Analysis of Actin-Based Intracellular Trafficking in Pollen Tubes
Underlying rapid and directional pollen tube growth is the active intracellular trafficking system that carries materials necessary for cell wall synthesis and membrane expansion to the expanding point of the pollen tube. The actin cytoskeleton has been shown to control various intracellular trafficking events in the pollen tube, but the underlying cellular and molecular mechanisms remain poorly understood. To better understand how the actin cytoskeleton is involved in the regulation of intracellular trafficking events, we need to establish assays to visualize and quantify the distribution and dynamics of organelles, vesicles, or secreted proteins. In this chapter, we introduce methods regarding the visualization and quantification of the distribution and dynamics of organelles or vesicles in pollen tubes.
Key wordsArabidopsis thaliana Pollen tube Intracellular trafficking Cytoplasmic streaming Exocytosis Actin dynamics Actin-binding protein (ABP) Fluorescence recovery after photobleaching (FRAP)
We thank Prof. Yan Zhang (Shandong Agricultural University) for providing us the transgenic Arabidopsis seeds harboring Lat52::YFP-RabA4b and Lat52::YFP-ARA7. This work was supported by grants from the Ministry of Science and Technology of China (2013CB945100) and the National Natural Science Foundation of China (31671390 and 31471266).