The peptide-encoding MtRGF3 gene negatively regulates nodulation of Medicago truncatula

Leguminous root nodules specifically induced by rhizobium species fix nitrogen gas to gain nitrogen sources, which is important in sustainable agriculture and ecological balance. Several peptide signals are reported to be involved in regulation of legume nodule number and development. There are fifteen genes coding Root Meristem Growth Factor (RGF) peptide in Medicago truncatula, herein we find the expression of MtRGF3 is significantly induced by Sinorhizobium meliloti with production of Nod factors. The gene promoter is active in nodule primordia, young nodules and the meristem region of mature nodules. Knock-down (RNAi) roots of the gene (MtRGF3-RNAi) formed more root nodules than the empty vector control, and the nodule number decreased in MtRGF3-overexpressing (MtRGF3-OX) roots. Exogenous addition of the synthesized peptide significantly promoted primary root growth and inhibited lateral root emergence, in addition, the peptide application reduced the number of infection threads, nodule primordia and root nodules of M. truncatula. We also found that tyrosine sulfation determines the biological activity of MtRGF3 functioning in nodulation process, and MtRGF3 peptide negatively regulates nodulation in a dosage manner. These results demonstrate that the MtRGF3 peptide is a novel regulator during nodulation of Medicago trucatula.