Transcriptome Profiling of Melon Development

Total RNA was extracted with an ultrapure RNA Kit (Tiangen, Beijing, China) and template cDNA synthesis was performed using the PrimeScript™ RT Master Mix (Tiangen, Beijing, China) following the manufacturer’s instructions. A part of the template cDNA was used for RNA-Seq. Meanwhile, the template cDNAs amplified with a 20 μL of reaction solution by using TOROGreen^® qPCR Master Mix (Toroivd, Shanghai, China). The qRT-PCR program consisted of a preliminary step of 60 s at 95 °C, followed by 40 cycles at 95 °C for 10 s, and 60 °C for 30 s. The Actin7 from melon was used as an internal control [55 (link)]. The relative gene expression level was calculated via the 2^−ΔΔCt method. Three biological and three technical replicates were used for each sample. All the primers used in this study are listed in Table S2. In addition, the raw transcriptome sequencing data was used to analyze the tissue specificity of NF-Y members and their roles in fruit ripening, including five melon plant tissues, including roots, leaves, male flowers, female flowers, and fruits (PRJNA383830) [62 (link)], as well as fruit development of green-fleshed (Dulce) and orange-fleshed (Tam-Dew) at 10, 20, and 30 days after anthesis (DAA) and maturity, respectively (PRJNA286120) [37 (link)].