Soybean PHR1-regulated low phosphorus-responsive GmRALF22 promotes phosphate uptake by stimulating the expression of GmPTs

Phosphorus (P) is an essential macronutrient for plant growth and development. Rapid alkalisation factors (RALFs) play crucial roles in plant responses to nutrient stress. However, the functions of Glycine max RALFs (GmRALFs) under low P (LP) stress remain elusive. In this study, we first identified 27 GmRALFs in soybean and then revealed that, under LP conditions, GmRALF10, GmRALF11, and GmRALF22 were induced in both roots and leaves, whereas GmRALF5, GmRALF6, and GmRALF25 were upregulated in leaves. Furthermore, GmRALF22 was found to be the target gene of the transcription factor GmPHR1, which binds to the P1BS cis-element in the promoter of GmRALF22 via electrophoretic mobility shift assay and dual-luciferase experiments. Colonisation with Bacillus subtilis which delivers GmRALF22, increases the expression of the high-affinity phosphate (Pi) transporter genes GmPT2, GmPT11, GmPT13, and GmPT14, thus increasing the total amount of dry matter and soluble Pi in soybeans. RNA sequencing revealed that GmRALF22 alleviates LP stress by regulating the expression of jasmonic acid- (JA-), salicylic acid- (SA-), and immune-related genes. Finally, we verified that GmRALF22 was dependent on FERONIA (FER) to promote Arabidopsis primary root growth under LP conditions. In summary, the GmPHR1-GmRALF22 module positively regulates soybean tolerance to LP.