Caillaud Marie-Cécile, Favery Bruno
Laboratoire de Reproduction et Développement des Plantes, Centre National de la Recherche Scientifique, Institut National de la Recherche Agronomique, Ecole Normale Supérieure de Lyon, Université Claude Bernard Lyon 1, Lyon, France.
INRA, Institut Sophia Agrobiotech, UMR 1355, 400 route des Chappes, 06903, Sophia-Antipolis, France.
Methods Mol Biol. 2016;1370:137-44. doi: 10.1007/978-1-4939-3142-2_11.
Mitosis which is a major step during plant development can also be observed in physiopathological conditions. During the compatible interaction between the root-knot nematode Meloidogyne incognita and its host Arabidopsis, the pathogen induce through repeated divisions without complete cytokinesis the formation of hypertrophied and multinucleate feeding cells, named giant cells. Due to the presence of hypertrophied plant cell material surrounding the giant cells, classical live cell imaging gave therefore very poor resolution. Here, we describe a protocol which allows the in vivo observation of the mitotic apparatus in developing giant cells using confocal imaging of vibrosliced tissues. This approach can also be used to visualize in vivo other cellular processes occurring in different steps of giant cells.
有丝分裂是植物发育过程中的一个重要步骤,在生理病理条件下也可以观察到。在根结线虫南方根结线虫与其宿主拟南芥的亲和互作过程中,病原体通过重复分裂但不进行完全胞质分裂,诱导形成肥大的多核取食细胞,即巨型细胞。由于巨型细胞周围存在肥大的植物细胞物质,传统的活细胞成像分辨率非常低。在这里,我们描述了一种方案,该方案允许使用振动切片组织的共聚焦成像在发育中的巨型细胞中对有丝分裂装置进行体内观察。这种方法也可用于在体内可视化巨型细胞不同阶段发生的其他细胞过程。