利用活细胞成像技术可视化伸长棉纤维中的细胞骨架组织和动态变化。

Visualization of Cytoskeleton Organization and Dynamics in Elongating Cotton Fibers by Live-Cell Imaging.

机构信息

State Key Laboratory of Plant Genomics, Institute of Microbiology, Innovative Academy of Seed Design, Chinese Academy of Sciences, Beijing, China.

Shanxi Agricultural University, Taigu, China.

出版信息

Methods Mol Biol. 2023;2604:311-316. doi: 10.1007/978-1-0716-2867-6_25.

DOI:10.1007/978-1-0716-2867-6_25

PMID:36773245

Abstract

Cotton fibers are extremely elongated single cells and have long been regarded as an ideal model to investigate polarized plant cell elongation. Actin filaments (F-actin), as well as the cortical microtubules (CMTs), play vital roles in polarized cell growth and morphogenesis. We have generated stable transgenic cotton plants expressing fluorescent markers for the actin and microtubule cytoskeletons. Further live-cell imaging identified dynamic features of the F-actin and cortical microtubule (CMT) architectures and discovered that cotton fibers elongate in a unique tip-biased diffuse growth mode. Here, we describe methods for preparing growing cotton fiber samples, as well as the visualization of cytoskeletal organization and dynamics by live-cell imaging. Combined with comprehensive image analyses, these methods can be used to identify how cytoskeleton organization and dynamics determine cell morphogenesis in highly polarized cotton fibers.

摘要

棉纤维是极其细长的单细胞，长期以来一直被视为研究植物细胞极性伸长的理想模型。肌动蛋白丝（F-actin）和皮层微管（CMTs）在极性细胞生长和形态发生中起着至关重要的作用。我们已经生成了稳定表达荧光标记物的转基因棉花植物，用于肌动蛋白和微管细胞骨架。进一步的活细胞成像确定了 F-actin 和皮层微管（CMT）结构的动态特征，并发现棉花纤维以独特的尖端偏向弥散生长模式伸长。在这里，我们描述了制备生长中的棉花纤维样品的方法，以及通过活细胞成像观察细胞骨架组织和动态的方法。结合全面的图像分析，这些方法可用于确定细胞骨架组织和动态如何决定高度极性棉花纤维中的细胞形态发生。