Overexpression of the rice Osmyb4 gene increases chilling and freezing tolerance of Arabidopsis thaliana plants

The expression of the gene OsMyb4, detected at low level in rice (Oryza sativa) coleoptiles grown for three days at 29°C, is strongly induced by treatments at 4°C. At subletal temperatures of 10 and 15°C, its expression in rice seedlings is already evident, but this effect cannot be vicariated by other stresses or ABA treatment. We demonstrate by transient expression that Myb4 transactivates the PAL2, ScD9 SAD and COR15a cold-inducible promoters. The OsMyb4 function in vivo is demonstrated overexpressing its cDNA in Arabidopsis thaliana plants (ecotype Wassilewskija) under the control of the constitutive CaMV 35S promoter. Myb4 overexpressing plants show a significant increased cold and freezing tolerance, measured as membrane or Photosystem II (PSII) stability and as whole plant tolerance. Finally, in OsMyb4 transgenic plants, the expression of genes participating in different cold-induced pathways is affected, suggesting that Myb4 represent a master switch in cold tolerance.