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The TIFY family is a novel plant-specific protein family, and is characterized by a conserved TIFY motif (TIFF/YXG). Our previous studies indicated the potential roles of TIFY10/11 proteins in plant responses to alkaline stress. In the current study, we focused on the regulatory roles and possible physiological and molecular basis of the TIFY10 proteins in plant responses to alkaline stress. We demonstrated the positive function of TIFY10s in alkaline responses by using the  AtTIFY10a  and  AtTIFY10b  knockout Arabidopsis, as evidenced by the relatively lower germination rates of  attify10a  and  attify10b  mutant seeds under alkaline stress. We also revealed that ectopic expression of  GsTIFY10a  in  Medicago sativa  promoted plant growth, and increased the NADP-ME activity, citric acid content and free proline content but decreased the MDA content of transgenic plants under alkaline stress. Furthermore, expression levels of the stress responsive genes including  NADP-ME ,  CS ,  H + -ppase  and  P5CS  were also up-regulated in  GsTIFY10a  transgenic plants under alkaline stress. Interestingly,  GsTIFY10a  overexpression increased the jasmonate content of the transgenic alfalfa. In addition, we showed that neither GsTIFY10a nor GsTIFY10e exhibited transcriptional activity in yeast cells. However, through Y2H and BiFc assays, we demonstrated that GsTIFY10a, not GsTIFY10e, could form homodimers in yeast cells and in living plant cells. As expected, we also demonstrated that GsTIFY10a and GsTIFY10e could heterodimerize with each other in both yeast and plant cells. Taken together, our results provided direct evidence supporting the positive regulatory roles of the TIFY10 proteins in plant responses to alkaline stress.

The Positive Regulatory Roles of the TIFY10 Proteins in Plant Responses to Alkaline Stress