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.

Protein SUMOylation is a key modification for regulating the fate of proteins and it is widely involved in plant development and stress responses. The SUMO molecules are conjugated to the lysine residues of substrate proteins via isopeptide bonds by enzyme reaction. SUMOylation is mediated by an enzyme cascade composed of a SUMO activating enzyme complex (E1), a SUMO-conjugating enzyme (E2) and usually a SUMO ligase (E3). Here, we report an efficient in vitro detection system for SUMOylation of plant proteins. We established a system for SUMOylation detection of plant proteins by reconstructing the Arabidopsis SUMOylation enzyme cascade in Escherichia coli. Using this system, SUMOylation of several substrates were detected via immunoblotting. Therefore, this system simplifies the SUMOylation detection of plant protein substrates and provides a powerful tool for functional analysis of SUMOylation in plant cells.