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Tea [ Camellia sinensis  (L.) O. Kuntze] leaves are a major source of epicatechin (EC) and its gallolyl derivatives epicatechin gallate, epigallocatechin and epigallocatechin gallate, collectively known as epicatechins (ECs). Epicatechins are important factors determining tea quality, and they also possess many medicinal properties. To gain further information about the regulation of the biosynthesis of ECs, we cloned the gene encoding anthocyanidin reductase from tea ( CsANR ) by first quantifying changes in the concentrations of ECs in response to drought, gibberellic acid (GA 3 ), abscisic acid (ABA) and wounding treatments, followed by differential display of mRNAs and analysis of those bands exhibiting a change in expression paralleling the treatment-induced changes observed in the EC data. Analysis of 133 bands yielded a partial cDNA of  CsANR  that was later cloned to the full length by rapid amplification of the cDNA ends. The full-length  CsANR  (Accession No.  AY641729 ) comprised 1233 bp with an ORF of 1014 bp (from 79 to 1092 bp) encoding a polypeptide of 337 amino acids. Expression of  CsANR  in an  Escherichia coli  expression vector yielded a functional protein that catalyzed the conversion of cyanidin to EC in the presence of NADPH. Analysis of ECs and gene expression in leaves at different developmental stages and across five tea clones exhibiting variable concentrations of ECs revealed a positive correlation between concentration of ECs and  CsANR  expression. Expression of  CsANR  was down-regulated in response to drought, ABA and GA 3  treatments and up-regulated in response to wounding.

tree physiology

Differential display mediated cloning of  anthocyanidin reductase  gene from tea ( Camellia sinensis ) and its relationship with the concentration of epicatechins