The CIN-like transcription factor CsTCP2 positively regulates the theanine biosynthesis in Camellia sinensis

Theanine is a non-protein amino acid that accumulates only in the tea plant and has high economic value and unique health benefits. Recently some theanine synthases have been identified, but the transcriptional regulatory mechanism of theanine anabolism remains largely unclear. In this work, transcription factor CsTCP2, which belonged to the CIN subclass of the TCP gene family, was cloned from cDNA of Yinghong 9，and found that the CsTCP2 localized in nucleus and exhibited transcriptional activation activity in tobacco and yeast cells. Moreover, Dual luciferase, Y1H, and EMSA analysis revealed that CsTCP2 can promote CsGS1b expression by binding to the MeJA-responsive cis-acting element of proCsGS1b. Overexpression or knockdown of CsTCP2 significantly increased or decreased the expression of CsGS1b and the accumulation of theanine in callus, respectively. In addition, it was found that CsTCP2 is involved in the photosynthesis pathway and various free amino acid synthesis pathways, which can regulate theanine synthesis by influencing the photosynthesis, as well as the accumulation of free amino acids can positively determine the quality of tea plants.