Fifteen genotypes of faba bean (Vicia faba L.) were inoculated with salt-tolerant Rhizobium legum... more Fifteen genotypes of faba bean (Vicia faba L.) were inoculated with salt-tolerant Rhizobium leguminosarum biovar. viciae strain GRA 19 in solution culture with 0 (control) and 75 mM NaCl added immediately after transplanting. Genotypes varied in their tolerance of high levels of NaCl. Physiological parameters (dry weight of shoot and root, number and dry weight of nodules) were not affected by salinity in lines VF46, VF64 and VF112. Faba bean line VF60 was sensitive to salt stress. Host tolearance appeared to be a major requisite for nodulation and N2 fixation under salt stress. Tolerant line VF112 sustained nitrogen fixation under saline conditions. Activity of the ammonium assimilation enzymes glutamine synthetase and glutamate synthase, and soluble protein content, were reduced by salinity in all genotypes tested. Evidence presented here suggests a need to select faba bean genotypes that are tolerant to salt stress.
The effect of salinity on growth response, nitrogen (N) fixation and tissue mineral content was i... more The effect of salinity on growth response, nitrogen (N) fixation and tissue mineral content was investigated for four legumes: faba bean (Vicia faba L), pea (Pisum sativum L), soybean (Glycine max L), and common bean (Phaseolus vulgaris L). Plants were grown in a vermiculite culture system supplied with a N‐free nutrient solution with the addition of 0, 50, and 100 mM NaCl. Plants were harvested at the beginning of the flowering period and the dry weights of shoots and roots and acetylene reduction activity (ARA) were evaluated at the same time plant tissues were analysed for N, potassium (K), calcium (Ca), magnesium (Mg), and sodium (Na) contents.The depressive effect of saline stress on ARA of nodules was directely related to the salt induced decline in dry weight and N content in shoots. Growth inhibition by NaCl treatments was greater for the pea than for other legumes, whereas the soybean was the most salt‐tolerant Saline stress also affected the N content in shoots and roots. In general the N content accumulated in the shoot and Na in the roots of the four legumes tested, while K accumulated both organs. The acquisition of other macronutrients differed according to the legume species. The legumes most sensitive were P. sativum and V. faba which accumulated Ca in shoot and Mg both in the shoot and the roots. On the contrary, in G. max and P. vulgaris, the two most salt tolerant legumes, accumulated Mg in the roots and Ca in both vegetative organs. Our results suggest a relationship between the salt‐tolerant range in legumes and the macronutrient accumulation in vegetative organs.
Lotus japonicus and Medicago truncatula model legumes, which form determined and indeterminate no... more Lotus japonicus and Medicago truncatula model legumes, which form determined and indeterminate nodules, respectively, provide a convenient system to study plant–Rhizobium interaction and to establish differences between the two types of nodules under salt stress conditions. We examined the effects of 25 and 50 mM NaCl doses on growth and nitrogen fixation parameters, as well as carbohydrate content and carbon metabolism of M. truncatula and L. japonicus nodules. The leghemoglobin (Lb) content and nitrogen fixation rate (NFR) were approximately 10.0 and 2.0 times higher, respectively, in nodules of L. japonicus when compared with M. truncatula. Plant growth parameters and nitrogenase activity decreased with NaCl treatments in both legumes. Sucrose was the predominant sugar quantified in nodules of both legumes, showing a decrease in concentration in response to salt stress. The content of trehalose was low (less than 2.5% of total soluble sugars (TSS)) to act as an osmolyte in nodules, despite its concentration being increased under saline conditions. Nodule enzyme activities of trehalose-6-phosphate synthase (TPS) and trehalase (TRE) decreased with salinity. L. japonicus nodule carbon metabolism proved to be less sensitive to salinity than in M. truncatula, as enzymatic activities responsible for the carbon supply to the bacteroids to fuel nitrogen fixation, such as sucrose synthase (SS), alkaline invertase (AI), malate dehydrogenase (MDH) and phosphoenolpyruvate carboxylase (PEPC), were less affected by salt than the corresponding activities in barrel medics. However, nitrogenase activity was only inhibited by salinity in L. japonicus nodules.
Phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) was purified 56-fold from Vicia faba root nod... more Phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) was purified 56-fold from Vicia faba root nodules to a specific activity of 24.8 units mg-1 protein. Native molecular mass was determined to be 443 kDa by gel permeation chromatography, whereas a molecular mass of 113 kDa was obtained for the subunit by means of SDS-PAGE, indicating that the enzyme is a homotetramer. One peak of activity was obtained by ion-exchange chromatography or gel filtration, and thus there was no evidence of isoenzymes. The effect of pH on PEPC activity was studied, the pH optimum found at 8.25. The effect of substrate (phosphoenolpyruvate, PEP) on the enzyme activity was studied at five different pH values from 6.5 to 9.5. The Km(PEP) at pH 8.25 proved to be 0.064 mM. Inhibition by malate or activation by glucose-6-phosphate was dependent on the pH of the reaction mixture. Malate behaved as a non-competitive mixed-type inhibitor with a Ki of 0.76 mM, a Ki(s) of 1.15 mM and a Ki(i) of 0.72 mM, at pH 7.0 while at pH 8.25 Ki was about 140 mM. Activation by glucose-6-P was 70% with 4 mM PEP at pH 7, whereas no effect was found at pH 8.25. Experiments with mixed effectors at pH 7 and 1 mM PEP, showed that glucose-6-P can reverse the inhibition caused by L-malate on the PEPC activity.
Aims: The effect of some abiotic factors, dryness, heat and salinity on the growth and biological... more Aims: The effect of some abiotic factors, dryness, heat and salinity on the growth and biological activity of Gluconacetobacter diazotrophicus, and the influence of a salt stress on some enzymes involved in carbon metabolism of these bacteria is studied under laboratory conditions.Methods and Results: Strain PAL-5 of G. diazotrophicus was incubated under different conditions of drying, heat and salinity. Cells showed tolerance to heat treatments and salt concentrations, and sensitivity to drying conditions. Higher NaCl dosage of 150 and 200 mmol l −1 limited its growth and drastically affected the nitrogenase activity and the enzymes glucose dehydrogenase, alcohol dehydrogenase, fumarase, isocitrate dehydrogenase and malate dehydrogenase.Conclusions:Gluconacetobacter diazotrophicus, despite its endophytic nature, tolerated heat treatments and salinity stress, but its nitrogenase activity and carbon metabolism enzymes were affected by high NaCl dosage.Significance and Impact of the Study: The investigation of the biological activity of G. diazotrophicus in response to different abiotic factors led to more knowledge of this endophyte and may help to clarify pathways involved in its transmission into the host plant.
The effects of the interaction between sodium chloride, nitrate, and concentrations on growth and... more The effects of the interaction between sodium chloride, nitrate, and concentrations on growth and internal ion content of faba bean (Vicia faba L.) plants were studied, to understand the relationship between the above parameters and salt tolerance. Increased salinity substantially reduced the dry weight of roots and shoots and increased the root/shoot biomass ratio. Additional nitrate‐N considerably moderated the salinity effects on these parameters. The promotive effects of nitrate‐N were more pronounced on shoot dry weight. These results suggest that an exogenous supply of nitrate‐N would improve the vegetative growth of V. faba plants by moderating the suppresive effects of salinity. The evolution of the root and shoot content in potassium (K), sodium (Na), magnesium (Mg), calcium (Ca), and nitrogen (N) was monitored during vegetative growth. A high correspondence between total N and Ca content was found. The acquisition of Ca and K in response to salt and nitrate was similar in shoots and roots, whereas Mg uptake showed notable differences in the two organs. In salt‐affected plants, the roots were found to be high in accumulated Na while the shoots exhibited the lowest Na concentration. Potassium accumulation was higher in the shoots. In this way, there was an antagonistic effect between Na and K uptake. Analyses of the nutrient contents in plant organs have provided a data base on salt‐tolerance mechanisms of V. faba plants.
The effects of different carbon and nitrogen sources on the growth, nitrogenase activity, and car... more The effects of different carbon and nitrogen sources on the growth, nitrogenase activity, and carbon metabolism of Gluconacetobacter diazotrophicus were investigated. The amino acids asparagine, aspartic acid, and glutamic acid affected microbial growth and nitrogenase activity. Several enzymatic activities involved in the tricarboxylic acid cycle were affected by the carbon source used. In addition, glucose and gluconate significantly increased the oxygen consumption (respiration rate) of whole cells of G. diazotrophicus grown under aerobic conditions. Enzymes responsible for direct oxidation of glucose and gluconate were especially active in cells grown with sucrose and gluconate. The presence of amino acids in the apoplastic and symplastic sap of sugarcane stems suggests that these compounds might be of importance in the regulation of growth and nitrogenase activity during the symbiotic association. The information obtained from the plant-bacterium association together with the results of other biochemical studies could contribute to the development of biotechnological applications of G. diazotrophicus.
In this work the effect of abscisic acid (ABA) and 100 mM NaCl on common bean (Phaseolus vulgaris... more In this work the effect of abscisic acid (ABA) and 100 mM NaCl on common bean (Phaseolus vulgaris var. Coco) growth, nitrogenase activity, and nodule metabolism was studied. Experiments were carried out in a controlled environmental chamber and plants, at the vegetative growth stage (16 days old), were treated with ABA (1 μM and 10 μM) and 48 h later were exposed to saline treatment. Results revealed that plant dry weight, nodule dry weight, nitrogen fixation (acetylene reduction activity and ureides content), and most enzymes of ammonium and ureides metabolism were affected by both ABA and NaCl. The addition of 1 μM ABA to the nutrient solution before the exposure to salt stress reduced the negative effect of NaCl. Based on our results, we suggest that ABA application improves the response of Phaseolus vulgaris symbiosis under saline stress conditions, including the nitrogen fixation process and enzymes of ammonium assimilation and purine catabolism.
The main findings of a cooperative research group of agronomists, plant breeders, microbiologists... more The main findings of a cooperative research group of agronomists, plant breeders, microbiologists, physiologists and molecularists to improve the symbiotic nitrogen fixation (SNF) and N2-dependent yield of common bean under moderate salinity in the Mediterranean basin are summarised. Agronomic surveys in reference production areas show large spatial and temporal variations in plant nodulation and growth, and in efficiency of utilisation of the rhizobial symbiosis. The latter was associated with a large rhizobial diversity, including new bean nodulating species. Macrosymbiont diversity in SNF and adaptation to NaCl was found. However, contrasts between plant genotypes could be altered by specific interactions with some native rhizobia. Therefore, variations in soil rhizobial population, in addition to agronomic practices and environmental constraints, may have contributed to erratic results observed in field inoculations. At the mechanistic level, nodule C and N metabolisms, and abcissic acid content, were related to SNF potential and tolerance to NaCl. Their relation with nodule conductance to O2 diffusion was addressed by in situ hybridisation of candidate carbonic anhydrase and aquaporin genes in nodule cortex. The limits and prospects of the cooperative strategy are discussed.
Faba bean (Vicia faba L. var. minor cv. Alborea) and pea (Pisum sativum L. cv. Lincoln) plants, i... more Faba bean (Vicia faba L. var. minor cv. Alborea) and pea (Pisum sativum L. cv. Lincoln) plants, inoculated with Rhizobium leguminosarum biovar. viciae strain GRA19, were treated with salt (100 mM NaCl) and/or nitrate (8 mM KNO3) to test whether plants grown with inorganic-nitrogen are more tolerant to salinity than plants entirely reliant upon fixed nitrogen. According to the growth inhibition recorded, pea plants dependent on dinitrogen fixation proved more tolerant to salt stress than those N-fertilized, in contrast to results obtained for faba bean plants. This study therefore confirms that plants dependent on nitrogen fixation are not always more sensitive to salinity than are N-fertilized plants. Nitrate addition did not reduce the specific nitrogenase activity in pea, but did in faba bean. However, nodulation was inhibited in both legumes. The specific nitrogenase activity was more affected by salt treatment in N-fertilized plants for both legumes. The activity of the enzymes mediating ammonium assimilation in nodules (GS, NADH-GOGAT) was inhibited by salt stress both in N-fixing and in N-fertilized pea and faba bean plants.
Alterations of plant growth, nitrogenase activity and nutrient concentration as a consequence of ... more Alterations of plant growth, nitrogenase activity and nutrient concentration as a consequence of salt treatments were studied in five chickpea (Cicer arietinum L.) cultivars from Spain and Syria. Plants, in symbiosis with Mesorhizobium ciceri ch-191 strain, were grown under controlled conditions for 32 days and subjected to salinity stress. Parameters of growth and nitrogen fixation were affected under salt stress in all cultivars tested; plant dry weight decreased by about 15% in ILC1919; and in Sirio and Lechoso about 50% with the highest salt dosage (100 mM NaCl). ILC1919 showed a less growth accompanied by a lower dry matter formation under low salt conditions compared with most of the salt sensitive cultivar. Nitrogenase activity decreased by about 60% in the salt-resistant cultivar (cv. ILC1919) and more than 90% in salt-sensitive cultivars (cv. Sirio and Lechoso) with the highest salt dosage during the reproductive growth. We show that the higher NaCl tolerance of the ILC1919 cultivar is supported by the less N2 fixation inhibition, a higher root-to-shoot ratio, normalized nodule weight and shoot K/Na ratio; and a reduced foliar accumulation of Na+. Moreover, our results reveal the effectiveness of these nutritional and physiological indicators in the selection of salinity-tolerant chickpea plants growing under symbiotic conditions.
... Urban & Fischer Verlag Published by Elsevier GmbH, Cited By in Scopus (19). Permissio... more ... Urban & Fischer Verlag Published by Elsevier GmbH, Cited By in Scopus (19). Permissions & Reprints. Nitrogenase and antioxidant enzyme activities in Phaseolus vulgaris nodules formed by Rhizobium tropici isogenic strains with varying tolerance to salt stress. Noel A. Tejera a ...
A commercial cultivar (Alborea) of faba bean (Vicia faba L. var. minor) was inoculated with salt-... more A commercial cultivar (Alborea) of faba bean (Vicia faba L. var. minor) was inoculated with salt-tolerant Rhizobium leguminosarum biovar. viciae strain GRA19 in solution culture with different salt concentrations (0, 50, 75 and 100 mmoles l−1 NaCl) added immediately at the time of inoculation. The results indicated that Rhizobium leguminosarum strain GRA19 formed an infective and effective symbiosis with faba bean under saline and nonsaline conditions. Salinity significantly decreased shoot and root dry weight, nodule weight and mean nodule weight. Roots were more sensitive than shoots, and N2 fixation was more sensitive to salinity than was plant growth. Analyses of ammonium assimilating enzymes in the nodule showed that glutamine synthetase appeared to be more tolerant to salinity than glutamate synthase, and that it limits ammonium assimilation under saline stress.
Aims: To characterize the physiological and metabolic responses of Mesorhizobium ciceri strain ch... more Aims: To characterize the physiological and metabolic responses of Mesorhizobium ciceri strain ch-191 to salt stress, investigating the changes induced by salinity in protein and lipopolysaccharide profiles, as well as determining the accumulation of amino acids, glutamate and proline.Methods and Results: Strain ch-191 of M. ciceri was grown with different NaCl concentrations. Protein and lipopolysaccharide patterns were determined by electrophoresis. The strain ch-191 tolerated up to 200 mmol l−1 NaCl, although higher salt dosages limited its growth and induced changes in the protein profile. The most noteworthy change in the LPS-I pattern was the decrease in the slowest band and the appearance of an intermediate mobility band. The accumulation of proline in response to salt stress surpassed that of glutamate.Conclusions: The protein profile showed major alterations at salinity levels which inhibited growth. However, the alterations in the LPS profile and accumulation of compatible solutes were evident from the lowest levels, suggesting that these changes may constitute adaptative responses to salt, allowing normal growth.Significance and Impact of the Study: The selection and characterization of salt-tolerant strains, which also show efficient symbiotic performance under salinity, may constitute a strategy for improving Cicer arietinum–Mesorhizobium ciceri symbiosis in adverse environments.
Fifteen genotypes of faba bean (Vicia faba L.) were inoculated with salt-tolerant Rhizobium legum... more Fifteen genotypes of faba bean (Vicia faba L.) were inoculated with salt-tolerant Rhizobium leguminosarum biovar. viciae strain GRA 19 in solution culture with 0 (control) and 75 mM NaCl added immediately after transplanting. Genotypes varied in their tolerance of high levels of NaCl. Physiological parameters (dry weight of shoot and root, number and dry weight of nodules) were not affected by salinity in lines VF46, VF64 and VF112. Faba bean line VF60 was sensitive to salt stress. Host tolearance appeared to be a major requisite for nodulation and N2 fixation under salt stress. Tolerant line VF112 sustained nitrogen fixation under saline conditions. Activity of the ammonium assimilation enzymes glutamine synthetase and glutamate synthase, and soluble protein content, were reduced by salinity in all genotypes tested. Evidence presented here suggests a need to select faba bean genotypes that are tolerant to salt stress.
The effect of salinity on growth response, nitrogen (N) fixation and tissue mineral content was i... more The effect of salinity on growth response, nitrogen (N) fixation and tissue mineral content was investigated for four legumes: faba bean (Vicia faba L), pea (Pisum sativum L), soybean (Glycine max L), and common bean (Phaseolus vulgaris L). Plants were grown in a vermiculite culture system supplied with a N‐free nutrient solution with the addition of 0, 50, and 100 mM NaCl. Plants were harvested at the beginning of the flowering period and the dry weights of shoots and roots and acetylene reduction activity (ARA) were evaluated at the same time plant tissues were analysed for N, potassium (K), calcium (Ca), magnesium (Mg), and sodium (Na) contents.The depressive effect of saline stress on ARA of nodules was directely related to the salt induced decline in dry weight and N content in shoots. Growth inhibition by NaCl treatments was greater for the pea than for other legumes, whereas the soybean was the most salt‐tolerant Saline stress also affected the N content in shoots and roots. In general the N content accumulated in the shoot and Na in the roots of the four legumes tested, while K accumulated both organs. The acquisition of other macronutrients differed according to the legume species. The legumes most sensitive were P. sativum and V. faba which accumulated Ca in shoot and Mg both in the shoot and the roots. On the contrary, in G. max and P. vulgaris, the two most salt tolerant legumes, accumulated Mg in the roots and Ca in both vegetative organs. Our results suggest a relationship between the salt‐tolerant range in legumes and the macronutrient accumulation in vegetative organs.
Lotus japonicus and Medicago truncatula model legumes, which form determined and indeterminate no... more Lotus japonicus and Medicago truncatula model legumes, which form determined and indeterminate nodules, respectively, provide a convenient system to study plant–Rhizobium interaction and to establish differences between the two types of nodules under salt stress conditions. We examined the effects of 25 and 50 mM NaCl doses on growth and nitrogen fixation parameters, as well as carbohydrate content and carbon metabolism of M. truncatula and L. japonicus nodules. The leghemoglobin (Lb) content and nitrogen fixation rate (NFR) were approximately 10.0 and 2.0 times higher, respectively, in nodules of L. japonicus when compared with M. truncatula. Plant growth parameters and nitrogenase activity decreased with NaCl treatments in both legumes. Sucrose was the predominant sugar quantified in nodules of both legumes, showing a decrease in concentration in response to salt stress. The content of trehalose was low (less than 2.5% of total soluble sugars (TSS)) to act as an osmolyte in nodules, despite its concentration being increased under saline conditions. Nodule enzyme activities of trehalose-6-phosphate synthase (TPS) and trehalase (TRE) decreased with salinity. L. japonicus nodule carbon metabolism proved to be less sensitive to salinity than in M. truncatula, as enzymatic activities responsible for the carbon supply to the bacteroids to fuel nitrogen fixation, such as sucrose synthase (SS), alkaline invertase (AI), malate dehydrogenase (MDH) and phosphoenolpyruvate carboxylase (PEPC), were less affected by salt than the corresponding activities in barrel medics. However, nitrogenase activity was only inhibited by salinity in L. japonicus nodules.
Phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) was purified 56-fold from Vicia faba root nod... more Phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) was purified 56-fold from Vicia faba root nodules to a specific activity of 24.8 units mg-1 protein. Native molecular mass was determined to be 443 kDa by gel permeation chromatography, whereas a molecular mass of 113 kDa was obtained for the subunit by means of SDS-PAGE, indicating that the enzyme is a homotetramer. One peak of activity was obtained by ion-exchange chromatography or gel filtration, and thus there was no evidence of isoenzymes. The effect of pH on PEPC activity was studied, the pH optimum found at 8.25. The effect of substrate (phosphoenolpyruvate, PEP) on the enzyme activity was studied at five different pH values from 6.5 to 9.5. The Km(PEP) at pH 8.25 proved to be 0.064 mM. Inhibition by malate or activation by glucose-6-phosphate was dependent on the pH of the reaction mixture. Malate behaved as a non-competitive mixed-type inhibitor with a Ki of 0.76 mM, a Ki(s) of 1.15 mM and a Ki(i) of 0.72 mM, at pH 7.0 while at pH 8.25 Ki was about 140 mM. Activation by glucose-6-P was 70% with 4 mM PEP at pH 7, whereas no effect was found at pH 8.25. Experiments with mixed effectors at pH 7 and 1 mM PEP, showed that glucose-6-P can reverse the inhibition caused by L-malate on the PEPC activity.
Aims: The effect of some abiotic factors, dryness, heat and salinity on the growth and biological... more Aims: The effect of some abiotic factors, dryness, heat and salinity on the growth and biological activity of Gluconacetobacter diazotrophicus, and the influence of a salt stress on some enzymes involved in carbon metabolism of these bacteria is studied under laboratory conditions.Methods and Results: Strain PAL-5 of G. diazotrophicus was incubated under different conditions of drying, heat and salinity. Cells showed tolerance to heat treatments and salt concentrations, and sensitivity to drying conditions. Higher NaCl dosage of 150 and 200 mmol l −1 limited its growth and drastically affected the nitrogenase activity and the enzymes glucose dehydrogenase, alcohol dehydrogenase, fumarase, isocitrate dehydrogenase and malate dehydrogenase.Conclusions:Gluconacetobacter diazotrophicus, despite its endophytic nature, tolerated heat treatments and salinity stress, but its nitrogenase activity and carbon metabolism enzymes were affected by high NaCl dosage.Significance and Impact of the Study: The investigation of the biological activity of G. diazotrophicus in response to different abiotic factors led to more knowledge of this endophyte and may help to clarify pathways involved in its transmission into the host plant.
The effects of the interaction between sodium chloride, nitrate, and concentrations on growth and... more The effects of the interaction between sodium chloride, nitrate, and concentrations on growth and internal ion content of faba bean (Vicia faba L.) plants were studied, to understand the relationship between the above parameters and salt tolerance. Increased salinity substantially reduced the dry weight of roots and shoots and increased the root/shoot biomass ratio. Additional nitrate‐N considerably moderated the salinity effects on these parameters. The promotive effects of nitrate‐N were more pronounced on shoot dry weight. These results suggest that an exogenous supply of nitrate‐N would improve the vegetative growth of V. faba plants by moderating the suppresive effects of salinity. The evolution of the root and shoot content in potassium (K), sodium (Na), magnesium (Mg), calcium (Ca), and nitrogen (N) was monitored during vegetative growth. A high correspondence between total N and Ca content was found. The acquisition of Ca and K in response to salt and nitrate was similar in shoots and roots, whereas Mg uptake showed notable differences in the two organs. In salt‐affected plants, the roots were found to be high in accumulated Na while the shoots exhibited the lowest Na concentration. Potassium accumulation was higher in the shoots. In this way, there was an antagonistic effect between Na and K uptake. Analyses of the nutrient contents in plant organs have provided a data base on salt‐tolerance mechanisms of V. faba plants.
The effects of different carbon and nitrogen sources on the growth, nitrogenase activity, and car... more The effects of different carbon and nitrogen sources on the growth, nitrogenase activity, and carbon metabolism of Gluconacetobacter diazotrophicus were investigated. The amino acids asparagine, aspartic acid, and glutamic acid affected microbial growth and nitrogenase activity. Several enzymatic activities involved in the tricarboxylic acid cycle were affected by the carbon source used. In addition, glucose and gluconate significantly increased the oxygen consumption (respiration rate) of whole cells of G. diazotrophicus grown under aerobic conditions. Enzymes responsible for direct oxidation of glucose and gluconate were especially active in cells grown with sucrose and gluconate. The presence of amino acids in the apoplastic and symplastic sap of sugarcane stems suggests that these compounds might be of importance in the regulation of growth and nitrogenase activity during the symbiotic association. The information obtained from the plant-bacterium association together with the results of other biochemical studies could contribute to the development of biotechnological applications of G. diazotrophicus.
In this work the effect of abscisic acid (ABA) and 100 mM NaCl on common bean (Phaseolus vulgaris... more In this work the effect of abscisic acid (ABA) and 100 mM NaCl on common bean (Phaseolus vulgaris var. Coco) growth, nitrogenase activity, and nodule metabolism was studied. Experiments were carried out in a controlled environmental chamber and plants, at the vegetative growth stage (16 days old), were treated with ABA (1 μM and 10 μM) and 48 h later were exposed to saline treatment. Results revealed that plant dry weight, nodule dry weight, nitrogen fixation (acetylene reduction activity and ureides content), and most enzymes of ammonium and ureides metabolism were affected by both ABA and NaCl. The addition of 1 μM ABA to the nutrient solution before the exposure to salt stress reduced the negative effect of NaCl. Based on our results, we suggest that ABA application improves the response of Phaseolus vulgaris symbiosis under saline stress conditions, including the nitrogen fixation process and enzymes of ammonium assimilation and purine catabolism.
The main findings of a cooperative research group of agronomists, plant breeders, microbiologists... more The main findings of a cooperative research group of agronomists, plant breeders, microbiologists, physiologists and molecularists to improve the symbiotic nitrogen fixation (SNF) and N2-dependent yield of common bean under moderate salinity in the Mediterranean basin are summarised. Agronomic surveys in reference production areas show large spatial and temporal variations in plant nodulation and growth, and in efficiency of utilisation of the rhizobial symbiosis. The latter was associated with a large rhizobial diversity, including new bean nodulating species. Macrosymbiont diversity in SNF and adaptation to NaCl was found. However, contrasts between plant genotypes could be altered by specific interactions with some native rhizobia. Therefore, variations in soil rhizobial population, in addition to agronomic practices and environmental constraints, may have contributed to erratic results observed in field inoculations. At the mechanistic level, nodule C and N metabolisms, and abcissic acid content, were related to SNF potential and tolerance to NaCl. Their relation with nodule conductance to O2 diffusion was addressed by in situ hybridisation of candidate carbonic anhydrase and aquaporin genes in nodule cortex. The limits and prospects of the cooperative strategy are discussed.
Faba bean (Vicia faba L. var. minor cv. Alborea) and pea (Pisum sativum L. cv. Lincoln) plants, i... more Faba bean (Vicia faba L. var. minor cv. Alborea) and pea (Pisum sativum L. cv. Lincoln) plants, inoculated with Rhizobium leguminosarum biovar. viciae strain GRA19, were treated with salt (100 mM NaCl) and/or nitrate (8 mM KNO3) to test whether plants grown with inorganic-nitrogen are more tolerant to salinity than plants entirely reliant upon fixed nitrogen. According to the growth inhibition recorded, pea plants dependent on dinitrogen fixation proved more tolerant to salt stress than those N-fertilized, in contrast to results obtained for faba bean plants. This study therefore confirms that plants dependent on nitrogen fixation are not always more sensitive to salinity than are N-fertilized plants. Nitrate addition did not reduce the specific nitrogenase activity in pea, but did in faba bean. However, nodulation was inhibited in both legumes. The specific nitrogenase activity was more affected by salt treatment in N-fertilized plants for both legumes. The activity of the enzymes mediating ammonium assimilation in nodules (GS, NADH-GOGAT) was inhibited by salt stress both in N-fixing and in N-fertilized pea and faba bean plants.
Alterations of plant growth, nitrogenase activity and nutrient concentration as a consequence of ... more Alterations of plant growth, nitrogenase activity and nutrient concentration as a consequence of salt treatments were studied in five chickpea (Cicer arietinum L.) cultivars from Spain and Syria. Plants, in symbiosis with Mesorhizobium ciceri ch-191 strain, were grown under controlled conditions for 32 days and subjected to salinity stress. Parameters of growth and nitrogen fixation were affected under salt stress in all cultivars tested; plant dry weight decreased by about 15% in ILC1919; and in Sirio and Lechoso about 50% with the highest salt dosage (100 mM NaCl). ILC1919 showed a less growth accompanied by a lower dry matter formation under low salt conditions compared with most of the salt sensitive cultivar. Nitrogenase activity decreased by about 60% in the salt-resistant cultivar (cv. ILC1919) and more than 90% in salt-sensitive cultivars (cv. Sirio and Lechoso) with the highest salt dosage during the reproductive growth. We show that the higher NaCl tolerance of the ILC1919 cultivar is supported by the less N2 fixation inhibition, a higher root-to-shoot ratio, normalized nodule weight and shoot K/Na ratio; and a reduced foliar accumulation of Na+. Moreover, our results reveal the effectiveness of these nutritional and physiological indicators in the selection of salinity-tolerant chickpea plants growing under symbiotic conditions.
... Urban & Fischer Verlag Published by Elsevier GmbH, Cited By in Scopus (19). Permissio... more ... Urban & Fischer Verlag Published by Elsevier GmbH, Cited By in Scopus (19). Permissions & Reprints. Nitrogenase and antioxidant enzyme activities in Phaseolus vulgaris nodules formed by Rhizobium tropici isogenic strains with varying tolerance to salt stress. Noel A. Tejera a ...
A commercial cultivar (Alborea) of faba bean (Vicia faba L. var. minor) was inoculated with salt-... more A commercial cultivar (Alborea) of faba bean (Vicia faba L. var. minor) was inoculated with salt-tolerant Rhizobium leguminosarum biovar. viciae strain GRA19 in solution culture with different salt concentrations (0, 50, 75 and 100 mmoles l−1 NaCl) added immediately at the time of inoculation. The results indicated that Rhizobium leguminosarum strain GRA19 formed an infective and effective symbiosis with faba bean under saline and nonsaline conditions. Salinity significantly decreased shoot and root dry weight, nodule weight and mean nodule weight. Roots were more sensitive than shoots, and N2 fixation was more sensitive to salinity than was plant growth. Analyses of ammonium assimilating enzymes in the nodule showed that glutamine synthetase appeared to be more tolerant to salinity than glutamate synthase, and that it limits ammonium assimilation under saline stress.
Aims: To characterize the physiological and metabolic responses of Mesorhizobium ciceri strain ch... more Aims: To characterize the physiological and metabolic responses of Mesorhizobium ciceri strain ch-191 to salt stress, investigating the changes induced by salinity in protein and lipopolysaccharide profiles, as well as determining the accumulation of amino acids, glutamate and proline.Methods and Results: Strain ch-191 of M. ciceri was grown with different NaCl concentrations. Protein and lipopolysaccharide patterns were determined by electrophoresis. The strain ch-191 tolerated up to 200 mmol l−1 NaCl, although higher salt dosages limited its growth and induced changes in the protein profile. The most noteworthy change in the LPS-I pattern was the decrease in the slowest band and the appearance of an intermediate mobility band. The accumulation of proline in response to salt stress surpassed that of glutamate.Conclusions: The protein profile showed major alterations at salinity levels which inhibited growth. However, the alterations in the LPS profile and accumulation of compatible solutes were evident from the lowest levels, suggesting that these changes may constitute adaptative responses to salt, allowing normal growth.Significance and Impact of the Study: The selection and characterization of salt-tolerant strains, which also show efficient symbiotic performance under salinity, may constitute a strategy for improving Cicer arietinum–Mesorhizobium ciceri symbiosis in adverse environments.
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