GENE ACTIVATION BY CYTOPLASMIC ACIDIFICATION IN SUSPENSION-CULTURED RICE CELLS IN RESPONSE TO THE POTENT ELICITOR, N-ACETYLCHITOHEPTAOSE

N-Acetylchitoheptaose strongly induces a set of defense reactions in s uspension-cultured rice cells including cytoplasmic acidification (K. Kuchitsu, Y. Yazaki, K. Sakano, and N. Shibuya, Plant Cell Physiol. 38 :1012-1018, 1997) and the accumulation of mRNAs for two rapidly activa ted genes, EL2 and EL3 (E. Minami, K. Kuchitsu, D.-Y. He, H. Kouchi, N . Midoh, Y. Ohtsuki, and N. Shibuya, Plant Cell Physiol. 37:563-567, 1 996), as well as phenylalanine ammonia-lyase (PAL), chitinase and beta -1,3-glucanase. Treatment of cells with propionic acid resulted in the accumulation of the mRNAs for EL2, EL3, and PAL in a manner similar t o the accumulation induced by N-acetylchitoheptaose. Concomitantly, th ere was a rapid decrease in the cytoplasmic pH as detected with in viv o P-31-nuclear magnetic resonance (NMR) spectroscopy. Interestingly, K -252a, a potent inhibitor of Ser/Thr protein kinases, strongly inhibit ed gene induction by N-acetylchitoheptaose, but showed much less inhib ition of gene induction caused by propionic acid. Calyculin A, a prote in phosphatase inhibitor, induced mRNA accumulations for EL2, EL3 and PAL, with concomitant acidification of the cytoplasm. On the other han d, chitinase and P-glucanase mRNA levels did not change after addition of propionic acid or calyculin A. Treatment of the cells with propion ic acid did not induce the production of reactive oxygen species. Thes e results strongly suggest that cytoplasmic acidification at the early stage of elicitor action could be a key step in the signal transducti on events leading to the expression of elicitor-responsive genes. A hy pothetical model of elicitor signal pathway is proposed based on these results.