Imanifard Zahra, Vandelle Elodie, Bellin Diana
Department of Biotechnology, University of Verona, Verona, Italy.
Methods Mol Biol. 2018;1743:39-50. doi: 10.1007/978-1-4939-7668-3_4.
The hypersensitive response is one of the most powerful and complex defense reactions to survive to pathogen attacks during an incompatible plant-pathogen interaction. Local programmed cell death accompanies the hypersensitive response at the site of infection to prevent pathogen growth and spread. A precise quantitative assessment of this form of programmed cell death is essential to unravel the genetic and molecular mechanisms underlying the process. Here, we first describe the optimization of a Trypan Blue staining protocol for quantitatively measuring the HR-cell death in Arabidopsis. Furthermore, we provide an electrolyte leakage protocol based on pathogen vacuum infiltration, which allows its simultaneous application to a large number of plants as well as to Arabidopsis mutants affected by small size phenotype.
过敏反应是植物与病原菌非亲和互作过程中抵御病原菌侵袭的最有效且最复杂的防御反应之一。在感染部位,过敏反应伴随着局部程序性细胞死亡,以阻止病原菌的生长和扩散。对这种程序性细胞死亡形式进行精确的定量评估,对于揭示该过程潜在的遗传和分子机制至关重要。在此,我们首先描述了一种用于定量检测拟南芥中过敏反应细胞死亡的台盼蓝染色方法的优化。此外,我们提供了一种基于病原菌真空渗透的电解质渗漏检测方法,该方法可同时应用于大量植株以及受小尺寸表型影响的拟南芥突变体。