Gas Exchanges and Biological Nitrogen Fixation in Soybean under Water Restriction

Biological nitrogen fixation (BNF) in soybean is vulnerable to drought;however, there are genotypic variations among soybean cultivars regarding the ability to keep BNF under moderate water restriction. The aim of this study was to evaluate parameters related to gas exchanges and regulation of BNF in soybean genotypes BNF drought-tolerant (R01-581F and R01-416F), or drought-susceptible (CD 215 and BRS 317), submitted to adequate water supply or restriction between 45 and 55 days after emergence. We searched for traits associated with tolerance/susceptibility to drought, which might be useful in the selection of drought-tolerant soybean genotypes. Plant biomass was not affected under water restriction, but the number and dry weight of nodules reduced by 33% and 12%, respectively, in the average of genotypes. Drought-tolerant genotypes were more effective in maintaining gas exchanges under water restriction. Under water restriction, all genotypes increased the concentration of ureides in nodules, but only the susceptible genotypes showed that in leaves. The maintenance of gas exchanges and N metabolism regulation under water restriction in genotype R01-581F suggests that these parameters may be used to characterize soybean genotypes that can be sources of drought tolerance in genetic breeding programs.

A high-resolution transcriptomic atlas depicting nitrogen fixation and nodule development in soybean

Although root nodules are essential for biological nitrogen fixation in legumes,the cell types and molecular regulatory mechanisms contributing to nodule development and nitrogen fixation in determinate nodule legumes,such as soybean(Glycine max),remain incompletely understood.Here,we generated a single-nucleus resolution transcriptomic atlas of soybean roots and nodules at 14 days post inoculation(dpi)and annotated 17 major cell types,including six that are specific to nodules.We identified the specific cell types responsible for each step in the ureides synthesis pathway,which enables spatial compartmentalization of biochemical reactions during soybean nitrogen fixation.By utilizing RNA velocity analysis,we reconstructed the differentiation dynamics of soybean nodules,which differs from those of indeterminate nodules in Medicago truncatula.Moreover,we identified several putative regulators of soybean nodulation and two of these genes,GmbHLH93 and GmSCL1,were as-yet uncharacterized in soybean.Overexpression of each gene in soybean hairy root systems validated their respective roles in nodulation.Notably,enrichment for cytokinin-related genes in soybean nodules led to identification of the cytokinin receptor,GmCRE1,as a prominent component of the nodulation pathway.GmCRE1 knockout in soybean resulted in a striking nodule phenotype with decreased nitrogen fixation zone and depletion of leghemoglobins,accompanied by downregulation of nodule-specific gene expression,as well as almost complete abrogation of biological nitrogen fixation.In summary,this study provides a comprehensive perspective of the cellular landscape during soybean nodulation,shedding light on the underlying metabolic and developmental mechanisms of soybean nodule formation.