Isolation of Plant Nuclei Compatible with Microfluidic Single-nucleus ATAC-sequencing

Area of Science:

• Plant molecular biology
• Epigenetics
• Genomics

Background:

• Gene expression is regulated by transcription factor binding to DNA.
• Chromatin accessibility is crucial for this binding and varies across cell types.
• Previous ATAC-seq methods had limitations in plant single-cell resolution.

Purpose of the Study:

• To develop and present a robust protocol for single-nucleus ATAC-seq (sNucATAC-seq) in plants.
• To enable high-resolution analysis of chromatin accessibility in individual plant cells.
• To facilitate deeper understanding of gene regulation in plants.

Main Methods:

• Isolation of nuclei from plant root and green tissues.
• Nuclear membrane permeabilization for Tn5 transposase access.
• Library preparation using 10x Genomics Chromium technology for sNucATAC-seq.

Main Results:

• A detailed protocol for plant sNucATAC-seq is described.
• The method successfully generated finely tuned chromatin accessibility profiles.
• Successful application on *Arabidopsis thaliana* and *Glycine max* root nuclei.

Conclusions:

• The developed sNucATAC-seq protocol is effective for plant tissues.
• This method enhances the study of plant epigenomes at single-cell resolution.
• It opens new avenues for investigating gene regulation in diverse plant species.