Quantifying Gene Expression in Lilium Cultivars

The sequences of candidate genes were retrieved from the L. japonica genome database. Specific primer pairs used in qRT-PCR were designed using NCBI Primer-Blast¹⁴ (Supplementary Table 1). Total RNA of LJFs at the silver stage of ‘Yujin2’ and ‘Fengjin1’ was isolated using the OminiPlant RNA Kit (Cwbiotech, Beijing, China) and then reverse-transcribed using a HiScript® II Q RT SuperMix for qPCR kit (+ gDNA wiper; Vazyme Biotech, Nanjing, China). qRT-PCR was performed on a LightCycler® 96 real-time PCR system (Roche, Hong Kong, China) using SYBR Green Master Mix (Vazyme Biotech, Nanjing, China; Yu et al., 2021 (link)). Three biological and three technical replicates were performed. Relative expression levels were calculated using the 2^–ΔΔCt method (Liu et al., 2019 (link)). The Lonja.ACT2/7 and Lonja.27738 genes were used as internal references (Liu, 2017 ).