Common Bean Sprout Transcriptomics

Different tissues (cotyledon, hypocotyl, and radicle) of WR were selected on the fifth day during the sprout stage, and hypocotyls of the four samples (WR, WN, SR, and SN) were selected on the fourth, fifth and sixth days. Each sample had three biological repeats. Total RNA was extracted using RNA isolater Total RNA Extraction Reagent (Vazyme Biotech, China). After the extraction, the RNA samples were electrophoresed in 1% agarose gel for detection and quality control. Additionally, a NanoDrop spectrophotometer was used to check the concentration and purity of the RNAs. Samples of poor quality were discarded. Premier 5.0 (Pi et al., 2018 (link)) was used to design the RT-qPCR primers (Supplementary Table 2). PvC3Hs were selected from the GO enrichment and KEGG pathways, and some PvC3Hs were selected as candidate genes for verification. ACTIN-11 of common bean served as an internal control gene (Borges et al., 2012 (link)). The RNA was transcribed to cDNA, and then, RT-qPCR was performed using 2 × ChamQ Universal SYBR qPCR Master Mix Kit (Vazyme Biotech, China). The RT-qPCR was performed in triplicate. The relative gene expression was determined as follows (Livak and Schmittgen, 2001 (link)):
Relative Expression = 2^ΔΔCt, Where ^ΔΔCt = [Ct(Pv target genes) −Ct(PvACTIN-11)].