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Nitric oxide (NO), an inorganic gas, acts as a potent second messenger in most animal and plant tissues. NO diffuses out of the cells that produce it and enters the neighboring cells to generate a downstream response. NO synthase (NOS) catalyzes NO production by the deamination of the amino acid arginine. There are three isoforms of NOS. Endothelial cells have endothelial NOS (eNOS), nerve and muscle cells have neuronal NOS (nNOS), and macrophages produce inducible NOS (iNOS) upon exposure...

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Nitric Oxide Detection Using a Chemical Trap Method for Applications in Bacterial Systems.

Marilene Silva Oliveira^1,2,3, Karina F D N Santos², Railane Monteiro de Paula²

¹Instituto Federal de Educação, Ciência e Tecnologia Goiano, Departamento de Agroquímica, Campus Rio Verde, Rio Verde 75901-970, GO, Brazil.

|September 28, 2023

Summary

Plant growth-promoting bacteria (PGPB) produce nitric oxide (NO), vital for plant-microbe interactions. This study quantifies NO production in PGPB strains, identifying key NO-producing species for biofertilizer development.

Keywords:

Greiss reaction N2-fixing bacteria chromatography fluorescence nitric oxide

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Published on: December 25, 2016

Area of Science:

Microbiology
Plant Science
Biochemistry

Background:

Plant growth-promoting bacteria (PGPB) enhance plant development through mechanisms like nitrogen fixation and phytohormone production.
Nitric oxide (NO), a free radical, plays a crucial role in plant and bacterial growth and defense responses.
Accurate detection of NO is essential for understanding PGPB functions and optimizing biofertilizer applications.

Purpose of the Study:

To evaluate and quantify nitric oxide (NO) production in various agronomically important bacterial strains.
To identify PGPB strains with significant NO production capabilities for potential biofertilizer formulations.
To explore novel methods for NO detection and characterization in bacterial systems.

Main Methods:

Eight bacterial strains were cultured in the presence of KNO3 for 1-3 days.
Nitric oxide (NO) production was quantified using the Griess reaction and monitored via 2,3-diaminonaphthalene (DAN) trapping.
Analysis of NO byproducts was performed using fluorescence spectroscopy, gas chromatography-mass spectrometry, and high-performance liquid chromatography.

Main Results:

Azospirillum brasilense (HM053 and FP2), Rhizobium tropici (Br322), and Gluconacetobacter diazotrophicus (Pal 5) exhibited the highest NO production within 24 hours.
Nitrospirillum amazonense (Y2) and Herbaspirillum seropedicae (SmR1) showed no detectable NO production under the tested conditions.
A novel NO adduct, 1,2,3,4-naphthotetrazole (NTT) with a molecular mass of 182 Da, was identified, differing from the typical 2,3-naphthotriazole (NAT).

Conclusions:

Specific PGPB strains, including Azospirillum brasilense and Gluconacetobacter diazotrophicus, are significant producers of nitric oxide (NO).
The study presents an effective strategy for monitoring NO production in PGPB, aiding in the selection of strains for biofertilizers.
Understanding NO production mechanisms in PGPB offers new avenues for improving plant-microbe interactions and agricultural productivity.