Abstract: Induced cold inactivation is a general characteristic of V-type H⁺-ATPase.In this paper, fluorescence spectrum technique has been used to thoroughly study the mechanism of cold inactivation of H⁺-ATPase on rice tonoplast.It shows that the chemicals that can cause the cold inactivation considerably usually cause the destruction of the order of tonoplast lipid first.The capability of inducing cold inactivation and changing the order of membrane lipid in cations, anions and chelation agents has been testified and compared.The result proved the conclusion above.We consider that the changing of membrane lipid in tonoplast plays an important role in cold inactivation and is one of the reasons of early cold pet of plants.