RESUMO
BACKGROUND: Soybean establishes a mutualistic interaction with nitrogen-fixing rhizobacteria, acquiring most of its nitrogen requirements through symbiotic nitrogen fixation. This crop is susceptible to water deficit; evidence suggests that its nodulation status-whether it is nodulated or not-can influence how it responds to water deficit. The translational control step of gene expression has proven relevant in plants subjected to water deficit. RESULTS: Here, we analyzed soybean roots' differential responses to water deficit at transcriptional, translational, and mixed (transcriptional + translational) levels. Thus, the transcriptome and translatome of four combined-treated soybean roots were analyzed. We found hormone metabolism-related genes among the differentially expressed genes (DEGs) at the translatome level in nodulated and water-restricted plants. Also, weighted gene co-expression network analysis followed by differential expression analysis identified gene modules associated with nodulation and water deficit conditions. Protein-protein interaction network analysis was performed for subsets of mixed DEGs of the modules associated with the plant responses to nodulation, water deficit, or their combination. CONCLUSIONS: Our research reveals that the stand-out processes and pathways in the before-mentioned plant responses partially differ; terms related to glutathione metabolism and hormone signal transduction (2 C protein phosphatases) were associated with the response to water deficit, terms related to transmembrane transport, response to abscisic acid, pigment metabolic process were associated with the response to nodulation plus water deficit. Still, two processes were common: galactose metabolism and branched-chain amino acid catabolism. A comprehensive analysis of these processes could lead to identifying new sources of tolerance to drought in soybean.
Assuntos
Glycine max , Raízes de Plantas , Transcriptoma , Glycine max/genética , Glycine max/fisiologia , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , Regulação da Expressão Gênica de Plantas , Nodulação/genética , Redes Reguladoras de Genes , Perfilação da Expressão Gênica , DesidrataçãoRESUMO
The actinobacterium Arthrobacter sp. UMCV2 promotes plant growth through the emission of N,N-dimethylhexadecilamine (DMHDA). The Medicago-Sinorhizobium nodulation has been employed to study symbiotic nitrogen fixation by rhizobia in nodulating Fabaceae. Herein, we isolated three Sinorhizobium medicae strains that were used to induce nodules in Medicago truncatula. The co-inoculation of M. truncatula with Arthrobacter sp. strain UMCV2 produced a higher number of effective nodules than inoculation with only Sinorhizobium strains. Similarly, the exposure of inoculated M. truncatula to DMHDA produced a greater number of effective nodules compared to non-exposed plants. Thus, we conclude that Arthrobacter sp. UMCV2 promotes nodulation, and propose that this effect is produced, at least partly, via DMHDA emission.
Assuntos
Arthrobacter , Medicago truncatula , Medicago truncatula/microbiologia , Arthrobacter/efeitos dos fármacos , Arthrobacter/fisiologia , Sinorhizobium/fisiologia , Sinorhizobium/efeitos dos fármacos , Nodulação/efeitos dos fármacos , Simbiose , Fixação de Nitrogênio/efeitos dos fármacosRESUMO
Legume plants develop two types of root postembryonic organs, lateral roots and symbiotic nodules, using shared regulatory components. The module composed by the microRNA390, the Trans-Acting SIRNA3 (TAS3) RNA and the Auxin Response Factors (ARF)2, ARF3, and ARF4 (miR390/TAS3/ARFs) mediates the control of both lateral roots and symbiotic nodules in legumes. Here, a transcriptomic approach identified a member of the Lateral Organ Boundaries Domain (LBD) family of transcription factors in Medicago truncatula, designated MtLBD17/29a, which is regulated by the miR390/TAS3/ARFs module. ChIP-PCR experiments evidenced that MtARF2 binds to an Auxin Response Element present in the MtLBD17/29a promoter. MtLBD17/29a is expressed in root meristems, lateral root primordia, and noninfected cells of symbiotic nodules. Knockdown of MtLBD17/29a reduced the length of primary and lateral roots and enhanced lateral root formation, whereas overexpression of MtLBD17/29a produced the opposite phenotype. Interestingly, both knockdown and overexpression of MtLBD17/29a reduced nodule number and infection events and impaired the induction of the symbiotic genes Nodulation Signaling Pathway (NSP) 1 and 2. Our results demonstrate that MtLBD17/29a is regulated by the miR390/TAS3/ARFs module and a direct target of MtARF2, revealing a new lateral root regulatory hub recruited by legumes to act in the root nodule symbiotic program.
Assuntos
Medicago truncatula , Proteínas de Plantas , Nodulação , Raízes de Plantas , Fatores de Transcrição , Regulação da Expressão Gênica de Plantas , Técnicas de Silenciamento de Genes , Ácidos Indolacéticos/metabolismo , Medicago truncatula/genética , Medicago truncatula/crescimento & desenvolvimento , Medicago truncatula/microbiologia , MicroRNAs/genética , MicroRNAs/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Nodulação/genética , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Regiões Promotoras Genéticas/genética , Nódulos Radiculares de Plantas/genética , Nódulos Radiculares de Plantas/crescimento & desenvolvimento , Simbiose/genética , Fatores de Transcrição/metabolismo , Fatores de Transcrição/genéticaRESUMO
All non-Mimosoid nodulated genera in the legume subfamily Caesalpinioideae confine their rhizobial symbionts within cell wall-bound 'fixation threads' (FTs). The exception is the large genus Chamaecrista in which shrubs and subshrubs house their rhizobial bacteroids more intimately within symbiosomes, whereas large trees have FTs. This study aimed to unravel the evolutionary relationships between Chamaecrista growth habit, habitat, nodule bacteroid type, and rhizobial genotype. The growth habit, bacteroid anatomy, and rhizobial symbionts of 30 nodulated Chamaecrista species native to different biomes in the Brazilian state of Bahia, a major centre of diversity for the genus, was plotted onto an ITS-trnL-F-derived phylogeny of Chamaecrista. The bacteroids from most of the Chamaecrista species examined were enclosed in symbiosomes (SYM-type nodules), but those in arborescent species in the section Apoucouita, at the base of the genus, were enclosed in cell wall material containing homogalacturonan (HG) and cellulose (FT-type nodules). Most symbionts were Bradyrhizobium genotypes grouped according to the growth habits of their hosts, but the tree, C. eitenorum, was nodulated by Paraburkholderia. Chamaecrista has a range of growth habits that allow it to occupy several different biomes and to co-evolve with a wide range of (mainly) bradyrhizobial symbionts. FTs represent a less intimate symbiosis linked with nodulation losses, so the evolution of SYM-type nodules by most Chamaecrista species may have (i) aided the genus-wide retention of nodulation, and (ii) assisted in its rapid speciation and radiation out of the rainforest into more diverse and challenging habitats.
Assuntos
Chamaecrista , Filogenia , Floresta Úmida , Simbiose , Chamaecrista/fisiologia , Chamaecrista/genética , Chamaecrista/crescimento & desenvolvimento , Brasil , Ecossistema , Rhizobium/fisiologia , Nodulação/fisiologia , Evolução Biológica , Fixação de NitrogênioRESUMO
Rhizobial phosphatidylcholine (PC) is thought to be a critical phospholipid for the symbiotic relationship between rhizobia and legume host plants. A PC-deficient mutant of Sinorhizobium meliloti overproduces succinoglycan, is unable to swim, and lacks the ability to form nodules on alfalfa (Medicago sativa) host roots. Suppressor mutants had been obtained which did not overproduce succinoglycan and regained the ability to swim. Previously, we showed that point mutations leading to altered ExoS proteins can reverse the succinoglycan and swimming phenotypes of a PC-deficient mutant. Here, we report that other point mutations leading to altered ExoS, ChvI, FabA, or RpoH1 proteins also revert the succinoglycan and swimming phenotypes of PC-deficient mutants. Notably, the suppressor mutants also restore the ability to form nodule organs on alfalfa roots. However, nodules generated by these suppressor mutants express only low levels of an early nodulin, do not induce leghemoglobin transcript accumulation, thus remain white, and are unable to fix nitrogen. Among these suppressor mutants, we detected a reduced function mutant of the 3-hydoxydecanoyl-acyl carrier protein dehydratase FabA that produces reduced amounts of unsaturated and increased amounts of shorter chain fatty acids. This alteration of fatty acid composition probably affects lipid packing thereby partially compensating for the previous loss of PC and contributing to the restoration of membrane homeostasis.
Assuntos
Ácidos Graxos , Medicago sativa , Fosfatidilcolinas , Nodulação , Sinorhizobium meliloti , Simbiose , Sinorhizobium meliloti/fisiologia , Sinorhizobium meliloti/genética , Medicago sativa/microbiologia , Medicago sativa/genética , Nodulação/genética , Ácidos Graxos/metabolismo , Ácidos Graxos/biossíntese , Fosfatidilcolinas/metabolismo , Fosfatidilcolinas/biossíntese , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Nódulos Radiculares de Plantas/microbiologia , Nódulos Radiculares de Plantas/genética , Nódulos Radiculares de Plantas/metabolismo , Mutação , Polissacarídeos Bacterianos/metabolismo , Polissacarídeos Bacterianos/biossíntese , Fixação de NitrogênioAssuntos
Medicago truncatula , Rhizobium , Sinorhizobium meliloti , Rhizobium/fisiologia , Nodulação , Proteínas de Plantas/metabolismo , Medicago truncatula/metabolismo , Simbiose , Nódulos Radiculares de Plantas/metabolismo , Regulação da Expressão Gênica de Plantas , Sinorhizobium meliloti/fisiologia , Fixação de NitrogênioRESUMO
Legumes associate with Gram-negative soil bacteria called rhizobia, resulting in the formation of a nitrogen-fixing organ, the nodule. Nodules are an important sink for photosynthates for legumes, so these plants have developed a systemic regulation mechanism that controls their optimal number of nodules, the so-called autoregulation of nodulation (AON) pathway, to balance energy costs with the benefits of nitrogen fixation. In addition, soil nitrate inhibits nodulation in a dose-dependent manner, through systemic and local mechanisms. The CLE family of peptides and their receptors are key to tightly controlling these inhibitory responses. In the present study, a functional analysis revealed that PvFER1, PvRALF1, and PvRALF6 act as positive regulators of the nodule number in growth medium containing 0 mM of nitrate but as negative regulators in medium with 2 and 5 mM of nitrate. Furthermore, the effect on nodule number was found to be consistent with changes in the expression levels of genes associated with the AON pathway and with the nitrate-mediated regulation of nodulation (NRN). Collectively, these data suggest that PvFER1, PvRALF1, and PvRALF6 regulate the optimal number of nodules as a function of nitrate availability.
Assuntos
Phaseolus , Nodulação , Nodulação/genética , Nódulos Radiculares de Plantas/metabolismo , Phaseolus/genética , Phaseolus/metabolismo , Nitratos/farmacologia , Nitratos/metabolismo , Peptídeos/metabolismo , Simbiose , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
MAIN CONCLUSION: PvSYMRK-EGFP undergoes constitutive and rhizobia-induced endocytosis, which rely on the phosphorylation status of T589, the endocytic YXXØ motif and the kinase activity of the receptor. Legume-rhizobia nodulation is a complex developmental process. It initiates when the rhizobia-produced Nod factors are perceived by specific LysM receptors present in the root hair apical membrane. Consequently, SYMRK (Symbiosis Receptor-like Kinase) becomes active in the root hair and triggers an extensive signaling network essential for the infection process and nodule organogenesis. Despite its relevant functions, the underlying cellular mechanisms involved in SYMRK signaling activity remain poorly characterized. In this study, we demonstrated that PvSYMRK-EGFP undergoes constitutive and rhizobia-induced endocytosis. We found that in uninoculated roots, PvSYMRK-EGFP is mainly associated with the plasma membrane, although intracellular puncta labelled with PvSymRK-EGFP were also observed in root hair and nonhair-epidermal cells. Inoculation with Rhizobium etli producing Nod factors induces in the root hair a redistribution of PvSYMRK-EGFP from the plasma membrane to intracellular puncta. In accordance, deletion of the endocytic motif YXXØ (YKTL) and treatment with the endocytosis inhibitors ikarugamycin (IKA) and tyrphostin A23 (TyrA23), as well as brefeldin A (BFA), drastically reduced the density of intracellular PvSYMRK-EGFP puncta. A similar effect was observed in the phosphorylation-deficient (T589A) and kinase-dead (K618E) mutants of PvSYMRK-EGFP, implying these structural features are positive regulators of PvSYMRK-EGFP endocytosis. Our findings lead us to postulate that rhizobia-induced endocytosis of SYMRK modulates the duration and amplitude of the SYMRK-dependent signaling pathway.
Assuntos
Phaseolus , Rhizobium , Nódulos Radiculares de Plantas/metabolismo , Phaseolus/metabolismo , Nodulação , Rhizobium/fisiologia , Simbiose , Proteínas de Transporte/metabolismo , Endocitose , Raízes de Plantas/metabolismo , Proteínas de Plantas/metabolismoRESUMO
Legumes form a symbiotic association with rhizobia and fix atmospheric nitrogen in specialized root organs known as nodules. It is well known that salt stress inhibits root nodule symbiosis by decreasing rhizobial growth, rhizobial infection, nodule number, and nitrogenase activity in diverse legumes. Despite this knowledge, the genetic and molecular mechanisms governing salt stress's inhibition of nodulation and nitrogen fixation are still elusive. In this Viewpoint, we summarize the most recent knowledge of the genetic mechanisms that shape this symbiosis according to the salt levels in the soil. We emphasize the relevance of modulating the activity of the transcription factor Nodule Inception to properly shape the symbiosis with rhizobia accordingly. We also highlight the knowledge gaps that are critical for gaining a deeper understanding of the molecular mechanisms underlying the adaptation of the root nodule symbiosis to salt-stress conditions. We consider that filling these gaps can help to improve legume nodulation and harness its ecological benefits even under salt-stress conditions.
Assuntos
Fabaceae , Rhizobium , Nódulos Radiculares de Plantas , Simbiose/genética , Salinidade , Fabaceae/genética , Fixação de Nitrogênio/genética , Rhizobium/fisiologia , Estresse Salino/genética , Nodulação/genéticaRESUMO
Although several studies have demonstrated that biological nitrogen fixation can provide the nitrogen required by soybean, doubts regarding the need for nutrient application at sowing, specifically "starter N," inearly soybean cultivars persist, particularly in high-yielding crop fields. Thus, this study aimed to evaluate the effect of starter nitrogen fertilization on soybean cultivars with different maturity cycles. The experiment was conducted at Fazenda Recanto, in Carmo do Rio Claro, Minas Gerais state, during two crop seasons (2016/2017 and 2017/2018), using a randomized complete block designwith split-plot arrangement and four replications. In the 2016/17 croppingseason, the plots were comprised of three soybean cultivars (M6410 IPRO, NS7300 IPRO, and TEC7849 IPRO). In the 2017/18 cropping season, the plots consisted of four cultivars (M5917 IPRO, M6410 IPRO, AS3680 IPRO, andNS7670 RR). The subplots were assignedfour nitrogen doses at sowing (0, 10, 20, 40 kg ha-1). Grain yield, number of nodules per plant, mass of dry nodules per plant, number of grains and pods per plant, plant height and insertion of the first legume were evaluated. Experimental data were subjected to analysis of variance, and means were compared using the Scott-Knott test at 5% probability. The nitrogen rates at sowing did not influence soybean yields and agronomic characters of cultivars evaluated in the cropping 2016/17 and 2017/18. Increasing doses of nitrogen inhibited soybean nodulation.(AU)
Apesar de vários estudosjá terem demostrado quea fixação biológica de nitrogênioé capaz de fornecer o nitrogênio (N) requerido pela soja, dúvidassobre a necessidade deaplicação no nutriente na semeadura "N dearranque", em cultivares precoces de sojaainda são recorrentes, principalmente em lavouras com alto potencial de produtividade. Dessa forma, objetivou-se com esse estudo avaliar o efeito da adubação nitrogenada de arranque em cultivares de soja com diferentes ciclos. O experimento foi realizado na Fazenda Recanto, emCarmo do Rio Claro, Estado de Minas Gerais, em duas safras (2016/2017 e 2017/2018), em delineamento em blocos casualizados, em esquema de parcelas subdivididas, com quatro repetições. As parcelas foram compostas na safra 2016/17 por três cultivares de soja (M6410 IPRO, NS7300 IPRO e TEC7849 IPRO), e na safra 2017/18 por quatro cultivares (M5917IPRO, M6410IPRO, AS3680IPROeNS7670RR). A subparcelas foram constituídas por quatro doses de N na semeadura (0, 10, 20, 40 kg ha-1). Foi avaliada a produtividade de grãos, número de nódulos por planta, massa de nódulos secos por planta, número de grãos e vagens por planta, altura de plantas e de inserção do primeiro legume. Os dados experimentais foram submetidos à análise de variância, e as médias comparadas através do teste de Scott-Knott a 5% de probabilidade. As doses de N em semeadura não influenciaram a produtividade e os caracteres agronômicos das cultivares avaliadas nassafras2016/17 e 2017/18. Doses crescentes de nitrogênio inibiram a nodulação da soja.(AU)
Assuntos
Glycine max/fisiologia , 24444 , Fertilizantes/análise , Brasil , Nodulação/fisiologia , Nitrogênio/efeitos adversos , Fixação de NitrogênioRESUMO
Nitrogen-fixing symbiosis is globally important in ecosystem functioning and agriculture, yet the evolutionary history of nodulation remains the focus of considerable debate. Recent evidence suggesting a single origin of nodulation followed by massive parallel evolutionary losses raises questions about why a few lineages in the N2 -fixing clade retained nodulation and diversified as stable nodulators, while most did not. Within legumes, nodulation is restricted to the two most diverse subfamilies, Papilionoideae and Caesalpinioideae, which show stable retention of nodulation across their core clades. We characterize two nodule anatomy types across 128 species in 56 of the 152 genera of the legume subfamily Caesalpinioideae: fixation thread nodules (FTs), where nitrogen-fixing bacteroids are retained within the apoplast in modified infection threads, and symbiosomes, where rhizobia are symplastically internalized in the host cell cytoplasm within membrane-bound symbiosomes (SYMs). Using a robust phylogenomic tree based on 997 genes from 147 Caesalpinioideae genera, we show that losses of nodulation are more prevalent in lineages with FTs than those with SYMs. We propose that evolution of the symbiosome allows for a more intimate and enduring symbiosis through tighter compartmentalization of their rhizobial microsymbionts, resulting in greater evolutionary stability of nodulation across this species-rich pantropical legume clade.
Assuntos
Fabaceae , Rhizobium , Ecossistema , Fabaceae/genética , Nitrogênio , Fixação de Nitrogênio , Nodulação/genética , Nódulos Radiculares de Plantas , SimbioseRESUMO
The development of a symbiotic nitrogen-fixing nodule in legumes involves infection and organogenesis. Infection begins when rhizobia enter a root hair through an inward structure, the infection thread (IT), which guides the bacteria towards the cortical tissue. Concurrently, organogenesis takes place by inducing cortical cell division (CCD) at the infection site. Genetic analysis showed that both events are well-coordinated; however, the dynamics connecting them remain to be elucidated. To visualize the crossroads between IT and CCD, we benefited from the fact that, in Phaseolus vulgaris nodulation, where the first division occurs in subepidermal cortical cells located underneath the infection site, we traced a Rhizobium etli strain expressing DsRed, the plant cytokinesis marker YFP-PvKNOLLE, a nuclear stain and cell wall auto-fluorescence. We found that the IT exits the root hair to penetrate an underlying subepidermal cortical (S-E) cell when it is concluding cytokinesis.
Assuntos
Phaseolus , Rhizobium , Divisão Celular , Phaseolus/microbiologia , Proteínas de Plantas/genética , Nodulação , Raízes de Plantas/genética , Rhizobium/genética , Nódulos Radiculares de Plantas/microbiologia , Simbiose/genéticaRESUMO
Bacteria from the rhizobia group are able to associate symbiotically with bean crop, forming nodules in the root, in which the biological nitrogen-fixing process occurs. However, the efficiency of this process has been low and it can be attributed to genetic and environmental factors. Thus, the objective of this study was to evaluate the capacity of nodulation of local varieties and commercial common bean cultivars inoculated with a Rhizobium tropici strain used in commercial inoculants and rhizobia isolates from common bean root nodules. The experiment was carried out in a factorial scheme (2x4), in a randomized block design with four replicates. It was tested two local varieties and two commercial cultivars, inoculated with the reference Rhizobium tropicistrain CIAT899 and the RBZ14 strain isolated from common bean nodules grown in soils of Southern Brazil, in adapted Leonard-type pots. The CIAT899 strain promoted either higher mass of viable nodules and higher nitrogen accumulation in the aerial part. The black group local variety showed better response than the cultivar TAA Dama for nodule viability, suggesting more efficiency for nodulation. The interaction between genotypes (local varieties and commercial cultivars) and bacteria showed the specificity of the complex symbiotic relationship of biological nitrogen fixation in common bean, requiring further studies of these interactions.(AU)
As bactérias do grupo rizóbios estão associadas simbioticamente à cultura do feijão, formando nódulos na raiz, nos quais ocorre o processo biológico de fixação do nitrogênio. No entanto, a eficiência desse processo tem sido baixa e pode ser atribuída a fatores genéticos e ambientais. Assim, este estudo teve como objetivo avaliar a capacidade de nodulação de variedades locais e cultivares comerciais de feijão comum inoculadas com uma estirpe de Rhizobium tropiciusada em inoculantes comerciais e isolados de nódulos da raiz do feijão comum. O experimento foi realizado em esquema fatorial (2x4), em delineamento de blocos ao acaso, com quatro repetições. Foram testadas duas variedades locais e duas cultivares comerciais, inoculadascom a estirpe de referência Rhizobium tropiciCIAT899 e o isolado RBZ14 obtido de nódulos de feijão cultivados em solos do sul do Brasil, em vasos adaptados do tipo Leonard. A estirpe CIAT899 promoveu maior massa de nódulos viáveis e maior acúmulo de nitrogênio na parte aérea. A variedade regional do grupo preto apresentou melhor resposta que a cultivar TAA Dama, quanto à viabilidade dos nódulos, sugerindo resposta mais eficiente à nodulação. A interação entre genótipos e bactérias mostrou a especificidade da complexa relação simbiótica da fixação biológica de nitrogênio no feijoeiro, sendo necessário aprofundar os estudos nessas interações.(AU)
Assuntos
Fenômenos Biológicos , Phaseolus/genética , Rhizobium tropici/genética , Nodulação , Fixação de NitrogênioRESUMO
Ethylene has been implicated in nitrogen fixing symbioses in legumes, where rhizobial invasion occurs via infection threads (IT). In the symbiosis between peanut (Arachis hypogaea L.) and bradyrhizobia, the bacteria penetrate the root cortex intercellularly and IT are not formed. Little attention has been paid to the function of ethylene in the establishment of this symbiosis. The aim of this article is to evaluate whether ethylene plays a role in the development of this symbiotic interaction and the participation of Nod Factors (NF) in the regulation of ethylene signalling. Manipulation of ethylene in peanut was accomplished by application of 1-aminocyclopropane-1-carboxylic acid (ACC), which mimics applied ethylene, or AgNO3, which blocks ethylene responses. To elucidate the participation of NF in the regulation of ethylene signalling, we inoculated plants with a mutant isogenic rhizobial strain unable to produce NF and evaluated the effect of AgNO3 on gene expression of NF and ethylene responsive signalling pathways. Data revealed that ethylene perception is required for the formation of nitrogen-fixing nodules, while addition of ACC does not affect peanut symbiotic performance. This phenotypic evidence is in agreement with transcriptomic data from genes involved in symbiotic and ethylene signalling pathways. NF seem to modulate the expression of ethylene signalling genes. Unlike legumes infected through IT formation, ACC addition to peanut does not adversely affect nodulation, but ethylene perception is required for establishment of this symbiosis. Evidence for the contribution of NF to the modulation of ethylene-inducible defence gene expression is provided.
Assuntos
Bradyrhizobium , Fabaceae , Arachis , Etilenos , Nodulação , Raízes de Plantas , Nódulos Radiculares de Plantas , SimbioseRESUMO
Phaseolus vulgaris is a grain cultivated in vast areas of different countries. It is an excellent alternative to the other legumes in the Venezuelan diet and is of great agronomic interest due to its resistance to soil acidity, drought, and high temperatures. Phaseolus establishes symbiosis primarily with Rhizobium and Ensifer species in most countries, and this rhizobia-legume interaction has been studied in Asia, Africa, and the Americas. However, there is currently no evidence to show that rhizobia nodulate the endemic cultivars of P. vulgaris in Venezuela. Therefore, we herein investigated the phylogenetic diversity of plant growth-promoting and N2-fixing nodulating bacteria isolated from the root nodules of P. vulgaris cultivars in a different agroecosystem in Venezuela. In comparisons with other countries, higher diversity was found in isolates from P. vulgaris nodules, ranging from α- and ß-proteobacteria. Some isolates belonging to several new phylogenetic lineages within Bradyrhizobium, Ensifer, and Mesorhizobium species were also specifically isolated at some topographical regions. Additionally, some isolates exhibited tolerance to high temperature, acidity, alkaline pH, salinity stress, and high Al levels; some of these characteristics may be related to the origin of the isolates. Some isolates showed high tolerance to Al toxicity as well as strong plant growth-promoting and antifungal activities, thereby providing a promising agricultural resource for inoculating crops.
Assuntos
Bactérias/genética , Bactérias/isolamento & purificação , Variação Genética , Phaseolus/microbiologia , Nódulos Radiculares de Plantas/microbiologia , Simbiose , Bactérias/classificação , Fenômenos Fisiológicos Bacterianos , DNA Bacteriano/genética , Fixação de Nitrogênio , Phaseolus/crescimento & desenvolvimento , Filogenia , Nodulação , Microbiologia do Solo , VenezuelaRESUMO
BACKGROUND: Rhizobium-legume symbiosis is a specific, coordinated interaction that results in the formation of a root nodule, where biological nitrogen fixation occurs. NADPH oxidases, or Respiratory Burst Oxidase Homologs (RBOHs) in plants, are enzymes that generate superoxide (O2 â¢-). Superoxide produces other reactive oxygen species (ROS); these ROS regulate different stages of mutualistic interactions. For example, changes in ROS levels are thought to induce ROS scavenging, cell wall remodeling, and changes in phytohormone homeostasis during symbiotic interactions. In common bean (Phaseolus vulgaris), PvRbohB plays a key role in the early stages of nodulation. RESULTS: In this study, to explore the role of PvRbohB in root nodule symbiosis, we analyzed transcriptomic data from the roots of common bean under control conditions (transgenic roots without construction) and roots with downregulated expression of PvRbohB (by RNA interference) non-inoculated and inoculated with R. tropici. Our results suggest that ROS produced by PvRBOHB play a central role in infection thread formation and nodule organogenesis through crosstalk with flavonoids, carbon metabolism, cell cycle regulation, and the plant hormones auxin and cytokinin during the early stages of this process. CONCLUSIONS: Our findings provide important insight into the multiple roles of ROS in regulating rhizobia-legume symbiosis.
Assuntos
Carbono/metabolismo , Ciclo Celular , NADPH Oxidases/metabolismo , Phaseolus/enzimologia , Nodulação , Raízes de Plantas/enzimologia , Simbiose/fisiologia , Phaseolus/genética , Phaseolus/microbiologia , Raízes de Plantas/microbiologia , Espécies Reativas de Oxigênio/metabolismo , Rhizobium/fisiologia , TranscriptomaRESUMO
Legume plants from Fabaceae family (phylogenetic group composed by three subfamilies: Caesalpinioideae, Mimosoideae, and Papilionoideae) can fix atmospheric nitrogen (N2) into ammonia (NH3) by the symbiotic relationship with rhizobia bacteria. These bacteria respond chemotactically to certain compounds released by plants such as sugars, amino acids and organic acids. Root secretion of isoflavonoids acts as inducers for nod genes in rhizobia and ABC transporters and ICHG (isoflavone conjugates hydrolyzing beta-glucosidase) at apoplast are related to the exudation of genistein and daidzein in soybean roots. Biological nitrogen fixation (BNF) occurs inside the nodule by the action of nitrogenase enzyme, which fixes N2 into NH3, which is converted into ureides (allantoin and allantoic acid). In this review, we bring together the latest findings on flavonoids biosynthesis and ureide metabolism in several legume plant species. We emphasize how flavonoids induce nod genes in rhizobia, affecting chemotaxis, nodulation, ureide production, growth and yield of legume plants. Mainly, isoflavonoids daidzein and genistein are responsible for nod genes activation in the rhizobia bacteria. Flavonoids also play an important role during nodule organogenesis by acting as auxin transporter inhibitors in root cells, especially in indeterminate nodules. The ureides are the main N transport form in tropical legumes and they are catabolized in leaves and other sink tissues to produce amino acids and proteins needed for plant growth and yield.
Assuntos
Fabaceae , Rhizobium , Flavonoides , Fixação de Nitrogênio , Filogenia , Nodulação , SimbioseRESUMO
When subjected to nutritional stress, bacteria modify their amino acid metabolism and cell division activities by means of the stringent response, which is controlled by the Rsh protein in alphaproteobacteria. An important group of alphaproteobacteria are the rhizobia, which fix atmospheric N2 in symbiosis with legume plants. Although nutritional stress is common for rhizobia while infecting legume roots, the stringent response has scarcely been studied in this group of soil bacteria. In this report, we obtained a mutant with a kanamycin resistance insertion in the rsh gene of Bradyrhizobium diazoefficiens, the N2-fixing symbiont of soybean. This mutant was defective for type 3 secretion system induction, plant defense suppression at early root infection, and nodulation competition. Furthermore, the mutant produced smaller nodules, although with normal morphology, which led to lower plant biomass production. Soybean (Glycine max) genes GmRIC1 and GmRIC2, involved in autoregulation of nodulation, were upregulated in plants inoculated with the mutant under the N-free condition. In addition, when plants were inoculated in the presence of 10 mM NH4NO3, the mutant produced nodules containing bacteroids, and GmRIC1 and GmRIC2 were downregulated. The rsh mutant released more auxin to the culture supernatant than the wild type, which might in part explain its symbiotic behavior in the presence of combined N. These results indicate that the B. diazoefficiens stringent response integrates into the plant defense suppression and regulation of nodulation circuits in soybean, perhaps mediated by the type 3 secretion system.IMPORTANCE The symbiotic N2 fixation carried out between prokaryotic rhizobia and legume plants performs a substantial contribution to the N cycle in the biosphere. This symbiotic association is initiated when rhizobia infect and penetrate the root hairs, which is followed by the growth and development of root nodules, within which the infective rhizobia are established and protected. Thus, the nodule environment allows the expression and function of the enzyme complex that catalyzes N2 fixation. However, during early infection, the rhizobia find a harsh environment while penetrating the root hairs. To cope with this nuisance, the rhizobia mount a stress response known as the stringent response. In turn, the plant regulates nodulation in response to the presence of alternative sources of combined N in the surrounding medium. Control of these processes is crucial for a successful symbiosis, and here we show how the rhizobial stringent response may modulate plant defense suppression and the networks of regulation of nodulation.
Assuntos
Bradyrhizobium/genética , Glycine max/microbiologia , Farmacorresistência Bacteriana/genética , Fertilizantes , Resistência a Canamicina/genética , Proteínas Associadas aos Microtúbulos/genética , Proteínas Monoméricas de Ligação ao GTP/genética , Mutação , Nitratos , Fixação de Nitrogênio , Proteínas de Plantas/genética , Nodulação , Glycine max/genética , Simbiose , Sistemas de Secreção Tipo IIIRESUMO
The aim of this work was to characterize and identify some bacteria isolated from the root nodules of Retama monosperma grown in Sidi Boubker lead and zinc mine tailings. Very few root nodules were obtained on the root nodules of R. monosperma grown in these soils. The three bacteria isolated from the root nodules were tolerant in vitro to different concentrations of heavy metals, including lead and zinc. The rep-PCR experiments showed that the three isolates have different molecular fingerprints and were considered as three different strains. The analysis of their 16S rRNA gene sequences proved their affiliation to the genus Bradyrhizobium. The analysis and phylogeny of the housekeeping genes atpD, glnII, gyrB, recA, and rpoB confirmed that the closest species was B. valentinum with similarity percentages of 95.61 to 95.82%. The three isolates recovered from the root nodules were slow-growing rhizobia capable to renodulate their original host plant in the presence of Pb-acetate. They were able to nodulate R. sphaerocarpa and Lupinus luteus also but not Glycine max or Phaseolus vulgaris. The phylogeny of the nodA and nodC nodulation genes as well as the nifH gene of the three strains showed that they belong to the symbiovar retamae of the genus Bradyrhizobium. The three strains isolated could be considered for use as inoculum for Retama plants before use in phytoremediation experiments.