Expression Characteristics of CHS Gene in Tea and Its Relationship with Catechins Accumulation

In this study,the content of catechins in two kinds of tea were detected with high performance liquid chromatography(HPLC),and the open reading frame(ORF)of CHS genes were cloned by RT-PCR from the Yinghong NO.9(YH)and Camellia ptilophylla Chang(MY)and presented comparison analysis. The subcellular localization of CHS gene expressed protein was carried out by tobacco transient expression technique. The CHS specific antibody was obtained with synthetic peptide segment on the basis of bioinformatics analysis. The expression characteristics of CHS conducted by fluorescence quantitative real-time PCR(qRT-PCR)at the level of transcription and by Western blot assays at the level of translation,respectively. Moreover,and the relationship of CHS expression with catechins contents was analyzed. Results showed that the content of catechins in YH and MY was 181.51 mg/g and 106.02 mg/g,the former was significantly higher than the latter and reached 1.71-fold.The CHS genes isolated from the two tea materials exhibited only one nucleotide difference but shared the same amino acid sequence. The protein encoded by CHS gene localized in cytoplasm,cell membrane,and nucleus based on tobacco transient expression system. The titer of the developed CHS antibody reached 1:80000,and the CHS expression at transcription and translation level in YH were both higher than that in MY which reached 3.9 and 1.7-fold,the expression of CHS gene both at transcription and translation level consistent with the accumulation of tea catechins. Results verified the accumulation of catechins was positively correlated with the expression of CHS gene.