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 ��M 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 (), 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 () expression, ferric leghaemoglobin reductases (), and S-nitrosoglutathione reductase (). 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 , pivotal for the reduction of Lb to Lb. 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.
