Characterization of NtChitIV, a class IV chitinase induced by beta-1,3-, 1,6-glucan elicitor from Alternaria alternata 102: Antagonistic effect of salicylic acid and methyl jasmonate on the induction of NtChitIV.

Tobacco BY-2 class IV chitinases (TBC-1, TBC-3) were rapidly and transiently induced by the beta-1,3-, 1,6-glucan elicitor from Alternaria alternata 102 (AaGlucan). The full-length cDNA and 5'-flanking region of a gene encoding class IV chitinases were isolated on the basis of the amino acid sequence of TBC-1. Sequence analysis indicated that NtChitIV encoded TBC-1, TBC-3, or both. Since purified TBC-1 and TBC-3 from BY-2 cells lack a chitin binding domain in the N-terminal region, these enzymes suggested to be derived from NtChitIV by post-translational proteolytic processing. The transcripts of NtChitIV accumulated rapidly within 1h after treatment with AaGlucan. Accumulation was maximal 3h after treatment. Reporter gene assays were used to analyze the promoter regions involved in the transcriptional control of NtChitIV, and these assays revealed that the 1.89-kb NtChitIV promoter was activated by AaGlucan but not by salicylic acid (SA) or methyl jasmonate (MeJA). The AaGlucan-induced transcriptional activation via 1.89-kb NtChitIV promoter was attenuated by pretreatment with SA or MeJA. These results suggest that NtChitIV expression is particularly induced by AaGlucan and that the AaGlucan-dependent signaling pathway is different from the SA- and MeJA-dependent signaling pathways.