| Microscopic Observation of Subcellular GFP-tagged Protein Localization in Rice Anthers
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Author:
Date:
2018-08-05
[Abstract] This protocol demonstrates a simple method to determine the subcellular localization of fluorescence-tagged proteins on the vibratome sections of rice developing anthers. If a cell type-specific promoter is used to drive the tagged protein-encoding gene, the method enables to clearly distinguish the cells retaining fluorescent signals from other anther cells. It is applicable to both live and fixed samples, and presumably to other plant tissues.
[摘要] 该方案演示了一种确定荧光标记蛋白在水稻发育花药的振动切片上的亚细胞定位的简单方法。 如果使用细胞类型特异性启动子来驱动标记的蛋白质编码基因，该方法能够清楚地区分保留荧光信号的细胞与其他花药细胞。 它适用于活的和固定的样品，并且可能适用于其他植物组织。
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| Vacuole Structure Analysis during Cell Death Subsequent to Application of Erwinia carotovora Culture Filtrates to Cell Cultures of Nicotiana tabacum
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Author:
Date:
2015-10-20
[Abstract] We recently established an experimental model system for efficient defense-related cell death using tobacco BY-2 cultured cells treated with culture filtrates of the pathogenic bacterium Erwinia carotovora (E. carotovora) (Hirakawa et al., 2015). Applying this experimental system to transgenic BY-2 cells stably expressing the vacuolar membrane marker GFP-VAM3 (Kutsuna and Hasezawa, 2002) allowed us to monitor changes in vacuolar membrane structures including a decrease of transvacuolar strands during cell death (Hirakawa et al., 2015). Our model system can ...
[摘要] 我们最近建立了使用烟草BY-2培养细胞的有效防御相关细胞死亡的实验模型系统，所述培养细胞用致病性细菌欧文氏菌（Erwinia carotovora）（ E。carotovora （Hirakawa ，，2015）。将该实验系统应用于稳定表达液泡膜标记物GFP-VAM3的转基因BY-2细胞（Kutsuna和Hasezawa，2002），使我们能够监测液泡膜结构的变化，包括细胞死亡期间跨血管链的减少（Hirakawa et al。 al。，2015）。我们的模型系统可以帮助调查细胞器动力学在国防相关的细胞死亡。在这里，我们显示应用 E的协议。 carotovora过滤到BY-2细胞和共聚焦观察液泡膜动力学和随后的细胞死亡。我们使用细胞周期同步的BY-2细胞来有效地监测内陷的液泡膜，例如在我们最近的报告中的跨血管链（Hirakawa等人，2015）;然而，我们不描述本文中的细胞周期同步的协议。对于BY-2细胞同步的逐步方案，请参考先前的方案论文（Nagata和Kumagai，1999; Kumagai-Sano等人，2006）。
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