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Media type:
E-Article
Title:
Functional dissection of a strong and specific microbe‐associated molecular pattern‐responsive synthetic promoter
Contributor:
Lehmeyer, Mona;
Kanofsky, Konstantin;
Hanko, Erik K. R.;
Ahrendt, Sarah;
Wehrs, Maren;
Machens, Fabian;
Hehl, Reinhard
Published:
Wiley, 2016
Published in:
Plant Biotechnology Journal, 14 (2016) 1, Seite 61-71
Language:
English
DOI:
10.1111/pbi.12357
ISSN:
1467-7644;
1467-7652
Origination:
Footnote:
Description:
SummarySynthetic promoters are important for temporal and spatial gene expression in transgenic plants. To identify novel microbe‐associated molecular pattern (MAMP)‐responsive cis‐regulatory sequences for synthetic promoter design, a combination of bioinformatics and experimental approaches was employed. One cis‐sequence was identified which confers strong MAMP‐responsive reporter gene activity with low background activity. The 35‐bp‐long cis‐sequence was identified in the promoter of the Arabidopsis thaliana DJ1E gene, a homologue of the human oncogene DJ1. In this study, this cis‐sequence is shown to be a tripartite cis‐regulatory module (CRM). A synthetic promoter with four copies of the CRM linked to a minimal promoter increases MAMP‐responsive reporter gene expression compared to the wild‐type DJ1E promoter. The CRM consists of two WT‐boxes (GGACTTTT and GGACTTTG) and a variant of the GCC‐box (GCCACC), all required for MAMP and salicylic acid (SA) responsivity. Yeast one‐hybrid screenings using a transcription factor (TF)‐only prey library identified two AP2/ERFs, ORA59 and ERF10, interacting antagonistically with the CRM. ORA59 activates reporter gene activity and requires the consensus core sequence GCCNCC for gene expression activation. ERF10 down‐regulates MAMP‐responsive gene expression. No TFs interacting with the WT‐boxes GGACTTTT and GGACTTTG were selected in yeast one‐hybrid screenings with the TF‐only prey library. In transgenic Arabidopsis, the synthetic promoter confers strong and specific reporter gene activity in response to biotrophs and necrotrophs as well as SA.