利用流式细胞仪分选拟南芥根尖发育早期非根毛细胞

建立了应用流式细胞仪分选植物特定类型细胞的方法。以拟南芥(Arabidopsis thaliana)Wer::GFP转基因株系为材料,用激光共聚焦显微镜鉴定GFP的表达位置,采用酶解法制备拟南芥根尖原生质体,应用流式细胞仪荧光激活细胞分选技术(FACS)分选收集GFP阳性细胞,并提取细胞的RNA。结果表明,Wer::GFP转基因株系仅在根表皮发育早期的非根毛细胞中表达GFP;利用酶解法制备的根尖原生质体数目较多;从FACS分选收集的细胞中提取的RNA质量较好,可用于研究特定类型细胞的基因表达谱。应用流式细胞仪分选拟南芥非根毛细胞的方法为研究植物特定类型细胞的基因表达谱及基因功能奠定了技术基础。

orting of early developmental non-hair cells in root by flow cytometry in Arabidopsis thaliana

We have developed a method for sorting specific cells in plant. Here, WER::GFP transgenic lines in Arabidopsis thaliana were as the experimental material; laser scanning confocal microscopy was used to identify the expression location of GFP; enzymatic digestion was applied for preparation of root tip protoplasts; fluorescence activated cell sorting (FACS) was applied to collect GFP positive cells; and then the total RNA was extracted from these cells. The results showed that only the early developmental non-root hair cells in the root epidermis express GFP in Wer::GFP transgenic lines; larger number of root tip protoplasts were obtained by using enzymatic digestion; and then GFP positive cells were collected successfully by FACS. The RNA extracted from these specific cells can be used for gene expression profiling. The method for sorting specific cells had been established successfully.