Plant Total RNA Extraction and qPCR Analysis

Total RNA of each sample was extracted using a plant RNA extraction kit (TIANGEN DP441), and the sequences were used for cDNA library construction. qPCR SYBR Green Premix (Vazyme, China) was used to conduct qPCR analysis in a CFX™ real-time PCR detection system (Bio-Rad, USA). The primer sequences used were designed by Primer 5.0 (Additional File 6: Table S6). We used the Actin gene, which was stably expressed at each growth stage in almost all tissues, as the internal control [76 (link)]. The ACTIN gene was used as calibration to detect three technical repeats of the three biological repeats, and 2^-ΔΔCT method was used to analyze the expression [77 (link)].