TY  - EJOU
AU  - Chen, Enyan 
AU  - Jia, Jingyu 
AU  - Sun, Jiaju 
AU  - Wang, Jie 
AU  - Chen, Xinxin 
AU  - Li, Xin 

TI  - Trilobatin Acts as a Marker Metabolite Involved in Flavonoid Accumulation Regulated by CsWRKY28-MYC2 with Trypsin Activation in <i>Cucumber (Cucumis sativus)</i>
T2  - Phyton-International Journal of Experimental Botany

PY  - 2024
VL  - 93
IS  - 11
SN  - 1851-5657

AB  - During post-harvest storage of <i>Cucumis sativus</i> fruit, the application of trypsin treatment could increase flavonoid compound levels and reduce oxidative damage. To investigate the mechanism of trypsin-induced flavonoid biosynthesis in <i>C. sativus</i>, we conducted a combined analysis of transcriptomics and widely targeted metabolomics. One hundred and seventy-five significantly different metabolites were obtained from metabolomics data. The kyoto encyclopedia of genes and genomes (KEGG) functional enrichment results indicated that these metabolites were mainly involved in the phenylpropanoid biosynthesis pathway. By combining the results of the weighted gene co-expression network analysis (WGCNA) with the 130 upregulated phenylpropanoid metabolites, 22 significantly upregulated phenylpropanoid metabolites were identified. Trilobatin was identified as the most prominent metabolite through cluster analysis and variable importance in projection (VIP) analysis. High performance liquid chromatography (HPLC) experiments confirmed that trilobatin was the key metabolite induced by trypsin. The transcriptomic results showed that 1068 genes in the brown module of WGCNA were highly positively correlated with flavonoid biosynthesis. The gene set enrichment analysis (GSEA) identified leading edges in 4 key KEGG pathways. Finally, combined with WGCNA and GSEA analysis results, 35 core genes were obtained. The co-expression network of transcriptomics and metabolomics suggested that <i>CsWRKY28</i> and <i>CsMYC2</i> regulated the biosynthesis of trilobatin. The quantitative real-time polymerase chain reaction (RT-qPCR) and dual luciferase experiments confirmed the activation effect of <i>CsWRKY28</i> on <i>CsMYC2</i> and downstream target genes. This study revealed the key transcription factors involved in the trypsin-controlled biosynthesis of trilobatin in <i>C. sativus</i> and provided a new theoretical basis for elucidating the molecular mechanism of trypsin preservation.
KW  - <i>CsWRKY28</i>; transcriptomics; trilobatin; trypsin; widely targeted metabolomics

DO  - 10.32604/phyton.2024.057932