Silencing of tomato RBOH1 and MPK2 abolishes brassinosteroid-induced H2O2 generation and stress tolerance

Abstract

Brassinosteroids (BRs) are involved in the regulation of plant growth, development and stress responses. While the signalling pathways for BR-regulated plant growth and development are well studied, the mechanisms by which BRs regulate plant stress tolerance remain largely unclear. Here we showed that 24-epibrassinolide (EBR), which induced tolerance to oxidative and heat stress in tomato, was also capable of elevating the transcript levels of RBOH1, MPK1 and MPK2, increasing apoplastic H2O2 accumulation, and enhancing activation of MPK1/2. Virus-induced gene silencing of RBOH1, MPK1, MPK2 and MPK1/2 resulted in reduced stress tolerance. Silencing of RBOH1 had no effect on the transcripts of MPK1 and MPK2 but inhibited MPK1/2 activation and H2O2 accumulation. Silencing of either MPK1 or MPK2, on the other hand, reduced RBOH1 transcript, H2O2 accumulation and MPK1/2 activity. BR-induced tolerance and MPK1/2 activation were compromised in RBOH1-, MPK2- and MPK1/2-silenced plants but not in MPK1-silenced plants. These results suggested that MPK2 played a more critical role than MPK1 in EBR-induced apoplastic H2O2 accumulation. RBOH1, MPK1 and MPK2 were involved in the stress tolerance and BR-induced stress tolerance likely involved a positive feedback loop among RBOH1, H2O2and MPK2, leading to sustained apoplastic accumulation of H2O2and related signalling processes. BRs are involved in the regulation of stress responses and the mechanisms remain largely unclear. BRs upregulated transcript of RBOH1, MPK1 and MPK2, apoplastic H2O2 accumulation and activation of MPK1/2. MPK2 played a more critical role than MPK1 in BR-induced apoplastic H2O2 accumulation. BR-induced tolerance and MPK1/2 activation were compromised in RBOH1-, MPK2- and MPK1/2-silenced plants but not in MPK1-silenced plants. BR-induced stress tolerance likely involved a positive feedback loop among RBOH1, H2O2and MPK2, leading to sustained apoplastic accumulation of H2O2and related signaling processes. © 2012 Blackwell Publishing Ltd.