Trizol 117 reagent

Total RNAs were extracted from various tomato tissues using the TRIzol^® 117 reagent (Invitrogen). A reverse transcription kit (Vazyme, Nanjing, China) was utilized to generate single-stranded cDNA using 2 μg of total RNA. Gene expression assays by qPCR analysis were performed using the ChamQ SYBR Color qPCR Master Kit (Vazyme, Nanjing, China). All data had three biological replicates and were analyzed. The internal control was the expression of the Actin gene (Solyc11g005330). Supplementary Data Table S1 lists the primer sequences used in real-time PCR.

RNA from various tissues of wild-type and transgenic plants were isolated using TRIzol^® 117 reagent (Invitrogen, USA). For cDNA synthesis, 3 µg RNA was used with M-MLV reverse transcriptase (Toyobo, Japan) according to the manufacturer’s instructions. The cDNA concentrations were normalized to actin expression levels for RT-PCR analysis. Primers of the ripening-related genes are listed in Supplementary Table S1 at JXB online. The actin gene was used as an internal control for quantitative real-time PCR (qPCR), which was performed using the Power SYBR Premix Ex Taq kit and the TaKaRa two-step method (TaKaRa, Japan). PCR products were quantified using the Roche LightCycler 480 Real-Time PCR Detection System and the SYBR Green I Master Kit (Roche, Switzerland). The wild-type and transgenic plants were represented by three biological replicates for each sample.