Abstract：
Macroautophagy, hereafter autophagy, plays a crucial role in the degradation of harmful or unwanted cellular components through a double-membrane autophagosome. Upon autophagosome fusion with the vacuole, the degraded materials are subsequently recycled to generate macromolecules, contributing to cellular homeostasis, metabolism, and stress tolerance in plants. A hallmark during autophagy is the formation of isolation membrane structure named as phagophore, which undergoes multiple steps to become as a complete double-membrane autophagosome. Methodologies have been developed in recent years to observe and quantify the autophagic process, which greatly advance knowledge of autophagosome biogenesis in plant cells. In this chapter, we will introduce two methods to dissect the autophagosome-related structures in the Arabidopsis plant cells, including the correlative light and electron microscopy, to map the ultrastructural feature of autophagosomal structures, and time-lapse imaging to monitor the temporal recruitment of autophagy machinery during autophagosome formation.
摘要：
巨自噬,自噬以后，在通过双膜自噬体降解有害或不需要的细胞成分中起着至关重要的作用。自噬体与液泡融合后，降解的材料随后被回收以产生大分子，有助于细胞内稳态，新陈代谢，和植物的胁迫耐受性。自噬过程中的一个标志是形成称为吞噬团的隔离膜结构，它经历多个步骤成为一个完整的双膜自噬体。近年来已经开发了观察和量化自噬过程的方法，这极大地促进了植物细胞中自噬体生物发生的知识。在这一章中,我们将介绍两种方法来解剖拟南芥植物细胞中的自噬体相关结构，包括相关的光学和电子显微镜，绘制自噬体结构的超微结构特征，和延时成像来监测自噬体形成过程中自噬机制的时间募集。
