Medicago Laciniata Research Articles

Qualitative Phytochemical Screening and Anti-Inflammatory Evaluation of Aerial Part Extracts of Medicago laciniata (L.) Mill

Inflammation is a live tissue's immunological reaction to an allergen or injury to a bodily component. The majority of anti-inflammatory medicines are synthetic, and long-term usage of these medications can have serious side effects and harm the human biological system. Medicago laciniata, sometimes known as cutleaf medick traditionally used as an abortifacient, anti-inflammatory, antidepressant, and antidiabetic. The goal of this study was to scientifically document the anti-inflammatory activity of the aerial portions of Medicago laciniata. For this purpose, extraction was carried out using solvents ethyl acetate and n-hexane. The preliminary phytochemical screening was performed to check the presence of important secondary metabolites followed by applying the carrageenan-induced edema hind paw method to test the anti-inflammatory efficacy of both extracts. The result showed the presence of secondary flavonoids steroids, terpenoids, saponins, and tannins. In vivo anti-inflammatory activity showed that after five hours, the maximum reduction of 70.90% and 69.29% were observed for ethyl acetate and n-hexane extracts respectively, each at a dose of 100 mg/kg, respectively. This study suggests that Medicago laciniata has an excellent ability to protect against inflammation due to the presence of secondary metabolites.

Editor's Summaries of the Articles Published in This Issue of Phytopharmacological Communications

In this issue, Iram and colleagues described their study on the anti-inflammatory potential of Medicago laciniata, Afzal and her colleagues rationalized the traditional use of Peganum harmala in cardiovascular diseases, Aisha and her colleagues summarized studies showing inhibition of tau by medicinal plants and phytocompounds isolated from them for the treatment of Alzheimer’s disease, Zia-Ul-Hq et al, compiled investigations in which adenosine diphosphate-induced platelet aggregation is shown to be inhibited by medicinal plants, and Ullah and his colleagues reviewed studies showing potential calcium channel blockers from medicinal plants and provided their insights and recommended their further investigation in clinical trials.

A taxonomic study of twelve wild forage species of Fabaceae

Twelve species of wild leguminosae were studied to determine similarities in the coat details of the seeds using a Scanning Electron Microscope (SEM). The numerical cluster analysis method was used to examine the morphological characteristics (98 characteristics) and to clarify the taxonomic relationship between the studied species (6 genera and 3 tribes) belonging to the Fabaceae family. The relevant wild species were: Lotus edulis L, Lotus ornithopodioides L., Tetragonolobus purpureus Moench, Medicago laciniata (L.) Mill., Gard. Dict., M.orbicularis (L.) Bart., M.turbinata (L.) All, M.polymorpha L., Ononis vaginalis Vahl, Lathyrus aphaca L., Vicia sativa L., V. peregrine L., and V.tetrasperma (L.) Schreb. The aim of this study was to produce a taxonomy reflecting the relations between these twelve forage species of Fabaceae by using the morphological and SEM features to provide a details about and clarify the relations between the examined taxa. The taxonomic histories of the Fabaceae family were reviewed. The results of the morphological description and SEM showed that it was possible to distinguish between the taxa using the cluster analysis attributes for the differences in characteristic correlation between the groups under study. This study will help researchers better grasp the classification of these species of legumes which were chosen because of the difficulty of differentiating between them, their environment benefits, their use for human consumption and pasture. The SEM is a suitable tool for this analysis, owing to the similarities exhibited by the seeds.

Variation in Morphological Traits and Yield Evaluation among Natural Populations of Medicago truncatula and Medicago laciniata

Variation in Morphological Traits and Yield Evaluation among Natural Populations of Medicago truncatula and Medicago laciniata

Specificity in Legume-Rhizobia Symbioses.

Most species in the Leguminosae (legume family) can fix atmospheric nitrogen (N2) via symbiotic bacteria (rhizobia) in root nodules. Here, the literature on legume-rhizobia symbioses in field soils was reviewed and genotypically characterised rhizobia related to the taxonomy of the legumes from which they were isolated. The Leguminosae was divided into three sub-families, the Caesalpinioideae, Mimosoideae and Papilionoideae. Bradyrhizobium spp. were the exclusive rhizobial symbionts of species in the Caesalpinioideae, but data are limited. Generally, a range of rhizobia genera nodulated legume species across the two Mimosoideae tribes Ingeae and Mimoseae, but Mimosa spp. show specificity towards Burkholderia in central and southern Brazil, Rhizobium/Ensifer in central Mexico and Cupriavidus in southern Uruguay. These specific symbioses are likely to be at least in part related to the relative occurrence of the potential symbionts in soils of the different regions. Generally, Papilionoideae species were promiscuous in relation to rhizobial symbionts, but specificity for rhizobial genus appears to hold at the tribe level for the Fabeae (Rhizobium), the genus level for Cytisus (Bradyrhizobium), Lupinus (Bradyrhizobium) and the New Zealand native Sophora spp. (Mesorhizobium) and species level for Cicer arietinum (Mesorhizobium), Listia bainesii (Methylobacterium) and Listia angolensis (Microvirga). Specificity for rhizobial species/symbiovar appears to hold for Galega officinalis (Neorhizobium galegeae sv. officinalis), Galega orientalis (Neorhizobium galegeae sv. orientalis), Hedysarum coronarium (Rhizobium sullae), Medicago laciniata (Ensifer meliloti sv. medicaginis), Medicago rigiduloides (Ensifer meliloti sv. rigiduloides) and Trifolium ambiguum (Rhizobium leguminosarum sv. trifolii). Lateral gene transfer of specific symbiosis genes within rhizobial genera is an important mechanism allowing legumes to form symbioses with rhizobia adapted to particular soils. Strain-specific legume rhizobia symbioses can develop in particular habitats.

Growth, photosynthesis and water relations as affected by different drought regimes and subsequent recovery in Medicago laciniata (L.) populations

The aims of this study were to investigate the effects of water deficit and recovery on growth, photosynthesis and water relations in four Medicago laciniata populations from saharian (Ml-90), inferior arid (Ml-204), superior arid (Ml-306) and semi-arid (Ml-376) Tunisian regions. After 28 d of sowing with ample irrigation, the plants were subjected to 4 water regimes: optimal irrigation (100% of field capacity, FC), moderate drought (75% FC), severe drought (35% FC) and rewatering (plants submitted to 35% FC during 7 d, afterwards the plants were rewatered to 100% FC). Harvest was carried out after 28 d of treatments. The drought tolerance in M. laciniata populations was found to be increased particularly with increasing temperatures of collection site of the population. The Ml-204 and Ml-90 populations used mainly physiological strategies for survival under moderate water shortage. Higher severe drought tolerance in both signaled populations would be related to their lower photosynthesis metabolic impairment, relatively higher leaf RWC and greater osmotic potential decrease. The results suggest that plants with low values of leaf features are likely to maintain higher leaf RWC under sever drought. The largest decrease of osmotic potential was found associated with the solute accumulations such as proline and K+.

Chromosome number, genome size, seed storage protein profile and competence for direct somatic embryo formation in Algerian annual Medicago species

The genome size of eight species from different wild Algerian populations of diploid and one tetraploid Medicago L., initially analysed for chromosome numbers and seed storage proteins, has been assessed by flow cytometry. Genome size ranged from 2C = 0.94 pg in Medicago orbicularis (L.) Bartal. to 2C = 1.80 pg in Medicago laciniata (L.) Miller. The competence for direct somatic embryo formation in liquid medium was studied for 4 of these species having distinct genome sizes: M. orbicularis (2C = 0.94 pg); M. truncatula Gaertn. (2C = 1.08 pg); M. scutellata (L.) Miller (2C = 1.11 pg); M. arabica (L.) Huds. (2C = 1.22 pg). M. orbicularis, with the smallest genome size, formed somatic embryos most quickly, with a high frequency of reactive explants and with numerous somatic embryos per explant. It was followed by M. truncatula, M. scutellata and M. arabica, which in fact represents the order of increasing genome size.

Inoculation of Phaseolus vulgaris, Medicago laciniata and Medicago polymorpha with Agrobacterium sp. strain 10C2 may enhance nodulation and shoot dry weight but does not affect host range specificity

The aim of this work was to assess the effect of inoculation with the non-symbiotic Agrobacterium strain 10C2, previously isolated from root nodules of Phaseolus vulgaris, on nodulation, effectiveness and host range specificity. Two rhizobial strains, Ensifer meliloti 2011 and Ensifer medicae A321, were used in this study on the basis of their differential sensitivity to the in vitro antagonistic activity exercised by Agrobacterium sp. 10C2. Three host legumes, P. vulgaris, Medicago laciniata and M. polymorpha, were also selected for this experiment on the basis of their variable symbiotic specificity towards strains 2011 and A321. Cross inoculation experiments were conducted with or without co-inoculation with Agrobacterium sp. 10C2. Results showed that both rhizobial strains induced ineffective nodules on P. vulgaris. Nevertheless, inoculation with Agrobacterium sp. 10C2 slightly increased shoot dry weight with both strains and enhanced nodule number with strain 2011 only. Inoculation of M. polymorpha with strain 10C2 significantly increased the nodule number induced by E. medicae A321 but did not affect biomass production. However, inoculation of M. laciniata by strain 10C2 enhanced nodule number and shoot dry weight with strain 2011 only. Agrobacterium sp. 10C2 did not affect the non-inoculated control, nor the contrasted host range of both rhizobial strains towards M. polymorpha and M. laciniata. The potential negative effect of the in vitro antagonistic activity of strain 10C2 on reduction of nodule number or symbiotic effectiveness was not proved. No evidence of symbiotic gene transfer from the rhizobial strains to Agrobacterium sp. 10C2 was also proved. The effect of strain 10C2 is probably mediated by nodules since no result was observed in the non-inoculated control or when the rhizobial strain failed to induce nodules. It would be interesting to investigate the implications on nodule functioning, mainly the impact on nitrogenase activity, antioxidant enzymes and nodule senescence.

Phenology, leaf gas exchange, growth, and seed yield in contrasting Medicago truncatula and Medicago laciniata populations during prolonged water deficit and recovery

The aims of this study were to investigate the effect of prolonged water stress and recovery on phenology, growth, and seed yield in Tunisian contrasting populations of Medicago truncatula and Medicago laciniata . After ample irrigation for 24 days, the plants of each population were divided into two lots: the first lot was irrigated at 100% field capacity (FC), and the second at only 45% FC. After 24 days of treatment, one lot of dehydrated plants was rewatered at 100% FC, while the other was maintained at 45% FC. Interspecific and intraspecific differences were found in phenological responses to water deficit. All growth parameters were more reduced in M. truncatula populations than in M. laciniata populations. The water shortage tolerance of M. laciniata populations was associated with a lower metabolic impairment of photosynthesis and maintenance of relatively higher leaf relative water content. Seed yield was also more reduced in M. truncatula populations compared with M. laciniata populations. In M. laciniata, seed mass was a compensation mechanism to sustain seed yield under drought conditions. Seed yield variation between populations under water deficit was explained mainly by variation in seed number per plant.

Comparative effects of water deficit on Medicago laciniata and Medicago truncatula lines sampled from sympatric populations

We evaluated the responses to water deficit of twelve lines of Medicago laciniata and Medicago truncatula including eight lines from four Tunisian sympatric populations of both species, and four references lines of M. truncatula. They were exposed to two water treatments, well irrigated and drought-stress (33% of field capacity) for a period of 45 days. At harvest, we measured five quantitative traits including the length of stems (LS), aerial dry weight (ADW), root dry weight (RDW), RDW/ADW ratio and leaf area (LA). Analysis of variance showed that the variation of measured traits among studied lines was significantly explained by the effects of species, line, treatment, and their interactions. Treatment and species had the largest effects. We also analyzed the broad-sense heritability of the drought response index (DRI), defined as the ratio between the observed values with and without water deficit treatment. DRI of most measured traits had high broad-sense heritability ( H²). The length of stems (LS) was the trait most affected by drought stress, while RDW/ADW ratio was not affected by drought. Generally, few significant differences were observed between DRI values of measured traits between lines within species as well as between lines of M. laciniata and M. truncatula collected from sympatric populations. Most of the correlations established between DRI values were positive. The environmental factors that most influenced variation of DRI values among populations were available phosphorus (P) and mean annual rainfall.

Symbiotic diversity of Ensifer meliloti strains recovered from various legume species in Tunisia

Ensifer meliloti (formerly Sinorhizobium meliloti) was first considered as a specific microsymbiont of Medicago, Melilotus and Trigonella. However, strains of E. meliloti were recovered from root nodules of various legume species and their symbiotic status still remains unclear. Here, we further investigate the specificity of these strains. A collection of 47 E. meliloti strains isolated in Tunisia from root nodules of Medicago truncatula, Medicago sativa, Medicago ciliaris, Medicago laciniata, Medicago marina, Medicago scutellata, Phaseolus vulgaris, Cicer arietinum, Argyrolobium uniflorum, Lotus creticus, Lotus roudairei, Ononis natrix, Retama raetam, Genista saharae, Acacia tortilis, Hedysarum carnosum and Hippocrepis bicontorta were examined by REP-PCR fingerprinting, PCR-RFLPs of the 16S-23S rDNA IGS, the nifH gene and nifD-K intergenic spacer, and sequencing of 16S rRNA and nodA genes. Their nodulation range was also assessed by cross-inoculation experiments. No clear correlation was found between chromosomal backgrounds and host plants of origin. The nodulation polyvalence of the species E. meliloti was associated with a high symbiotic heterogeneity. On the basis of PCR-RFLP data from the nifH gene and nifD-K intergenic spacer, E. meliloti strains isolated from non- Medicago legumes harboured distinct genes and possessed wider host ranges. Some strains did not nodulate Medicago species. On the basis of nodA phylogeny, the majority of the Tunisian strains, including strains from Medicago, harboured distinct nodA alleles more related to those found in E. medicae than those found in E. meliloti. However, more work is still needed to characterize this group further. The diversity observed among M. laciniata isolates, which was supported by nodA phylogeny, nifH typing and the efficiency profile on M. ciliaris, indicated that what was thought to be bv. medicaginis is certainly heterogeneous.

Ensifer meliloti bv. lancerottense establishes nitrogen-fixing symbiosis with Lotus endemic to the Canary Islands and shows distinctive symbiotic genotypes and host range

Eleven strains were isolated from root nodules of Lotus endemic to the Canary Islands and they belonged to the genus Ensifer, a genus never previously described as a symbiont of Lotus. According to their 16S rRNA and atpD gene sequences, two isolates represented minority genotypes that could belong to previously undescribed Ensifer species, but most of the isolates were classified within the species Ensifer meliloti. These isolates nodulated Lotus lancerottensis, Lotus corniculatus and Lotus japonicus, whereas Lotus tenuis and Lotus uliginosus were more restrictive hosts. However, effective nitrogen fixation only occurred with the endemic L. lancerottensis. The E. meliloti strains did not nodulate Medicago sativa, Medicago laciniata Glycine max or Glycine soja, but induced non-fixing nodules on Phaseolus vulgaris roots. nodC and nifH symbiotic gene phylogenies showed that the E. meliloti symbionts of Lotus markedly diverged from strains of Mesorhizobium loti, the usual symbionts of Lotus, as well as from the three biovars (bv. meliloti, bv. medicaginis, and bv. mediterranense) so far described within E. meliloti. Indeed, the nodC and nifH genes from the E. meliloti isolates from Lotus represented unique symbiotic genotypes. According to their symbiotic gene sequences and host range, the Lotus symbionts would represent a new biovar of E. meliloti for which bv. lancerottense is proposed.

Morphological and microsatellite diversity associated with ecological factors in natural populations of Medicago laciniata Mill. (Fabaceae)

Genetic variability in 10 natural Tunisian populations of Medicago laciniata were analysed using 19 quantitative traits and 12 polymorphic microsatellite loci. A large degree of genetic variability within-populations and among-populations was detected for both quantitative characters and molecular markers. High genetic differentiation among populations for quantitative traits was seen, with Q(ST) = 0.47, and F(ST) = 0.47 for microsatellite markers. Several quantitative traits displayed no statistical difference in the levels of Q(ST) and F(ST). Further, significant correlations between quantitative traits and eco-geographical factors suggest that divergence in the traits among populations may track environmental differences. There was no significant correlation between genetic variability at quantitative traits and microsatellite markers within populations. The site-of-origin of eco-geographical factors explain between 18.13% and 23.40% of genetic variance among populations at quantitative traits and microsatellite markers, respectively. The environmental factors that most influence variation in measured traits among populations are assimilated phosphorus (P(2)0(5)) and mean annual rainfall, followed by climate and soil texture, altitude and organic matter. Significant associations between eco-geographical factors and gene diversity, He, were established in five microsatellite loci suggesting that these simple sequence repeats (SSRs) are not necessarily biologically neutral.

Medicago species affect the community composition of arbuscular mycorrhizal fungi associated with roots

Sequencing of the 5' end of the large ribosomal subunit (LSU rDNA) and quantitative polymerase chain reaction (qPCR) were combined to assess the impact of four annual Medicago species (Medicago laciniata, Medicago murex, Medicago polymorpha and Medicago truncatula) on the genetic diversity of arbuscular mycorrhizal (AM) fungi, and on the relative abundance of representative AM fungal genotypes, in a silty-thin clay soil (Mas d'Imbert, France). Two hundred and forty-six Glomeromycete LSU rDNA sequences from the four plant species and the bulk soil were analysed. The high bootstrap values of the phylogenetic tree obtained allowed the delineation of 12 operational taxonomic units (OTUs), all belonging to Glomus. Specific primers targeting Glomeromycetes and major OTUs were applied to quantify their abundance by qPCR. Glomeromycetes and targeted OTUs were significantly more abundant in the root tissues than in the bulk soil, and the frequencies of three of them differed significantly in the root tissues of the different plant species. These differences indicate that, despite the absence of strict host specificity in mycorrhizal symbiosis, there was a preferential association between some AM fungal and plant genotypes.

Comparison of rhizobia that nodulate Medicago laciniata and Medicago truncatula present in a single Tunisian arid soil

The rhizobia present in a single arid region Tunisian soil that nodulate Medicago laciniata and Medicago truncatula were compared. All isolates, 40 from each host, were Sinorhizobium meliloti based on 16S rRNA polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) patterns and subsequent confirmation by sequence analysis of the 16S rRNA genes in four representatives from each host species. There was no apparent relationship between Medicago host species of isolation and the nodulating rhizobial genome as determined by repetitive extragenic palandromic PCR. The isolates of M. laciniata were distinguished from those of M. truncatula present in the same soil by variation in PCR-RFLP of nifDK, indicating that this dissimilarity is originally genetic and not geographic. While forming effective symbioses with their own respective isolates, both M. laciniata and M. truncatula formed ineffective true nodules, nodule-like structures, or no nodules at all in cross-inoculation tests, as confirmed by the histological observations.

Nitrogen-fixing sinorhizobia with Medicago laciniata constitute a novel biovar (bv. medicaginis) of S. meliloti

Sixty-eight new rhizobial isolates were obtained from root-nodules of Medicago laciniata and from Mediterranean soils in Tunisia and France. All of them were identified as Sinorhizobium meliloti on the basis of PCR-RFLP analyses of 16S rDNA and the intergenic spacer sequence between 16S and 23S rDNAs. DNA/DNA hybridization, phenotypic characterization and 16S rRNA gene sequencing led to the conclusion that they belong the same taxon. All new isolates shared the ability to nodulate and fix nitrogen with M. laciniata except 11 of them not capable of fixing nitrogen with this plant and originating from French soils containing no efficiently adapted symbionts with M. laciniata. The nitrogen-fixing rhizobia on M. laciniata differed markedly from the other S. meliloti or Sinorhizobium medicae isolates and references in their symbiotic traits such as nifDK RFLP diversity, nodA sequences and nitrogen effectiveness with tree other different annual Medicago species ( M. truncatula, M. polymorpha and M. sauvagei). Two infrasubspecific (biovar) divisions are therefore proposed within S. meliloti: bv. medicaginis for Sinorhizobium efficient on M. laciniata and bv. meliloti for the classically known S. meliloti group represented by the strains ATCC9930 T and RCR 2011 efficient on M. sativa.

Spot inoculation of Medicago laciniata root by Sinorhizobium meliloti CI-4 or Glomus sp. S043 induces local accumulation of flavonoids

Medicago laciniata roots were in vitro inoculated with Sinorhizobium meliloti or the arbuscular mycorrhizal fungus Glomus sp. S043. Inocula were applied on well-defined parts of the root system. We studied the chemical response of root cells in presence of the bacteria or the fungus, using visible and fluorescent microscopy. The inoculation spot induced localised accumulation of flavonoids in the root tissue for both Sinorhizobium meliloti and Glomus sp. S043. There was no flavonoid accumulation in other areas of the inoculated root. No tannin accumulation was observed in the roots of the control as well as in the inoculated tissues. The role of these flavonoids is discussed.

Flow Cytometric Analysis in Diploid Medicago Species from Algeria: Relationship Between Genome Size and Competence for Direct Somatic Embryo Formation

ABSTRACTThe genome sizes of eight species from different wild population of diploid Medicago with origin from Algeria have been assessed by flow cytometry. This parameter vary from 2C=0.94 pg in Medicago orbicularis (the smallest genome) to 2C=1.80 pg in Medicago laciniata (the largest one). The relationship between genome size and competence for direct somatic embryo formation in liquid medium for four of the tested species (M. orbicularis Orbi DZ19 C 2C=0.94; M. truncatula Tru DZ 2B 2C=1.08; M.scutelata Scu 274 2C=1.11; M. arabica Ara 46 Ina 2C=1.22) was studied. It was found that Medicago orbicularis with the smallest genome size formed somatic embryos for a shortest period of time, with a high percent of reacting explants and number of somatic embryos per explant. It was followed by M. truncatula, M. scutelata and M. arabica.

Screening of some indigenous Qatari medicinal plants for antimicrobial activity.

Aqueous, ethanol and butanol crude extracts of the aerial parts of ten plants exhibited variable degrees of antimicrobial activity against four bacterial and two fungal species. Aqueous extracts had low antimicrobial activity against E.coli, P.aeruginosa, B. cerreus, S.aureus, C.albicans and A.flavus. Avicennia marina (AM) aqueous extract exhibited a moderate antifungal activity. Ethanol and butanol crude extracts exhibited an improved antimicrobial activity. However, butanol exhibited a superior antimicrobial activity compared with aqueous and ethanol crudes. Compared with the standard antibiotics tested the butanol extract had the highest activity. Butanol extracts at 2000 microg/disc of AM, Lotus halophilus (LA), Pulicaria gnaphaloides (PG) and Capparis spinosa (CS) had a very good antibacterial activity against both gram-positive and -negative bacteria as well as moderate to good antifungal activity against C. albicans and A. flavus. Medicago laciniata (ML), Limonium axillare (LA) and (PG) butanol crude extract compared with standard chloramphenicol, tetracycline and nalidixic acid exhibited a superior antifungal activity.

Identification and cloning of the bacterial nodulation specificity gene in the Sinorhizobium meliloti--Medicago laciniata symbiosis.

Medicago laciniata (cut-leaf medic) is an annual medic that is highly nodulation specific, nodulating only with a restricted range of Sinorhizobium meliloti. e.g., strain 102L4, but not with most strains that nodulate Medicago sativa (alfalfa), e.g., strains RCR2011 and Rm41. Our aim was to identify and clone the S. meliloti 102L4 gene implicated in the specific nodulation of M. laciniata and to characterize the adjacent nodulation (nod) region. An 11-kb EcoRI DNA fragment from S. meliloti 102L4 was shown to complement strain RCR2011 for nodulation of M. laciniata. Nucleotide sequencing revealed that this fragment contained nodABCIJ genes whose overall arrangement was similar to those found in strains RCR2011 and Rm41, which do not nodulate M. laciniata. Data for Tn5 mutagenesis of the nodABCIJ region of strain 102L4 suggested that the nodC gene was involved in the specific nodulation of M. laciniata. Tn5 insertions in the nodIJ genes gave mutants with nodulation delay phenotypes on both M. laciniata and M. sativa. Only subclones of the 11-kb DNA fragment containing a functional nodC gene from strain 102L4 were able to complement strain RCR2011 for nodulation of M. laciniata. The practical implications of these findings are discussed in the context of the development of a specific M. sativa - S. meliloti combination that excludes competition for nodulation by bacterial competitors resident in soil.