Identification and expression profiling of MYB transcription factors related to l-theanine biosynthesis in Camellia sinensis

The MYB proteins belong to a large family of transcription factors in plant genomes and play significant roles in primary and secondary metabolism. Although several CsMYB genes have been identified in Camellia sinensis, few CsMYBs involved in l-theanine biosynthesis have been analyzed. In this study, we screened and identified 20 CsMYBs related to l-theanine biosynthesis. Transcriptomic analysis revealed that the expression profiles of the CsMYBs were positively or negatively related to dynamic changes in the l-theanine content. Validation of selected l-theanine biosynthetic and CsMYB genes was conducted by qRT-PCR. The results illustrated that most of the structural and CsMYB genes were downregulated with a decrease in the l-theanine levels. Protein–protein interaction networks of CsMYB5, CsMYB12 and CsMYB94 proteins demonstrated that they might form complexes with bHLH and WD 40 proteins. Multiple DNA-binding sites of the R2R3-MYB protein were observed in promoter regions of structural genes, indicating CsMYB family proteins might be involved in l-theanine metabolism via the attachment of AC elements. Moreover, CsMYB73 demonstrated binding specificity to the promoter region of CsGDH2 (CsGDH2-pro). These findings provide fundamental understanding of specific members of the CsMYBs related to the l-theanine biosynthesis pathway.