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Phylogenetic diversity analysis reveals Bradyrhizobium yuanmingense and Ensifer aridi as major symbionts of mung bean (Vigna radiata L.) in Pakistan.

ABSTRACT: The present study was carried out to evaluate the diversity of rhizobia associated with nodules of mung bean in Pakistan, because this information is necessary for inoculum development. Based on sequence analysis of 16S rRNA gene of thirty-one bacteria, 11 were assigned to genus Bradyrhizobium, 17 to Ensifer, and 3 to Rhizobium. Phylogenetic analyses on the basis of 16S-23S ITS region, atpD, recA, nifH, and nodA of representative strains revealed that B. yuanmingense is the predominant species distributed throughout different mung bean-growing areas. Among the fast-growing rhizobia, Ensifer aridi was predominant in Faisalabad, Layyah, and Rawalpindi, while E. meliloti in Thal desert. Sequence variations and phylogeny of nifH and nodA genes suggested that these genes might have been co-evolved with the housekeeping genes and maintained by vertical gene transfer in rhizobia detected in the present study. Host infectivity assay revealed the successful nodulation of host by rhizobia related to genera Bradyrhizobium, Ensifer and Rhizobium. Among all, Bradyrhizobium and Ensifer spp. inoculation exhibited a significantly higher number of nodules (11-34 nodules plant^-1) and nitrogenase activity (nodule ARA 60-110 μmol g^-1 h^-1). Contrary to the previous studies, our data reveal that B. yuanmingense and E. aridi are predominant species forming effective nodules in mung bean in Pakistan. Furthermore, to the best of our knowledge, this is the first report showing the effective symbiosis of E. aridi, E. meliloti, and Rhizobium pusense with mung bean. The diversity of rhizobia in different habitats revealed in the present study will contribute towards designing site-specific inocula for mung bean.

SUBMITTER: Hakim S

PROVIDER: S-EPMC7966693 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundMung bean is a pulse crop principally grown in the tropic and subtropic parts of the world for its nutrient-rich seeds. Seven mung beans accessions from Eastern Kenya were evaluated using thirteen phenotypic traits. In addition, 10 SSR markers were used to determine their genetic diversity and population structure. This aimed at enhancing germplasm utilization for subsequent mung bean breeding programs.ResultsAnalysis of variance for most of the phenology traits showed significant variation, with the yield traits recording the highest. The first three principal components (PC) explained 83.4% of the overall phenotypic variation, with the highest (PC1) being due to variation of majority of the traits studied such as pod length, plant height, and seeds per pod. The dendogram revealed that the improved genotypes had common ancestry with the local landraces. The seven mung beans were also genotyped using 10 microsatellite markers, eight of which showed clear and consistent amplification profiles with scorable polymorphisms in all the studied genotypes. Genetic diversity, allele number, and polymorphic information content (PIC) were determined using powermarker (version 3.25) and phylogenetic tree constructed using DARWIN version 6.0.12. Analysis of molecular variance (AMOVA) was calculated using GenALEx version 6.5. A total of 23 alleles were detected from the seven genotypes on all the chromosomes studied with an average of 2.875 across the loci. The PIC values ranged from 0.1224 (CEDG056) to 0.5918 (CEDG092) with a mean of 0.3724. Among the markers, CEDG092 was highly informative while the rest were reasonably informative except CEDG056, which was less informative. Gene diversity ranged from 0.1836 (CEDG050) to 0.5102 (CDED088) with an average of 0.3534. The Jaccards dissimilarity matrix indicated that genotypes VC614850 and N26 had the highest level of dissimilarity while VC637245 and N26 had lowest dissimilarity index. The phylogenetic tree grouped the genotypes into three clusters as revealed by population structure analysis (K = 3), with cluster III having one unique genotype (VC6137B) only. AMOVA indicated that the highest variation (99%) was between individual genotype. In addition, marker traits association analysis revealed 18 significant associations (P < 0.05).ConclusionThese findings indicate sufficient variation among the studied genotypes that can be considered for germplasm breeding programs.

Project description:Bradyrhizobium guangxiense CCBAU53363 efficiently nodulates peanut but exhibits incompatible interaction with mung bean. By comparing the common nod region with those of other peanut bradyrhizobia efficiently nodulating these two hosts, distinctive characteristics with a single nodD isoform (nodD1) and a truncated nolA were identified. However, the regulatory roles of NodD1 and NolA and their coordination in legume-bradyrhizobial interactions remain largely unknown in terms of explaining the contrasting symbiotic compatibility. Here, we report that nolA was important for CCBAU53363 symbiosis with peanut but restricted nodulation on mung bean, while nodD1 was dispensable for CCBAU53363 symbiosis with peanut but essential for nodulation on mung bean. Moreover, nolA exerted a cumulative contribution with nodD1 to efficient symbiosis with peanut. Additionally, mutants lacking nolA delayed nodulation on peanut, and both nolA and nodD1 were required for competitive nodule colonization. It is noteworth that most of the nodulation genes and type III secretion system (T3SS)-related genes were significantly downregulated in a strain 53ΔnodD1nolA mutant compared to wild-type strain CCBAU53363, and the downregulated nodulation genes also had a greater impact than T3SS-related genes on the symbiotic defect of 53ΔnodD1nolA on peanut, which was supported by a more severe symbiotic defect induced by 53ΔnodC than that with the 53ΔnodD1nopP, 53ΔnodD1rhcJ, and 53ΔnodD1ttsI mutants. NolA did not regulate nod gene expression but did regulate the T3SS effector gene nopP in an indirect way. Meanwhile, nolA, nodW, and some T3SS-related genes besides nopP were also demonstrated as new "repressors" that seriously impaired CCBAU53363 symbiosis with mung bean. Taken together, the roles and essentiality of nolA and nodD1 in modulating symbiotic compatibility are sophisticated and host dependent. IMPORTANCE The main findings of this study were that we clarified that the roles and essentiality of nodD1 and nolA are host dependent. Importantly, for the first time, NolA was found to positively regulate T3SS effector gene nopP to mediate incompatibility on mung bean. Additionally, NolA does not regulate nod genes, which are activated by NodD1. nolA exerts a cumulative effect with nodD1 on CCBAU53363 symbiosis with peanut. These findings shed new light on our understanding of coordinated regulation of NodD1 and NolA in peanut bradyrhizobia with different hosts.

Project description:IntroductionDespite the development of oral hypoglycemic medications, diabetes and its associated complications continue to be significant clinical issues. The purpose of this study was to examine the antidiabetic effects of Vigna radiata (L.) Wilczek seeds in mice that had been given alloxan to cause diabetes.MethodsIn Swiss albino mice, diabetes was brought on by a single intraperitoneal injection of the drug alloxan (150 mg/kg). For 14 days, glibenclamide (5 mg/kg) and methanol extract of V. radiata seeds (100, 200, and 400 mg/kg) were given orally. Following oral administration of V. radiata to mice, the blood glucose levels (BGL) and body weight were measured at 7 and 14 days. The mice were sacrificed at the end of the trial, and blood samples were taken for the evaluation of insulin, glycated hemoglobin, aspartate aminotransferase (AST), alanine aminotransferase (ALT), high-density lipoprotein (HDL), total cholesterol (TC), and triglyceride (TG) levels. It was determined how much glycogen was present in the liver. Additionally, the total phenolic and flavonoid contents of V. radiata were determined, along with the in vitro DPPH (2, 2 diphenyl-1-picrylhrazyl) free radical-scavenging activity. P < 0.05 was chosen as the cutoff for statistical significance.ResultsFollowing oral administration of V. radiata for 14 days, diabetic mice's BGL and bad cholesterol (TC and TG) levels significantly decreased, while HDL levels increased. Treatment with V. radiata significantly decreased the levels of AST, ALT, and glycated hemoglobin when compared with diabetes control. On the other hand, it raised insulin levels and the amount of liver glycogen. V. radiata underwent phytochemical analysis, which identified the presence of tannins, saponins, phenols, alkaloids, terpenoids, steroids, flavonoids, and glycosides. Per gram of V. radiata seed extract, the total phenolic content was 43.12 ± 3.14 mg of gallic acid equivalents, while the total flavonoid content was 38.35 ± 2.6 mg of quercetin equivalents. Ascorbic acid was shown to have an IC50 value of 18.64 µg/ml during a DPPH-scavenging assay, while V. radiata had an IC50 value of 73.35 µg/ml.ConclusionAccording to the findings of the current study, the methanolic extract of the seeds from the plant V. radiata possesses significant antidiabetic characteristics that are on par with those of the commonly used drug glibenclamide. Hence, V. radiata seems to be effective as a natural antidiabetic.

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Phylogenetic diversity analysis reveals Bradyrhizobium yuanmingense and Ensifer aridi as major symbionts of mung bean (Vigna radiata L.) in Pakistan.

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