Introduction: WRKY proteins belong to a plant-specific class of transcription factors. Seventy-four WKRY genes have been identified in Arabidopsis and many WRKY proteins are known to be involved in responses to stress, especially to biotic stress. They may act either as transcriptional activators or as repressors of genes that play roles in the stress response. A number of studies have proposed the connection of Arabidopsis WRKY transcription factors in induced pathogenesis-related (PR) gene expression, although no direct evidence has been presented for specific WRKY-PR promoter interactions.
Objective: We previously identified AtWRKY50 as a transcriptional activator of SAR gene PR1. Although PR1 accumulates to high levels in plants after attack by pathogens, its function is still elusive. Here we investigated the effects of overexpression of several WRKY proteins, including AtWRKY50, on the metabolome of Arabidopsis thaliana.
Methods: The influence of overexpression of WRKY proteins on the metabolites of Arabidopsis was investigated by using an NMR spectroscopy-based metabolomic approach. The 1H NMR data was analysed using the multivariate data analysis methods, such as principal component analysis, hierarchical cluster analysis and partial least square-discriminant analysis.
Results: The results showed that the metabolome of transgenic Arabidopsis seedlings overexpressing AtWRKY50 was different from wild type Arabidopsis and transgenic Arabidopsis overexpressing other WRKY genes. Amongst other metabolites, sinapic acid and 1-O-sinapoyl-β-D-glucose especially appeared to be the most prominent discriminating metabolites, accumulating to levels 2 to 3 times higher in the AtWRKY50 overexpressor lines.
Conclusion: Our results indicate a possible involvement of AtWRKY50 in secondary metabolite production in Arabidopsis, in particular of hydroxycinnamates such as sinapic acid and 1-O-sinapoyl-β-D-glucose.
Keywords: 1-O-sinapoyl-β-D-glucose; Arabidopsis; AtWRKY; NMR spectroscopy; Overexpression; Sinapic acid.