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Systematic Methods for Isolating High Purity Nuclei from Ten Important Plants for Omics Interrogation.

ABSTRACT: Recent advances in developmental biology have been made possible by using multi-omic studies at single cell resolution. However, progress in plants has been slowed, owing to the tremendous difficulty in protoplast isolation from most plant tissues and/or oversize protoplasts during flow cytometry purification. Surprisingly, rapid innovations in nucleus research have shed light on plant studies in single cell resolution, which necessitates high quality and efficient nucleus isolation. Herein, we present efficient nuclei isolation protocols from the leaves of ten important plants including Arabidopsis, rice, maize, tomato, soybean, banana, grape, citrus, apple, and litchi. We provide a detailed procedure for nucleus isolation, flow cytometry purification, and absolute nucleus number quantification. The nucleus isolation buffer formula of the ten plants tested was optimized, and the results indicated a high nuclei yield. Microscope observations revealed high purity after flow cytometry sorting, and the DNA and RNA quality extract from isolated nuclei were monitored by using the nuclei in cell division cycle and single nucleus RNA sequencing (snRNA-seq) studies, with detailed procedures provided. The findings indicated that nucleus yield and quality meet the requirements of snRNA-seq, cell division cycle, and likely other omic studies. The protocol outlined here makes it feasible to perform plant omic studies at single cell resolution.

SUBMITTER: Yang MC

PROVIDER: S-EPMC9740283 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature

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Systematic Methods for Isolating High Purity Nuclei from Ten Important Plants for Omics Interrogation.

Yang Ming-Chao MC Wu Zi-Chen ZC Huang Liang-Liang LL Abbas Farhat F Wang Hui-Cong HC

Cells 20221203 23

Recent advances in developmental biology have been made possible by using multi-omic studies at single cell resolution. However, progress in plants has been slowed, owing to the tremendous difficulty in protoplast isolation from most plant tissues and/or oversize protoplasts during flow cytometry purification. Surprisingly, rapid innovations in nucleus research have shed light on plant studies in single cell resolution, which necessitates high quality and efficient nucleus isolation. Herein, we ...[more]

PMID: 36497177

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Dataset Information

Systematic Methods for Isolating High Purity Nuclei from Ten Important Plants for Omics Interrogation.

Publications

Systematic Methods for Isolating High Purity Nuclei from Ten Important Plants for Omics Interrogation.

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