Abstract
Nitrogen fixation in legume nodules is crucial for plant growth and development. Therefore, this study aims to investigate the effects of nitric oxide [S-nitrosoglutathione (GSNO)] and silicon [sodium metasilicate (Si)], both individually and in combination, on soybean growth, nodule formation, leghaemoglobin (Lb) synthesis, and potential post-translational modifications. At the V1 stage, soybean plants were treated for 2 weeks with 150 µM GSNO, and Si at concentrations of 1 mM, 2 mM, and 4 mM. The results showed that NO and Si enhance the nodulation process by increasing phenylalanine ammonia-lyase activity and Nod factors (NIP2-1), attracting rhizobia and accelerating nodule formation. This leads to a greater number and larger diameter of nodules. Individually, NO and Si support the synthesis of Lb and leghaemoglobin protein (Lba) expression, ferric leghaemoglobin reductases (FLbRs), and S-nitrosoglutathione reductase (GSNOR). However, when used in combination, NO and Si inhibit these processes, leading to elevated levels of S-nitrosothiols in the roots and nodules. This combined inhibition may potentially induce post-translational modifications in FLbRs, pivotal for the reduction of Lb(3+) to Lb(2+). These findings underscore the critical role of NO and Si in the nodulation process and provide insight into their combined effects on this essential plant function.