Imaging nuclear morphology and organization in cleared plant tissues treated with cell cycle inhibitors
2016
Antonino de Souza Junior, Jose Dijair | Grossi de Sa, Maria Fatima | Engler, Gilbert | Engler, Janice de Almeida | Laboratório de Interação Molecular Planta-Praga ; Embrapa Recursos Genéticos e Biotecnologia = Embrapa Genetic Resources & Biotechnology (CENARGEN) ; Brazilian Agricultural Research Corporation = Empresa Brasileira de Pesquisa Agropecuária (Embrapa)-Brazilian Agricultural Research Corporation = Empresa Brasileira de Pesquisa Agropecuária (Embrapa) | Institut Sophia Agrobiotech (ISA) ; Institut National de la Recherche Agronomique (INRA)-Université Nice Sophia Antipolis (1965 - 2019) (UNS)-Centre National de la Recherche Scientifique (CNRS)
International audience
Mostrar más [+] Menos [-]Inglés. 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.
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