2.5.3 Validation of RNA-Seq data
RNA was extracted using the plant RNA kit (OMEGA) with three replications. cDNA was obtained using PrimeScript™ RT Reagent Kit following the manufacturer’s protocol (TaKaRa, Japan) and then utilized for RT‐qPCR reactions performed in Bio‐Rad CFX96 (Bio‐Rad, USA) using SYBR® Green Realtime PCR Master Mix (TaKaRa, Japan). Gene‐specific primers utilized in this study are listed in Table S1. Relative gene expression levels were normalized utilizing SlUbi3 as a reference gene and data analyzed using 2−ΔΔCt method (Livak and Schmittgen, 2001).