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The legume–rhizobia symbiosis is a unique natural phenomenon, which supplies the plant with the necessary mineral nitrogen via fixation of atmospheric dinitrogen. This interaction involves two partners: the legume plant and root nodule bacteria (rhizobia). In the wild, members of the Fabaceae family enter into symbiosis with a polymorphic group of rhizobia specific to them; the mechanism and reasons for the formation of heterogeneity of rhizobia are currently the subject of active research. In the present work, a Rhizobium leguminosarum strain strictly specific to Phaseolus vulgaris L. was used to show that within 30 days upon its introduction into soil, genetic rearrangements occurred in the cells, as was revealed by changes in the pattern of its genetic profile. It was also found that recombination activity of the cells was also affected by the root exudates produced during seed germination, which may indicate involvement of the plant in the formation of polymorphism of its microsymbionts. These findings suggest interpretation of this process not as a spontaneous event, but rather as an event controlled by the plant. [ABSTRACT FROM AUTHOR]

Aims: To assess the symbiotic effectiveness, tolerance to abiotic stress factors and phosphate solubilizing ability of new chickpea root-nodule bacteria. Methods: Symbiotic effectiveness, abiotic stress tolerance and phosphate solubilizing ability of ten new chickpea rhizobial strains collected from soils were evaluated using laboratory and glasshouse experiments. Results: Six of the ten strains were symbiotically efficient with diversity between those and the commercial chickpea strain CC1192. High temperatures significantly affected strain survival in liquid and peat carrier. Peat carrier offered greater protection. Above 37 °C, strain infectivity decreased with no correlation between strain origin and their infectivity after exposure. Three of the new strains and CC1192 produced optimum growth and survival at pH 6.8 and at two lower pH's of 4.4 and 5.4 were able to neutralize the growth medium while the other seven strains at high pH were able to either neutralize or acidify the growth medium. Strain survival was significantly higher at increased salt concentrations of NaCl compared to CaCl2. At 3% NaCl concentration, 8 strains survived while at 3% CaCl2 only 3 survived. Many strains were resistant to more than one antibiotic. All strains were able to solubilize phosphate. The ratio between the most efficient strain and the least was 3:1. Conclusions: Chickpea rhizobia strains sourced from soils in Narrabri New South Wales and Kununurra in Western Australia differed in expressed traits from the commercial strain CC1192. These unique traits could provide additional tools for rhizobial strain selection to benefit chickpea production in different soil environments. [ABSTRACT FROM AUTHOR]

The legume-Rhizobium symbiosis is a unique natural phenomenon responsible for providing a legume plant with mineral nitrogen fixed from the atmosphere. In the temperate climatic zone, due to seasonal cycles, plants have to form new interactions with rhizobia at the beginning of each round of vegetation with the formation of specialized structures on the roots, called root nodules, where molecular nitrogen is actually fixed. Nitrogen requirements of the host vary depending on the period of plant growth and development, which may affect its trophic relationships with the microsymbionts. In the present work, using the perennial leguminous plant Lotus corniculatus L. as an example, we assessed the influence of the macrosymbiont vegetation stage on the composition of its root nodule bacteria and carried out the characterization of these microbial communities. The highest microbial genetic diversity was found in the communities developing at the beginning of L. corniculatus vegetation. At the medium stage of plant development, the diversity decreased, while by the end of vegetation it increased again. Analysis of the nitrogen-fixing activity revealed the highest values for microbial strains isolated from the root nodules collected at the early stages of the macrosymbiont development, while it was lower in the strains isolated at the end of vegetation. At different stages of their development legumes may probably use different strategies for selection of microsymbionts. [ABSTRACT FROM AUTHOR]

Legume plants rely upon multipartite interactions between rhizobia and bacterial endophytes within root nodules to facilitate plant growth. This study aimed to isolate and identify indigenous endophytic bacteria from root nodules of Sulla aculeolata L. in Northeast Morocco. Based on their tri-calcium phosphate (TCP) solubilization capacity, five endophytes were chosen for further evaluation of their plant growth traits. All isolates were hydrogen cyanide (HCN) and siderophore producers, while only BCH24 tested positive for ACC deaminase activity. Indole-3-acetic acid (IAA) synthesis ranged from 1.27 mgL− 1 to 2.89 mgL− 1, while soluble phosphate concentrations was between 7.99 mg L− 1 and 110.58 mg L− 1. Additionally, all the endophytes were able to produce more than two lytic enzymes. Based on the analysis of 16 S rRNA gene sequences five isolates were identified as Enterobacter sp (BCH13, BCH2), Pseudomonas sp (BCH16, BCH24), and Serratia sp (BCH10). The strains inhibited the growth of three phytopathogenic fungi, with BCH13 exhibiting the highest rate against Aspergillus ochraceus (45%), followed by BCH24 against Fusarium oxysporum (40%) and Botrytis cinerea (35%), respectively. In vivo inoculation of halotolerant strains Enterobacter hormaechei (BCH13) and Pseudomonas moraviensis (BCH16) under gnotobiotic conditions revealed that co-inoculation with Rhizobium sullae KS6 improved plant development compared to single inoculation, making it a promising eco-friendly bio-inoculant for legume Sulla flexuosa L. production. [ABSTRACT FROM AUTHOR]

The functional activity of different phylogenetic variants of the transcriptional activator of nitrogen fixation genes in the cells of root nodule bacteria (rhizobia) of different taxa was analyzed. For this purpose, recombinant strains of root nodule bacteria of the genera Mesorhizobium, Ensifer (Sinorhizobium), and Rhizobium were created using genetic engineering constructs based on the plasmid pJN105. This plasmid contains different phylogenetic variants of the nifA gene typical for bacteria of the genera Mesorhizobium, Ensifer, and Rhizobium under the control of the ParaBAD bacterial promoter. It was shown that in free-living recombinant cells of all three bacterial genera, the introduction of an additional active copy of the nifA gene to their genome resulted in emergence of detectable nitrogenase activity. The level of nitrogenase activity was not directly related to the phylogeny of the introduced nifA gene, which is most likely explained by the universality of the product of this gene within the studied group of bacteria. The data obtained indicate the possibility of combinatorial processes in the evolution of nitrogen fixation genes. [ABSTRACT FROM AUTHOR]

A molecular phylogenetic analysis of salt-tolerance-related genes was carried out using complete genome sequencing data available for 26 Sinorhizobium meliloti strains and for 25 bacterial strains belonging to 17 genera. It was revealed that the genes of the first and the second stages of the response to salt stress (aqpZ, trkH, and trkA, and betICBA) have copies of many of the above- indicated genes on pSymA. Data obtained can provide evidence that this replicon, known to be essential for nitrogen fixation rhizobia activity, also has a significant role in the formation of a stress-related gene pool. The closest putative phylogenetic relatives were identified for all 14 tested genes and these are the first insights into the evolutionary pathways for the formation of a stress-related gene pool in root nodule nitrogen-fixing bacteria. [ABSTRACT FROM AUTHOR]

Mungbean (Vigna radiata L. Wilczek) is an economically important legume crop grown across India. The growth and yield of the crop have been declining over the recent years due to climate change which leads to decrease in soil moisture. Applications of drought-tolerant plant growth-promoting (PGP) rhizobial strains have been found to enhance crop productivity under water deficiency. Therefore, the objective of this study was to screen the bacterial strains from the mungbean root nodule that may help in improving the plant growth under drought condition. Bacteria were isolated from mungbean root nodules and screened for in vitro drought-tolerance using different concentrations of PEG 6000 at 10, 20, 30 and 40 %, and temperature-tolerance at 30, 35, 40 and 45 °C. In primary screening, out of 98 root nodule bacterial isolates tested, only 25 % showed drought-tolerance at 40 % polyethylene glycol (PEG)-6000 while 22 % of bacteria survived at 45 °C. During secondary screening on combined stress-tolerance, only 8 % of isolates showed tolerance of 40 % PEG-6000 at 45 °C. The in vitro drought-tolerance of bacteria varied significantly according to their sampling region/district. Various PGP traits, i.e., nitrogen fixation, phosphate solubilization, and production of indole acetic acid, ammonia, and 1-aminocyclopropane-1-carboxylate (ACC) deaminase, were analyzed in isolated stress-tolerant bacteria, which may contribute towards stress-tolerance and crop productivity in mungbean. The most promising drought-tolerant nodule bacterial isolates were identified as Rhizobium sp. and Pseudomonas indica , respectively by 16S rRNA sequencing. Moreover, bacterial isolates MuJs52b, MuJs53b, MuJs72a and MuBk32b, which exhibited drought-tolerance of 40 % PEG-6000 and different PGP activities, were used as bioinoculant on mungbean plants grown under moderate to severe drought at 50 and 25 % of field capacity (FC) in pot experiment. Bacteria inoculated plants showed maximum increase in nodule dry weight (56.5 %) and shoot dry weight (87.5 %) per plant even under severe drought at 25 % FC. The results indicated that application of four potential nodule bacterial isolates, which were able to tolerate drought stress of 40 % PEG-6000, and having multiple PGP traits, showed stimulation of the mungbean growth even up to 25 % FC. These plant growth-promoting nodule bacterial strains may be used for the enhancement of mungbean crop productivity under drought stress and could be used as biofertilizer. • Drought-tolerance of mungbean nodule bacteria varied with sampling regions. • About 25 % of isolates showed drought-tolerance at 40 % polyethylene glycol-6000. • Only 8 % of bacterial isolates showed tolerance of 40 % PEG-6000 at 45 °C. • 87.5 % increase in shoot biomass under 25 % field capacity in bacteria inoculation. • Promising drought-tolerant bacteria were Rhizobium sp. and Pseudomonas indica. [ABSTRACT FROM AUTHOR]

Purpose: To investigate the role nutrients play in arbuscular mycorrhizal fungal improvement of biological nitrogen fixation (BNF) by Bradyrhizobium sp. and increases in Pratylenchus thornei (Sher & Allen) population density in mung bean (Vigna radiata (L.) R. Wilczek). Methods: A glasshouse experiment was conducted on mung bean with 26 factorial treatments of AMF, rhizobia, P. thornei, nitrogen (N), phosphorus (P) and zinc (Zn) in a pasteurised vertisol. Variates of biomass, yield, nodulation, natural abundance δ15N, mycorrhizal colonisation, nutrients in the plant shoot and P. thornei multiplication were assessed at 6 and 11 weeks. Results: The combination of AMF and P improved BNF in the shoots at 6 weeks, while AMF alone improved BNF and nodulation greater than the addition of P at 11 weeks. Inoculation with AMF increased the shoot concentrations of P and Zn greater than fertilisation with either nutrient alone. Seed yield and biomass were similar when AMF or P was each applied alone with no further increase when combined. Rhizobia increased seed yield greater than the addition of N. Inoculation with AMF and rhizobia increased yield and biomass greater than rhizobia alone, and to a higher level than inoculation with AMF and fertiliser N. Pratylenchus thornei populations in the roots increased with AMF, but the addition of N, P and Zn decreased them. Conclusions: AMF increase supply of P to mung bean, improving BNF by Bradyrhizobium, yield and crop nutrition while reducing fertiliser inputs. Active management to ensure effective AMF colonisation and reduced P. thornei reproduction is warranted. [ABSTRACT FROM AUTHOR]

Background: Phosphorus is one of the limiting plant nutrients in most soil due to its fixation with metal ions both in acidic and alkaline soil. However, some soil bacteria have potential to solubilize the cation-fixed phosphorus and make it available to plants. Methods: Sixty-six cowpea root nodule bacterial species were tested for inorganic phosphate solubilization (PS). PS potential of the isolates from Ca3 (PO4)2, AlPO4 and FePO4 were tested on four different kinds of agar media. Solublization efficiency was determined as Solubilization Index (SI) on agar media and quantity of bioavailable phosphorous on broth media. Result: About 30% of the bacterial isolates, out of which 60% endophytes were Ca3 (PO4)2 solubilizers on Pikovskaya medium but did not grow on media containing AlPO4 and FePO4 as sole P sources. Isolates showed significant variation (p<0.05) in Ca3 (PO4)2 solubilization efficiency on the different culture media and ECE-21 (Pseudomonas putida) was versatile in solubilizing Ca3(PO4)2 from the four test media. However, ECE-26A (Bacillus subtilis) showed the highest PS in broth media (140.8 µg ml-1) indicating the moderate correlation (r=0.5578; p<0.05) of PS between solid and liquid media. Amounts of P solubilized and pH change of the culture filtrate was inversely correlated (r=-0.731; p<0.01). In general, ECE-21 was the most efficient P solubilizer (SI=2.67±0.1; P=87.74±7.4µg ml-1) solublizers and hence can be an ideal candidate as inoculants for the crop production in Ethiopia after field evaluation. [ABSTRACT FROM AUTHOR]

Compared to free-living relatives, bacterial symbionts of plants exhibit higher rates of population diversification (microevolution), as well as of speciation and formation of super-species taxa (macroevolution). Analysis of rhizobia, the N2-fixing symbionts of leguminous plants, suggests that both evolutionary modes are determined by the hosts, which have mutagenic and recombinogenic effects on their microsymbionts, apart from activating the selective processes that determine the fate of newly emerged genotypes in their populations. Migration induced by leguminous plants plays a key role in the evolution of rhizobia, including: (1) joint movement of the symbionts and their hosts into new ecological zones and (2) local circulation of the symbionts in plant‒soil systems. In the first case, allopatric speciation occurs in rhizobia due to transfer of the symbiotically specialized (sym) genes from the introduced strains into local bacteria, which are transformed into novel symbionts. In the second case, a sympatric divergence of Rhizobium populations occurs, resulting in formation of intraspecies taxa (biovars and symbiovars) contrasting in their host ranges. In the evolutionarily advanced rhizobia, Rhizobium leguminosarum and Neorhizobium galegae interacting with galegoid legumes (tribes Fabeae, Galegae, and Trifolieae), these processes are associated with diversification of the sym genes representing the accessory genome parts due to the interaction between bacteria and the hosts differing in their symbiotic affinities. In parallel, formation of new cryptic (genomic) species was revealed in local rhizobia populations, which is associated with divergence of the core genome parts. It may be caused either by genetic drift or by selection for resistance against local edaphic stresses, which in combination with disruptive selection can sufficiently enhance the rhizobia diversity and change the structures of their pangenomes. [ABSTRACT FROM AUTHOR]

The genetic diversity and phylogeny of root nodule bacteria, microsymbionts of plants of the genus Lupinaster Adans. (L. albus Link and L. pentaphyllus Moench), were studied, and also their symbiotic genes were analyzed. The bacterial strains studied were shown to be phylogenetically different; however, all of them are related to the Mezorhizobium genus with the exception of the single strain which is related to the Rhizobium genus. Analysis of the symbiotic genes nifH and nodC revealed their high homology among all strains independently of strain phylogeny, as well as the phylogenetic relation of these genes to those of Mezorhizobium. Mezorhizobium bacteria are likely specific microsymbionts of these plants, while the Rhizobium strain acquired its symbiotic genes and became capable of nodule formation in Lupinaster plants through horizontal gene transfer. Thus, the genetic composition of nodule bacteria inhabiting Lupinaster plants represents additional support for the idea that they do not belong to the Trifolium genus. [ABSTRACT FROM AUTHOR]

The present study aims to address the problem of chromium (Cr) toxicity by providing important insights into the mechanisms involved in its bioremediation. Among the 22 Rhizobium and Sinorhizobium isolates obtained from Sesbania sesban root nodules, Sinorhizobium sp. SAR1 (JX174035.1) tolerated the maximum Cr concentration (1mM) and hence was used for further studies. The excess secretion of extra polymeric substances, as seen from scanning electron micrographs, could be a probable mechanism of adaptation to the Cr stress. The Energy dispersive X-ray spectroscopy data did not show any peaks of Cr. The biosorption studies done on the isolate gave maximum adsorption capacity as 285.71mg/g. The isotherm studies showed a better fit to Langmuir isotherm. The Weber and Morris plot established that the phenomenon of adsorption was governed by film diffusion mechanism. The FTIR analysis suggested the role of cell wall components and extracellular polymeric substances in Cr adsorption to the biomass of Sinorhizobium. On the basis of these results a compiled mechanism of Cr (VI) adsorption and its biotransformation into Cr (III) by Sinorhizobium sp. SAR1 is explained. This work outlines a comprehensive detail for the exact phenomenon of Cr biotransformation by Sinorhizobium sp. SAR1. These results may further help in developing and enhancing effective bioremediation approaches. [ABSTRACT FROM AUTHOR]

The root nodule bacteria were isolated from three different leguminous plants belonging to the Fabaceae family such as Cajanus cajan, Lablab purpureus and Vigna unguiculata using YEMA medium and identified. The isolates were submitted to 16S rRNA and PCR- RFLP typing. A representative sample was further submitted to sequence analysis of 16S rRNA. Isolates were assigned to two genera and were related to Rhizobium spp. (75%) and Bacillus spp. (25%) respectively. This study opens the doors to researchers to do specific studies on this root nodule bacteria which are symbiotic with these legume plants in this area for their antibacterial activity, medicine and food application, and sustainable agriculture. Rhizobium and other beneficial microbes may be used as biofertilizers as a substitute for chemical fertilizers. [ABSTRACT FROM AUTHOR]

Background:Biserrula pelecinusis an annual legume native to the Mediterranean basin, found in pastureland, alone or in association with other legumes (Leguminosae) and grasses (Poaceae). It has been used in revegetation programmes of mining wastes showing phytoremediation potentials and thus becoming potentially highly attractive for plant ecology and restoration management of natural ecosystems. Aims:To characterise native root-nodule bacteria isolated fromB. pelecinusfrom the Iberian Peninsula, and to select suitable N fixers for field-application and soil rehabilitation. Methods:Strains were isolated and molecularly identified by 16S rRNA amplification and sequencing. Strains were phenotypically characterised in different abiotic conditions (acidity, salinity and heavy metals) and tested for their ability to fix atmospheric N2. The most suitable N fixers were applied in greenhouse experiments withB. pelecinusunder different fertilization levels to assess their tolerance to fertilized and polluted soils, commonly encountered in restoration projects. Results:B. pelecinusroot-nodule isolates tolerated pH from 4.5 to 9.5 grew in saline conditions (2.5% of NaCl), and tolerated 50 µM of Al3+and Mn2+. Three isolates efficient in N2fixation, relative to the referenceMesorhizobiumstrain, were considered excellent candidates for the amelioration of nutrient poor sites. Conclusions:These results provide valuable information for the potential use in soil restoration ofB. pelecinusin a wide-range of conditions, exploiting the natural variability of its root-nodule bacteria. [ABSTRACT FROM AUTHOR]

The processes of speciation and macroevolution of root nodule bacteria (rhizobia), based on deep rearrangements of their genomes and occurring in the N-fixing symbiotic system, are reconstructed. At the first stage of rhizobial evolution, transformation of free-living diazotrophs (related to Rhodopseudomonas) to symbiotic N-fixers ( Bradyrhizobium) occurred due to the acquisition of the fix gene system, which is responsible for providing nitrogenase with electrons and redox potentials, as well as for oxygen-dependent regulation of nitrogenase synthesis in planta, and then of the nod genes responsible for the synthesis of the lipo-chitooligosaccharide Nod factors, which induce root nodule development. The subsequent rearrangements of bacterial genomes included (1) increased volume of hereditary information supported by species, genera (pangenome), and individual strains; (2) transition from the unitary genome to a multicomponent one; and (3) enhanced levels of bacterial genetic plasticity and horizontal gene transfer, resulting in formation of new genera-of which Mesorhizobium, Rhizobium, and Sinorhizobium are the largest-and of over 100 species. Rhizobial evolution caused by development and diversification of the Nod factor-synthesizing systems may result in either relaxed host specificity range (transition of Bradyrhizobium from autotrophic to symbiotrophic carbon metabolism in interaction with a broad spectrum of legumes) or narrowed host specificity range (transition of Rhizobium and Sinorhizobium to 'altruistic' interaction with legumes of the galegoid clade). Reconstruction of the evolutionary pathway from symbiotic N-fixers to their free-living ancestors makes it possible to initiate the studies based on up-to-date genome screening technologies and aimed at the issues of genetic integration of organisms into supraspecies complexes, ratios of the macro- and microevolutionary mechanisms, and development of cooperative adaptations based on altruistic interaction between the symbiotic partners. [ABSTRACT FROM AUTHOR]

Root-nodulating bacteria are intimate associates of legumes. From a pool of rhizobia isolated from root nodules of Mucuna pruriens (Velvet bean/Kaunch), RMP66 and BMP17 were found to be capable of promoting siderophore and IAA production and phosphate solubilization (insoluble tri-calcium). Both symbionts were studied further to determine their abilities to promote plant growth and to control root-rot in Mucuna pruriens caused by the pathogenic plant fungus Macrophomina phaseolina. RMP66 and BMP17 were selected based on their excellent inhibitory activities against M. phaseolina (by 78% and 71%, respectively) in dual culture and in agar-well assays using cell-free culture filtrate (CFCF) (by 76% and 62%, respectively). Both strains inhibited fungal growth to a greater extent in iron-deficient medium (51% and 69%) than in iron-supplemented medium (37% and 0%), respectively. CFCFs of RMP66 and BMP17 obtained from Pikovskaya's broth and tryptophan-amended YEM broth inhibited fungal growth by 80%-55% and 70%-43%, respectively, and were identified as Sinorhizobium meliloti RMP66 and Bradyrhizobium diazoefficiens BMP17 by 16S rDNA sequencing. Centrifuged and pelleted cells harvested from exponentially grown cultures of S. meliloti RMP66 and B. diazoefficiens BMP17 were used to bacterize seeds of M. pruriens, which then showed enhanced seed germination (by up to 17% and 12%, respectively), and subsequent increases in other plant growth parameters in field trials. Considerable increases in seedling vigour indices (62%: 53% and 110%: 130%) and biomass (8%: 13% and 25%: 28%) were also observed for bacterial treatments. Tn5-mediated antibiotic-resistant marker strains showed enhanced nodule occupancy by up to 72% and 68%, respectively. This study describes a multifunctional legume nodule rhizobia that could be utilized in multicropping systems under different agroclimatic conditions as a bioinoculant and alternative to fertilizers. [ABSTRACT FROM AUTHOR]

The theoretical and experimental data on salt tolerance of root nodule bacteria Sinorhizobium meliloti (Ensifer meliloti), an alfalfa symbiont, and on genetic determination of this feature are reviewed. Extensive data are provided on the genes affecting adaptation of proteobacteria and on the groups of genes with activity depending on the osmolarity of the medium. Structural and functional polymorphism of the bet genes involved in betaine synthesis and transport in S. meliloti is discussed. The phenotypic and genotypic polymorphism in 282 native rhizobial strains isolated from the centers of alfalfa diversity affected by aridity and salinity is discussed. The isolates from the Aral Sea area and northern Caucasus were shown to possess the betC gene represented by two types of alleles: the dominant Atype allele found in Rm1021 and the less common divergent Etype allele, which was revealed in regions at the frequencies of 0.35 and 0.48, respectively. In the isolates with the salt-tolerant phenotype, which were isolated from root nodules and subsequently formed less effective symbioses with alfalfa, the frequency of Etype alleles was 2.5 times higher. Analysis of the nucleotide and amino acid sequences of the Etype allele of the betC gene revealed that establishment of this allele in the population was a result of positive selection. It is concluded that diversification of the functionally diverse bet genes occurring in S. meliloti affects the salt tolerance and symbiotic effectiveness of rhizobia. [ABSTRACT FROM AUTHOR]

In this study, bacteria (8 species and 5 genera) belonging to the classes Betaproteobacteria, Gammaproteobacteria, and Sphingobacteria were isolated from root nodules of the multipurpose legume Clitoria ternatea L. and identified on the basis of partial 16S rRNA sequencing. The root nodule bacteria were subjected to phenotypic clustering and diversity studies using biochemical kits, including Hi-Media Carbokit™, Enterobacteriaceae™ identification kit, ERIC-PCR, and 16S ARDRA. All the strains showed growth on Ashby's N-free media over 7 generations, indicative of presumptive nitrogen fixation and further confirmed by amplification of the nifH gene. None of the strains showed the capability to renodulate the host plant, neither alone nor in combination with standard rhizobial strains, which was further confirmed by the absence of nodC bands in PCR assay. The results clearly indicate the common existence of nonrhizobial microflora inside the root nodules of legumes, which were thought to be colonized only by rhizobia and were responsible for N2 fixation in leguminous crops. However, with the recent discovery of nodule endophytes from a variety of legumes, as also observed here, it can be assumed that symbiotic rhizobia are not all alone and that these invasive endophytes belonging to various bacterial genera are more than just opportunistic colonizers of specialized nodule niche. [ABSTRACT FROM AUTHOR]

In the present study, a total of 353 nodule-associated bacteria were isolated from 220 legume plant samples belonging to Cicer arietinum (85), Glycine max (74), Vigna radiata (21) and Cajanus cajan (40). A total of 224 bacteria were identified as fast-growing Rhizobium spp. on the basis of differential staining (Gram staining and carbol fuchsin staining) and biochemical tests. All the isolates were tested for indole acetic acid production (IAA), phosphate solubilization and siderophore production on plate assay. To examine the effect of volatile organic metabolites (VOM) and water soluble soil components (WSSC) on nodule bacteria, culture conditions were optimized by observing the effects of various parameters such as pH, salt content and temperatures on the growth of bacteria. Selected rhizobia were subjected to random amplified polymorphic DNA (RAPD) and amplified ribosomal DNA restriction analysis (ARDRA) analysis to identify their species. On the basis of RAPD and ARDRA, 10 isolates were identified as Rhizobium meliloti. In this study, Rhizobium GO4, G16, G20, G77, S43, S81, M07, M37, A15 and A55 were observed as the best candidates among the tested bacteria and can be further used as potent bioinoculants. [ABSTRACT FROM AUTHOR]

Summary Inoculation with beneficial soil microorganisms has the potential to enhance success of restoration, particularly in harsh Mediterranean-type ecosystems (MTEs). We investigated the effects of microorganisms (mycorrhizal fungi and root nodule bacteria) and planting material (seed and nursery-raised seedlings) on early establishment and growth of two key postdisturbance colonizing species with different life histories, life forms and functional types ( Eucalyptus gomphocephala and Acacia saligna) under field conditions. Establishment and growth were monitored at 13 months, following the first MTE drought period. For E. gomphocephala, establishment was higher for seedlings (81%) than for seeding (7.5%). Inoculation with ectomycorrhizal fungal spores was not beneficial. For A. saligna, establishment was also higher for seedlings (84%) than for seeding (42.5%). Mycorrhizal fungal inoculum had no effect on establishment or growth. This study has shown that in harsh MTE conditions, the use of seedlings is more effective than seeding in degraded woodlands even when attempting to reintroduce key colonizing species. The microorganism treatments tested did not result in significant improvement in establishment or growth. [ABSTRACT FROM AUTHOR]

The genetic diversity and phylogeny of root nodule bacteria entering into symbiosis with bitter peavine Lathyrus vernus (L.) Bernh. ( Fabaceae) growing in various regions of the Republic of Bashkortostan were studied. RAPD analysis revealed a high degree of polymorphism of the DNA of the isolated strains giving evidence of the heterogeneity of the microorganisms in question. The study of the phylogeny of microsymbionts based on comparative analysis of the nucleotide sequences of 16S rRNA genes showed that the bacteria isolated from the plant nodules of L. vernus growing on the territory of Ufa and Beloretsk raions belonged to the species Rhizobium leguminosarum, whereas the microsymbionts of L. vernus growing on the territory of Tatyshly raion belonged to the species Rhizobium tropici,@ except for several strains of Rhizobium leguminosarum [ABSTRACT FROM AUTHOR]

Plant growth experiments were conducted to assess symbiotic efficiency, photosynthetic rates, and the development of soybean ( Glycine max (L.) Merrill) seedlings after seed inoculation with active and inactive strains of root nodule bacteria Bradyrhizobium japonicum preincubated in the presence homologous and heterologous proteins. The properties of active and inactive symbiotic strains were differentially modulated by homologous soybean lectin, which had a marked influence on plant physiological condition. The incubation of active rhizobia with a homologous lectin, i.e., lectin of the respective plant, increased the nitrogen-fixing activity of nodules and, consequently, elevated photosynthetic rates and weight increments in soybean plants. At the same time, the homologous lectin suppressed the symbiotic properties of inactive strain of nodule bacteria. The preincubation of rhizobia with a heterologous pea lectin had virtually no effect on functioning of symbiotic apparatus and photosynthetic rate, whereas the preincubation of root nodule bacteria with human albumin exerted an effect similar to that induced by a homologous lectin on symbiotic productivity. [ABSTRACT FROM AUTHOR]

Bacteria were isolated from root-nodules collected from indigenous legumes at 38 separate locations in the Gascoyne and Pilbara regions of Western Australia. Authentication of cultures resulted in 31 being ascribed status as root-nodule bacteria based upon their nodulation of at least one of eight indigenous legume species. The authenticated isolates originated from eight legume genera from 19 sites. Isolates were characterised on the basis of their growth and physiology; 20 isolates were fast-growing and 11 were slow-growing (visible growth within 3 and 7 d, respectively). Fast-growers were isolated from Acacia, Isotropis, Lotus and Swainsona, whilst slow-growers were from Muelleranthus, Rhynchosia and Tephrosia. Indigofera produced one fast-growing isolate and seven slow-growing isolates. Three indigenous legumes (Swainsona formosa, Swainsona maccullochiana and Swainsona pterostylis) nodulated with fast-growing isolates and four species (Acacia saligna, Indigofera brevidens, Kennedia coccinea and Kennedia prorepens) nodulated with both fast- and slow-growing isolates. Swainsona kingii did not form nodules with any isolates. Fast-growing isolates were predominantly acid-sensitive, alkaline- and salt-tolerant. All slow-growing isolates grew well at pH 9.0 whilst more than half grew at pH 5.0, but all were salt-sensitive. All isolates were able to grow at 37 °C. The fast-growing isolates utilised disaccharides, whereas the slow-growing isolates did not. Symbiotic interactions of the isolates were assessed on three annual, one biennial and nine perennial exotic legume species that have agricultural use, or potential use, in southern Australia. Argyrolobium uniflorum, Chamaecytisus proliferus, Macroptilium atropurpureum, Ononis natrix, Phaseolus vulgaris and Sutherlandia microphylla nodulated with one or more of the authenticated isolates. Hedysarum coronarium, Medicago sativa, Ornithopus sativus, Ornithopus compressus, Trifolium burchellianum, Trifolium polymorphum and Trifolium uniflorum did not form nodules. Investigation of the 31 authenticated isolates by polymerase chain reaction with three primers resulted in the RPO1 primer distinguishing 20 separate banding patterns, while ERIC and PucFor primers distinguished 26 separate banding patterns. Sequencing the 16S rRNA gene for four fast- and two slow-growing isolates produced the following phylogenetic associations; WSM1701 and WSM1715 (isolated from Lotus cruentus and S. pterostylis, respectively) displayed 99% homology with Sinorhizobium meliloti, WSM1707 and WSM1721 (isolated from Sinorhizobium leeana and Indigofera sp., respectively) displayed 99% homology with Sinorhizobium terangae, WSM1704 (isolated from Tephrosia gardneri) shared 99% sequence homology with Bradyrhizobium elkanii, and WSM1743 (isolated from Indigofera sp.) displayed 99% homology with Bradyrhizobium japonicum. [Copyright &y& Elsevier]

Bacterial strains isolated from the nodules, tissues, and root surface of wild legumes growing in the Southern Urals related to the tribes Galegeae, Hedysareae, Genisteae, Trifolieae, and Loteae were examined for the presence in their genomes of symbiotic ( sym) genes. It was found that the sym-genes are present in microorganisms isolated only from the nodules of the analyzed plants ( sym strains). Phylogenetic analysis of sym strains on the basis of a comparative analysis of 16S rRNA gene sequences showed that sym strains belong to five families of nodule bacteria: Mesorhizobium, Bradyrhizobium, Sinorhizobium, Rhizobium, and Phyllobacterium. A study the phylogeny of the sym-genes showed that the nodule bacteria of leguminous plants of the Southern Urals at the genus level are mainly characterized by a parallel evolution of symbiotic genes and the 16S rRNA gene. Thus, cases of horizontal transfer of sym genes, which sometimes leads to the formation of certain types of atypical rhizobial strains of leguminous plants, are detected in nodule bacteria populations. [ABSTRACT FROM AUTHOR]

Competitiveness and genetic variation of the Rhizobium galegae strains from the collection of the All-Russia Institute of Agricultural Microbiology, Russian Academy of Agricultural Sciences, causing nodulation of oriental goat’s rue under conditions of Bashkortostan soils (lacking this rhizobial species) were studied. It was demonstrated that of all the tested strains, the strains CIAM 0702 and CIAM 0704, each carrying two megaplasmids of 1500 and 2000 MDa, were the most competitive. RAPD (random amplified polymorphic DNA) analysis showed that R. galegae strains were able to intensively exchange the genetic material in the host plant rhizosphere. We did not succeed in detecting the local root nodule bacteria that were either initially able to infect oriental goat’s rue or had adapted to infecting this species due to various genetic rearrangements. [ABSTRACT FROM AUTHOR]

1. Extensive clearing of native vegetation in Australia has contributed to major environmental problems, including land degradation, dryland salinity, soil erosion and loss of biodiversity. Re-establishing cover with deep-rooted perennial species is a major focus for conservation and sustainable land management, particularly with regard to hydrological control of recharge and saline discharge areas. However, considerable expense is involved in large-scale revegetation programmes and cost effectiveness is a real concern. 2. Low-cost revegetation approaches are needed that require little maintenance yet can substantially enhance reliable establishment and growth of native trees and shrubs. We evaluated results from direct-seeding field trials that examined the benefits of using native Australian Acacia species inoculated with effective strains of nitrogen-fixing root-nodule bacteria to revegetate degraded landscapes. 3. On average, inoculation led to a 118% increase in establishment of acacia seedlings, indicating that the use of elite strains of native bacteria can substantially reduce seed requirements. This is a major benefit given the expense of collecting sufficient native seed and the impacts of this activity on remnant population viability. 4. Particularly at sites experiencing harsher climatic conditions, subsequent survival of inoculated seedlings was significantly greater than for uninoculated controls. Moreover, inoculated acacias grew 10–58% faster than uninoculated controls during the critical early phase of establishment, although this varied among species and sites. 5. Synthesis and applications. Inoculation of Acacia species or other native leguminous shrubs and trees with elite strains of native rhizobia as part of direct-seeding techniques has the potential to increase the scope, rate and success of land restoration world-wide. Re-establishment of important plant–soil interactions in degraded soils can contribute significantly to the development of biodiverse self-regenerating native ecosystems in agricultural landscapes. Journal of Applied Ecology (2005) 42, 740 –751 doi: 10.1111/j.1365-2664.2005.01058.x [ABSTRACT FROM AUTHOR]

The genetic diversity and phylogeny of root module bacteria entering into symbiotic relations with Caragana arborescens introduced on the territory of the Republic of Bashkortostan (RB) were studied. Analysis of the 85 strains isolated from root-nodules showed that, according to the DNA polymorphism revealed by RAPD analysis, they can be divided into 12 groups. It was found that, both in natural habitats and on the territory of the RB, Caragana arborescens is primarily nodulated by Mesorhizobium bacteria whose 16S rRNA gene sequences differ to some degree from those of the bacteria described earlier by Chinese authors. Bacteria assigned to the genus Phyllobacterium based on their 16S rRNA gene sequences were also revealed in plants growing on the territory of the RB. [ABSTRACT FROM AUTHOR]

The South African legumes Lotononis bainesii, L. listii and L. solitudinis are specifically nodulated by highly effective, pink-pigmented bacteria that are most closely related to Methylobacterium nodulans on the basis of 16S rRNA gene homology. Methylobacterium spp. are characterized by their ability to utilize methanol and other C1 compounds, but 11 Lotononis isolates neither grew on methanol as a sole carbon source nor were able to metabolize it. No product was obtained for PCR amplification of mxaF, the gene encoding the large subunit of methanol dehydrogenase. Searches for methylotrophy genes in the sequenced genome of Methylobacterium sp. 4-46, isolated from L. bainesii, indicate that the inability to utilize methanol may be due to the absence of the mxa operon. While methylotrophy appears to contribute to the effectiveness of the Crotalaria/M. nodulans symbiosis, our results indicate that the ability to utilize methanol is not a factor in the Lotononis/ Methylobacterium symbiosis. [ABSTRACT FROM AUTHOR]

Abstract: The leguminous tree genus Inga is thought to be critical for providing N to certified organic coffee shrubs that commonly grow beneath its shade in the state of Chiapas, Mexico. Organic certification prohibits use of synthetic N fertilizers, necessitating understanding of the Inga–rhizobia symbiosis. The objectives of this study were to: (1) establish a collection of isolates from Inga sp. nodules found on organic coffee farms in Chiapas, Mexico; (2) use phenotypic and genotypic approaches to characterize these isolates, and (3) examine cross-inoculation patterns of strains isolated from Inga sp. with other Inga species and N2-fixing trees. Eighty-three strains were isolated from root nodules of Inga oerstediana, Inga pavoniana and an unknown species of Inga. Aside from isolates of I. pavoniana, Inga isolates demonstrate characteristics consistent with Bradyrhizobium. Dendrograms generated using phylogenetic and phenotypic traits showed strains to exhibit significant diversity, even among strains originating from the same farm. Strains isolated from I. pavoniana were 95% similar, and displayed strikingly different characteristics from strains isolated from other species. Cross-inoculation experiments used three Inga strains (cowpea miscellany CB756, and isolates from I. oerstediana and I. sp.) as inoculants with I. edulis, I. fuellii, Acacia mangium, Acacia koa, Vigna unguculata and Gliricidia sepium. All three strains formed nodules containing leghaemoglobin with V. unguculata, while isolates from I. oerstediana, Inga sp. and cowpea miscellany formed nodules with I. feulli and I. edulis, but not with A. mangium and G. sepium. A. koa formed nodules containing leghaemoglobin when inoculated by cowpea miscellany strain CB756 and the strain from Inga sp., but nodules formed with the strain isolated from I. oerstediana did not contain leghaemoglobin. One and two-way ANOVA results showed that no mean biomass of inoculated seedlings was significantly greater than the −N control for each of A. koa, G. sepium, A. mangium (p &#60;0.001), supporting our hypothesis that inoculation with non-Inga strains would have no effect on biomass production. We suggest that great spatial variation exists among Inga isolates from organic coffee farms in the Chiapas highland region, both between and within farms, and that more research is needed to identify reasons for the genetic distribution and function of Inga symbionts. [Copyright &y& Elsevier]

Twenty-six root nodule bacteria from two native forage legumes namely Sulla aculeolata subsp. aculeolata and Sulla aculeolata subsp. mauritanica were isolated and analyzed using a polyphasic approach comprising phenotypic traits, ERIC-PCR, and 16S rRNA gene sequencing. This is the first time a study has been performed to determine the diversity of bacteria associated with Sulla aculeolata spp. Phenotypically, all the isolates were identified as fast-growing bacteria and shows high tolerance toward various stressed conditions, particularly those derived from S. aculeolata subsp. mauritanica. On the other hand, the genotypic characterization revealed high diversity among the isolated bacteria and clustered into 14 clusters at the similarity index of 90% based on ERIC-PCR analysis. Furthermore, the 16S rRNA gene sequencing of representatives strains indicates that all the strains share 99 to 100% identity with bacteria belonging to Pseudomonas, Enterobacter, Serratia, and Paenibacillus genera with a clear relation to their host plant. In conclusion, the findings of the present study suggested the inoculation of plants with appropriate bacteria to enhance plant growth and quality of Sulla aculeolata under semi-arid conditions of the Mediterranean area. [ABSTRACT FROM AUTHOR]

Aims: Milletia pinnata is a leguminous tropical tree that produces seed oil suitable for biodiesel and is targeted to be planted on marginal land associated with nitrogen poor soil. This study aimed to identify effective rhizobia species for M. pinnata. Methods: Soil samples were collected from M. pinnata grown in Kununurra, Australia. Rhizobia were trapped, characterised and sequenced for 16S rRNA, atpD, dnaK and recA genes. Results: Forty isolates tolerated pH 7 - 9, temperatures 29 - 37 °C, salinity below 1 % NaCl, and had optimal growth on mannitol, arabinose or glutamate as a single carbon source, a few grew on sucrose and none grew on lactose. Inoculation of isolates increased shoot dry weight of M. pinnata's seedlings in nitrogen minus media. Slow-growing isolates were closely related to Bradyrhizobium yuanmingense, Bradyrhizobium sp. DOA10, Bradyrhizobium sp. ORS305 and B. liaoningense LMG 18230. The fast-growing isolates related to Rhizobium sp. 8211, R. miluonense CCBAU 41251, R miluonense CC-B-L1, Rhizobium sp. CCBAU 51330 and Rhizobium sp. 43015 Conclusions: Millettia pinnata was effectively nodulated by slow-growing isolates related to Bradyrhizobium yuanmingense, Bradyrhizobium sp. DOA10 Bradyrhizobium sp. ORS305, B. liaoningense LMG 18230 and fast-growing isolates related Rhizobium sp. 8211, R. miluonense, Rhizobium sp. CCBAU 51330 and Rhizobium sp. 43015 [ABSTRACT FROM AUTHOR]

A collection of 104 isolates from root-nodules of Vicia faba was submitted to 16S rRNA PCR-RFLP typing. A representative sample was further submitted to sequence analysis of 16S rRNA. Isolates were assigned to 12 genera. All the nodulating isolates (45 %) were closely related to Rhizobium leguminosarum USDA2370 (99.34 %). The remaining isolates, including potential human pathogens, failed to nodulate their original host. They were checked for presence of symbiotic genes, P-solubilization, phytohormone and siderophore production, and then tested for their growth promoting abilities. Results indicated that 9 strains could induce significant increase (41-71 %) in shoot dry yield of faba bean. A Pseudomonas strain was further assessed in on-farm trial in combination with a selected rhizobial strain. This work indicated that nodule-associated bacteria could be a valuable pool for selection of effective plant growth promoting isolates. Nevertheless, the possible involvement of nodules in increasing risks related to pathogenic bacteria should not be neglected and needs to be investigated further. [ABSTRACT FROM AUTHOR]

The review summarizes the results of studies on the comigration of tubercular bacteria and bean plants to new habitats, which is often accompanied by a decrease in the symbiosis efficiency due to a loss of the diversity of genes responsible for the interaction. This migration may lead to a rise in new symbionts as a result of gene transfers from initial symbionts to local bacteria. It was demonstrated that typically new symbionts lack an ability for N fixation but are highly competitive, blocking the inoculation of bean cultures by industrial strains. The design of coadapted systems of recognition and signal interaction of partners is a perspective approach to ensure competitive advantages of efficient rhizobia strains introduced into agrocenoses, together with host plants, over inactive local strains. [ABSTRACT FROM AUTHOR]

The genus Psoralea (tribe Psoraleae, family Leguminosae) is indigenous to the Cape fynbos of South Africa and little is known about its symbiosis and/or adaptation. The aim of this study was to assess root nodulation and N2 fixation in eight of the 50 Psoralea species, as well as the biodiversity of their asso-ciated nodulating microsymbionts. The eight species studied (namely, Psoralea pinnata, Psoralea aphylla, Psoralea aculeata, Psoralea monophylla, Psoralea repens, Psoralea laxa, Psoralea asarina and Psoralea res-tioides) all had round-shaped, determinate type (desmodioid) nodules, and data from 15N natural abundance showed that they obtained 60-88% of their N nutrition from symbiotic fixation. These Psoralea species also transported their fixed-N as ureides (allantoin and allantoic acid) in the xylem stream, a symbiotic trait that links them very closely to the tribe Phaseoleae. Bacteria isolated from root nodules of the eight Psoralea species varied in phenotypic characteristics, nodulation promiscuity, and N2-fixing efficacy. Furthermore, 16S rDNA gene sequence data showed that Psoralea species can form root nodules with different soil bacteria, including Rhizobium, Mesorhizobium and Burkholderia strains. This is not only evidence of nodulation promiscuity, but also an indication of the species' adaptation to the nutrient-poor, low-N, sandy acidic soils of the Cape fynbos. [ABSTRACT FROM AUTHOR]

Aims: Aim of this study is to determine the genetic variation of rhizobia associated with horse gram [ Macrotyloma uniflorum (Lam.) Verdc.] plants grown in different regions of Andhra Pradesh, India. Methods and Results: Four representative isolates having most representative characters from the previous characterization were selected for 16S rRNA sequence. The sequences were submitted to the NCBI GenBank and Ribosomal Database Project (RDP). The isolates HGR-4, 6 and 13 showed more than 99% homology between them and they were grouped with Rhizobium reference strains where as the isolate HGR-25 showed 87·1, 87·4 and 87·2% homology with the isolates HGR-4, 6 and 13, respectively, and were grouped with reference strains for Caulobacter. The nodulation ability of these isolates on horse gram was confirmed by inoculation tests. Conclusions: The isolate HGR-25 was identified as Caulobacter isolated from the plants growing in soil samples collected from Khareemnagar district, Andhra Pradesh, India. Inoculation tests revealed that Caulobacter formed nodules on horse gram. It was also confirmed by RDP. Significance and Impact of the Study: This is the first report that a legume was nodulated by a member of the genus Caulobacter, which belongs to the family Caulobacteriaceae in the order Caulobacterales of Alphaproteobacteria. [ABSTRACT FROM AUTHOR]

Abstract: Many fast growing tree species have been introduced to promote biodiversity rehabilitation on degraded tropical lands. Although it has been shown that plant productivity and stability are dependent on the composition and functionalities of soil microbial communities, more particularly on the abundance and diversity of soil symbiotic micro-organisms (mycorrhizal fungi and rhizobia), the impact of tree introduction on soil microbiota has been scarcely studied. This research has been carried in a field plantation of Acacia holosericea (Australian Acacia species) inoculated or not with an ectomycorrhizal fungus isolate, Pisolithus albus IR100. After 7 year''s plantation, the diversity and the symbiotic properties of Bradyrhizobia isolated from the plantation soil or from the surrounding area (Faidherbia albida (Del.) a. Chev. parkland) and able to nodulate F. albida, a native Sahelian Acacia species, have been studied. Results clearly showed that A. holosericea modified the structure of Bradyrhizobia populations and their effectiveness on F. albida growth. This negative effect was counterbalanced by the introduction of an ectomycorrhizal fungus, P. albus, on A. holosericea root systems. In conclusion, this study shows that exotic plant species can drastically affect genotypic and symbiotic effectiveness of native Bradyrhizobia populations that could limit the natural regeneration of endemic plant species such as F. albida. This effect could be counterbalanced by controlled ectomycorrhization with P. albus. These results have to be considered when exotic tree species are used in afforestation programs that target preservation of native plants and soil ecosystem rehabilitation. [Copyright &y& Elsevier]

The effect of various biopreparations of the root nodule bacterium Bradyrhizobium japonicum, modified with a homologous lectin, on the virulence of rhizobia, the nitrogen-fixing activity of root nodules, and the productivity of the soybean ( Glycine max (L.) Merr.) was studied. It was shown that a homologous lectin, added to a bacterial suspension when manufacturing biopreparations on a liquid and solid support, increases the efficiency of the soybean symbiotic system and the productivity of the host plant. The potentialities of using bacterial preparations modified with a homologous lectin are discussed. [ABSTRACT FROM AUTHOR]

Hydrogen peroxide (H2O2) content and catalase activity were studied in pea ( Pisum sativum L.) seedlings with a normal (cultivar Marat) and disrupted (pea mutants) process of nodulation that were inoculated with the nitrogen-fixing bacterium Rhizobium leguminosarum strain CIAM 1026. Differences in hydrogen peroxide content and catalase activity in pea seedlings with different ability for nodulation that were inoculated with rhizobia were found. It was assumed that H2O2 and catalase are involved in defensive and regulatory mechanisms in the host plant. [ABSTRACT FROM AUTHOR]

This study investigated the ability of Rhizobium soil inoculants isolated from cadmium (Cd)-contaminated soils to form effective symbioses with peas in the presence of Cd. With Cd present, certain isolates induced high pea dry matter production; an isolate, G, from 399-ppm Cd soil induced high nitrogen uptake and one from 33-ppm Cd, C, soil accelerated Cd uptake. Promising isolates regarding Cd response were used to inoculate various pea selects. Transmission electron microscope analyses showed differences in Cd tolerance between bacterial isolates and pea selects. [ABSTRACT FROM AUTHOR]

Field beans of indeterminate (cv. Nadwiślański) and determinate (cv. Tibo) growth habits were grown on field plots with a density of 20, 40 and 80 plants m-2 at two levels of nitrogen fertilization: low (20 kg N ha-1) and very high (150 kg N ha-1). At the phase of intense pod growth the number and the dry matter weight of root nodules as well as their nitrogenase activity and some features of the plants growth were determined, and in the period of ripeness the components of seed yield were established. It was found that increased density of sowing as well as the high level of nitrogen fertilization inhibited the growth and development of root nodules and limited their nitrogenase activity in both cultivars. Plants of both varieties were characterized by a similar potential of forming the root nodules, however, in plants of the cv. Tibo the nitrogenase activity of nodules was much lower than in the cv. Nadwiślański. Increased population density of the plants has in both varieties given increased seed yield when calculated per m² of the soil, compensating in excess the depression of the seed yield from one plant. Such compensation did not occur in the case of the depression of nitrogen fixation under intense nitrogen fertilization. [ABSTRACT FROM AUTHOR]

The plants of field bean var. Nadwiślański were detopped after about 1 month since the beginning of flowering and after that all flower buds were removed from some plants. The effect of a change in the trophic balance on the setting and subsequent growth of root nodules was studied. The removal of pods had the greater effect on the increase in the weight of the remaining organs than the detopping of plants Detopping retarded the ageing of the plants and delayed the phase of the greatest reduction of the number of pods. The detopping of plants revealed their considerable compensative possibilities in terms of production of the pod weight and the potential of leaves for biomass production. The data obtained indicate that plant detopping may disturb the root--bacteria exchange of signals, necessary to initiate the nodule development, while the further growth of nodules depends, above all, on the amount of available photosynthates. A very high correlation between the dry weight of the whole plant to the dry weight of leaves ratio and the weight of nodules was observed. It proves the importance of the trophic balance of the plant for the root nodule weight production. [ABSTRACT FROM AUTHOR]

Root nodule bacteria (RNB) or 'rhizobia' are a type of plant growth promoting bacteria, typified by their ability to fix nitrogen for their plant host, fixing nearly 65% of the nitrogen currently utilized in sustainable agricultural production of legume crops and pastures. In this study, we sequenced the genomes of 110 RNB from diverse hosts and biogeographical regions, and undertook a global exploration of all available RNB genera with the aim of identifying novel genetic determinants of symbiotic association and plant growth promotion. Specifically, we performed a subtractive comparative analysis with non-RNB genomes, employed relevant transcriptomic data, and leveraged phylogenetic distribution patterns and sequence signatures based on known precepts of symbiotic- and host-microbe interactions. A total of 184 protein families were delineated, including known factors for nodulation and nitrogen fixation, and candidates with previously unexplored functions, for which a role in host-interaction, -regulation, biocontrol, and more, could be posited. These analyses expand our knowledge of the RNB purview and provide novel targets for strain improvement in the ultimate quest to enhance plant productivity and agricultural sustainability. [ABSTRACT FROM AUTHOR]

The diversity of phage-related sequences (PRSs) and their site-specific integration into the genomes of nonpathogenic, agriculturally valuable, nitrogen-fixing root nodule bacteria, such as Sinorhizobium meliloti, were evaluated in this study. A total of 314 PRSs, ranging in size from 3.24 kb to 88.98 kb, were identified in the genomes of 27 S. meliloti strains. The amount of genetic information foreign to S. meliloti accumulated in all identified PRSs was 6.30 Mb. However, more than 53% of this information was contained in prophages (Phs) and genomic islands (GIs) integrated into genes encoding tRNAs (tRNA genes) located on the chromosomes of the rhizobial strains studied. It was found that phiLM21-like Phs were predominantly abundant in the genomes of S. meliloti strains of distant geographical origin, whereas RR1-A- and 16-3-like Phs were much less common. In addition, GIs predominantly contained fragments of phages infecting bacteria of distant taxa, while rhizobiophage-like sequences were unique. A site-specific integration analysis revealed that not all tRNA genes in S. meliloti are integration sites, but among those in which integration occurred, there were "hot spots" of integration into which either Phs or GIs were predominantly inserted. For the first time, it is shown that at these integration "hot spots", not only is the homology of attP and attB strictly preserved, but integrases in PRSs similar to those of phages infecting the Proteobacteria genera Azospirillum or Pseudomonas are also present. The data presented greatly expand the understanding of the fate of phage-related sequences in host bacterial genomes and also raise new questions about the role of phages in bacterial–phage coevolution. [ABSTRACT FROM AUTHOR]

Abstract—The importance of root nodule bacteria in biotechnology is determined by their distinctive feature: symbiotic nitrogen fixation resulting in the production of organic nitrogen-containing compounds. While interacting with host legume plants, the cells of these bacteria undergo global changes at all levels of expression of genetic information leading to the formation in root nodules of so-called bacteroids functioning as nitrogen fixation factories. The molecular mechanisms underlying plant-microbial symbiosis are actively investigated, and one of the most interesting and poorly studied aspects of this problem is the species-specificity of interaction between root nodule bacteria and host plants. In this work we have performed the proteomic analysis of the Sinorhizobium meliloti bacteroids isolated from two legume species: alfalfa (Medicago sativa L.) and yellow sweet clover (Melilotus officinalis L.). It has been shown that the S. meliloti bacteroids produce a lot of proteins (many of them associated with symbiosis) in a host-specific manner, i.e., only in certain host plant species. It has been demonstrated for the first time that the levels of expression in bacteroids of the genes encoding the ExoZ and MscL proteins responsible for the synthesis of surface lipopolysaccharides and formation of a large conductance mechanosensitive channel, respectively, depend on a host plant species that confirms the results of proteomic analysis. Overall, our data show that the regulation of bacteroid development by the host plant has species-specific features. [ABSTRACT FROM AUTHOR]

Soil Sinorhizobium phage AP-16-3, a strain phylogenetically close to Rhizobium phage 16-3, was isolated in a mountainous region of Dagestan, belonging to the origin of cultivated plants in the Caucasus, according to Vavilov N.I. The genome of phage AP-16-3 is 61 kbp in size and contains 62 ORFs, of which 42 ORFs have homologues in the genome of Rhizobium phage 16-3, which was studied in the 1960s–1980s. A search for Rhizobium phage 16-3-related sequences was performed in the genomes of modern strains of root nodule bacteria belonging to different species, genera, and families. A total of 43 prophages of interest were identified out of 437 prophages found in the genomes of 42 strains, of which 31 belonged to Sinorhizobium meliloti species. However, almost all of the mentioned prophages contained single ORFs, and only two prophages contained 51 and 39 ORFs homologous to phages related to 16-3. These prophages were detected in S. meliloti NV1.1.1 and Rh. leguminosarum OyaliB strains belonging to different genera; however, the similarity level of these two prophages did not exceed 14.7%. Analysis of the orphan genes in these prophages showed that they encoded predominantly virion structural elements, but also enzymes and an extensive group of hypothetical proteins belonging to the L, S, and E regions of viral genes of phage 16-3. The data obtained indicate that temperate phages related to 16-3 had high infectivity against nodule bacteria and participated in intragenomic recombination events involving other phages, and in horizontal gene transfer between rhizobia of different genera. According to the data obtained, it is assumed that the repetitive lysogenic cycle of temperate bacteriophages promotes the dissolution of the phage genetic material in the host bacterial genome, and radical updating of phage and host bacterial genomes takes place. [ABSTRACT FROM AUTHOR]

Background: Soybean [Glycie max (L.) Merril] is a very important oil seed and protein rich crop. It has the specific character to fix atmospheric nitrogen through root nodule bacteria in symbiotic relationship. A field experiment was conducted during kharif, 2019 at ARS farm, College of Agriculture, Ummedganj, Kota (Rajasthan). To find out the effect of organic and inorganic sources of nutrients on growth, yield attributes and nutrient uptake of soybean in Vertisols of Rajasthan. The soil of the experimental field comes under Vertisols with clay loam texture having low to medium fertility status and good porosity. Methods: The experiment consisted of 10 treatments viz. T0- Control (Absolute), T1- RDF (20-40-40), T2- 75% RDF + FYM (2.0 t ha-1), T3- 50% RDF + FYM (4.0 t ha-1), T4- 75% RDF + FYM (2.0 t ha-1)+Rhizobium, T5- 50% RDF + FYM (4.0 t ha-1) + Rhizobium, T6- 75% RDF + Vermicompost (1.0 t ha-1), T7- 50% RDF + Vermicompost (2.0 t ha-1), T8- 75% RDF+Vermicompost (1.0 t ha-1) + Rhizobium, T9- 50% RDF + Vermicompost (2.0 t ha-1) + Rhizobium. The experiment was laid out in randomised block design and was replicated thrice. Result: Among the nutrient management treatment, application of 75% RDF + Vermicompost (1.0 t ha-1) + Rhizobium (T8) resulted into maximum plant height (58.70 cm), branches plant-1 (5.60), chlorophyll content (2.84 mg g-1), total nodules per plant (47.40), effective nodules (31.59) and dry weight (84.20 mg), pods plant-1 (41.90), seeds pod-1 (3.47), grain yield (1380 kg ha-1), haulm yield (1885 kg ha-1), biological yield (3325 kg ha-1), protein (42.25%) and oil content (19.92%), N, P, K content and their uptake, with maximum gross return (53042.71 Rs ha-1), net return (28980.71 Rs ha-1) and B:C ratio (2.20) in soybean crop. [ABSTRACT FROM AUTHOR]

Microvirga lotononidis is a recently described species of root-nodule bacteria that is an effective nitrogen- (N) fixing microsymbiont of the symbiotically specific African legume Listia angolensis (Welw. ex Bak.) B.-E. van Wyk & Boatwr. M. lotononidis possesses several properties that are unusual in root-nodule bacteria, including pigmentation and the ability to grow at temperatures of up to 45°C. Strain WSM3557 is an aerobic, motile, Gram-negative, non-spore-forming rod isolated from a L. angolensis root nodule collected in Chipata, Zambia in 1963. This is the first report of a complete genome sequence for the genus Microvirga. Here we describe the features of Microvirga lotononidis strain WSM3557, together with genome sequence information and annotation. The 7,082,538 high-quality-draft genome is arranged in 18 scaffolds of 104 contigs, contains 6,956 protein-coding genes and 84 RNA-only encoding genes, and is one of 20 rhizobial genomes sequenced as part of the DOE Joint Genome Institute 2010 Community Sequencing Program. [ABSTRACT FROM AUTHOR]

Rhizobial nitrogen-fixing bacteria are the main inhabitants of the root nodules of legume plants. Studying the bacterial community of legume nodules is important in understanding plant growth and nutrient requirements. Culture-based technique was used to examine the bacterial community of these underground organs from Vigna subterranea L. Verdc (Bambara nut), an underutilized legume in Africa, for plant growth-promoting traits. In this study, Bambara nuts were planted to trap root-nodule bacteria, and the bacteria were morphologically, biochemically, and molecularly characterized. Five selected isolates were screened in vitro for their plant growth-promoting traits and exhibited differences in their phenotypic traits. The polymerase chain reaction (PCR) products were subjected to partial 16S rRNA gene sequencing for phylogenetic analysis. Based on 16S rRNA gene sequence, the isolates were identified as BA1 (Stenotrophomonas maltophilia), BA2 (Chryseobacterium sp.), BA3 (Pseudomonas alcaligenes), BA4 (Pseudomonas plecoglossicida), and BA5 (Pseudomonas hibiscicola). Results showed that four of the five isolates could produce IAA. The capability to solubilize phosphate in Pikovskaya's agar plates was positively shown by four isolates (BA2, BA3, BA4, and BA5). Three isolates could produce hydrogen cyanide while isolates BA1, BA3, BA4, and BA5 were found to have ammonia-production traits. The results suggest that these plant growth-promoting isolates can be used as inoculants for plant growth and productivity. [ABSTRACT FROM AUTHOR]

This review evaluates oilseed crop soybean endophytic bacteria, their prospects, and challenges for sustainable agriculture. Soybean is one of the most important oilseed crops with about 20–25% protein content and 20% edible oil production. The ability of soybean root-associated microbes to restore soil nutrients enhances crop yield. Naturally, the soybean root endosphere harbors root nodule bacteria, and endophytic bacteria, which help increase the nitrogen pool and reclamation of another nutrient loss in the soil for plant nutrition. Endophytic bacteria can sustain plant growth and health by exhibiting antibiosis against phytopathogens, production of enzymes, phytohormone biosynthesis, organic acids, and secondary metabolite secretions. Considerable effort in the agricultural industry is focused on multifunctional concepts and bioprospecting on the use of bioinput from endophytic microbes to ensure a stable ecosystem. Bioprospecting in the case of this review is a systemic overview of the biorational approach to harness beneficial plant-associated microbes to ensure food security in the future. Progress in this endeavor is limited by available techniques. The use of molecular techniques in unraveling the functions of soybean endophytic bacteria can explore their use in integrated organic farming. Our review brings to light the endophytic microbial dynamics of soybeans and current status of plant microbiome research for sustainable agriculture. [ABSTRACT FROM AUTHOR]