Reverse transcription quantitative PCR

RNA extraction, cDNA synthesis, and reverse transcription quantitative PCR (RT-qPCR) were performed as described previously (Yonekura-Sakakibara et al., 2014). The developmental stages of leaves, stems, roots, floral buds, and flowers used for analyses were as described previously (Yonekura-Sakakibara et al., 2007). The developmental stages of siliques were as follows: stage 3, 24–36 h after flowering (HAF); stage 4, 36–48 HAF; stage 5, 48–72 HAF; stage 6, 72–96 HAF; stage 7, 96–108 HAF; stage 8, 108–120 HAF; stage 9, 120–132 HAF; stage 10, 132–144 HAF; stage 11, 144–192 HAF. Primers used are described in Supplemental Table 2 (At4g11180cDNA-RT101f and At4g11180cDNA-RT186r for AtDP1/AtDIR12, At1g61720pda14333_RT107f andAt1g61720pda14333_RT179r for BANYULS, At5g48100pda12625_RT478f and At5g48100pda12625_RT552r for AtLAC15/TT10, and At2g40370qPCRNo2_1090F and At2g40370qPCRNo2_1107R for AtLAC5) and were checked for specific product formation using a dissociation program. Plasmid DNAs containing the corresponding genes were used as templates for calibration. RT-qPCR was performed in triplicate.