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Insights and Pitfalls of the Methodology for Free Radical Research in Legume Nodules: Hemoglobins as a Case Study

AuthorsBecana Ausejo, Manuel ; Rubio Luna, María Carmen ; Calvo-Begueria, Laura; Martínez, , Jesús I.; Duanmu, Deqiang; Pérez-Rontomé, Carmen
legume nodules
reactive oxygen/nitrogen species
Issue DateOct-2019
PublisherHuazhong Agricultural University
Citation21st International Congress of Nitrogen Fixation (Wuhan, China. October 10-15 2019)
AbstractReactive oxygen (ROS; superoxide radicals and H2O2) and nitrogen (RNS; NO and nitrosothiols) species are of paramount importance in biology. In plants, ROS and RNS are generated in most cellular compartments. At the light microscopy level, the production of superoxide radicals, H2O2 and NO may be detected based on their reactions with nitroblue tetrazolium, diaminobenzidine and 4,5-diaminofluorescein diacetate, respectively. Additional techniques are the HyPer bioprobe for H2O2 and electron paramagnetic resonance (EPR) spectroscopy for free radicals. However, strict controls must be used to ensure specificity, discard interferences and minimize artifacts during tissue extraction. In the rhizobia-legume symbiosis, all three reactive species were detected in the infection threads and in mature and senescent nodules. Leghemoglobins (Lbs) are heme proteins found at millimolar concentrations in the cytoplasm of infected cells, where they transport and deliver O2 to the symbiosomes. We have used two experimental approaches involving Lbs to localize ROS and RNS in nodules and identify potential pitfalls of the methodology.
Description1 Pag.
Appears in Collections:(EEAD) Comunicaciones congresos
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