DNA-Seq of Paspalum rufum sexual genotype : full spikelets samples at premeiotic stage, digested with AciI/MseI.
A detailed comparative methylation analysis was carried out in full spikelets of highly sexual plants (showing 1.22% and 0% of ovules bearing Aposporous Embryo Sacs), and highly aposporic plants (showing 32.7% and 35.8% of ovules bearing one or more AES), at different developmental stages (premeiosis, postmeiosis). Three specific enzyme combinations were chosen including each of the three methylation-sensitive enzymes, combined with methylation-insensitive MseI, to infer CG (AciI/MseI), CHG (PstI/MseI) and CHH (EcoT22I/MseI) methylation contexts, respectively. Triplicate samples were extracted for each genotype at each developmental stage (total number of samples: 36). The library set-up protocol was performed according to Marconi et al. (2019) and Di Marsico et al. (2020).Each library was quantified using qPCR (Light Cycler 480 Roche) and normalized to a 3 nM concentration. The grouped libraries were pooled in an equimolar fashion, and were Illumina-sequenced by HiSeq 1500 Illumina, using 150-bp single-end chemistry.