Elicitation of defense reactions in tobacco by cryptogein, triggered a production of active oxygen species (AOS) via the NADPH oxidase, NtrbohD, and an accumulation of beta1din, a defense induced beta-type subunit of 20S proteasome. The proteasome inhibitor, MG132, stimulated this AOS production. Tobacco cells transformed with sense constructs of beta1din showed an inhibition of the AOS production following elicitin treatment, whereas the antisense transformed cells showed a strongly enhanced AOS production. In cells transformed with sense construct of beta1din, the NtrbohD transcripts failed to be induced by cryptogein as observed in control and antisense transformed cells. Conversely, in tobacco cells transformed with antisense constructs for NtrbohD, beta1din transcripts remained at a low level after elicitation. These results constitute the first demonstration of proteasome comprising beta1din acting as a negative regulator of NtrbohD and contributes to the regulation of AOS generation during plant defense reactions.
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