Bioluminescence reporter assay system to monitor Arabidopsis MPK3 gene expression in response to infection by Botrytis cinerea

Abstract

The Arabidopsis MPK3 gene product participates in disease resistance mediated by the MAP kinase cascade. The expression of the MPK3 gene is induced by pathogen inoculation and treatment with chemicals such as salicylic acid (SA) and methyl jasmonate (JA), but the detailed expression pattern of the MPK3 gene has been largely unknown. To investigate MPK3 gene expression in response to disease stress, we fused the MPK3 promoter to the firefly luciferase gene to create a real-time monitoring system for regulated gene expression in planta. The results of an in vivo reporter assay using transgenic Arabidopsis plants harboring MPK3::Fluc showed that the MPK3 promoter activity was induced by treatment with chemicals such as SA and benzo(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester (BTH), that induce defense gene expression. Inoculation with the fungal pathogen Botrytis cinerea resulted in systemic induction of MPK3::Fluc.

OsMKK3, a Stress-Responsive Protein Kinase, Positively Regulates Rice Resistance to Nilaparvata lugens via Phytohormone Dynamics

“…Additionally, our results indicated that SA elicited the expression of OsMKK3 which was a negative regulator of SA. Similarly, AtMPK3 could be induced by SA but negatively regulated SA accumulation in Arabidopsis [57,60]. We speculate OsMKK3 might have a negative feedback to SA via its downstream MAPKs.…”

Evaluation of the Use of the TobaccoPR-1aPromoter to Monitor Defense Gene Expression by the Luciferase Bioluminescence Reporter System

“…9,20) In this study, we found that induction of the PR-1a promoter was easily detectable even in Arabidopsis seedlings germinated in multi-well plates. This is particularly important in considering the utilization of transgenic seedlings in high-throughput screening using multi-well plates.…”

Non-destructive bioluminescence detection system for monitoring defense gene expression in tobacco BY-2 cells

“…As shown in Figure 3, induction by less than 5 mM of SA or BTH treatment was readily detectable in this assay system. Compared with the previously described induction experiments in response to treatment with SA or BTH, the sensitivity of this assay system was about 10 to 100 times higher Tanaka et al 2006).…”