragali samples were sequenced, aligned and compared. Eight polymorphic point mutations were identified which separated the Radix Astragali samples into three groups. These results indicate that the chemical and genetic variability that exists among Radix Astragali medicinal products is still a consistency and quality issue for this herbal. Two-way ANOVA analysis showed significant effects on the contents of the seven tested compounds when both phylogenetic and geographic (i.e., point of purchase) factors were considered. Therefore, chemical profiles determined by LC-MS and DNA profiles in ITS spacer domains could serve as barcode markers for quality control of Radix Astragali.

Phylogenetic origin of Phyllolobium with a further implication for diversification of Astragalus in China.

Astragalus is a species-rich genus occurring in the western arid habitats in China and its diversification and infrageneric relationships in this region remain unclear. In the present study, based on molecular data, we aim to (i) test whether Phyllolobium (previously treated as a subgenus Pogonophace in Astragalus) should be warranted; and (ii) date the origin of Phyllolobium and probable diversification of Astragalus sensu stricto (s.s.). We sequenced five species from Phyllolobium first and collected all related sequences from the genus, Astragalus s.s and their close relatives (Oxytropis and Caragana etc.). Our phylogenetic analyses suggested that all species of Phyllolobium comprise a monophyletic sister-group to genera of the subtribe Coluteinae. Molecular dating suggested that Phyllolobium and Astragalus s.s. originated around 8 and 10 million years ago. These two estimates are highly consistent with the intense uplifts of the Qinghai-Tibetan Plateau inferred from geological evidence. In addition, one section of Pogonophace (Sect. Robusti) was estimated to originate 2.5 million years ago and this section with a tendency for dry habitats seems to be evidence of Asian intensified aridity resulting from the intense uplift of the Qinghai-Tibetan Plateau.

Discrimination of different genera Astragalus samples via quantitative symmetry analysis of two-dimensional hetero correlation spectra.

It has been proved to be a very useful method to distinguish similar samples by two-dimensional correlation spectroscopy when they are hardly distinguished by the conventional one-dimensional spectroscopy. To acquire the quantitative description of the differences between samples, the similarity of the series dynamic spectra, which reflects the similarity of the samples themselves if obtained under the same perturbation condition, is evaluated by the symmetry of hetero 2DCOS map. Two parameters, the Euclidian distance and correlation coefficient between the upper left and lower right triangular parts of a hetero 2DCOS map, are introduced for the quantitative measure of the symmetry, which in turn characterizes the similarity of the responses of samples to a given perturbation. The above method is used to discriminate one genus of Astragalus from the others to ensure the medicinal efficacy and safety of the herb. Hypothesis tests show that the inter-distances between samples from different genera are significantly larger than the intra-ones within the same genera, while the inter-correlation coefficients are smaller than the intra-ones. The excellent result of the identification for all samples carried out by a t-test based on the distances indicates that this method provides an efficient technique for the quantitative evaluation of similarity between samples.

[Study on genetic relationship of Astragalus membranaceus var mongholicus in different producing area using SRAP].

OBJECTIVE: To study the genetic relationship of Astragalus membranaceus var. mongholicus in different producing area and provide theoretical basis for the evaluation of Astragalus germplasm resources. METHOD: Through quence-related amplified polymorphism (SRAP) analysis, the systematic diagram of genetic relationship was constructed by UPGMA method. RESULT: A total of 141 SRAP markers were scored. By the use of UPGMA cluster analysis with genetic distance, Astragalus could be divided into two provenance plots of Gansu and Shanxi. CONCLUSION: The genetic differentiation among populations of A. membranaceus var. mongholicus is remarkable. SRAP marker could be efficiently used for the study of the genetic relationship of Astragalus.

[A taxonomic study on the original plant of radix astragali].

There are dispute about the status of taxonomy among Astragalus membranaceus (Fisch.) Bge, A. membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao. and A. pallidipurpureus stat. nov. The varieties and taxa of the complex are still in need of revision. With molecular biology study used trnH-psbA intergenic region, the taxonomic revision of Radix Astragali has been made. A. pallidipurpureus stat. nov is suggested as a new species.

[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.

Determination and discrimination of intraspecific diversity of Astragalus gossypinus by eco-phytosociological method from west of Iran.

Astragalus gossypinus Fischer with wide distribution in Iran belongs to the genus Astragalus (Fabaceae). According to existing references and information, individuals of this species are present in many stations with different ecological conditions. This study carried out for determination and discrimination of intraspecific diversity of Astragalus gossypinus by Eco-phytosociological method from west of Iran. In this method, the principle of data collecting and analyzing based on floristical composition (as floristical markers) of each Endogenous milieu (as the unit of study in Eco-phytosociological method). In this order, application of Endogenous milieu (special station) for data collecting and then their analyzing permit us only determine existence of inter and intraspecific diversity. Then for determinating kind and level of intraspecific diversity (Ecophene, Chemotype, Cytotype, Ecotype ...), can use other studies such as: morphological, anatomical, phytochemical, cytological and etc. In this survey, 29 special stations were studied and 195 species distinguished as companions for Astragalus gossypinus. Then floristic-ecologic data collected from each 29 special stations and analyzed by Anaphyto software (F.C.A, A.H.C, B.O, Marquag methods). Comparison of obtained results on multiple coordinate axes from F.C.A method with results from B.O, Marquag and A.H.C methods led to determination of 7 main groups of Endogenous milieus (special station). Flavonoid analyses were used for determinating kind and level of intraspecific diversity in 7 discriminated groups. Leaves flavonoid components of all collected individuals of Astragalus gossypinus were investigated by TLC method. Obtained data from flavonoid survey analyzed by SPSS and MVSP package with WARD and UPGMA methods. Finally, the results of flavonoid studies confirmed the same 7 groups that identified by floristical composition study and showed intraspecific diversity in chemotype level. So according to these results, we can introduce 7 chemotypes for Astragalus gossypinus from west of Iran. These chemotypes exist in different stations with various ecological conditions.