Single‐nucleus RNA sequencing of plant tissues using a nanowell‐based system

• DOI: 10.1111/tpj.15458
• Publication Date: 2021-08-13T09:08:31Z
• Authors: Daniele Y. Sunaga...; Jose M. Muino; Caroline Braeuning; Xiaocai Xu; Minglei Zong; Cezary Smaczniak; Wenhao Yan; Cornelius Fischer; Ramon Vidal; Magdalena Kliem; Kerstin Kaufmann; Sascha Sauer
• Keywords: Genetic Markers; snRNA‐seq; Arabidopsis; 570 Biologie; Flowers; 03 medical and health sciences; Gene Expression Regulation, Plant; RNA, Small Nuclear; flower development; ddc:580; Inflorescence; Cell Nucleus; 0303 health sciences; Sequence Analysis, RNA; Reproducibility of Results; 580 Pflanzen (Botanik); RNA, Plant; Seedlings; Arabidopsis thaliana seedlings; anther; single‐nucleus isolation; ddc:570; Technology Platforms; Genes, Cells and Cell-Based Medicine [Topic 1]; technical advance

SUMMARY Single‐cell genomics provides unprecedented potential for research on plant development and environmental responses. Here, we introduce a generic procedure nucleus isolation combined with nanowell‐based library preparation. Our method enables the transcriptome analysis of thousands individual nuclei. It serves as an alternative to use protoplast isolation, which is currently standard methodology single‐cell genomics, although it can be challenging some tissues. We show applicability our by using different materials from species. The single‐nucleus RNA sequencing shown through characterization transcriptomes seedlings developing flowers Arabidopsis thaliana . evaluated dynamics during early stages anther development, identified stage‐specific activities transcription factors regulating this process, predicted target genes these factors. applied in species tissues, thus expanding toolkit experiments.