Acetylation participation in theanine biosynthesis: insights from transcriptomics, proteomics, and acetylomics

Acetylation, a crucial post-translational modification, regulates transcriptional activation, enzymatic activity, and protein interactions, playing vital roles in plant physiology and metabolism. However, the regulatory mechanism of acetylation in the biosynthesis of theanine remains unexplored. This study aims to elucidate the regulatory role of acetylation on the biosynthesis of theanine using transcriptomics, proteomics, and acetylomics in tea leaves from three tea plant cultivars with markedly different theanine content. Nineteen theanine biosynthesis-related genes were identified in the transcriptome, with ten showing significant correlation with theanine content. Proteomic analysis revealed elevated expression levels of proteins associated with the biosynthesis of theanine precursor glutamate in leaves with high theanine content, such as GOGAT and GDH. Unexpectedly, the expression level of TS was inversely correlated with the theanine content in leaves. Several highly expressed acetylated proteins and sites, such as TS, GS, and GOGAT, were found in the acetylome of leaves with high theanine content. Acetylation at lysine 304 (K304) of the TS protein may significantly contribute to the abundant accumulation of theanine in leaves. Our findings indicate that acetylation modification may play a pivotal role in theanine biosynthesis, thereby offering novel insights into the development of high-theanine tea plant germplasm resources.