Quantitative Analysis of Sesame Gene Expression

The gene sequences for these proteins were obtained from the genome database of Yuzhi 11 (data unpublished) and quantitative real-time PCR (qRT-PCR) analysis. Total RNA was extracted from sesame leaves of all three phenotypes with an RNA extraction and purification kit (Sangon Biotech, Shanghai, China) according to the manufacturer’s instructions. The total RNA was reverse transcribed into cDNA with a Revert Aid First Strand cDNA Synthesis Kit (Thermo Scientific, Vilnius, Lithuania). qRT-PCR was performed with FastStart Essential DNA Green Master (Roche, Mannheim, Germany) using the Realplex 4 MasterCycler real-time PCR System (Eppendorf AG, Hamburg, Germany). qRT-PCR was conducted in accordance with the manufacturer’s instructions. The reaction conditions were as follows: 1 cycle of 50 °C for 2 min; 1 cycle of 95 °C for 10 min; 40 cycles of 95 °C for 15 s; 40 cycles of 60 °C for 20 s; and 40 cycles of 72 °C for 20 s. Three independent biological replicates per sample were performed. SiTUB was used as an internal reference gene to normalize the relative gene expression (Wei et al. 2013 (link)). The relative gene expression was calculated using the 2^−∆∆Ct method (Livak and Schmittgen 2001 (link)). The primer sequences of the genes of DEPs for qRT-PCR are shown in Table 1S.