Isolation of high purity guard cell protoplasts of Arabidopsis thaliana for omics research

摘要: Stomata are unique plant structures responsible for photosynthesis, transpiration and innate immunity to pathogens, guard cells one of the most studied cell types with respect functioning, signalling, stress responses. The ability easily purify large quantities high purity protoplasts (GCPs) would facilitate further studies, as current methods GCP isolation barely sufficient omics research. Here, we report a new procedure isolating GCPs. For GCPs, detached epidermal peels were used extract GCPs instead whole leaves. mesophyll (MCPs) found have diameters 2.5–9.1 µm 6.5–43.5 µm, respectively. overlap in sizes MCPs suggests that blending filtering leaves previous may not necessarily avoid contamination during extraction. There were, on average, 8.4 ± 0.18 chloroplasts 34.6 ± 1.5 MCPs. similar sizes, there fourfold more MCPs, which made readily distinguishable from by microscopic inspection. protocol enabled over 1.44 × 106 about 150 cm2Arabidopsis abaxial epidermis 97% purity. These protocols provide an advance sufficient, purity, viable RNA extraction transcriptomic analysis. application this other species accelerate research development cell-type specific omics.