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植物学报 >

2024 , Vol. 59 >Issue 5: 774 - 782

DOI: https://doi.org/10.11983/CBB24107

特邀专家方法

植物基因组学中的流式细胞分析及分选技术

  • 夏春皎 ,
  • 李运广 ,
  • 夏姝 ,
  • 庞伟 ,
  • 陈春丽
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  • 华中农业大学生命科学技术学院, 武汉 430070
*陈春丽, 华中农业大学生命科学技术学院, 教授, 博士生导师。从事植物干细胞生物学、植物染色体生物学研究以及本科生课程(植物学、植物学实验、植物学基础)和研究生课程(植物显微技术、高级植物学)的教学工作。先后主持国际科学青年基金、国家自然科学基金以及科技部“十四五”重点研发计划等项目, 在植物干细胞与再生以及植物染色体研究中取得系列成果。以第一作者或通讯作者身份在Nature Plants、Molecular Plant、New Phytologist和Journal Genetics and Genomics等国内外主流期刊发表论文20多篇。其研究团队聚焦国际前沿, 与英国和日本等国的知名大学及研究所长期合作, 发起成立植物干细胞国际学术联盟和植物干细胞国际联合实验室, 组织召开植物干细胞与再生国际研讨会(2022年)、首届全国植物干细胞生物学大会(2023年)及植物干细胞国际讲坛(2022年)。E-mail: chenchunli@mail.hzau.edu.cn

收稿日期: 2024-07-18

  录用日期: 2024-08-20

  网络出版日期: 2024-08-22

基金资助

国家重点研发计划(2023ZD04073);华中农业大学研究生培养条件建设(2024SC03)

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Flow Cytometric Analysis and Sorting in Plant Genomics

  • Chunjiao Xia ,
  • Yunguang Li ,
  • Shu Xia ,
  • Wei Pang ,
  • Chunli Chen
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  • College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China

Received date: 2024-07-18

  Accepted date: 2024-08-20

  Online published: 2024-08-22

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摘要

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。

关键词： 植物; 流式细胞术; 细胞核; 分选; ATAC-seq; RNA-seq

本文引用格式

夏春皎 , 李运广 , 夏姝 , 庞伟 , 陈春丽 . 植物基因组学中的流式细胞分析及分选技术[J]. 植物学报, 2024 , 59(5) : 774 -782 . DOI: 10.11983/CBB24107

Abstract

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.

Key words： plants; flow cytometry; nuclei; sorting; ATAC-seq; RNA-seq

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