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).