RESUMEN
The bean yellow mosaic virus (BYMV) is one of the most serious economic diseases affecting faba bean crop production. Rhizobium spp., well known for its high nitrogen fixation capacity in legumes, has received little study as a possible biocontrol agent and antiviral. Under greenhouse conditions, foliar application of molecularly characterized Rhizobium leguminosarum bv. viciae strain 33504-Borg201 to the faba bean leaves 24 h before they were infected with BYMV made them much more resistant to the disease while also lowering its severity and accumulation. Furthermore, the treatment promoted plant growth and health, as evidenced by the increased total chlorophyll (32.75 mg/g f.wt.) and protein content (14.39 mg/g f.wt.), as well as the improved fresh and dry weights of the plants. The protective effects of 33504-Borg201 greatly lowered the levels of hydrogen peroxide (H2O2) (4.92 µmol/g f.wt.) and malondialdehyde (MDA) (173.72 µmol/g f.wt.). The antioxidant enzymes peroxidase (1.58 µM/g f.wt.) and polyphenol oxidase (0.57 µM/g f.wt.) inhibited the development of BYMV in plants treated with 33504-Borg201. Gene expression analysis showed that faba bean plants treated with 33504-Borg201 had higher amounts of pathogenesis-related protein-1 (PR-1) (3.28-fold) and hydroxycinnamoyl-CoA quinate hydroxycinnamoyltransferase (4.13-fold) than control plants. These findings demonstrate the potential of 33,504-Borg201 as a cost-effective and eco-friendly method to protect faba bean plants against BYMV. Implementing this approach could help develop a simple and sustainable strategy for protecting faba bean crops from the devastating effects of BYMV.
Asunto(s)
Enfermedades de las Plantas , Hojas de la Planta , Rhizobium leguminosarum , Vicia faba , Rhizobium leguminosarum/genética , Rhizobium leguminosarum/crecimiento & desarrollo , Rhizobium leguminosarum/fisiología , Vicia faba/virología , Vicia faba/microbiología , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/virología , Enfermedades de las Plantas/prevención & control , Hojas de la Planta/microbiología , Hojas de la Planta/virología , Resistencia a la Enfermedad , Peróxido de Hidrógeno/metabolismoRESUMEN
Bacterial persistence in the rhizosphere and colonization of root niches are critical for the establishment of many beneficial plant-bacteria interactions including those between Rhizobium leguminosarum and its host legumes. Despite this, most studies on R. leguminosarum have focused on its symbiotic lifestyle as an endosymbiont in root nodules. Here, we use random barcode transposon sequencing to assay gene contributions of R. leguminosarum during competitive growth in the rhizosphere and colonization of various plant species. This facilitated the identification of 189 genes commonly required for growth in diverse plant rhizospheres, mutation of 111 of which also affected subsequent root colonization (rhizosphere progressive), and a further 119 genes necessary for colonization. Common determinants reveal a need to synthesize essential compounds (amino acids, ribonucleotides, and cofactors), adapt metabolic function, respond to external stimuli, and withstand various stresses (such as changes in osmolarity). Additionally, chemotaxis and flagella-mediated motility are prerequisites for root colonization. Many genes showed plant-specific dependencies highlighting significant adaptation to different plant species. This work provides a greater understanding of factors promoting rhizosphere fitness and root colonization in plant-beneficial bacteria, facilitating their exploitation for agricultural benefit.
Asunto(s)
Raíces de Plantas , Rhizobium leguminosarum , Rizosfera , Simbiosis , Raíces de Plantas/microbiología , Rhizobium leguminosarum/genética , Rhizobium leguminosarum/crecimiento & desarrollo , Rhizobium leguminosarum/fisiología , Fabaceae/microbiología , Fabaceae/crecimiento & desarrollo , Microbiología del SueloRESUMEN
KEY MESSAGE: Accurate genomic prediction of yield within and across generations was achieved by estimating the genetic merit of individual white clover genotypes based on extensive genetic replication using cloned material. White clover is an agriculturally important forage legume grown throughout temperate regions as a mixed clover-grass crop. It is typically cultivated with low nitrogen input, making yield dependent on nitrogen fixation by rhizobia in root nodules. Here, we investigate the effects of clover and rhizobium genetic variation by monitoring plant growth and quantifying dry matter yield of 704 combinations of 145 clover genotypes and 170 rhizobium inocula. We find no significant effect of rhizobium variation. In contrast, we can predict yield based on a few white clover markers strongly associated with plant size prior to nitrogen fixation, and the prediction accuracy for polycross offspring yield is remarkably high. Several of the markers are located near a homolog of Arabidopsis thaliana GIGANTUS 1, which regulates growth rate and biomass accumulation. Our work provides fundamental insight into the genetics of white clover yield and identifies specific candidate genes as breeding targets.
Asunto(s)
Genes de Plantas , Fijación del Nitrógeno , Rhizobium leguminosarum/fisiología , Trifolium/genética , Variación Genética , Genotipo , Modelos Genéticos , Desarrollo de la Planta/genética , Rhizobium leguminosarum/clasificación , Rhizobium leguminosarum/aislamiento & purificación , Trifolium/crecimiento & desarrollo , Trifolium/metabolismo , Trifolium/microbiologíaRESUMEN
Rhizobia - nitrogen-fixing, root-nodulating bacteria - play a critical role in both plant ecosystems and sustainable agriculture. Rhizobia form intracellular infections within legumes roots where they produce plant accessible nitrogen from atmospheric nitrogen and thus reduce the reliance on industrial inputs. The rhizobia-legume symbiosis is often treated as a pairwise relationship between single genotypes, both in research and in the production of rhizobial inoculants. However in nature individual plants are infected by a high diversity of rhizobia symbionts. How this diversity affects productivity within the symbiosis is unclear. Here, we use a powerful statistical approach to assess the impact of diversity within the Rhizobium leguminosarum - clover symbiosis using a biodiversity-ecosystem function framework. Statistically, we found no significant impact of rhizobium diversity. However this relationship was weakly positive - rather than negative - indicating that there is no significant cost to increasing inoculant diversity. Productivity was influenced by the identity of the strains within an inoculant; strains with the highest individual performance showed a significant positive contribution within mixed inoculants. Overall, inoculant effectiveness was best predicted by the individual performance of the best inoculant member, and only weakly predicted by the worst performing member. Collectively, our data suggest that the Rhizobium leguminosarum - clover symbiosis displays a weak diversity-function relationship, but that inoculant performance can be improved through the inclusion of high performing strains. Given the wide environmental dependence of rhizobial inoculant quality, multi-strain inoculants could be highly successful as they increase the likelihood of including a strain well adapted to local conditions across different environments.
Asunto(s)
Medicago/microbiología , Rhizobium leguminosarum/fisiología , Simbiosis , Ecosistema , Interacciones Microbiota-Huesped , Medicago/crecimiento & desarrollo , Medicago/fisiología , Rhizobium leguminosarum/clasificación , Rhizobium leguminosarum/genéticaRESUMEN
Heavy metals polluting the 100-year-old waste heap in Boleslaw (Poland) are acting as a natural selection factor and may contribute to adaptations of organisms living in this area, including Trifolium repens and its root nodule microsymbionts-rhizobia. Exopolysaccharides (EPS), exuded extracellularly and associated with bacterial cell walls, possess variable structures depending on environmental conditions; they can bind metals and are involved in biofilm formation. In order to examine the effects of long-term exposure to metal pollution on EPS structure and biofilm formation of rhizobia, Rhizobium leguminosarum bv. trifolii strains originating from the waste heap area and a non-polluted reference site were investigated for the characteristics of the sugar fraction of their EPS using gas chromatography mass-spectrometry and also for biofilm formation and structural characteristics using confocal laser scanning microscopy under control conditions as well as when exposed to toxic concentrations of zinc, lead, and cadmium. Significant differences in EPS structure, biofilm thickness, and ratio of living/dead bacteria in the biofilm were found between strains originating from the waste heap and from the reference site, both without exposure to metals and under metal exposure. Received results indicate that studied rhizobia can be assumed as potentially useful in remediation processes.
Asunto(s)
Biopelículas , Metales Pesados/metabolismo , Polisacáridos Bacterianos/metabolismo , Rhizobium leguminosarum/fisiología , Contaminantes del Suelo/metabolismo , Trifolium/microbiología , Biopelículas/efectos de los fármacos , Biopelículas/crecimiento & desarrolloRESUMEN
Peat is the standard carrier material used for commercial microbial inoculants produced in Canada and the United States. Peat is a slowly renewable resource and its production is extremely vulnerable to variable weather conditions. Furthermore, it may not be widely available in all countries. We investigated the potential to develop biochar as a carrier material. Our goal was to evaluate if different biochars perform comparably in supporting rhizobial survival, and what characteristics contribute to their ability to support rhizobial survival. Evaluation included characterization of the biochars, assessment of biochar phytotoxicity, survival of Rhizobium on biochars, and growth chamber evaluation of two biochars as Rhizobium carriers for inoculating pea. Of the original nine biochars evaluated, six supported Rhizobium leguminosarum for 84 days at 4 °C; of this six, two supported numbers >1 × 106 cfu·(g biochar)-1. The only characteristics that correlated with survival were C/N ratio and percent C. The two biochars evaluated delivered R. leguminosarum to pea that initiated nodulation, biomass production, and biomass N at levels higher than a noninoculated control and heat-killed inoculated biochars. We demonstrate that there is considerable potential to develop biochar as a carrier for rhizobial inoculants.
Asunto(s)
Inoculantes Agrícolas/fisiología , Carbón Orgánico/química , Rhizobium leguminosarum/fisiología , Biomasa , Canadá , Viabilidad Microbiana , Pisum sativum/crecimiento & desarrollo , Pisum sativum/microbiología , Nodulación de la Raíz de la Planta , Suelo/química , Microbiología del SueloRESUMEN
BACKGROUND AND OBJECTIVE: Using the foliar application in development agriculture became a new strategy in the production system to decrease the cost of nutrition. This study amid to investigate the effect of yeast and algae on yield and quality of soybean under fertigation in newly soils at Wadi El-Natroun, El-Behaira Governorate, Egypt, during the two seasons 2018 and 2019. MATERIALS AND METHODS: The experimental design was a split-plot design in a randomized complete block arrangement with three replications. Yeast and algae were allocated to the main plots, while rates treatments were distributed at random in the sub-plots and control treatment foliar water. RESULTS: The results showed that either the foliar application with algae and yeast stimulates many growth aspects individually, such as plant length, number of seeds/plant and seed weight/plant compared to the control treatment. In addition, foliar spraying with algae at 8 g L-1 increased NPK content, crude protein content and total carbohydrate in seed. CONCLUSION: This study concluded that the foliar application influences at algae have provided high yields and chemical constituents under fertigation in sandy soil.
Asunto(s)
Cianobacterias/fisiología , Fertilizantes , Glycine max/microbiología , Hojas de la Planta/microbiología , Semillas/microbiología , Microbiología del Suelo , Levaduras/fisiología , Aerosoles , Egipto , Hojas de la Planta/crecimiento & desarrollo , Rhizobium leguminosarum/fisiología , Semillas/crecimiento & desarrollo , Glycine max/crecimiento & desarrolloRESUMEN
Rhizobium leguminosarum bv. trifolii produces exopolysaccharide (EPS) composed of glucose, glucuronic acid, and galactose residues at a molar ratio 5:2:1. A majority of genes involved in the synthesis, modification, and export of exopolysaccharide are located in the chromosomal Pss-I region. In the present study, a ΔpssJ deletion mutant was constructed and shown to produce EPS lacking terminal galactose in the side chain of the octasaccharide subunit. The lack of galactose did not block EPS subunit translocation and polymerization. The in trans delivery of the pssJ gene restored the production of galactose-containing exopolysaccharide. The mutant was compromised in several physiological traits, e.g., motility and biofilm production. An impact of the pssJ mutation and changed EPS structure on the symbiotic performance was observed as improper signaling at the stage of molecular recognition, leading to formation of a significant number of non-infected empty nodules. Terminal galactosyltransferase PssJ was shown to display a structure typical for the GT-A class of glycosyltransferases and interact with other GTs and Wzx/Wzy system proteins. The latter, together with PssJ presence in soluble and membrane protein fractions indicated that the protein plays its role at the inner membrane interface and as a component of a larger complex.
Asunto(s)
Proteínas Bacterianas/genética , Galactosiltransferasas/genética , Mutación , Polisacáridos Bacterianos/metabolismo , Rhizobium leguminosarum/genética , Proteínas Bacterianas/metabolismo , Biopelículas , Galactosa/química , Galactosa/metabolismo , Galactosiltransferasas/metabolismo , Interacciones Huésped-Patógeno , Nodulación de la Raíz de la Planta/genética , Polisacáridos Bacterianos/química , Rhizobium leguminosarum/enzimología , Rhizobium leguminosarum/fisiología , Nódulos de las Raíces de las Plantas/genética , Nódulos de las Raíces de las Plantas/microbiología , Simbiosis/genética , Trifolium/microbiologíaRESUMEN
By analyzing successive lifestyle stages of a model Rhizobium-legume symbiosis using mariner-based transposon insertion sequencing (INSeq), we have defined the genes required for rhizosphere growth, root colonization, bacterial infection, N2-fixing bacteroids, and release from legume (pea) nodules. While only 27 genes are annotated as nif and fix in Rhizobium leguminosarum, we show 603 genetic regions (593 genes, 5 transfer RNAs, and 5 RNA features) are required for the competitive ability to nodulate pea and fix N2 Of these, 146 are common to rhizosphere growth through to bacteroids. This large number of genes, defined as rhizosphere-progressive, highlights how critical successful competition in the rhizosphere is to subsequent infection and nodulation. As expected, there is also a large group (211) specific for nodule bacteria and bacteroid function. Nodule infection and bacteroid formation require genes for motility, cell envelope restructuring, nodulation signaling, N2 fixation, and metabolic adaptation. Metabolic adaptation includes urea, erythritol and aldehyde metabolism, glycogen synthesis, dicarboxylate metabolism, and glutamine synthesis (GlnII). There are 17 separate lifestyle adaptations specific to rhizosphere growth and 23 to root colonization, distinct from infection and nodule formation. These results dramatically highlight the importance of competition at multiple stages of a Rhizobium-legume symbiosis.
Asunto(s)
Rhizobium leguminosarum , Rizosfera , Simbiosis/genética , Fabaceae/microbiología , Genes Bacterianos/genética , Fijación del Nitrógeno/genética , Rhizobium leguminosarum/genética , Rhizobium leguminosarum/fisiología , Nódulos de las Raíces de las Plantas/genética , Nódulos de las Raíces de las Plantas/microbiologíaRESUMEN
Establishment of the symbiotic relationship that develops between rhizobia and their legume hosts is contingent upon an interkingdom signal exchange. In response to host legume flavonoids, NodD proteins from compatible rhizobia activate expression of nodulation genes that produce lipochitin oligosaccharide signaling molecules known as Nod factors. Root nodule formation commences upon legume recognition of compatible Nod factor. Rhizobium leguminosarum was previously considered to contain one copy of nodD; here, we show that some strains of the Trifolium (clover) microsymbiont R. leguminosarum bv. trifolii contain a second copy designated nodD2. nodD2 genes were present in 8 out of 13 strains of R. leguminosarum bv. trifolii, but were absent from the genomes of 16 R. leguminosarum bv. viciae strains. Analysis of single and double nodD1 and nodD2 mutants in R. leguminosarum bv. trifolii strain TA1 revealed that NodD2 was functional and enhanced nodule colonization competitiveness. However, NodD1 showed significantly greater capacity to induce nod gene expression and infection thread formation. Clover species are either annual or perennial and this phenological distinction is rarely crossed by individual R. leguminosarum bv. trifolii microsymbionts for effective symbiosis. Of 13 strains with genome sequences available, 7 of the 8 effective microsymbionts of perennial hosts contained nodD2, whereas the 3 microsymbionts of annual hosts did not. We hypothesize that NodD2 inducer recognition differs from NodD1, and NodD2 functions to enhance competition and effective symbiosis, which may discriminate in favor of perennial hosts.IMPORTANCE Establishment of the rhizobium-legume symbiosis requires a highly specific and complex signal exchange between both participants. Rhizobia perceive legume flavonoid compounds through LysR-type NodD regulators. Often, rhizobia encode multiple copies of nodD, which is one determinant of host specificity. In some species of rhizobia, the presence of multiple copies of NodD extends their symbiotic host-range. Here, we identified and characterized a second copy of nodD present in some strains of the clover microsymbiont Rhizobium leguminosarum bv. trifolii. The second nodD gene contributed to the competitive ability of the strain on white clover, an important forage legume. A screen for strains containing nodD2 could be utilized as one criterion to select strains with enhanced competitive ability for use as inoculants for pasture production.
Asunto(s)
Proteínas Bacterianas/genética , Interacciones Microbianas , Nodulación de la Raíz de la Planta , Rhizobium leguminosarum/fisiología , Trifolium/microbiología , Proteínas Bacterianas/metabolismo , Raíces de Plantas/microbiologíaRESUMEN
Lotus species develop infection threads to guide rhizobia into nodule cells. However, there is evidence that some species have a genetic repertoire to allow other modes of infection. By conducting confocal and electron microscopy, quantification of marker gene expression, and phenotypic analysis of transgenic roots infected with mutant rhizobia, we elucidated the infection mechanism used by Rhizobium leguminosarum Norway to colonize Lotus burttii. Rhizobium leguminosarum Norway induces a distinct host transcriptional response compared with Mesorhizobium loti. It infects L. burttii utilizing an epidermal and transcellular infection thread-independent mechanism at high frequency. The entry into plant cells occurs directly from the apoplast and is primarily mediated by 'peg'-like structures, the formation of which is dependent on the production of Nod factor by the rhizobia. These results demonstrate that Lotus species can exhibit duality in their infection mechanisms depending on the rhizobial strain that they encounter. This is especially relevant in the context of interactions in the rhizosphere where legumes do not encounter single strains, but complex rhizobial communities. Additionally, our findings support a perception mechanism at the nodule cell entry interface, reinforcing the idea that there are successive checkpoints during rhizobial infection.
Asunto(s)
Lotus/microbiología , Lotus/fisiología , Nodulación de la Raíz de la Planta , Rhizobium leguminosarum/fisiología , Nódulos de las Raíces de las Plantas/microbiología , Nódulos de las Raíces de las Plantas/fisiología , SimbiosisRESUMEN
Coexistence requires that stabilizing niche differences, which cause species to limit themselves more than others, outweigh relative fitness differences, which cause competitive exclusion. Interactions with shared mutualists, which can differentially affect host fitness and change in magnitude with host frequency, can satisfy these conditions for coexistence, yet empirical tests of mutualist effects on relative fitness and stabilizing niche differences are largely lacking within the framework of coexistence theory. Here, we show that N-fixing rhizobial mutualists mediate coexistence in four naturally co-occurring congeneric legume (Trifolium) species. Using experimental greenhouse communities, we quantified relative fitness and stabilizing niche differences for each species in the presence of rhizobia originating from conspecific or congeneric hosts. Rhizobia stabilized coexistence by increasing the self-limitation of Trifolium species grown with rhizobia isolated from conspecifics, thus allowing congeners to increase when rare. Greenhouse-measured invasion growth rates predicted natural, unmanipulated coexistence dynamics of Trifolium species over 2 years at our field sites. Our results demonstrate that interactions with shared mutualists can stabilize the coexistence of closely related species.
Asunto(s)
Nodulación de la Raíz de la Planta , Rhizobium leguminosarum/fisiología , Trifolium/microbiología , Especificidad de la Especie , Simbiosis , Trifolium/crecimiento & desarrolloRESUMEN
Despite their ubiquitous distribution and significant ecological roles, soil microorganisms have long been neglected in investigations addressing parasitic plant-host interactions. Because nutrient deprivation is a primary cause of host damage by parasitic plants, we hypothesized that beneficial soil microorganisms conferring nutrient benefits to parasitized hosts may play important roles in alleviating damage. We conducted a pot cultivation experiment to test the inoculation effect of an arbuscular mycorrhizal fungus (Glomus mosseae), a rhizobium (Rhizobium leguminosarum) and their interactive effects, on alleviation of damage to a legume host (Trifolium repens) by two root hemiparasitic plants with different nutrient requirements (N-demanding Pedicularis rex and P-demanding P. tricolor). Strong interactive effects between inoculation regimes and hemiparasite identity were observed. The relative benefits of microbial inoculation were related to hemiparasite nutrient requirements. Dual inoculation with the rhizobium strongly enhanced promotional arbuscular mycorrhizal effects on hosts parasitized by P. rex, but reduced the arbuscular mycorrhizal promotion on hosts parasitized by P. tricolor. Our results demonstrate substantial contribution of arbuscular mycorrhizal and rhizobial symbioses to alleviating damage to the legume host by root hemiparasites, and suggest that soil microorganisms are critical factors regulating host-parasite interactions and should be taken into account in future studies.
Asunto(s)
Interacciones Huésped-Parásitos/fisiología , Micorrizas/fisiología , Pedicularis/fisiología , Rhizobium leguminosarum/fisiología , Trifolium/microbiología , Trifolium/parasitología , Inoculantes Agrícolas , Glomeromycota/fisiología , Nitrógeno/metabolismo , Fósforo/metabolismo , Nodulación de la Raíz de la Planta/fisiología , Raíces de Plantas/microbiología , Raíces de Plantas/parasitología , Brotes de la Planta/química , Brotes de la Planta/metabolismo , Simbiosis/fisiología , Trifolium/fisiologíaRESUMEN
MAIN CONCLUSION: The LysM receptor-like kinase K1 is involved in regulation of pea-rhizobial symbiosis development. The ability of the crop legume Pisum sativum L. to perceive the Nod factor rhizobial signals may depend on several receptors that differ in ligand structure specificity. Identification of pea mutants defective in two types of LysM receptor-like kinases (LysM-RLKs), SYM10 and SYM37, featuring different phenotypic manifestations and impaired at various stages of symbiosis development, corresponds well to this assumption. There is evidence that one of the receptor proteins involved in symbiosis initiation, SYM10, has an inactive kinase domain. This implies the presence of an additional component in the receptor complex, together with SYM10, that remains unknown. Here, we describe a new LysM-RLK, K1, which may serve as an additional component of the receptor complex in pea. To verify the function of K1 in symbiosis, several P. sativum non-nodulating mutants in the k1 gene were identified using the TILLING approach. Phenotyping revealed the blocking of symbiosis development at an appropriately early stage, strongly suggesting the importance of LysM-RLK K1 for symbiosis initiation. Moreover, the analysis of pea mutants with weaker phenotypes provides evidence for the additional role of K1 in infection thread distribution in the cortex and rhizobia penetration. The interaction between K1 and SYM10 was detected using transient leaf expression in Nicotiana benthamiana and in the yeast two-hybrid system. Since the possibility of SYM10/SYM37 complex formation was also shown, we tested whether the SYM37 and K1 receptors are functionally interchangeable using a complementation test. The interaction between K1 and other receptors is discussed.
Asunto(s)
Pisum sativum/enzimología , Proteínas de Plantas/fisiología , Proteínas Quinasas/fisiología , Rhizobium leguminosarum/fisiología , Simbiosis , Western Blotting , Ingeniería Genética/métodos , Pisum sativum/microbiología , Pisum sativum/fisiología , Hojas de la Planta/enzimología , Hojas de la Planta/metabolismo , Plantas Modificadas Genéticamente , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Nicotiana/genética , Técnicas del Sistema de Dos HíbridosRESUMEN
Biological nitrogen fixation through the legume-rhizobia symbiosis is important for sustainable pastoral production. In New Zealand, the most widespread and valuable symbiosis occurs between white clover (Trifolium repens L.) and Rhizobium leguminosarum bv. trifolii (Rlt). As variation in the population size (determined by most probable number assays; MPN) and effectiveness of N-fixation (symbiotic potential; SP) of Rlt in soils may affect white clover performance, the extent in variation in these properties was examined at three different spatial scales: (1) From 26 sites across New Zealand, (2) at farm-wide scale, and (3) within single fields. Overall, Rlt populations ranged from 95 to >1 x 108 per g soil, with variation similar at the three spatial scales assessed. For almost all samples, there was no relationship between rhizobia population size and ability of the population to fix N during legume symbiosis (SP). When compared with the commercial inoculant strain, the SP of soils ranged between 14 to 143% efficacy. The N-fixing ability of rhizobia populations varied more between samples collected from within a single hill country field (0.8 ha) than between 26 samples collected from diverse locations across New Zealand. Correlations between SP and calcium and aluminium content were found in all sites, except within a dairy farm field. Given the general lack of association between SP and MPN, and high spatial variability of SP at single field scale, provision of advice for treating legume seed with rhizobia based on field-average MPN counts needs to be carefully considered.
Asunto(s)
Medicago/microbiología , Rhizobium leguminosarum/fisiología , Simbiosis , Nueva Zelanda , Microbiología del SueloRESUMEN
Biological nitrogen fixation (BNF) can be improved by optimizing the interaction between the rhizobial inoculant and pea (Pisum sativum L.), leading to increased productivity and reduced nitrogen (N) fertilizer use. Eight Rhizobium leguminosarum bv. viciae strains were used to inoculate the super-nodulating pea mutant Rondo-nod3 (fix+), the hyper-nodulating pea mutant Frisson P88 Sym29, CDC Meadow commercial control, and the non-nodulating mutant Frisson P56 (nod-) to evaluate BNF in a greenhouse assay. Significant differences in strain × cultivar interactions were detected for shoot and root dry masses, which ranged from 1.8 to 4.7 g and from 0.27 to 0.73 g per plant, respectively; for nodule number on lateral roots, which ranged from 25 to 430 per plant; for amount of fixed N2, which ranged from 15 to 67 mg and from 4 to 15 mg per plant for shoot and root tissues, respectively; and for percentage of N derived from atmosphere (%Ndfa), which ranged from 37% to 61% and from 35% to 65% for shoot and root tissue, respectively. Strain × cultivar interactions in this study could contribute to identification of superior strains and pea breeding lines with genetic superiority in BNF. Nodule production in pea plants was not necessarily correlated with the amount of fixed N2, suggesting nodule activity is more important to BNF than is nodule number.
Asunto(s)
Fijación del Nitrógeno/fisiología , Pisum sativum/microbiología , Rhizobium leguminosarum/fisiología , Simbiosis/fisiología , Metabolismo de los Hidratos de Carbono , Genotipo , Nitrógeno/metabolismo , Raíces de PlantasRESUMEN
Trifolium rubens L., commonly known as the red feather clover, is capable of symbiotic interactions with rhizobia. Up to now, no specific symbionts of T. rubens and their symbiotic compatibility with Trifolium spp. have been described. We characterized the genomic diversity of T. rubens symbionts by analyses of plasmid profiles and BOX-PCR. The phylogeny of T. rubens isolates was inferred based on the nucleotide sequences of 16S rRNA and two core genes (atpD, recA). The nodC phylogeny allowed classification of rhizobia nodulating T. rubens as Rhizobium leguminosarum symbiovar trifolii (Rlt). The symbiotic efficiency of the Rlt isolates was determined on four clover species: T. rubens, T. pratense, T. repens and T. resupinatum. We determined that Rlt strains formed mostly inefficient symbiosis with their native host plant T. rubens and weakly effective (sub-optimal) symbiosis with T. repens and T. pratense. The same Rlt strains were fully compatible in the symbiosis with T. resupinatum. T. rubens did not exhibit strict selectivity in regard to the symbionts and rhizobia closely related to Rhizobium grahamii, Rhizobium galegae and Agrobacterium radiobacter, which did not nodulate Trifolium spp., were found amongst T. rubens nodule isolates.
Asunto(s)
Rhizobium leguminosarum/clasificación , Rhizobium leguminosarum/fisiología , Nódulos de las Raíces de las Plantas/microbiología , Simbiosis , Trifolium/microbiología , Genes Bacterianos , Variación Genética , Genoma Bacteriano , Tipificación de Secuencias Multilocus , Filogenia , Plásmidos/genética , Rhizobium leguminosarum/aislamiento & purificaciónRESUMEN
In legume-Rhizobium symbioses, specialised soil bacteria fix atmospheric nitrogen in return for carbon. However, ineffective strains can arise, making discrimination essential. Discrimination can occur via partner choice, where legumes prevent ineffective strains from entering, or via sanctioning, where plants provide fewer resources. Several studies have inferred that legumes exercise partner choice, but the rhizobia compared were not otherwise isogenic. To test when and how plants discriminate ineffective strains we developed sets of fixing and non-fixing strains that differed only in the expression of nifH - essential for nitrogen fixation - and could be visualised using marker genes. We show that the plant is unable to select against the non-fixing strain at the point of entry, but that non-fixing nodules are sanctioned. We also used the technique to characterise mixed nodules (containing both a fixing and a non-fixing strain), whose frequency could be predicted using a simple diffusion model. We discuss that sanctioning is likely to evolve in preference to partner choice in any symbiosis where partner quality cannot be adequately assessed until goods or services are actively exchanged.
Asunto(s)
Pisum sativum/microbiología , Rhizobium leguminosarum/fisiología , Simbiosis , Fijación del Nitrógeno , Pisum sativum/fisiología , Nódulos de las Raíces de las Plantas/microbiología , Nódulos de las Raíces de las Plantas/fisiologíaRESUMEN
Plants engineer the rhizosphere to their advantage by secreting various nutrients and secondary metabolites. Coupling transcriptomic and metabolomic analyses of the pea (Pisum sativum) rhizosphere, a suite of bioreporters has been developed in Rhizobium leguminosarum bv viciae strain 3841, and these detect metabolites secreted by roots in space and time. Fourteen bacterial lux fusion bioreporters, specific for sugars, polyols, amino acids, organic acids, or flavonoids, have been validated in vitro and in vivo. Using different bacterial mutants (nodC and nifH), the process of colonization and symbiosis has been analyzed, revealing compounds important in the different steps of the rhizobium-legume association. Dicarboxylates and sucrose are the main carbon sources within the nodules; in ineffective (nifH) nodules, particularly low levels of sucrose were observed, suggesting that plant sanctions affect carbon supply to nodules. In contrast, high myo-inositol levels were observed prior to nodule formation and also in nifH senescent nodules. Amino acid biosensors showed different patterns: a γ-aminobutyrate biosensor was active only inside nodules, whereas the phenylalanine bioreporter showed a high signal also in the rhizosphere. The bioreporters were further validated in vetch (Vicia hirsuta), producing similar results. In addition, vetch exhibited a local increase of nod gene-inducing flavonoids at sites where nodules developed subsequently. These bioreporters will be particularly helpful in understanding the dynamics of root exudation and the role of different molecules secreted into the rhizosphere.
Asunto(s)
Técnicas Biosensibles , Pisum sativum/metabolismo , Exudados de Plantas/metabolismo , Raíces de Plantas/metabolismo , Rhizobium leguminosarum/fisiología , Recuento de Colonia Microbiana , Regulación de la Expresión Génica de las Plantas , Hesperidina/análisis , Procesamiento de Imagen Asistido por Computador , Luminiscencia , Metaboloma , Fijación del Nitrógeno , Pisum sativum/genética , Pisum sativum/microbiología , Nodulación de la Raíz de la Planta , Raíces de Plantas/genética , Raíces de Plantas/microbiología , Rhizobium leguminosarum/crecimiento & desarrollo , Rizosfera , Nódulos de las Raíces de las Plantas/microbiología , Simbiosis , Factores de Tiempo , Vicia/microbiologíaRESUMEN
A change in the contents of endogenous salicylic and jasmonic acids in the roots of the host plant at the preinfectious stage of interaction with symbiotic (Rhizobium leguminosarum) and pathogenic (Agrobacterium rizogenes) bacteria belonging for to the family Rhizobiaceae was studied. It was found that the jasmonic acid content increased 1.52 times 5 min after inoculation with these bacterial species. It was shown that dynamics of the change in the JA and SA contents depends on the type of infection. Thus, the JA content decreased in the case of pathogenesis, while the SA content increased. At the same time, an increased JA content was observed during symbiosis. The observed regularities could indicate the presence of different strategies of hormonal regulation for interaction with symbiotic and pathogenic bacteria belonging to the family Rhizobiaceae in peas plants.