Objective Glycerol-3-phosphate acyltransferase (CsGPAT) involving the glycerolipid metabolic pathway was cloned from fresh leaves of Tieguanyin tea (Camellia sinensis) plants for bioinformatics analysis and expression analysis of different withering temperature. To understand the significance of GPAT gene in the process of tea withering, in order to provide a theoretical basis for temperature regulation in the process of tea withering.
Method By screening the transcriptome data on withering oolong tea, the homologous GPAT sequence was obtained. Software including ExPASy Protparam, SMART, SignalP 4.1 Server, PSORT, prediction protein, etc. were used to obtain its bioinformatics; the SWISS-MODEL online tool applied to arrive at its 3D protein structure; and, NCBI Blastp employed to analyze its homologous alignment on the amino acid sequence. RNA from the tea leaves was extracted for qRT-PCR to determine the real-time expression as well as for cloning the full length of GPAT.
Result The cloned CsGPAT (IDcsa:CSA000941.1/ IDcss:TEA019813.1) had a full length of 1 554 bp encoding a predicted protein of 497 amino acids. The bioinformatics showed CsGPAT not a signal peptide but a stable hydrophobic protein with a PlsC domain of phospholipid biosynthetic function that closely related to Camellia oleifera. Its expression peaked at the withering temperature of 20℃.
Conclusion The expression of CsGPAT in the tea leaves was up-regulated when the withering temperature was relatively low. It suggested a crucial role the gene played in oolong tea processing.
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