Real-time quantitative PCR (qRT-PCR) analysis

Twenty DEGs involved in plant-pathogen interaction and plant hormone signal transduction pathways were selected for qRT-PCR, and primers were designed using Premier 5 (Table S1). qRT-PCR was performed using ChamQ Universal SYBR qPCR Master Mix (Vazyme, Nanjing, China) on a CFX96™ Real-Time System (Bio-Rad, USA) with the cycle steps of pre-degeneration at 95 °C, 3min, and 1 cycle; followed by 40 cycles of 95 °C for 10s, and 60 °C for 40s; and melting curve analysis at 95 °C for 10s, 65 °C for 5s, and 95 °C for 5s. The relative expression level of each gene was calculated by the 2^−ΔΔCt method (Livak & Schmittgen, 2001), and the correlation coefficients between RNA-seq data and qRT-PCR were evaluated using GraphPad Prism 9 (San Diego, CA, USA).