Onobrychis Species Research Articles

Species of the OnobrychisGenus as Promising Species of Azerbaijan Flora

One of the tasks of pharmacy is to find affordable raw materials for herbal medicines. The studies were conducted in Azerbaijan. The results of the studies indicate that Onobrychis species are a rich source of β-arbutin. Arbutin (4-hydroxyphenyl-β-D-glucopyranoside), a monoglycoside form of hydroxyquinone, has been used in the field of cosmetology for several decades due to its effectiveness in the treatment of hyperpigmentation and removal of freckles. Onobrychis species are of significant ecological and economic importance, and also demonstrate intriguing therapeutic properties that have attracted attention in the field of medicine. The presence of bioactive compounds such as flavonoids and phenols, which are known for their antioxidant properties, is another noteworthy aspect. Antioxidants play a crucial role in neutralizing harmful free radicals in the body, which are involved in a wide range of diseases and the aging process. Onobrychis has anti-inflammatory properties. It is well known that inflammation is a common factor in many chronic diseases. Plants have antimicrobial, immunostimulating, hypoglycemic, hypocholesterolemic, gastrointestinal tract normalizing and vascular wall strengthening properties. The study of Onobrychis opens up potential prospects in the field of creating natural products.

In vitro Evaluation of Skin Related Enzyme Inhibitory Effect and Emulgel Formulation Development Studies of Onobrychis Argyrea subsp. Argyrea with Phytochemical Analysis.

Species of Onobrychis have been used to treat skin disorders such as wounds and cuts in folk medicine and Onobrychis argyrea subsp. argyrea (OA) commonly known as 'silvery sainfoin', is a member of this genus. In this study, it was aimed to investigate the skin-related biological activities and phytochemical characterization of OA. Moreover, an emulgel formulation was developed from the main methanolic extract of the plant (OAM). Initially, to identifiy of the active fractions, aerial parts of the plant material was extracted with methanol and fractionated by n-hexane, chloroform, ethyl acetate and n-butanol, respectively. Antioxidant activity was determined by CUPRAC, TOAC, FRAP and DPPH assays. Thereafter, the inhibition potential of OAM, novel formulation and all fractions was measured against elastase, collagenase, tyrosinase and hyaluronidase enzymes. OAM was analyzed and characterized by LC/MS-MS. The major bioactive flavonoids which are rutin and isoquercetin were measured and compared as qualitative and quantitative via high performance thin layer chromatography (HPTLC) analysis in OAM and fractions. The results showed that extracts of OA can be a potential cosmeceutical agent for skin related problems.

First Trials of Genome Analyses in Some Onobrychis Species using Dot-Blot and Genomic in situ Hybridization Techniques

The origin and genome composition of tetraploid cultivated Onobrychis viciifolia (2n = 4x = 28) were analyzed using dot-blot and genomic in situ hybridization (GISH) techniques. Dot-blot hybridization was used to find a genomic affinity between O. viciifolia and 16 diploid Onobrychis species. The hypothesis on the origin of the O. viciifolia was tested using GISH. Dot-blot analyses suggested a genomic affinity between O. viciifolia and four diploid Onobrychis species (O. kachetica, O. supina, O. pallasii, and O. vaginalis). Hybridization signals were observed on O. viciifolia chromosomes when gDNA of O. kachetica, O. supina, O. pallasi, and O. hypargyrea were used as probes. However, the observed chromosomal distribution of hybridization signals did not resemble GISH results. The observed signals colocalized with 35S rDNA or dispearse signals on all chromosomes were observed depending on the probe. Further investigations using more comprehensive and comparative analysis with both coding and repetitive DNA regions may provide a better understanding of the genome composition and evolution of O. viciifolia.

Phytochemical profile by LC-MS/MS analysis and evaluation of antioxidant, antidiabetic, anti-Alzheimer, and anticancer activity of Onobrychis argyrea leaf extracts

IntroductionStudies on different species of Onobrychis, a member of the Fabaceae family, have revealed a number of biological activities with potential applications in alternative medicine. The phytochemical content of the plant Onobrychis argyrea subsp. argyrea and some of the important biological activities linked to its metabolism are quite limited. Therefore, this study was the first to determine the content of secondary metabolites in this species and to elucidate important biological activities related to metabolism. MethodsLC-MS/MS was used for the quantitative analysis of bioactive compounds in the aerial part of O. argyrea extracts. The antioxidant properties of the extracts were assessed based on their ability to scavenge DPPH radicals and reduce iron ions. The anti-diabetic effect of the extracts was tested by the inhibition of α-amylase and α-glycosidase enzymes, and the anti-Alzheimer's ability was tested by the inhibition of the enzymes AChE and BChE. The cytotoxic effects of the extracts were tested on four different cancer lines, namely A-549, HT-29, MCF-7 and MDA-MB 453 using the XTT assay. The mechanism of the anti-cancer effect was determined by means of flow cytometry analysis on HT-29 cells. ResultsTwenty-nine phytochemicals were identified via the LC-MS/MS analysis in the plant extracts. The most abundant phytochemicals in the extracts were quinic acid, isoquercitrin, epicatechin, and routine, respectively. Antioxidant analysis showed that among all extracts, methanol extract (ME) was most effective in scavenging DPPH (IC50: 23.93 ± 0.96 µg/mL) and in reducing iron ions (53.24 ± 2.14 mgTEAC/g). Enzyme activity studies demonstrated that ME is a potent inhibitor of α-glycosidase (IC50: 15.06±0.64 µg/mL), AChE (IC50: 24.75±0.76 µg/mL), and BChE (IC50: 3.36±0.14 µg/mL). XTT assay results disclosed the strongest anti-proliferative activity of ME against the human colorectal adenocarcinoma HT-29 cell line with an IC50 value of 33.35±0.92 µg/mL. Flow cytometry analysis revealed that ME caused mitochondrial membrane damage in HT-29 cells (**p<0.01) and subsequently induced apoptosis (**p<0.01) by activating the caspase pathway (**p<0.001). ConclusionO. argyrea, rich in bioactive compounds, showed remarkable antioxidant, antidiabetic, Alzheimer's, and anticancer properties in vitro, demonstrating that this plant contains secondary metabolites that may be potential drug candidates for the treatment of metabolic diseases.

Potential use of wild Onobrychis species for climate change mitigation and adaptation

AbstractClimate change is threatening environmental and nutritional security. In this scenario, forage crops and their wild relatives may contribute to maintain ecosystem services and minimize the effects of global warming. We reviewed the literature regarding the ecological, agronomic, and nutraceutical values of wild Onobrychis Miller species (sainfoins, Family Fabaceae). We also discussed their application prospects and the existing problems hampering their widespread domestication. Wild species of the genus Onobrychis have some interesting agronomic and ecological features such as perenniality, deep root system, and high‐stress tolerance, which make them suitable for future cultivation under climate change. Wild Onobrychis species are multipurpose species, which can be utilized for fodder and honey production under harsh environmental conditions. In addition, wild Onobrychis species water, ethanol, and methanol extracts exhibit antioxidant, antidiabetic, anti‐inflammatory, antitumor, antimicrobial, anti‐stressor, and many other pharmacological effects. Wild Onobrychis species have not been the subject of detailed studies. Compared with the cultivated forage legumes, wild Onobrychis species are still underexploited despite their potential to alleviate forage and food insecurity, either by domestication or by providing novel alleles for breeding purposes of the cultivated species Onobrychis viciifolia. Our review aims to increase scientific awareness toward exploring the potential uses of wild Onobrychis species to tackle current climate change.

Evaluation of the Genetic Structure of Some Accessions Belonging to Onobrychis spp. Using Microsatellite DNA Markers

Turkey is in a very convenient position for animal husbandry in terms of both natural resources and ecological conditions. Forage crops, which has a very important place in agricultural activities, is the insurance of plant and animal production. Sainfoin is a perennial forage legume species that grown in the northern temperate regions of the world from the Mediterranean region and the Caucasus, and to Central Asia. In this study the genetic diversity of 100 genotypes representing 44 accessions from 18 different Onobrychis species (O. arenaria subsp. arenaria, O. inermis, O. petraea, O. cyri, O. iberica, O. altissima, O. vassilczenkoi, O. conferta subsp. argentea, O. alba subsp. laconica, O. biebersteinii, O. grandis, O. kachetica, O. kemulariae, O. oxyodonta, O. megataphros, O. pallasii, Onobrychis spp., and O. viciifolia) were evaluated using 8 simple sequence repeat (microsatellite) markers. Based on the results, OVK036, OVK094, OVK125, OVM033, OVK161, OVK046, OVM061, and OVK174 loci were polymorphic. The observed number of alleles per SSR locus ranged from 6 to 21 alleles (mean of 11.625). Maximum allele frequency ranged from 0.51 to 0.93 with a mean value of 0.73. The PIC value ranged from 0.124 to 0.244. The mean polymorphism information content of loci was 0.188. Genetic diversity coefficients according to the UPGMA ranged from 0.000 to 0.9375. Cluster analysis divided the 100 sainfoin genotypes into two main groups (Cluster-I and Cluster-II). All diploid genotypes (except for 1 diploid genotype) used in the study formed a separate group within Cluster-I. The results revealed that SSR markers used in this study are useful for molecular characterization and assessing genetic diversity of sainfoin accessions. The obtained SSR alleles and genetic variability in a studied certain loci provided significant information about the genetic structure of sainfoin accessions that could be used as parental lines in sainfoin breeding programs.

The Chromosome Number and rDNA Loci Evolution in Onobrychis (Fabaceae).

The evolution of chromosome number and ribosomal DNA (rDNA) loci number and localisation were studied in Onobrychis Mill. Diploid and tetraploid species, as well as two basic chromosome numbers, x = 7 and x = 8, were observed among analysed taxa. The chromosomal distribution of rDNA loci was presented here for the first time using fluorescence in situ hybridisation (FISH) with 5S and 35S rDNA probes. Onobrychis species showed a high polymorphism in the number and localisation of rDNA loci among diploids, whereas the rDNA loci pattern was very similar in polyploids. Phylogenetic relationships among the species, inferred from nrITS sequences, were used as a framework to reconstruct the patterns of basic chromosome number and rDNA loci evolution. Analysis of the evolution of the basic chromosome numbers allowed the inference of x = 8 as the ancestral number and the descending dysploidy and polyploidisation as the major mechanisms of the chromosome number evolution. Analyses of chromosomal patterns of rRNA gene loci in a phylogenetic context resulted in the reconstruction of one locus of 5S rDNA and one locus of 35S rDNA in the interstitial chromosomal position as the ancestral state in this genus.

The complete chloroplast genome of Onobrychis gaubae (Fabaceae-Papilionoideae): comparative analysis with related IR-lacking clade species

BackgroundPlastome (Plastid genome) sequences provide valuable markers for surveying evolutionary relationships and population genetics of plant species. Papilionoideae (papilionoids) has different nucleotide and structural variations in plastomes, which makes it an ideal model for genome evolution studies. Therefore, by sequencing the complete chloroplast genome of Onobrychis gaubae in this study, the characteristics and evolutionary patterns of plastome variations in IR-loss clade were compared.ResultsIn the present study, the complete plastid genome of O. gaubae, endemic to Iran, was sequenced using Illumina paired-end sequencing and was compared with previously known genomes of the IRLC species of legumes. The O. gaubae plastid genome was 122,688 bp in length and included a large single-copy (LSC) region of 81,486 bp, a small single-copy (SSC) region of 13,805 bp and one copy of the inverted repeat (IRb) of 29,100 bp. The genome encoded 110 genes, including 76 protein-coding genes, 30 transfer RNA (tRNA) genes and four ribosome RNA (rRNA) genes and possessed 83 simple sequence repeats (SSRs) and 50 repeated structures with the highest proportion in the LSC. Comparative analysis of the chloroplast genomes across IRLC revealed three hotspot genes (ycf1, ycf2, clpP) which could be used as DNA barcode regions. Moreover, seven hypervariable regions [trnL(UAA)-trnT(UGU), trnT(GGU)-trnE(UUC), ycf1, ycf2, ycf4, accD and clpP] were identified within Onobrychis, which could be used to distinguish the Onobrychis species. Phylogenetic analyses revealed that O. gaubae is closely related to Hedysarum. The complete O. gaubae genome is a valuable resource for investigating evolution of Onobrychis species and can be used to identify related species.ConclusionsOur results reveal that the plastomes of the IRLC are dynamic molecules and show multiple gene losses and inversions. The identified hypervariable regions could be used as molecular markers for resolving phylogenetic relationships and species identification and also provide new insights into plastome evolution across IRLC.

&lt;p&gt;&lt;strong&gt;&lt;em&gt;Onobrychis silvanensis sp. nov.&lt;/em&gt;&lt;/strong&gt;&lt;strong&gt;, a new Fabaceae (sect. &lt;em&gt;Hymenobrychis&lt;/em&gt;) taxon from Turkey&lt;/strong&gt;&lt;/p&gt;

Onobrychis silvanensis sp. nov., is described from Silvan, Diyarbakır province, in southeastern Anatolia, Turkey. The new species is closely related to O. galegifolia and O. radiata. The white corolla feature of O. silvanensis is uncommon in Turkish Onobrychis species and is otherwise known only in O. haussknecthii and O. hypargyrea. In addition to the diagnosis and taxonomic description, morphological characters of both flowering and fruiting O. silvanensis plants are presented. Distribution maps for O. silvanensis, O. galegifolia and O. radiata in Turkey are also provided. Although currently Data Deficient by IUCN status criteria, we anticipate O. silvanensis constitutes a candidate for Endangered status.

Micropropagation of some Onobrychis species through in vitro shoot regeneration

Onobrychis species have an extensive spread in Turkey and adapted to various environmental conditions. In this study, it was aimed to propagate with tissue culture methods of endemic Onobrychis fallax Freyn &amp; Sint. ex Freyn var. longifolia Aktoklu, Onobrychis stenostachya subsp. sosnowskyi, Onobrychis elata Boiss. &amp; Balansa and worldwide commonly cultivated Onobrychis viciifolia Scop. species. For this purpose, cotyledon node explants were cultured in Murashige and Skoog (MS) media containing various concentration of 6-benzylaminopurine (BAP), thidiazuron (TDZ) or meta-Topolin (mT) alone or in combination with 0.5 mg L–1 NAA. Significant variations in number of shoots per explant was observed depending on the species and growth regulators. The greatest number of shoots per explant was obtained from 1.2 mg L–1 TDZ treatments in O. stenostachya, O. fallax and O. elata and from 0.3 mg L–1 TDZ treatments in O. viciifolia, whereby 10.58, 9.50, 5.5, 6.42 shoots were recorded. Resultant shoots were rooted in half strength MS nutrient media containing 1 or 2 mg L–1 indole-3-butyric acid (IBA). The rooting ratio was 86.11% in O. viciifolia, 50.00% in O. stenostachya and 36.11% in O. fallax and O. elata.

Influence of Peg-induced Drought Stress on Antioxidant Components of Callus Tissue of Sainfoin (Onobrychis viciifolia Scop.) Ecotypes

Background: Drought is one of the important abiotic stress factors that restrict plant development. Sainfoin is known to be relatively tolerant of drought. However, there are limited reports on the effects of drought stress on antioxidant components of Onobrychis species and/or ecotypes. Methods: To determine drought stress effects on antioxidant components of 4 sainfoin ecotypes (“Koçaþ”, “Malya”, “Altýnova” and “Ulaþ”), callus tissue was grown on MS medium enriched with 200 g l-1 PEG-6000. Result: Callus of the sainfoin ecotypes, cultured on a medium having 200 g l-1 PEG-6000, showed significant increases in antioxidant enzyme activities (SOD, CAT, GR (except in “Ulaþ” ecotype) and APX). However, the PEG induced increase in the accumulation of MDA and proline in callus tissue of all sainfoin ecotypes. The findings of the present study show that in terms of the increasing rate of antioxidant components under drought stress, the “Koçaþ” ecotype seemed to be the best.

Analysis of pollen morphological characteristics in Iranian Onobrychis Miller (Fabaceae) taxa

We verified the taxonomic importance of pollen grain characteristics, based on pollen morphology of 44 populations of 36 Onobrychis Miller taxa belonging to two subgenera and six sections, chiefly for delimitation of taxa, sections, and subgenera. We evaluated 1100 pollen grains (25 per taxon) and tested them by 13 qualitative and quantitative characteristics using scanning electron microscopy and light microscopy. For 19 Onobrychis species, pollen descriptions are presented for the first time. We used Image Tools, MVSP, and SPSS software for statistical analyses. Pollen grains of all the taxa were tricolpate, single, radially symmetrical, and isopolar. We identified three shape classes according to the polar/equatorial axes length ratio: prolate (the dominant), perprolate, and subprolate. The outline shapes in polar view were almost identical, while the taxa were different in the equatorial view of pollen grains. We found the following three exine sculpture types occurred in the species studied: reticulate, suprareticulate, and microreticulate. Our results suggest that Onobrychis pollen morphological characters were relatively homogeneous, which agreed with previous investigations. Nevertheless, our analyses revealed significant variation in pollen characters among the studied taxa, whereas the outputs of principal components analysis and hierarchical clustering vaguely confirmed the arrangement of the Onobrychis species into subgenera and sections according to Flora Iranica. Therefore, the morphological features of pollen grains in Onobrychis taxa have limited taxonomic importance for delimitation of subgenera, sections and species.

Antidiabetic activity evaluation of Onobrychis species on alloxan-induced diabetic mice

In the current study, four Onobrychis species, O. albiflora Hub.-Mor., O. argyrea Boiss. subsp. argyrea Boiss., O. galegifolia Boiss., and O. tournefortii (Willd.) Desv. were collected from Anatolia to be evaluated for their antidiabetic activities. Methanol water extracts of the aerial parts were used for experiments. An alloxan-induced diabetic mice test model was used. Phytochemical analysis of the tested extracts was investigated using the HPLC method. The highest activity was observed with treatment of O. albiflora aerial part extract. Significant decrements were detected in the blood glucose levels as follows: 180.83±47.48 and 252.83±50.47mg/dL at 100 mg/kg and 200 mg/kg doses of O. albiflora, respectively, when compared to the isotonic saline solution control group, eliciting a blood glucose level of 494.20±27.32. Among the tested standard compounds, rutin and isoquercetin were detected in the examined species. The highest amount of rutin (1.1981±0.0017%) and isoquercetin (0.7318±0.0197%) were found in O. albiflora and O. argyrea subsp. argyrea, respectively. Antidiabetic activities of the tested Onobrychis species seem to indicate a possible correlation with their rutin and isoquercetin contents. Therefore, rutin and isoquercetin may be the antidiabetic compounds that contribute to the antidiabetic activity of the tested Onobrychis species.

Determination of the Fatty Acid Compositions of Four Onobrychis Species from Turkey

Determination of the Fatty Acid Compositions of Four Onobrychis Species from Turkey

The phylogeny and new classification of the genus Onobrychis (Fabaceae-Hedysareae): evidence from molecular data

The present study was conducted to analyze the phylogenetic status of the genus Onobrychis and to evaluate the monophyly of its subgenera and sections and relationship among them. We sequenced the nuclear ribosomal DNA internal transcribed spacer (nrDNA ITS) and three chloroplast regions trnL-F, rpl32/rpl32-trnL(UAG) and ndhF-rlp32 for phylogenetic reconstruction of 51 species of Onobrychis. In all of our analyses, Eversmannia subspinosa, Corethrodendron scoparium, Greuteria membranacea and G. argyrea were chosen as outgroups. Phylogenetic analyses were performed by maximum parsimony, maximum likelihood and Bayesian methods. Our molecular data indicate that Onobrychis is monophyletic and composed of two main clades, each corresponding to the redefined subgenus Onobrychis (including sections Onobrychis and Hemicyclobrychis) and subgenus Sisyrosema (including sections Afghanicae, Laxiflorae, Heliobrychis, Hymenobrychis, Insignes, Lipskyanae and Litvinovianae), respectively. Sections Lipskyanae and Litvinovianae are newly established and described, representing distinct lineages within the genus. Onobrychis splendida, a species hitherto without a sectional position, along with some members of sect. Anthyllium were retrieved representatives of section Lipskyanae. Sections Afghanicae, Insignes, Heliobrychis and Hymenobrychis (with the inclusion of two species of section Anthyllium) are monophyletic. Sections Dendrobrychis and Lophobrychis are reduced to synonymy of section Onobrychis and Anthyllium to synonymy of section Hymenobrychis. A taxonomic treatment for the genus is presented.

Antioxidant Activity, Total Phenolic and Flavonoid Contents of Three Onobrychis Species from Iran

Background: Plant phenolic compounds are a main group of plant natural products and flavonoids are the largest and best studied natural phenols. These substances possess a series of biological properties and act on biological systems as antioxidants. In present research, the aim is to determine in vitro total phenolics content and antioxidant activity of the methanolic extracts of three Onobrychis species belonging to the family Fabaceae, namely O. sosnovskyi Grossh., O. viciifolia Scop. and O. melanotricha Boiss. Furthermore, an attempt was made to identify any correlations between total phenolic content of the extracts with their antioxidant activities Methods: Total phenol and flavonoid contents of the extracts were determined by Folin-Ciocalteu and aluminum chloride methods, respectively. Antioxidant activity was evaluated by three different test systems, namely 2, 2-diphenyl-1-picrylhydrazyl free radical scavenging (DPPH), metal-chelation activity and s-carotene/linoleic acid model. Results: Results indicated that O. viciifolia extract contains the highest total phenolic content (10.38 ± 0.33mg GAE/g of dry extract). However, the species are not remarkable different (P < 0.05) in total flavonoid amount. The Onobrychis species represent strong antioxidant activity in DPPH radical scavenging (IC50; 0.121 ± 0.005 to 0.129 ± 0.001 mg/ml), metal chelating (IC50; 1.10 ± 0.006 to 1.80 ± 0.003 mg/ml) and also β-carotene bleaching methods. In addition, a high correlation was found between different antioxidant assays and total phenol content of the extracts (R2 > 0.55 to 0.98). Conclusion: Our results showed that the examined Onobrychis extracts represent strong antioxidant activity; hence, they can be suggested as antioxidant agents for special use in future.

Molecular phylogenetics of the Onobrychis genus (Fabaceae: Papilionoideae) using ITS and trnL–trnF DNA sequence data

Onobrychis, comprising more than 130 species, is a genus of the family Fabaceae. At this time, the interspecies relationship of this biologically important genus is still a subject of great discussion and debate. To help resolve this disagreement, we used molecular phylogeny to analyse internal transcribed spacer (ITS) and trnL–trnF sequences of 76 species of Onobrychis. Bayesian interference, maximum parsimony and maximum likelihood analyses of nuclear ITS and plastid trnL–trnF DNA sequence data generated trees with strong posterior probability for two groups: Onobrychis subgen. Sisyrosema (including: Heliobrychis, Hymenobrychis, Afghanicae and Anthyllium sections) along with Laxiflorae section in Group I and Onobrychis subgen. Onobrychis (except Laxiflorae section) in the other (Group II). The Laxiflorae section roots back to the ancestral node for Sisyrosema subgen. O. viciifolia (cultivated species), which is closely associated with O. cyri var. cyri, suggesting that the latter may be a wild progenitor of O. viciifolia. The present study supported the paraphyly of subgenera Onobrychis and Sisyrosema. The study proposed the paraphyletic nature of the sections Onobrychis, Dendrobrychis, Heliobrychis and Hymenobrychis. Together with our molecular phylogenetic analyses we present a review of Onobrychis morphology and discuss and compare our results with those of earlier morphological and molecular phylogenetic analyses.

Cytological characterisation of the underutilized forage crop Onobrychis viciifolia Scop. and other members of the Onobrychis genus

Sainfoin (Onobrychis viciifolia Scop.) is a perennial forage legume that possesses beneficial properties in the context of sustainable agriculture. A germplasm collection of O. viciifolia was assembled from many geographic regions, as well as a collection of Onobrychis species that might be crossed with O. viciifolia to improve its biological and agronomic properties. The objective of the present study was to obtain cytological information for a broad range of O. viciifolia varieties and related species, in order to confirm and extend previous findings. The study set was 143 accessions of O. viciifolia as well as 34 accessions of 17 other species from the Onobrychis genus. Three of the O. viciifolia accessions were diploid (2n = 2x = 14) and the remainder tetraploid (2n = 4x = 28). Chromosomes were <5 μm in length. Among the other Onobrychis species, roughly half were found to be diploid and the other half tetraploid, with two species, O. arenaria (Kit.) DC. and O. alba subsp. laconica (Orph. ex Boiss.) Hayek, including a mixture of diploid and tetraploid accessions. The basic number of chromosomes was seven, except for O. aequidentata (Sm.) d’Urv. and O. crista-galli Lam., where it was eight. The 2C value for O. viciifolia was estimated at 2.5 pg. These cytological data will be valuable as part of a pre-breeding programme aimed at the eventual production of improved O. viciifolia varieties for use in sustainable agriculture.

A phylogenetic analysis of genus Onobrychis and its relationships within the tribe Hedysareae (Fabaceae)

Results about a phylogenetic analysis of the genus Onobrychis Mill., tribe Hedysareae DC. are presented. The systematic knowledge of tribe Hedysareae is still incomplete, with difficult circumscription of genera and species. Analyses were undertaken using both nuclear (ITS) and chloroplast (matK) markers for a set of 78 accessions covering 41 Onobrychis species, besides previously sequenced Hedysareae accessions. The phylogenetic methods used were maximum parsimony, maximum likelihood, and Bayesian analyses to produce phylogenetic trees and robustness indices. The genus Onobrychis was resolved as paraphyletic, with species of the genera Eversmannia Bunge and Hedysarum L. nested within it. The position of the section Membranacea of genus Hedysarum was as a sister group to Onobrychis and Eversmannia, separated from other accessions of Hedysarum. Variation in the 2 markers was sufficient to resolve infrageneric groups in Onobrychis and Hedysarum, but we were unable to completely resolve certain species in Onobrychis, particularly those within the sect. Onobrychis. The cause of this difficult species delimitation may be related to recent speciation, hybridization, and introgression events, particularly between cultivated species and their wild relatives, and the presence of cryptospecies as suggested by intraspecific polyploid series.

Study of ploidy level of annual species ofOnobrychisin Iran

Abstract Iran has more than 63 species of genus Onobrychis. This genus has small chromosomes with different ploidy levels. Counting ploidy levels in somatic cells in the metaphase is difficult; therefore, this study aims to evaluate a method using counts of stomata guard cell chloroplasts, as well as using flow cytometry as methods for determining ploidy levels in annual species of Onobrychis genus. Three middle leaves of greenhouse-grown plants were randomly selected for chloroplast counts in 10 pairs of stomata guard cells of the lower surface of leaf cells. For determining ploidy levels, the usual method was counting chromosomes which determined in the metaphase cells of root tip meristems with having minimum of 10 metaphase plate mitosis for each species. All of the samples were also studied, to determine ploidy levels using flow cytometry, and to find the mode and ratio for each of them. The results indicated that numbers of chloroplast stomata guard cells and flow cytometry in the tetraploid samples...