Qu Xiaolu, Wang Qiannan, Wang Haiyan, Huang Shanjin
Key Laboratory of Horticultural Plant Biology (Ministry of Education), College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan, China.
Center for Plant Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China.
Bio Protoc. 2020 Jan 5;10(1):e3509. doi: 10.21769/BioProtoc.3509.
Although it is widely accepted that actin plays an important role in regulating pollen germination and pollen tube growth, how actin exactly performs functions remains incompletely understood. As the function of actin is dictated by its spatial organization, it is the key to reveal how exactly actin distributes in space in pollen cells. Here we describe the protocol of revealing and quantifying the spatial organization of actin using fluorescent phalloidin-staining in fixed pollen grains and pollen tubes. We also introduce the method of assessing the stability and/or turnover rate of actin filaments in pollen cells using the treatment of latrunculin B.
尽管人们普遍认为肌动蛋白在调节花粉萌发和花粉管生长中起重要作用，但肌动蛋白究竟如何发挥功能仍不完全清楚。由于肌动蛋白的功能取决于其空间组织，因此揭示肌动蛋白在花粉细胞中如何在空间上分布是关键。在这里，我们描述了使用荧光鬼笔环肽染色在固定的花粉粒和花粉管中揭示和量化肌动蛋白空间组织的方案。我们还介绍了使用拉特罗毒素B处理评估花粉细胞中肌动蛋白丝稳定性和/或周转率的方法。
