Imaging Nuclear Morphology and Organization in Cleared Plant Tissues Treated with Cell Cycle Inhibitors

Abstract

Synchronization of root cells through chemical treatment can generate a large number of cells blocked in specific cell cycle phases. In plants, this approach can be employed for cell suspension cultures and plant seedlings. To identify plant cells in the course of the cell cycle, especially during mitosis in meristematic tissues, chemical inhibitors can be used to block cell cycle progression. Herein, we present a simplified and easy-to-apply protocol to visualize mitotic figures, nuclei morphology, and organization in whole Arabidopsis root apexes. The procedure is based on tissue clearing, and fluorescent staining of nuclear DNA with DAPI. The protocol allows carrying out bulk analysis of nuclei and cell cycle phases in root cells and will be valuable to investigate mutants like overexpressing lines of genes disturbing the plant cell cycle.

Exploiting cell cycle inhibitor genes of the KRP family to control root‐knot nematode induced feeding sites in plants

“…4,6‐Diamidino‐2‐phenylindole (DAPI) staining of nuclei in cleared galls was performed and analysed as described by Antonino de Souza Junior et al (). DAPI stained samples were mounted in 90% glycerol on microscope slides.…”