Role of Brassinosteroids in plant growth promotion and stress tolerance during primingmediated by beneficial bacteria

Abstract: Brassinosteroids (BR) are plant hormones widely distributed throughout the plant kingdom inlow concentrations and with structural homology to animal and insect steroids. BR are involvedin numerous physiological processes, and they also fulfill an antagonistic role in anti-herbivorystructure formation in tomato (Campos et al., 2009). In order to characterize the role of BR uponpriming with B. amyloliquefasciens 5113, gene expression analysis of BR genes was assessed inArabidopsis thaliana. BAK1, BRI1 and DWF1 expression down-regulates, while DET2 upregulatesupon bacterial priming. CPD gene expression was not affected by priming. qPCRanalysis of VSP2 and PR1 were performed on BR mutants upon priming with B.amyloliquefasciens 5113. Basal levels of PR1 were higher in det2, bak1 and dwf1 compared toprimed samples. Primed bri1 displayed two-fold higher expression of PR1 compared to untreatedbri1. VSP2 level goes up on det2, bak1 and bri1 upon priming. No changes of VSP2 expressionwere observed in dwf1 upon priming. Methyl jasmonate treatment up-regulates VSP2 level twofoldin det2 and nine-fold in bak1. The role of BR genes in response to insect attack wasexamined. BR genes appear not to be responsive to herbivory by S. littoralis. However, S.littoralis larvae fed more on BR mutants compared to those that fed on Col-0 WT. In order tounderstand the role of BR in JA signaling pTRV-JAR1 and pTRV-LOX2 constructs weredeveloped and virus induced gene silencing were performed on Col-0 and BR mutants bak1 anddet2. Gene silencing was confirmed by qPCR analysis of the target genes in Col-0 and det2, butnot in bak1. Further insect feeding experiments are required to elucidate if BR play a role indefense responses to herbivory when JA signaling pathway is compromised.