| Detection of Nitric Oxide and Determination of Nitrite Concentrations in Arabidopsis thaliana and Azospirilum brasilense
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Author:
Date:
2016-03-20
[Abstract] There is now general agreement that nitric oxide (NO) is an important and almost ubiquitous signal in plants. Nevertheless, there are still many controversial observations and differing opinions on the importance and functions of NO in plants. Partly, this may be due to the difficulties in detecting and quantifying NO. Here, we summarize protocols for detecting NO and quantifying nitrite concentration in Arabidopsis seedlings. We also present a method to measure NO in biofilms formed by the plant growth promoting rhizobacteria Azospirillum brasilense (A. brasilense). NO in oxygen-containing aqueous solutions has a short half-life that is often attributed to a rapid oxidation to nitrite. Here we detail the use of the fluorescent probe DAF-FM DA and the ...
[摘要] 现在普遍认为一氧化氮（NO）是植物中重要的和几乎普遍存在的信号。然而，对于植物中NO的重要性和功能仍有许多有争议的观察和不同的观点。部分地，这可能是由于检测中的困难以及甚至更多的在定量NO。在这里，我们总结了在拟南芥幼苗中检测NO和定量亚硝酸盐浓度的方案以及在由植物生长促进根瘤菌（Azospirillum brasilense）形成的生物膜中的NO实时测量（ > A。brasilense ）。含氧水溶液中的NO具有短的半衰期，其通常归因于快速氧化成亚硝酸盐。在这里我们详细的使用荧光探针DAF-FM DA和电化学方法分别直接检测和定量NO，Griess试剂通过其氧化的亚硝酸盐形式间接检测NO。这些方案可用于多种细胞类型和植物的不同组织以及微生物。
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| A Protocol to Measure the Cytoplasmic Calcium in Arabidopsis Guard Cells
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Author:
Date:
2015-05-05
[Abstract] Cytoplasmic calcium ([Ca2+]cyt) acts as a stimulus-induced second messenger in multiple signal transduction cascades (Allen et al., 1999). In plant cells, a dramatic and readily assayed response to stimulus is the change of stomatal aperture. Changes in [Ca2+]cyt of stomatal guard cells were involved in stomatal movement in response to various stimuli and cellular processes. In general, there are two available ways to measure [Ca2+]cyt in guard cells, i.e., loading of calcium-sensitive fluorescence dyes such as fluo-3 AM and fura-2 or expressing genetically encoded calcium indicators such as yellow cameleon (Krebs et al., 2012). In this protocol, we aim at describing the experimental procedure to ...
[摘要] 细胞质钙（[Ca 2+] + cyt）在多个信号转导级联中用作刺激诱导的第二信使（Allen等人， 1999）。在植物细胞中，对刺激的戏剧性和容易测定的响应是气孔孔径的变化。气孔保卫细胞的[Ca + ] 细胞的变化参与响应于各种刺激和细胞过程的气孔运动。一般来说，有两种可用的方法来测量保护细胞中的钙离子荧光染料的加载量，即 例如fluo-3AM和fura-2或表达遗传编码的钙指示剂例如黄色卡梅伦（Krebs等人，2012）。在该方案中，我们旨在描述在ABA或PA处理时加载fluo-3AM的保卫细胞中记录[Ca 2 cyT 用共聚焦激光扫描显微镜进行荧光成像。
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