RNA extraction and qRT-PCR
Total RNA was extracted from inoculated leaves using TRIzol reagent (Invitrogen) following the manufacturer’s protocol. RNA samples were reverse transcripted to cDNA (Vazyme HiScript III All-in-one RT SuperMix Perfect for qPCR). The qRT-PCR reaction systems were prepared as manufacturer’s instruction (Vazyme AceQ Universal SYBR qPCR Master Mix) using a Real-Time PCR System (Applied Biosystems StepOnePlus™). The 2-ΔΔCt method was used to evaluate the gene expression levels. The rice β-actin (LOC_Os03g50885 ) was used as the control gene and three biological repeats were performed for qRT-PCR. The results were visualized with Prism (v8.0.1) or ChiPlot(Xie et al. , 2023).