GSM5605644: young inflorescences, S2-2; Oryza sativa Japonica Group; RNA-Seq
For protoplast isolation, samples were firstly cut into small pieces with a fresh sharp razor blade, then digested at room temperature in RNase-free enzyme solution [1.5 % (w/v) cellulase R-10, 1 % (w/v) macerozyme R-10 (Yakult), 0.4 M mannitol, 20 mM MES pH 5.7, 20 mM KCl, 10 mM CaCl2, 0.1 % BSA (Sigma-Aldrich)]. Protoplasts were filtered using a 45 μm cell strainer (Biologix), and centrifuged at 200 g for 3 min. After the supernatant was removed, the pellet was washed three times with 8 % (w/v) mannitol, and cell viability was determined by AO/PI (acridine orange/propidium iodide) staining (Countstar® Rigel, ALIT Life Science). Digestion progress was assayed every hour for 6 h, with the optimum time for cell viability and protoplast digestion determined to be 2 h. Sequencing libraries were conducted using the BD Rhapsody Single-Cell Analysis System (BD Biosciences), following the manufacture's protocol. scRNA-seq