植物基因组学中的流式细胞分析及分选技术

Flow Cytometric Analysis and Sorting in Plant Genomics

流式细胞术是一种高通量技术,可以同时快速检测单个颗粒物的多项物理及生物学特征。随着测序成本的大幅降低,流式细胞术在植物基因组学高通量样品获取中的作用日益凸显。该文以水稻(Oryza sativa)和大豆(Glycine max)为例,详细介绍了应用流式细胞术对植物细胞核进行精细分选以及后续的ATAC-seq和RNA-seq实验分析流程,为农业生物育种中基因的高效挖掘提供了优选工具。同时针对实验操作中的关键技术和常见问题,如细胞核制备注意事项、分选纯度与效率的平衡及单细胞分选调试方法进行分析并提出建议,为植物科学工作者应用流式细胞术开展基因组学研究提供参考。

Flow cytometry is a high-throughput technology that allows for the simultaneous and rapid detection of multiple physical and biological characteristics of individual particles.With the significant reduction in sequencing costs,flow cytometry is playing an increasingly prominent role in high-throughput sample acquisition for plant genomics.Taking rice and soybean as examples,this paper describes in detail the application of flow cytometry for fine sorting of plant cell nuclei and the subsequent ATAC-seq and RNA-seq experiments and analysis process,which provides a preferred tool for efficient mining of genes for agrobiological breeding.The key techniques and common problems in the experimental operation,such as the precautions for cell nuclei preparation,the balance between sorting purity and efficiency,and the debugging method for single-cell sorting,were also analyzed and suggested to provide references for the plant scientists in applying flow cytometry to carry out genomics research.