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The effects of salt stress on the growth, nodulation, and nitrogen (N) fixation of legumes are well known, but the relationship between symbiotic nitrogen fixation (SNF) driven by rhizobium–legume symbiosis and salt tolerance in Medicago truncatula is not well studied. The effects of the active nodulation process on salt stress tolerance of Medicago truncatula were evaluated by quantifying the compatible solutes, soluble sugars, and antioxidants enzymes, as well as growth and survival rate of plants. Eight weeks old plants, divided in three groups: (i) no nodules (NN), (ii) inactive nodules (IN), and (iii) active nodules (AN), were exposed to 150 mM of NaCl salt stress for 0, 8, 16, 24, 32, 40, and 48 h in hydroponic system. AN plants showed a higher survival rate (30.83% and 38.35%), chlorophyll contents (37.18% and 44.51%), and photosynthesis compared to IN and NN plants, respectively. Improved salt tolerance in AN plants was linked with higher activities of enzymatic and nonenzymatic antioxidants and higher K+ (20.45% and 39.21%) and lower Na+ accumulations (17.54% and 24.51%) when compared with IN and NN plants, respectively. Additionally, higher generation of reactive oxygen species (ROS) was indicative of salt stress, causing membrane damage as revealed by higher electrolyte leakage and lipid peroxidation. All such effects were significantly ameliorated in AN plants, showing higher compatible solutes (proline, free amino acids, glycine betaine, soluble sugars, and proteins) and maintaining higher relative water contents (61.34%). This study advocates positive role of Rhizobium meliloti inoculation against salt stress through upregulation of antioxidant system and a higher concentration of compatible solutes.

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Global warming negatively affects plant growth due to the detrimental effects of high temperature-induced heat stress. Rhizobium inoculation (RI) and exogenous melatonin (MT) have shown a positive role in resisting abiotic stress. However, their synergistic effect on avoiding heat-induced damages in Medicago truncatula has not been studied yet. Hence, the objective of the present study was to evaluate the impact of these amendments (RI and MT) to ameliorate the heat damages in Medicago truncatula. The study was comprised of two factors: (1) heat-induced stress: (i) optimum temperature (26 ± 1°C): (23 ± 1°C) (day: night), (ii) moderate heat (35 ± 1°C): (28 ± 1°C), and (iii) severe heat (41 ± 1°C): (35 ± 1°C) for 72 h, and (2) amendments: (i) no RI + no MT (NRI + NMT), (ii) Rhizobium inoculation (RI), (iii) 60 μM melatonin (MT), and (iii) RI + MT. Results showed that the combined application of RI and MT was better than their individual applications, as it prevented heat-induced membrane damages by declining the hydrogen peroxide (34.22% and 29.78%), superoxide anion radical (29.49% and 26.71%), malondialdehyde contents (26.43% and 21.96%), and lipoxygenase activity (44.75% and 25.51%) at both heat stress levels as compared to NRI + NMT. Moreover, RI + MT treated plants showed higher antioxidative and methylglyoxal detoxification enzymes (Gly I and Gly II) activities under heat stress. While, NRI + NMT treated plants showed a higher level of methylglyoxal contents (47.99% and 46.71%) under both levels of heat stress. Relative to NRI + NMT plants, RI + MT pretreated plants exhibited improved heat tolerance as indicated by higher chlorophyll (37.42% and 43.52%), carotenoid contents (32.41% and 47.08%), and photosynthetic rate (42.62% and 64.63%), under moderate and severe heat stress, respectively. Furthermore, RI + MT pretreated plants had considerably higher indole-3 acetic acid and abscisic acid concentrations under moderate (54.02% and 53.92%) and severe (68.36% and 64.61%) heat stress conditions. Similarly, plant dry biomass, NPK uptake, nitric oxide, and nitrate reductase activity were high in RI + MT treated plants, under both levels of stress. Therefore, this study advocates the positive synergistic effect of RI and MT pretreatment against moderate and severe heat-induced stress and for possible maintenance of plant growth under changing scenarios of global warming.

Biological nitrogen fixation (BNF) in soybean [Glycine max (L.) Merr.] is an important process to supply nitrogen (N) for the crop. However, the N fixation capacity of soybean varies widely among soils and climatic conditions. The N fixation potential is purportedly minimum in soils that do not have any prior history of soybean cultivation, and the application of inoculum is reported as necessary to facilitate this process. Rhizobium inocula can be applied at planting to promote BNF; however, their efficacy remains highly variable. A field experiment was conducted at irrigated and dryland sites in 2017 and 2018, where soybean was not cultivated in the preceding 10+ years. Bradyrhizobium inoculum was applied in liquid, dry, and a combination of liquid and dry formulations. The results showed that inoculum addition, in any formulation or combination, did not significantly increase soybean yield compared with the untreated control at any site. This study showed that inoculum application did not consistently increase yield in fields without soybean in the previous crop rotation for years. Future research is needed to identify soils, varieties, management practices, and environmental conditions for soybean inoculation which can increase yield and net farm profits.