RALF RNA-seq treatment rep2
In vitro culture of ovule was performed referring to a previous study (Qin et al., 2007). The peptide sequence of GhRALF1 after the canonical RRXL cleavage were synthesized (Dia-An Biotechnology, China). The peptide was added to the medium to a final concentration of 1 m as treatment. After 30 mins treatment, RNA of ovule tissue (1DPA) was extracted using our previous method (Wang et al., 2019a). The total RNA was reverse transcribed by using EasyScript One-step gDNA Removal and cDNA Synthesis SuperMix (AE311-03, Transgen Biotech, China) with oligo-dT and random primers. The DNBSEQ (BGI, China) with PE150 mode was used for sequencing the libraries.