2024-03-28T16:51:34Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/1672132020-09-30T08:40:32Zcom_10261_75com_10261_6com_10261_31com_10261_3com_10261_59col_10261_328col_10261_284col_10261_312
2018-06-29T09:55:37Z
urn:hdl:10261/167213
Redefining nitric oxide production in legume nodules through complementary insights from electron paramagnetic resonance spectroscopy and specific fluorescent probes
Calvo-Beguería, Laura
Rubio Luna, María Carmen
Martínez, Jesús I.
Pérez-Rontomé, Carmen
Delgado Igeño, María Jesús
Bedmar, Eulogio J.
Becana Ausejo, Manuel
Ministerio de Economía y Competitividad (España)
European Commission
Calvo-Begueria, Laura [0000-0001-5327-4046]
Rubio Luna, María Carmen [0000-0002-9378-7775]
Pérez-Rontomé, Carmen [0000-0002-9772-3170]
Bedmar, Eulogio J. [0000-0002-9421-1460]
Becana Ausejo, Manuel [0000-0002-1083-0804]
Denitrification
eElectron paramagnetic resonance
Leghemoglobin
Nitric oxide
Nitrogen fixation
Symbiosis
Nitric oxide (NO) is a signaling molecule with multiple functions in plants. Given its critical importance and reactivity as a gaseous free radical, we have examined NO production in legume nodules using electron paramagnetic resonance (EPR) spectroscopy and the specific fluorescent dye 4,5-diaminofluorescein diacetate. Also, in this context, we critically assess previous and current views of NO production and detection in nodules. EPR of intact nodules revealed that nitrosyl-leghemoglobin (Lb2+NO) was absent from bean or soybean nodules regardless of nitrate supply, but accumulated in soybean nodules treated with nitrate that were defective in nitrite or nitric oxide reductases or that were exposed to ambient temperature. Consequently, bacteroids are a major source of NO, denitrification enzymes are required for NO homeostasis, and Lb2+NO is not responsible for the inhibition of nitrogen fixation by nitrate. Further, we noted that Lb2+NO is artifactually generated in nodule extracts or in intact nodules not analyzed immediately after detachment. The fluorescent probe detected NO formation in bean and soybean nodule infected cells and in soybean nodule parenchyma. The NO signal was slightly decreased by inhibitors of nitrate reductase but not by those of nitric oxide synthase, which could indicate a minor contribution of plant nitrate reductase and supports the existence of nitrate- and arginine-independent pathways for NO production. Together, our data indicate that EPR and fluorometric methods are complementary to draw reliable conclusions about NO production in plants.
2018-06-29T09:55:37Z
2018-06-29T09:55:37Z
2018-06
artículo
Journal of Experimental Botany 69(15): 3703-3714 (2018)
0022-0957
http://hdl.handle.net/10261/167213
10.1093/jxb/ery159
1460-2431
http://dx.doi.org/10.13039/501100003329
http://dx.doi.org/10.13039/501100000780
29701804
eng
Publisher's version
https://doi.org/10.1093/jxb/ery159
Sí
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/AGL2014-53717-R
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/CTQ2015-64486-R
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/AGL2013-45087-R
http://creativecommons.org/licenses/by/4.0/
openAccess
Oxford University Press