Common Bean Nodules Research Articles

Responses of symbiotic nitrogen-fixing common bean to aluminum toxicity and delineation of nodule responsive microRNAs.

Aluminum (Al) toxicity is widespread in acidic soils where the common bean (Phaseolus vulgaris), the most important legume for human consumption, is produced and it is a limiting factor for crop production and symbiotic nitrogen fixation. We characterized the nodule responses of common bean plants inoculated with Rhizobioum tropici CIAT899 and the root responses of nitrate-fertilized plants exposed to excess Al in low pH, for long or short periods. A 43–50% reduction in nitrogenase activity indicates that Al toxicity (Alt) highly affected nitrogen fixation in common bean. Bean roots and nodules showed characteristic symptoms for Alt. In mature nodules Al accumulation and lipoperoxidation were observed in the infected zone, while callose deposition and cell death occurred mainly in the nodule cortex. Regulatory mechanisms of plant responses to metal toxicity involve microRNAs (miRNAs) along other regulators. Using a miRNA-macroarray hybridization approach we identified 28 (14 up-regulated) Alt nodule-responsive miRNAs. We validated (quantitative reverse transcriptase-PCR) the expression of eight nodule responsive miRNAs in roots and in nodules exposed to high Al for long or short periods. The inverse correlation between the target and miRNA expression ratio (stress:control) was observed in every case. Generally, miRNAs showed a higher earlier response in roots than in nodules. Some of the common bean Alt-responsive miRNAs identified has also been reported as differentially expressed in other plant species subjected to similar stress condition. miRNA/target nodes analyzed in this work are known to be involved in relevant signaling pathways, thus we propose that the participation of miR164/NAC1 (NAM/ATAF/CUC transcription factor) and miR393/TIR1 (TRANSPORT INHIBITOR RESPONSE 1-like protein) in auxin and of miR170/SCL (SCARECROW-like protein transcription factor) in gibberellin signaling is relevant for common bean response/adaptation to Al stress. Our data provide a foundation for evaluating the individual roles of miRNAs in the response of common bean nodules to Alt.

Regulatory nodD1 and nodD2 genes of Rhizobium tropici strain CIAT 899 and their roles in the early stages of molecular signaling and host-legume nodulation

BackgroundNodulation and symbiotic nitrogen fixation are mediated by several genes, both of the host legume and of the bacterium. The rhizobial regulatory nodD gene plays a critical role, orchestrating the transcription of the other nodulation genes. Rhizobium tropici strain CIAT 899 is an effective symbiont of several legumes—with an emphasis on common bean (Phaseolus vulgaris)—and is unusual in carrying multiple copies of nodD, the roles of which remain to be elucidated.ResultsPhenotypes, Nod factors and gene expression of nodD1 and nodD2 mutants of CIAT 899 were compared with those of the wild type strain, both in the presence and in the absence of the nod-gene-inducing molecules apigenin and salt (NaCl). Differences between the wild type and mutants were observed in swimming motility and IAA (indole acetic acid) synthesis. In the presence of both apigenin and salt, large numbers of Nod factors were detected in CIAT 899, with fewer detected in the mutants. nodC expression was lower in both mutants; differences in nodD1 and nodD2 expression were observed between the wild type and the mutants, with variation according to the inducing molecule, and with a major role of apigenin with nodD1 and of salt with nodD2. In the nodD1 mutant, nodulation was markedly reduced in common bean and abolished in leucaena (Leucaena leucocephala) and siratro (Macroptilium atropurpureum), whereas a mutation in nodD2 reduced nodulation in common bean, but not in the other two legumes.ConclusionOur proposed model considers that full nodulation of common bean by R. tropici requires both nodD1 and nodD2, whereas, in other legume species that might represent the original host, nodD1 plays the major role. In general, nodD2 is an activator of nod-gene transcription, but, in specific conditions, it can slightly repress nodD1. nodD1 and nodD2 play other roles beyond nodulation, such as swimming motility and IAA synthesis.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-1458-8) contains supplementary material, which is available to authorized users.

The role of nodules in the tolerance of common bean to iron deficiency

Iron is vital for the establishment and function of symbiotic root nodules of legumes. Although abundant in the environment, Fe is often a limiting nutrient for plant growth due to its low solubility and availability in some soils. We have studied the mechanism of iron uptake in the root nodules of common bean to evaluate the role of nodules in physiological responses to iron deficiency. Based on experiments using full or partial submergence of nodulated roots in the nutrient solution, our results show that the nodules were affected only slightly under iron deficiency, especially when the nodules were submerged in nutrient solution in the tolerant cultivar. In addition, fully submerged root nodules showed enhanced acidification of the nutrient solution and showed higher ferric chelate reductase activity than that of partially submerged roots in plants cultivated under Fe deficiency. The main results obtained in this work suggest that in addition to preferential Fe allocation from the root system to the nodules, this symbiotic organ probably develops some mechanisms to respond to iron deficiency. These mechanisms were implied especially in nodule Fe absorption efficiency and in the ability of this organ to take up Fe directly from the medium.

Diversity of sporadic symbionts and nonsymbiotic endophytic bacteria isolated from nodules of woody, shrub, and food legumes in Ethiopia

Fifty-five bacterial isolates were obtained from surface-sterilized nodules of woody and shrub legumes growing in Ethiopia: Crotalaria spp., Indigofera spp., and Erythrina brucei, and the food legumes soybean and common bean. Based on partial 16S rRNA gene sequence analysis, the majority of the isolates were identified as Gram-negative bacteria belonging to the genera Achromobacter, Agrobacterium, Burkholderia, Cronobacter, Enterobacter, Mesorhizobium, Novosphingobium, Pantoea, Pseudomonas, Rahnella, Rhizobium, Serratia, and Variovorax. Seven isolates were Gram-positive bacteria belonging to the genera Bacillus, Paenibacillus, Planomicrobium, and Rhodococcus. Amplified fragment length polymorphism (AFLP) fingerprinting showed that each strain was genetically distinct. According to phylogenetic analysis of recA, glnII, rpoB, and 16S rRNA gene sequences, Rhizobium, Mesorhizobium, and Agrobacterium were further classified into six different genospecies: Agrobacterium spp., Agrobacterium radiobacter, Rhizobium sp., Rhizobium phaseoli, Mesorhizobium sp., and putative new Rhizobium species. The strains from R. phaseoli, Rhizobium sp. IAR30, and Mesorhizobium sp. ERR6 induced nodules on their host plants. The other strains did not form nodules on their original host. Nine endophytic bacterial strains representing seven genera, Agrobacterium, Burkholderia, Paenibacillus, Pantoea, Pseudomonas, Rhizobium, and Serratia, were found to colonize nodules of Crotalaria incana and common bean on co-inoculation with symbiotic rhizobia. Four endophytic Rhizobium and two Agrobacterium strains had identical nifH gene sequences with symbiotic Rhizobium strains, suggesting horizontal gene transfer. Most symbiotic and nonsymbiotic endophytic bacteria showed plant growth-promoting properties in vitro, which indicate their potential role in the promotion of plant growth when colonizing plant roots and the rhizosphere.

Transcript profiling of common bean nodules subjected to oxidative stress

Several environmental stresses generate high amounts of reactive oxygen species (ROS) in plant cells, resulting in oxidative stress. Symbiotic nitrogen fixation (SNF) in the legume-rhizobia symbiosis is sensitive to damage from oxidative stress. Active nodules of the common bean (Phaseolus vulgaris) exposed to the herbicide paraquat (1,1'-dimethyl-4,4'-bipyridinium dichloride hydrate), which stimulates ROS accumulation, exhibited reduced nitrogenase activity and ureide content. We analyzed the global gene response of nodules subjected to oxidative stress using the Bean Custom Array 90K, which includes probes from 30,000 expressed sequence tags (ESTs). A total of 4280 ESTs were differentially expressed in stressed bean nodules; of these, 2218 were repressed. Based on Gene Ontology analysis, these genes were grouped into 42 different biological process categories. Analysis with the PathExpress bioinformatic tool, adapted for bean, identified five significantly repressed metabolic pathways related to carbon/nitrogen metabolism, which is crucial for nodule function. Quantitative reverse transcription (qRT)-PCR analysis of transcription factor (TF) gene expression showed that 67 TF genes were differentially expressed in nodules exposed to oxidative stress. Putative cis-elements recognized by highly responsive TF were detected in promoter regions of oxidative stress regulated genes. The expression of oxidative stress responsive genes and of genes important for SNF in bacteroids analyzed in stressed nodules revealed that these conditions elicited a transcriptional response.

Seeds enriched with phosphorus and molybdenum as a strategy for improving grain yield of common bean crop

Sowing seeds with high concentrations of P or Mo can improve growth and nodulation of common bean (Phaseolus vulgaris) plants. However, the performance of the bean crop originating from seeds enriched with P and Mo has not yet been assessed under field conditions. This study aimed to evaluate the effect of sowing seeds enriched with P and Mo, harvested from plants that received foliar sprayings of P or Mo or both, on growth and yield of common bean crop. Seeds with contrasting concentrations of P and Mo were sown in three field experiments in Brazil (in 2006 and 2008 in Rio de Janeiro State, and in 2009 in Goiás State) with different N sources (control without N, seed with rhizobia inoculation, rhizobia inoculation plus 40kgNha−1 side-dressed, mineral N with 60kgNha−1). In 2006, seeds enriched with P and Mo increased nodule mass at 32 days after emergence (DAE), shoot N and biomass at 48 DAE, and grain yield and N content, for most combinations of the three cultivars and N sources evaluated. In 2008, the highest shoot growth at 57 DAE and grain yield were obtained with low-P high-Mo seeds, which was partially due to the higher Mo concentration of these seeds. In 2009, seeds enriched with P and Mo increased nodule mass at 34 DAE and grain yield but only for plants receiving inoculation plus side-dressed N. The results demonstrate the benefits of seeds with high concentrations of P and Mo for improving bean yield, although such yield increases were more effective for plants inoculated with rhizobia than amended with N. Sowing enriched seeds, associated with rhizobia inoculation and side-dressed N, provided grain yields much higher than the average common bean yields in Brazil.

Sinorhizobium americanum symbiovar mediterranense is a predominant symbiont that nodulates and fixes nitrogen with common bean (Phaseolus vulgaris L.) in a Northern Tunisian field

A total of 40 symbiotic bacterial strains isolated from root nodules of common bean grown in a soil located in the north of Tunisia were characterized by PCR-RFLP of the 16S rRNA genes. Six different ribotypes were revealed. Nine representative isolates were submitted to phylogenetic analyses of rrs, recA, atpD, dnaK, nifH and nodA genes. The strains 23C40 and 23C95 representing the most abundant ribotype were closely related to Sinorhizobium americanum CFNEI 156T. S. americanum was isolated from Acacia spp. in Mexico, but this is the first time that this species is reported among natural populations of rhizobia nodulating common bean. These isolates nodulated and fixed nitrogen with this crop and harbored the symbiotic genes of the symbiovar mediterranense. The strains 23C2 and 23C55 were close to Rhizobium gallicum R602spT but formed a well separated clade and may probably constitute a new species. The sequence similarities with R. gallicum type strain were 98.7% (rrs), 96.6% (recA), 95.8% (atpD) and 93.4% (dnaK). The remaining isolates were, respectively, affiliated to R. gallicum, E. meliloti, Rhizobium giardinii and Rhizobium radiobacter. However, some of them failed to re-nodulate their original host but promoted root growth.

Effect of iron deficiency on the localization of phosphoenolpyruvate carboxylase in common bean nodules

Phosphoenolpyruvate carboxylase (PEPC) plays an important role in nodules, when there is an increase in the demand for energy. This enzyme provides carbon skeletons to sustain amino acid synthesis and malate to support energy required to fix nitrogen. Since PEPC is important for nodules, and there is lack of information about the effect of some nutrient deficiency in the expression and localization of this enzyme in legume nodules, this work focused on the localization of PEPC in nodules under iron deficiency of two common bean cultivars: Flamingo tolerant and Coco blanc sensitive to iron (Fe) deficiency. The results of immunolocalization using polyclonal antibody showed that this enzyme was detected in all regions of nodule sections; but the signal intensity was increased in Fe-deficient nodules as compared to Fe-sufficient ones in the tolerant cultivar, whereas the intensity was less pronounced in nodules of Fe-deficient plants than in those of Fe-sufficient plants for the sensitive cultivar Coco blanc. This work showed that the symbiotic tolerance of Flamingo to iron deficiency was linked to the increase of PEPC enzymes expression. However, the activity of these enzymes supported the energy required in bacteroids to maintain the nitrogenase activity. Keywords: Common bean, immunolocalization, iron deficiency, nodules, phosphoenol pyruvate carboxylase

Influence of integrated phosphorus supply and plant growth promoting rhizobacteria on growth, nodulation, yield and nutrient uptake in Phaseolus vulgaris

To guarantee a sufficient phosphorus supply for plants, a rapid and permanent mobilization of phosphorus from the labile phosphorus fractions is necessary, because phosphorus concentrations in soil solution are generally low. Several plant growth-promoting rhizobacteria (PGPR) have shown potential to enhance phosphorus solublization and nodulation of legumes when co-inoculated with Rhizobium . This investigation was undertaken to assess the feasibility and compatibility of two mineral phosphorus fertilizers; diammonium phosphate (DAP), triple super phosphate (TSP), poultry manure (PM) and two PGPR strains on the growth, nodulation, yield, nutrient uptake and protein content of common bean ( Phaseolus vulgaris ) under deficient phosphorus supply. Integrated application of mineral phosphorus (P), PM and PGPR significantly increased shoot height, shoot fresh weight, shoot dry weight and leaf chlorophyll content by 67, 160, 51 and 106%, respectively, while increase in root length, root fresh weight and root dry weight was 79, 161, and 187%, respectively, over unfertilized control without PGPR application. Integrated use of different P sources and PGPR also increased number of nodules per plant, nodule fresh weight and nodule dry weight by 158, 107 and 168% over the control. Treatment with PGPR significantly increased number of pods per plant and grain yield by 224 and 96%, respectively over the control. Co-inoculation with Bradyrhizobium sp. strain MN-S and Agrobacterium sp. strain Ca-18 demonstrated two-fold increase in the proportion of nitrogen (N) and P uptake as well as protein content of the common bean grain was increases by 48%. Therefore, application of PGPR with low P fertilizer rates and PM could be a viable supplementary strategy for maximum benefits in terms of cost of production and sustaining productivity. Key words: Phaseolus vulgaris L., plant growth promoting rhizobacteria (PGPR), nodulation, yield, phosphorus deficiency, protein.

Immunolocalization of H +-ATPase and IRT1 enzymes in N 2-fixing common bean nodules subjected to iron deficiency

The demand for iron in leguminous plants increases during symbiosis, as the metal is utilised for the synthesis of various Fe-containing proteins in both plant and bacteroids. However, the acquisition of this micronutrient is problematic due to its low bioavailability at physiological pH under aerobic conditions. Induction of root Fe(III)-reductase activity is necessary for Fe uptake and can be coupled to the rhizosphere acidification capacity linked to the H +-ATPase activity. Fe uptake is related to the expression of a Fe 2+ transporter (IRT1). In order to verify the possible role of nodules in the acquisition of Fe directly from the soil solution, the localization of H +-ATPase and IRT1 was carried out in common bean nodules by immuno-histochemical analysis. The results showed that these proteins were particularly abundant in the central nitrogen-fixing zone of nodules, around the periphery of infected and uninfected cells as well as in the vascular bundle of control nodules. Under Fe deficiency an over-accumulation of H +-ATPase and IRT1 proteins was observed especially around the cortex cells of nodules. The results obtained in this study suggest that the increase in these proteins is differentially localized in nodules of Fe-deficient plants when compared to the Fe-sufficient condition and cast new light on the possible involvement of nodules in the direct acquisition of Fe from the nutrient solution.

Effect of salinity on nodulation, nitrogen fixation and growth of common bean (Phaseolus vulgaris) inoculated with rhizobial strains isolated from the Haouz region of Morocco

This study compared growth, nodulation, nitrogen fixation, and nodular enzyme activities in response to salinity in some common bean-rhizobia symbiotic combinations. Seeds of Paulista and Efequince, two varieties of the common bean (Phaseolus vulgaris) were germinated and seedlings were transferred to pots containing vermiculite inoculated with the reference Rhizobium strain CIAT899 or with RhM11 or RhM14, two local strains. Plants were grown in a temperature-controlled glasshouse at 28°C and irrigated with a nutrient solution without NaCl (control) or supplemented with 25 mM NaCl (stressed). Plants were harvested at the flowering stage. The results showed that in controls, inoculation with RhM11 improved plant and nodule growth compared with those inoculated with RhM14 and CIAT 899. NaCl treatment generally had a negative affect on plant and nodule growth. Under the saline treatment, symbiotic nitrogen fixation was not significantly affected in the CIAT899-Paulista, CIAT899-Efequince and RhM11-Paulista combinations. Plant mineral nutrition was negatively affected under salt treatment for all of the tested symbiotic combinations. Inoculation with CIAT899 and RhM11 conferred more plant tolerance to salinity than inoculation with RhM14. The nodular phosphoenolpyruvate carboxylase (PEPC) and malate dehydrogenase (MDH) exhibited higher activities and were less affected by salinity in plants inoculated with the reference strain CIAT899 than those inoculated with local strains. We conclude that plants inoculated with CIAT899 and RhM11 showed more salinity stress tolerance than those inoculated with RhM14.

Genetic diversity and symbiotic effectiveness of rhizobia isolated from root nodules of Phaseolus vulgaris L. grown in soils of Iran

The common bean ( Phaseolus vulgaris L.) occupies more than 120,000 ha in Iran but information on the genetic structure and nitrogen fixation potential of the rhizobia nodulating this legume crop is scarce. Fifty-three bacterial isolates were sampled from root nodules of common bean cultivated in 12 different provinces of Iran. The genetic diversity of the isolates was analysed by PCR–RFLP of the 16S–23S rDNA internally transcribed spacer (ITS) region. At the similarity of 64%, common bean rhizobia were clustered into 10 groups, demonstrating a high degree of genetic variability among the isolates. Significant differences existed in the symbiotic potential of the isolates examined under greenhouse conditions. Ten isolates showed promising symbiotic characteristics in terms of shoot dry weight and symbiotic efficiency, and selected isolates revealed increased plant growth and yield under field conditions.

Metabolic changes of iron uptake in N 2-fixing common bean nodules during iron deficiency

Iron is an important nutrient in N 2-fixing legume nodules. The demand for this micronutrient increases during the symbiosis establishment, where the metal is utilized for the synthesis of various iron-containing proteins in both the plant and the bacteroid. Unfortunately, in spite of its importance, iron is poorly available to plant uptake since its solubility is very low when in its oxidized form Fe(III). In the present study, the effect of iron deficiency on the activity of some proteins involved in Strategy I response, such as Fe-chelate reductase (FC-R), H +-ATPase, and phospho enolpyruvate carboxylase (PEPC) and the protein level of iron regulated transporter (IRT1) and H +-ATPase proteins has been investigated in both roots and nodules of a tolerant (Flamingo) and a susceptible (Coco blanc) cultivar of common bean plants. The main results of this study show that the symbiotic tolerance of Flamingo can be ascribed to a greater increase in the FC-R and H +-ATPase activities in both roots and nodules, leading to a more efficient Fe supply to nodulating tissues. The strong increase in PEPC activity and organic acid content, in the Flamingo root nodules, suggests that under iron deficiency nodules can modify their metabolism in order to sustain those activities necessary to acquire Fe directly from the soil solution.

PLANT GROWTH PROMOTING RHIZOBACTERIA INCREASE GROWTH, YIELD AND NITROGEN FIXATION IN PHASEOLUS VULGARIS

Nitrogen (N) fixation by legume-Rhizobium symbiosis is important to agricultural productivity and is therefore of great economic interest. Growing evidence indicates that soil beneficial bacteria can positively affect symbiotic performance of rhizobia. The effect of co-inoculation with plant growth-promoting rhizobacteria (PGPR) and Rhizobium, on nodulation, nitrogen fixation, and yield of common bean (Phaseolus vulgaris L.) cultivars was investigated in two consecutive years under field conditions. The PGPR strains Pseudomonas fluorescens P-93 and Azospirillum lipoferum S-21 as well as two highly effective Rhizobium strains were used in this study. Common bean seeds of three cultivars were inoculated with Rhizobium singly or in a combination with PGPR to evaluate their effect on nodulation and nitrogen fixation. A significant variation of plant growth in response to inoculation with Rhizobium strains was observed. Treatment with PGPR significantly increased nodule number and dry weight, shoot dry weight, amount of nitrogen fixed as well as seed yield and protein content. Co-inoculation with Rhizobium and PGPR demonstrated a significant increase in the proportion of nitrogen derived from atmosphere. These results indicate that PGPR strains have potential to enhance the symbiotic potential of rhizobia.

Importance of ferric chelate reductase activity and acidification capacity in root nodules of N2-fixing common bean (Phaseolus vulgaris L.) subjected to iron deficiency

Iron is an essential nutrient for plants, especially in symbiotic N2-fixing legumes. Although abundant in the soil, iron is generally not available to plants as it is predominantly in an insoluble form (FeIII) . Mono- and dicotyledonous plants, except Grarnineae, have developed morphological and physiological responses, notably an increase in rhizosphere acidification (H+-ATPase) and an enhanced plasma membrane ferric chelate reductase activity (Fe-CR) in the roots. However, studies on the physiological responses of root nodules are lacking. The present study was initiated to investigate the acidification capacity and Fe-CR activity of nodulated roots, and intact nodules, in two contrasting common bean varieties, Coco blanc sensitive to iron deficiency and Flamingo tolerant to iron deficiency. The discovery of an induction of H+-ATPase and Fe-CR activities in root nodules of commonbean under iron deficiency, suggests that these organs participate in improving iron availability for the contained bacteroids.

Phosphatase and phytase activities in nodules of common bean genotypes at different levels of phosphorus supply

Plants grown at limited P supply can increase the activity of phosphatases in roots to hydrolyse organic-P compounds in the soil thus improving plant P acquisition, but little information is available about the role of these enzymes for internal plant metabolism at limited-P conditions. This work intended to measure the activities of acid phosphatases and phytases in nodules of common bean (Phaseolus vulgaris) genotypes at different levels of P supply. The experiment was carried out in a 5 × 5 factorial design with four replicates, comprising five bean genotypes and five P levels (20, 40, 80, 160 and 320 μmol P plant−1 week−1) in nutrient solution. Root seedlings were inoculated with Rhizobium tropici and plants were grown in 1-l bottles. Nodule samples were detached from 39-day-old plants and enzyme activities were determined in crude extracts. Plants were harvested at the stage of pod setting. Polynomial models fitted to data indicated maximal values at the level of 194 μmol P for shoot mass, at 206 μmol P for nodule mass and at 221 μmol P for shoot N. Whereas shoot mass was 1.7 times lower at 20 than at 160 μmol P, nodule mass was 7.5 times lower. Concentration of P in nodules increased from 40 to 320 μmol P but remained stable between 20 and 40 μmol P, suggesting a minimal threshold concentration of 3 mg P g−1 for nodule growth. Activities of phosphatases and phytases in nodules decreased strongly as P supply was raised from 20 to 80 μmol P, remaining almost stable at higher P levels. Phosphatase activity ranged from 1,154 to 406 nmol min−1 g−1 (nodule fresh mass) from 20 to 80 μmol P respectively, while the phytase activity ranged from 55 to 14 nmol min−1 g−1 from 20 to 80 μmol P. Bean genotypes differed in shoot and nodule mass at the levels of 80 and 160 μmol P, whilst they differed in nodule enzyme activities only at the lowest P level, the relationship between nodule enzyme activities and growth of different bean genotypes was not evident. It is concluded that bean plants at P-deficient conditions increase the activities of phosphatases and phytases in nodules. This may constitute an adaptive mechanism for N2-fixing legumes to tolerate P deficiency, by increasing the utilisation of the scarce P within the nodules.

Ascorbate and Homoglutathione Metabolism in Common Bean Nodules under Stress Conditions and during Natural Senescence

Ascorbate and glutathione are major antioxidants and redox buffers in plant cells but also play key functions in growth, development, and stress responses. We have studied the regulation of ascorbate and homoglutathione biosynthesis in common bean (Phaseolus vulgaris) nodules under stress conditions and during aging. The expression of five genes of the major ascorbate biosynthetic pathway was analyzed in nodules, and evidence was found that L-galactono-1,4-lactone dehydrogenase, the last committed step of the pathway, is posttranscriptionally regulated. Also, in nodules under stress conditions, gamma-glutamylcysteine synthetase was translationally regulated, but homoglutathione synthetase (mRNA and activity) and homoglutathione (content and redox state) were not affected. Most interestingly, in nodules exposed to jasmonic acid, dehydroascorbate reductase activity was posttranslationally suppressed, ascorbate oxidase showed strong transcriptional up-regulation, and dehydroascorbate content increased moderately. These changes were not due to a direct effect of jasmonic acid on the enzyme activities but might be part of the signaling pathway in the response of nodules to stress. We determined ascorbate, homoglutathione, and ascorbate-glutathione pathway enzyme activities in two senescing stages of nodules undergoing oxidative stress. When all parameters were expressed on a nodule fresh weight basis, we found that in the first stage ascorbate decreased by 60% and homoglutathione and antioxidant activities remained fairly constant, whereas in the second stage ascorbate and homoglutathione, their redox states, and their associated enzyme activities significantly decreased. The coexistence in the same plants of nodules at different senescence stages, with different ascorbate concentrations and redox states, indicates that the life span of nodules is in part controlled by endogenous factors and points to ascorbate as one of the key players.

Growth, nitrogen fixation and ammonium assimilation in common bean (Phaseolus vulgaris L) subjected to iron deficiency

A greenhouse experiment was carried out aiming to study the effect of iron deficiency on nitrogen fixation and ammonium assimilation in common bean nodules. Host-plant and nodule growth, symbiotic nitrogen fixation, glutamine synthetase (GS) and glutamate dehydrogenase (GDH) were analyzed in two common bean varieties subjected to iron deficiency. Results showed that host-plant and nodules growth, nitrogen fixation and GS activity decreased when under Fe-deficiency against an important increase of ammonium accumulation and GDH activity. Tolerant variety Flamingo is clearly less affected by iron deficiency than the sensitive one, Coco blanc. The allocation of iron to nodules and Fe use-efficiency for nodule growth and symbiotic nitrogen fixation were on the basis of the symbiotic performance of Flamingo under iron deprivation. Under Fe-deficiency, GDH take over GS the ammonium assimilation activity, particularly in the tolerant variety.

Composição de exopolissacarídeos produzidos por estirpes de rizóbios cultivados em diferentes fontes de carbono

O objetivo deste trabalho foi avaliar o efeito de diferentes fontes de carbono na composição dos exopolissacarídeos produzidos por Rhizobium tropici SEMIA 4077 e SEMIA 4080, e o efeito da inoculação dessas estirpes na nodulação em feijoeiro. As fontes de carbono utilizadas interferiram nas concentrações de galactose e glicose dos exopolissacarídeos, que foram maior na SEMIA 4080 em meio com sacarose. Embora a SEMIA 4080 tenha liberado grande quantidade de exopolissacarídeos na cultura, não houve diferença entre as estirpes quanto ao número de nódulos do feijoeiro, independentemente da fonte de carbono utilizada no preparo do inóculo. No parâmetro matéria seca de nódulos, houve diferença entre as estirpes, independentemente da fonte de carbono utilizada no preparo do inóculo.

BIOLOGICAL N2 FIXATION AND MINERAL N IN COMMON BEAN–MAIZE INTERCROPPING OR SOLE CROPPING IN SOUTHEASTERN BRAZIL

The aim of this work was to assess the effects of N levels and intercropping of two common bean (Phaseolus vulgaris) varieties (Carioca and Rio Tibagi) with maize (Zea mays) on accumulated N, grain yield and biomass of both crops, and nodulation of common bean inoculated or not with Rhizobium. Two field experiments were conducted simultaneously: common bean–maize intercropping and common bean in sole cropping. Intercropping increased common bean nodulation and biomass, mainly with Rhizobium, but mineral N was deleterious to nodulation. Inoculation also increased the Carioca cultivar yield most in sole cropping (+72%), but N levels did not affect common bean yield in either cultivar. Although intercropping reduced maize grain yield by 17%, the equivalent yield increased by 31%, whilst Rhizobium increased it by 11%. Despite the reduction of maize yield in intercropping, this system was shown to be more economically viable, in particular when common bean was inoculated with Rhizobium.