Abstract

Many functionally distinct plant tissues have relatively low numbers of cells that are embedded within complex tissues. For example, the shoot apical meristem (SAM) consists of a small population of pluripotent stem cells surrounded by developing leaves and/or flowers at the growing tip of the plant. It is technically challenging to collect enough high-quality SAM samples for molecular analyses. Isolation of Nuclei Tagged in specific Cell Types (INTACT) is an easily reproducible method that allows the enrichment of biotin-tagged cell-type-specific nuclei from the total nuclei pool using biotin-streptavidin affinity purification. Here, we provide a detailed INTACT protocol for isolating nuclei from the Arabidopsis SAM. One can also adapt this protocol to isolate nuclei from other tissues and cell types for investigating tissue/cell-type-specific transcriptome and epigenome and their changes during developmental programs at a high spatiotemporal resolution. Furthermore, due to its low cost and simple procedures, INTACT can be conducted in any standard molecular laboratory.

Keywords

Arabidopsis; Biotinylation; Cell-type-specific promoter; in situ streptavidin histochemistry; INTACT; low-input RT-qPCR; Shoot apical meristem

Published in

Methods in Molecular Biology
2023, Volume: 2686, number: 2686, pages: 313-328 Title: Flower Development : Methods and Protocols
ISBN: 978-1-0716-3298-7, eISBN: 978-1-0716-3299-4
Publisher: Humana Press Inc.

SLU Authors

• Schmid, Markus

  • Department of Plant Biology, Swedish University of Agricultural Sciences
    
  • Umeå University

UKÄ Subject classification

Cell Biology
Botany


Publication identifier

DOI: https://doi.org/10.1007/978-1-0716-3299-4_16

Permanent link to this page (URI)

https://res.slu.se/id/publ/129516