Detection of Nitric Oxide and Determination of Nitrite Concentrations in Arabidopsis thaliana and Azospirilum brasilense

FORESI NOELIA PAMELA; ARRUEBARRENA ANDRES; AMENTA MELINA; CORREA ARAGUNDE NATALIA; LAMATTINA LORENZO; CREUS CECILIA MÓNICA

Bio-protocol

Stanford University

Año: 2016 vol. 6

There is now general agreement that nitric oxide (NO) is an importantand almost ubiquitous signal in plants. Nevertheless, there are still many controversialobservations and different opinions on the importance and functions of NO inplants. Partly, this may be due to the difficulties in detecting and even morein quantifying NO. Here, we summarize protocols for detecting NO andquantifying nitrite concentration in Arabidopsis seedlings and for the NO real time measurement in biofilms formed by the plant growth promoting rhizobacteriaAzospirillum brasilense (A. brasilense). NO in oxygen-containing aqueoussolution has a short half-life that is often attributed to a rapid oxidation tonitrite. Here we detail the use of the fluorescent probe DAF-FM DA and theelectrochemical method for directly detecting and quantifying NO, respectively,and the Griess reagent to indirectly detect NO through its oxidized nitriteform. These protocols could be useful in a variety of cell types and differenttissues of plants,and for microorganisms.