Article

A new species of Astragalus L. (Leguminosae) from India based on morphological and molecular markers

Authors:
  • CSIR-National Botanical Research Institute - India
  • CSIR-National Botanical Research Institute, Lucknow, India
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Abstract

A new species, Astragalus himachalensis, is described and illustrated from the Himalaya in India. The species grows occasionally in different parts of the cold desert of Lahul-Spiti (Himachal Pradesh). It resembles A. himalayanus, but differs chiefly in the small size of the plants (length, 5–15 cm), diadelphous stamens, and small pods (length, 5–6 mm) with the stipe length (2–3 mm) more or less equal to the calyx length. The similarities and significant differences between A. himachalensis and A. himalayanus were confirmed by analyses of random amplification of polymorphic DNA (RAPD) and directed amplification of minisatellite DNA (DAMD) markers. RAPD and DAMD methods clearly distinguished between the two species and were in congruence with morphological markers. © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society, 2007, 154, 27–34.

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... Recently, A. himachalensis Chaudhary & Rana has been segregated from A. himalayanus Klotz. based on morphological and molecular markers (Chaudhary et al., 2007b). ...
... Apart from these major phylogenetic investigations, recently many genetic studies have also been carried out in Astragalus by using RAPD and AFLP and isoenzyme study to establish the correct identity and relationship of some closely related taxa belonging to different taxonomic complexes (Luo et al., 2000;Knaus et al., 2005;Mehrnia et al., 2005;Adiguzel et al., 2006;Zarre et al., 2007;Chaudhary et al., 2007b). ...
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Astragalus is considered one of the most diverse genera in the family Leguminosae (nom. alt. Fabaceae). Although a large number of works have been carried out on the genus, no monograph is available except some regional accounts and revisions chiefly at sectional level. It may be due to the sheer size of the genus (ca. 3000 spp.) and diverse nature, the genus is quite variable in habit and habitats, size of the plants, nature of indumentums, stipules, leaf rachis, types of inflorescence, relative length of petals, pods etc. Usually, genus is divided into eight to ten subgenera and more than 245 sections. In recently conducted molecular phylogenetic studies it has been shown that none of the subgenera and large sections are monophyletic. However, it has been clearly demonstrated that Astragalus is monophyletic except some outlier species. The chromosome numbers are also quite interesting and significant in Astragalus for its phylogenetic studies. There is a strong correlation between its geographic distribution and chromosome numbers. Currently about 80 species have been recorded from India chiefly from the Himalayas. Except some of our recent publications, not much studies have been carried out on the genus in India after 'The Flora of British India'. Astragalus is not of much economic importance, however, some of its species are well known for commercial gum tragacanth production especially in Iran and China. In India, A. candolleanus is a well known drug as 'Rudanti' or 'Rudravanti' used for tuberculosis, skin diseases, coughs and blood purifier. The aim of this article is to review the entire work carried out on Astragalus and to bring out scattered information at one place for better understanding of the subject and to find out the future prospective of the research in India on the genus.
... Astragalus, plusieurs recherches ont été entreprises à l'aide d'outils moléculaires tels que le polymorphisme de longueur des fragments de Restriction (RFLP) ciblant le locus des gènes codant pour les ARN polymérases chloroplastiques Rpoc 1 et Rpoc 2 (Sanderson, 1991;Liston, 1992;Sanderson et Doyle, 1993;Wojciechowski et al., 1993;Liston et Wheeler, 1994;Wojciechowski, 2005), le séquençage des gènes ribosomaux (nrDNA ITS) et des séquences simples internes répétées (ISSR) (Abdel Samad et al., 2014) mais également l'amplification aléatoire d'ADN polymorphe (RAPD), et l'analyse du polymophisme de séquences minisatellites (DAMD) (Chaudhary et al., 2007). ...
Thesis
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Astragalus gombo Bunge (Fabaceae) est une plante bien représentée dans le Sahara Septentrional Est Algérien (région de Ouargla, Algérie) et couramment utilisée comme fourrage pour les animaux ou en médecine traditionnelle. Malgré de nombreuses publications sur la richesse en polysaccharides d’autres espèces appartenant au genre Astragalus et la description de leurs propriétés biologiques putatives ou avérées, aucune étude ne s’est intéressée à l’espèce gombo. L’objectif de ce travail de thèse a donc été de réaliser des investigations sur plusieurs parties de cette plante en vue d’extraire, identifier et caractériser son contenu en polysaccharides. Dans un second temps les propriétés biologiques et rhéologiques de ces polymères ont été étudiées afin d’identifier d’éventuelles voies de valorisation. Les résultats obtenus ont permis de détecter la présence de composés pectiques et hémicellulosiques dans la tige d’Astragalus gombo et celle d’un galactomannane dans les graines. Le galactomannane est de haute masse moléculaire (1,1x106 Da) et est constitué d’une chaine principale de β-(1→4)-D-mannane ramifiée en α-(1→6) par des résidus Dgalactopyranoses avec un rapport M/G de 1,7. La caractérisation de ses propriétés rhéologiques a révélé un comportement typique rhéofluidifiant et des propriétés viscoélastiques. L’étude des activités biologiques associées à ce biopolymère a révélé son potentiel comme prébiotique et antioxydant.
... Astragalus is the largest genus in the Fabaceae family and represented by 445 species (224 are endemic) in the flora of Turkey and 2500-3000 species worldwide [1][2][3][4]. Astragalus roots are used in traditional chinese medicine due to their antidiabetic, antioxidant and antineoplastic properties [5][6]. Mainly three major classes of compounds, polysaccharides, saponins and isoflavonoids were isolated from Astragalus species [2]. ...
Article
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Astragalus species are medicinal plants which produce valuable secondary metabolites, especially cycloartane-type glycosides. In this study, stem and leaf explants of Astragalus trojanus were subjected to different plant growth regulators, environmental conditions and media compositions to identify their callus responses. Stem and leaf explants were cultured in Murashige and Skoog (MS) and woody plant (WPM) media supplemented with different concentrations of kinetin, naphthalene acetic acid, 2,4-dichlorophenoxyacetic acid, thidiazurone and indol acetic acid under two light intensities (1000 and 4000 lux) and also in dark conditions. Both MS and WPM media triggered callus regeneration. Although, callus regeneration was observed on both stem and leaf explants, callus biomass accumulation on stem explants were higher. Addition of 100 μg/L selenium and doubled concentration of WPM vitamins enhanced callus biomass on stem explants under dark conditions. Stem explants also regenerated shoots at high frequencies (up to 93%), especially in kinetin added media. Astragaloside IV and cycloastragenol accumulation efficiencies were determined in calli tissues. The highest astragaloside IV production (3.5 μg/mg) was found in callus tissue regenerated from stem explants in D1 medium, whereas the highest cycloastragenol accumulation (4.8 μg/mg) was detected in callus tissue regenerated from stem explants in N2 medium.
... RAPD-PCR revealed significant differences (45% polymorphism) in band pattern among longand short-term calli. When Chaudhary et al. (2007) compared two Himalayan species of Astragalus, the similarities and significant differences between A. himachalensis and A. himalayanus were confirmed by RAPD analysis. Even though these species share the same environment, the polymorphism percentage reached 100%. ...
Article
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The objective of this study was to investigate the phenotypic and genetic effects of long-term (12 years) in vitro culturing of Astragalus chrysochlorus callus tissue. Murashige and Skoog medium supplemented with 0.5 mg/l 2,4-dichlorophenoxyacetic acid was used for the production of young callus (2 months old) from hypocotyl explants. Morphological analysis showed that long-term cultures were well adapted, with greener and more friable calli, and a higher growth index (2.11 ± 0.39) was compared with that in short-term cultures (1.2 ± 0.73). Genetic variation between the two types of calli was investigated by randomly amplified polymorphic DNA analysis, which showed different amplification patterns with a polymorphism level of about 45%. We furthermore measured RNA transcript levels for two key enzymes of the important stress-related phenylpropanoid pathway, PAL and C4H, via quantitative polymerase chain reaction. The gene expression levels of C4H and PAL were 0.205- and 0.584-fold lower in old cultures than in young cultures. Metabolic analysis by high performance thin layer chromatography clearly showed differences in flavonoid type metabolic content. Together, these data show that long-term culturing of callus tissue leads to genetic heterogeneity and variations in secondary metabolite content.
... However, the use of RAPD has been criticized for revealing unreliable phylogenies owing to the occasional lack of reproducibility and a possible lack of homology in comigrating bands (8,9). Despite this limitation, the RAPD method has been successfully employed in several plant genera for revealing genetic relationships following the optimization of polymerase chain reaction (PCR) conditions (3,(10)(11)(12)(13)(14)(15)(16)(17). ...
Article
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Random amplified polymorphic DNA (RAPD) markers were used to determine genetic relationships among Velezia L. species from Turkey. A total of 432 amplified bands were obtained using 14 RAPD primers. The polymorphism in RAPD markers was high (98.61%) and was sufficient to distinguish each species. The degree of band-sharing was used for evaluating the genetic similarity between species and for constructing a dendrogram by the unweighted pair group method with arithmetic mean (UPGMA). Genetic relationships among the species were found to be fully consistent with those obtained by the use of morphological characters. Data obtained from our study demonstrated that the RAPD technique could be successfully used for the determination of genetic relationships among Velezia species and for species identification. Furthermore, it can be efficiently employed in future studies to provide preliminary data for conservation of endangered Velezia species.
... In recent years PCR based methods such as RAPD, ISSR and DAMD have also been used quite extensively due to their ease, rapidity and reliability for analysis of molecular differentiation and for resolving taxonomic problems in plants. These methods have also been applied to study intra and inter-specific relationship among different species of Astragalus (Luo et al., 2000;Mehrina et al., 2005;Alexander et al., 2004;Na et al., 2004;Chaudhary et al., 2007b) and in many other plant groups (Tautz and Renz, 1984;Nagaoka and Ogihara, 1997;Fang and Roose, 1997;Sant et al., 1999;Dangi et al., 2004). The RAPD analysis utilizes single arbitrary sequence of ten bases as oligonucleotide primers to amplify discrete fragments of DNA in low-stringency of polymerase chain reactions (Welsh and McClelland, 1990;Williams et al., 1990;Wolfe and Liston, 1998;Harris, 1999). ...
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Forty accessions belonging to four closely related species of the Astragalus rhizanthus complex (i. e., A. rhizanthus, A. candolleanus, A. malacophyllus and A. pindreensis) were collected from different parts of the Indian Himalaya and analyzed for intra-and inter-specific relationship using Random Amplified Polymorphic DNA (RAPD), Inter Simple Sequence Repeats (ISSR) and Directed Amplification of Minisatellite DNA (DAMD). Eighteen RAPD, 15 ISSR and 6 DAMD primers were used to study the genetic diversity between the species. A total of 449 bands for RAPD, 326 bands from ISSR and 179 bands from DAMD were obtained. Nearly 98.44% of RAPD, 99.38% of ISSR and 98.32% of DAMD bands were polymorphic. Pair-wise genetic similarity for a cumulative data was determined using Jaccard's similarity coefficient which varied from 0.23 to 0.82 and a Neighbor-joining (NJ) dendrogram was generated. The genetic similarity from the data matrix estimated by Jaccard's coefficient shows that all four species are distinct in accordance with our morphological findings as well as previous taxonomic treatment in which these species have been treated distinctly. The highest degree of genetic similarity was observed within A. rhizanthus followed by A. malacophyllus, A. candolleanus and A. pindreensis. This study has proved that these markers have the ability to distinguish the closely allied species as well as analyze the genetic diversity within and between the species.
... Coming year 2005 recorded 13 additions, including an orchid [68][69][70][71]. Herniaria incana [72] was added in 2006 and so were two new species viz., Stellaria pinvalliaca [73] and Astragalus himachalensis [74]. The year ended with further addition of an orchid [75]. ...
Article
Biodiversity information owes a lot to field surveys and floristic studies. This is the basis of monitoring and assessment exercises. The present paper provides temporal information on the history of floral surveys in Himachal Pradesh (HP) and the consequent statistics of species richness in HP. From 1984 (after the publication of Flora of HP—an analysis) till the end of 2010, estimates of species have increased by 14.85 %, of genera by 10.46 % and of families by 9.60 %. During the course, 16 new species were reported. The surveys have not only updated the knowledge but have also led to identification of localities and taxa that demand critical studies and conservation strategies. The paper argues for adequate support to survey based activities and an action plan in this regard.
... In recent years, PCR-based methods like ISSR, RAPD, and DAMD have been extensively used to assess the intraspecific genetic diversity as well as to establish the interspecific relationships in several plant groups like Cicer [30], Astragalus [31][32][33][34][35][36], Phyllanthus [37], and Trigonella [38]. In the present study, 20 genotypes of the A. rhizanthus ssp. ...
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Astragalus rhizanthus with three infraspecific taxa (i.e., A. rhizanthus ssp. rhizanthus var. rhizanthus, A. rhizanthus ssp. rhizanthus var. pindreensis, and A. rhizanthus ssp. candolleanus) is widely scattered in the Himalaya from Jammu and Kashmir to Uttarakhand provinces in India. Among them, A. rhizanthus ssp. rhizanthus var. rhizanthus exhibits enormous morphological diversity throughout its range of distribution. An assessment of genetic diversity studies was undertaken to understand the level and pattern of diversity, using Inter simple sequence repeats (ISSR), Random amplified polymorphic DNA (RAPD) and Directed amplification of minisatellite DNA (DAMD) profiles. Fifteen ISSR, 18 RAPD and 6 DAMD primers were used to unravel the diversity among 20 genotypes collected from the known localities in the Indian Himalaya. A total of 242 bands from ISSR, 352 from RAPD and 142 from DAMD were obtained with an average of 92.23% polymorphism in the species. Pair-wise genetic similarity for the cumulative data was determined using Jaccard's similarity coefficient which varied from 0.19 to 0.84. A combined UPGMA dendrogram was generated which revealed that different genotypes exhibited their affinity according to their geographical distribution. Tree topology suggests the existence of two distinct groups of the genotypes. Gene diversity (H) and Shannon's information index (I) were estimated and these values were found higher in the genotypes collected from Jammu and Kashmir than Himachal Pradesh.
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Ten new species of Astragalus are de- scribed here. They belong to the following sections: section Alopecuroidei: A. burqinensis; section Bra- chycarpus: A. barclayanus, A. golubojensis, A. mieherorum, A. nanshanicus, A. pseudojagnobicus, A. sagastaigolensis; section Hemiphaca: A. nigro- dentatus; section Polycladus: A. tibeticola; section Skythropos: A. griersonii. Furthermore, six taxa are raised in rank.
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Molecular genetic maps are commonly constructed by analyzing the segregation of restriction fragment length polymorphisms (RFLPs) among the progeny of a sexual cross. Here we describe a new DNA polymorphism assay based on the amplification of random DNA segments with single primers of arbitrary nucleotide sequence. These polymorphisms, simply detected as DNA segments which amplify from one parent but not the other, are inherited in a Mendellan fashion and can be used to construct genetic maps in a variety of species. We suggest that these polymorphisms be called RAPD markers, after Random Amplified Polymorphic DNA.
Flora of Lahul-Spiti District
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Flora of Lahul-Spiti District, Himachal Pradesh Dehra Dun: Bishen Singh & Mahendra Pal Singh Origin and genetic diversity of Spartina anglica (Poaceae) using nuclear DNA markers
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