Abstract To explore the effect of gene expression on the fate of root tip cells, different types of root cells need to be grouped for further analysis. In the present study, the protoplasts of young root tips of Zea mays were obtained by enzymatic hydrolysis, and analyzed and sorted by fluorescence staining and flow cytometry. The results showed that the optimum method for obtaining protoplasts of 4~5 d young maize root tip tissue by enzymatic hydrolysis included combination of 1.0% cellulase and 0.3% pectinase, optimum osmotic pressure with 10% mannitol, optimum enzymatic hydrolysis time with 4 h and optimum pH with 5.8. The protoplasts extracted under the above optimum conditions were purified and the impurities such as cell fragments in the protoplast suspension were effectively removed. The purified protoplasts were stained by Propidium iodide (PI) and Fluorescein diacetate (FDA), and the observation under fluorescence microscopy showed that the protoplasts were highly active. For further sorting by flow cytometry, different fluorescent dyes were used. The staining effect of Hoechst33342 on active protoplasts was better than that of 4', 6-diamidyl-2-phenylindole (DAPI), most protoplasts in the system were stained with high resolution. At the same time, for Hoechst33342, the optimum dyeing temperature was 38 ℃, and the optimum dyeing pH was 10. The study provides basic data for further related researches on root tip cells in plants.
KONG Yao,FENG Jiao-Jiao,ZHANG Zheng-Quan, et al. Isolation and Flow Analysis of Protoplasts from Maize (Zea mays) Root Tip[J]. , 2018, 26(12): 2160-2167.
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