Chemiluminescence Detection of the Oxidative Burst in Plant Leaf Pieces
化学发光检测植物叶片中的氧化迸发
发布: 2015年03月20日第5卷第6期 DOI: 10.21769/BioProtoc.1423 浏览次数: 19719
PDF 
Q&A
参见作者原研究论文
The authors used this protocol in:
May 2014
Advertisement
Abstract
The production of 'reactive oxygen species' (ROS), also termed oxidative burst, is a typical cellular response of animals and plants to diverse biotic and abiotic stresses. Here, we outline the detection of the ROS-burst in plant leaf pieces using a luminol-based bioassay which allows for the detection of chemiluminescence. The assay was originally described by Keppler et al. (1989) and subsequently adapted for other plant cells and tissues (Felix et al., 1999) and also used in recent publications (Albert et al., 2013; Albert et al., 2010; Butenko et al., 2014; Halter et al., 2014). In this protocol we outline a standardized version of this assay including remarks and recommendations for data evaluation and interpretation of results.
Materials and Reagents
- Plant leaf pieces (any plant of interest, e.g. Arabidopsis or N. benthamiana transiently expressing a construct of interest)
- Water
- 200 µM luminol L-012 (Wako Chemicals USA, catalog number: 075-05111 ; http://www.wako-chem.co.jp/english/labchem/journals/wpu_bio1/10.htm)
- 10 µg/ml peroxidase, horseradish peroxidase (AppliChem, catalog number: A3791 )
- MAMPs or any elicitors of interest
- Known MAMP as positive control (e.g. flg22, chitin-oligomers, etc.)
- 10x luminol master mix (see Recipes)
Equipment
- Scissors
- Petridishes
- Small spatula
- 96 well plates, white, flat-bottomed (e.g., LIA-plate, whit, 96-well, flat-bottom, Greiner Bio-One GmbH) or suitable cuvettes for single cell instruments
- Luminometer, either a 96 well plate reader (e.g., Centro LB 960, Microplate Luminometer, BERTHOLD TECHNOLOGIES) or a single cell instrument
Procedure
文章信息
版权信息
© 2015 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Albert, M., Butenko, M. A., Aalen, R. B., Felix, G. and Wildhagen, M. (2015). Chemiluminescence Detection of the Oxidative Burst in Plant Leaf Pieces. Bio-protocol 5(6): e1423. DOI: 10.21769/BioProtoc.1423.
Download Citation in RIS Format
分类
植物科学 > 植物免疫 > 病害生物测定
生物化学 > 其它化合物 > 活性氧
您对这篇实验方法有问题吗？
在此处发布您的问题，我们将邀请本文作者来回答。同时，我们会将您的问题发布到Bio-protocol Exchange，以便寻求社区成员的帮助。
发布问题
