Clone and quantification expression analysis of LaTPS7,LaTPS8 and LaCYP71D582
Glandular trichomes are tissues located in metabolism-specific sites (Tissieret al . 2017). Total RNAs were extracted from glandular trichomes of flowers and leaves in different developmental stages and reverse transcribed using oligo d(T) primers following the manufacturer’s directions (TSINGKE, China). After obtaining cDNA, target genes were amplified using cloning primer and Phanta Max Polymerase (Vazyme, China). To clone full-length open reading frames (ORFs), we used a PCR program at the following settings: 95°C for 5 min; 35 cycles at 95°C for 2 min; 58°C for 30 s and 72°C for 2 min; and final extension at 72°C for 5 min. The qRT-PCR was conducted using 2 × T5 Fast qPCR Mix (SYBR Green II) (TSINGKE, China) on an Mx3000P system (Agilent Stratagene), according to the manufacturer’s instructions and the following settings: 95°C for 1 min, followed by 40 cycles of 10 s at 95°C, then 5 s at 55°C, and 15 s at 72°C. Leaf trichomes at the blossoming stage were used as controls. Normalised expression values (2-△△CT) were calculated according to the expression of 18S rRNA used as reference gene. The primers used in this part of our study are shown in table S1.