Noel A. Tejera

Nitrogenase and antioxidant enzyme activities in Phaseolus vulgaris nodules formed by Rhizobium tropici isogenic strains with varying tolerance to salt stress

Common bean plants inoculated with salt-tolerant Rhizobium tropici wild-type strain CIAT899 formed a more active symbiosis than did its decreased salt-tolerance (DST) mutant derivatives (HB8, HB10, HB12 and HB13). The mutants formed partially effective (HB10, HB12) or almost ineffective (HB8, HB13) nodules (Fix(d)) under non-saline conditions. The DST mutant formed nodules that accumulated more proline than did the wild-type nodules, while soluble sugars were accumulated mainly in ineffective nodules. Under salt stress, plant growth, nitrogen fixation, and the activities of the antioxidant defense enzymes of nodules were affected in all symbioses tested. Overall, mutant nodules showed lower antioxidant enzyme activities than wild-type nodules. Levels of nodule catalase appeared to correlate with symbiotic nitrogen-fixing efficiency. Superoxide dismutase and dehydroascorbate reductase seem to function in the molecular mechanisms underlying the tolerance of nodules to salinity.

Isolation and characterization of Azotobacter and Azospirillum strains from the sugarcane rhizosphere

Bacteria with the ability to grow on nitrogen-free media and with nitrogenase activity under aerobic or microaerobic conditions were isolated from sugarcane roots collected from four different agricultural locations in Granada (Spain). Isolates were Gram negative rods and were identified as Azotobacter chroococcum and Azospirillum brasilense. Our results suggest that Azotobacter isolates do not have a particular affinity for sugarcane rhizospheres and that, on the contrary, Azospirillum isolates show specific association and perhaps endophytic colonization of sugarcane. However, obligate endophytes (Gluconacetobacter diazotrophicus) were not found in the apoplastic fluid of the stems and macerates extracts of sugarcane tissues with the procedure applied. Population of this microorganism might be in low number in the Spanish sugarcane varieties studied which is also discussed.

Alleviation of Salt Stress in Common Bean (Phaseolus vulgaris) by Exogenous Abscisic Acid Supply

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.

Trehalose and trehalase in root nodules of Medicago truncatula and Phaseolus vulgaris in response to salt stress.

Trehalose (alpha-D-glucopyranosyl-1,1-alpha-D-glucopyranoside), a non-reducing disaccharide, has been found in a wide variety of organisms playing an important role as an abiotic stress protectant. Plants may come into contact with trehalose from exogenous sources, such as in plant-rhizobia symbiosis in which the rhizobia have the capacity to produce trehalose. The aim of this work is to analyse how trehalose and trehalase respond to salt stress in root nodules of legumes. For this purpose, tissue expression of Medicago truncatula trehalase gene (MTTRE1) and the expression of MTTRE1 under salt stress were analysed by real-time quantitative reverse transcription-PCR method. Trehalase activity was determined and trehalose was also measured by gas chromatography. In addition, trehalase protein occurrence in different organs and at different developmental stages in Phaseolus vulgaris plants has been studied. MTTRE1 expression is induced in nodules compared with leaves and roots, indicating a transcriptional regulation of trehalase in the presence of the microsymbiont. Under salt stress conditions, trehalase activity is downregulated at the transcriptional level, allowing trehalose accumulation. The results found in this study led us to conclude that trehalase activity is induced in root nodules of legumes by the microsymbiont and that under salt stress conditions; trehalase activity is downregulated at the transcriptional level in M. truncatula nodules. This allows trehalose accumulation and supports the possible role of this disaccharide as a stabilizer against salt stress conditions.

Effect of Sodium Chloride on Growth, Nutrient Accumulation, and Nitrogen Fixation of Common Bean Plants in Symbiosis with Isogenic Strains

ABSTRACT The effect of sodium chloride (NaCl)-salinity on growth responses and tissues organic solutes and mineral content was investigated in common bean plants inoculated with salt-tolerant Rhizobium tropici wild-type strain CIAT899 and four mutant derivatives having decreased salt-tolerance (DST). Under non-saline conditions two mutants formed partially effective (HB10, HB12) and another two almost ineffective (HB8, HB13) nodules. A great variation of NaCl tolerance in the different symbiosis tested was observed at harvest, 32 day after planting. Common bean plant responded to salinity by decreasing the content of dry plant biomass, nodule number and the nitrogen fixation, and increasing the root to shoot ratio. The salt dose of 25 mM produced an increase of total soluble sugar and free amino acids content. This result suggest that these metabolites might be related with a nodule osmotic adjustment response under saline conditions, however cannot be excluded that the increase of amino acids content could be a consequence of protein degradation. In the other hand, sodium, calcium and phosphorus contents in shoot increased under the saline treatments. Potassium (K) and calcium (Ca) contents, unlike phosphorus (P) content, in shoot were not related with the symbiotic efficiency of mutant, however the decrease of P content suggest that these symbioses have limited their P absorption process independently of the saline treatment. NaCl tolerance associated with a retention of sodium and maintenance of potassium selectivity seem to be a strategy used for the salt stressed common bean plants in symbiosis assayed here.

Salicylic acid improves the salinity tolerance of Medicago sativa in symbiosis with Sinorhizobium meliloti by preventing nitrogen fixation inhibition

In this work we have investigated the contribution of pretreatment with 0.1 and 0.5mM salicylic acid (SA) to the protection against salt stress in root nodules of Medicago sativa in symbiosis with Sinorhizobium meliloti. SA alleviated the inhibition induced by salinity in the plant growth and photosynthetic capacity of M. sativa-S. meliloti symbiosis. In addition, SA prevented the inhibition of the nitrogen fixation capacity under salt stress since nodule biomass was not affected by salinity in SA pretreated plants. Antioxidant enzymes peroxidase (POX), superoxide dismutase (SOD), ascorbate peroxidase (APX), dehidroascorbate reductase (DHAR) and glutathione reductase (GR), key in the main pathway that scavenges H2O2 in plants, were induced by SA pretreatments which suggest that SA may participate in the redox balance in root nodules under salt stress. Catalase activity (CAT) was inhibited around 40% by SA which could be behind the increase of H2O2 detected in nodules of plants pretreated with SA. The accumulation of polyamines (PAs) synthesized in response to salinity was prevented by SA which together with the induction of 1-aminocyclopropane-l-carboxylic acid (ACC) content suggest the prevalence of the ethylene signaling pathway induced by SA in detriment of the synthesis of PAs. In conclusion, SA alleviated the negative effect of salt stress in the M. sativa-S. meliloti symbiosis through the increased level of nodule biomass and the induction of the nodular antioxidant metabolism under salt stress. The H2O2 accumulation and the PAs inhibition induced by SA in nodules of M. sativa suggest that SA activates a hypersensitive response dependent on ethylene.

Validamycin A improves the response of Medicago truncatula plants to salt stress by inducing trehalose accumulation in the root nodules

In this work, the role of trehalose as an osmoprotectant against salt stress conditions was examined in root nodules of Medicago truncatula. For this purpose, we used validamycin A, a potent trehalase inhibitor, in order to induce trehalose accumulation. Validamycin A induced an increase of trehalose concentration in root nodules of M. truncatula by inhibiting trehalase activity; no effect on trehalose concentration was observed in roots and leaves. Trehalose accumulation was accompanied by a decrease in sucrose and starch content, indicating interference with carbohydrate partitioning in the plants. Under salinity conditions, sucrose accumulation appears to be induced in M. truncatula to protect nodule functioning by the inhibition of sucrose catabolism by sucrose synthase and alkaline invertase activities. However, trehalose accumulation induced by val A in nodules improved the response to salinity by increasing plant dry weight (PDW), and no effects of validamycin A on nitrogenase activity and PDW were observed in nonsalinized plants.

Bioconcentration Factors and Trace Elements Bioaccumulation in Sporocarps of Fungi Collected from Quartzite Acidic Soils

The content of 19 metals (Al, V, Cr, Co, Ni, Cu, Zn, Ga, Rb, Sr, Zr, Nb, Cs, Ba, Ce, Pb, Th, U and Nd) was investigated in 15 edible species of phylum Basidiomycota collected in an area with quartzite acidic soils in a province of the central Spain. The study explores the differences in metal accumulation in relation to fungal species, and the results were related to metal content in soil through the determination of bioconcentration factors. Regarding the highest concentrations, Zn, Al, Cu and Rb were the metals more accumulated in the sporocarps. Notable concentrations were also found in Sr, Zr, Ba, Cs and Ce. The major bioconcentration factors were found for Cu and Zn in sporocarps of Agaricus silvicola and Lepista nuda. Regarding the different species, Tricholoma equestre and Cantharellus cibarius were those with the greatest capacity to absorb trace elements, and in contrast, Amanita caesarea and Agaricus campestris showed the lowest values. The cluster analysis shows that there are some species with the same nutritive physiology that share similarities in the absorptive behaviour. Lactarius sanguifluus and Lactarius deliciosus, both ectomycorrhizas of the genus Pinus, are closely related, and Clitocybe gibba, L. nuda and Marasmius oreades, all of them saprobes on soil organic matter, are very close too.

Effect of some abiotic factors on the biological activity of Gluconacetobacter diazotrophicus

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.

Involvement of abscisic acid in the response of Medicago sativa plants in symbiosis with Sinorhizobium meliloti to salinity.

Legumes are classified as salt-sensitive crops with their productivity particularly affected by salinity. Abcisic acid (ABA) plays an important role in the response to environmental stresses as signal molecule which led us to study its role in the response of nitrogen fixation and antioxidant metabolism in root nodules of Medicago sativa under salt stress conditions. Adult plants inoculated with Sinorhizobium meliloti were treated with 1 μM and 10 μM ABA two days before 200 mM salt addition. Exogenous ABA together with the salt treatment provoked a strong induction of the ABA content in the nodular tissue which alleviated the inhibition induced by salinity in the plant growth and nitrogen fixation. Antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR) were induced by ABA pre-treatments under salt stress conditions which together with the reduction of the lipid peroxidation, suggest a role for ABA as signal molecule in the activation of the nodular antioxidant metabolism. Interaction between ABA and polyamines (PAs), described as anti-stress molecules, was studied being detected an induction of the common polyamines spermidine (Spd) and spermine (Spm) levels by ABA under salt stress conditions. In conclusion, ABA pre-treatment improved the nitrogen fixation capacity under salt stress conditions by the induction of the nodular antioxidant defenses which may be mediated by the common PAs Spd and Spm that seems to be involved in the anti-stress response induced by ABA.