Hiscript 2 kit 117

Total RNA was extracted using an RNAprep Pure Plant Kit DP441 (Tiangen, Beijing, China), according to the manufacturer’s instructions. RNA integrity was evaluated using an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). Samples with an RNA integrity number of 7 to 10 were subjected to subsequent analysis. The cDNA libraries were constructed using a HiScript II kit 117 (Vazyme, Nanjing, China). The prepared libraries were sequenced on an Illumina HiSeq TM 2500 platform ((Illumina, San Diego, CA, USA), and 125 bp/150 bp paired-end reads were generated. The database can be found in National Center of Biotechnology Information (NCBI) database under the accession number PRJNA767274.

Total RNA isolated from the SZ of stage S6 cm to S9 cm treated with 0 or 50 μM MeJA (Bioduly, Nanjing, China) for 48 h. The cDNA was prepared using a HiScript II kit 117 (Vazyme, Nanjing, China). Quantitative RT-qPCR reactions were conducted using ChamQ Universal SYBR qPCR Master Mix (Vazyme, Nanjing, China) following the manufacturer’s protocol. Primers for the genes were designed using Primer express 3.0 software (Table S6). Gene expression levels were normalized using the reference gene 18S [5 (link)], and values were calculated using the 2^−ΔΔCt method [70 (link),71 (link)]. Expression values were obtained from three biological experiments. SPSS 17.0 statistical software was used to determine statistical significance.