Endogenous NO Is Involved in Dissimilar Responses to Rehydration and Pb(NO3)2 in Ramalina farinacea Thalli and Its Isolated Phycobionts

  • Joana R. ExpósitoEmail author
  • A. J. Coello
  • E. Barreno
  • L. M. Casano
  • M. Catalá
Environmental Microbiology


Lichens undergo desiccation/rehydration cycles and are permeable to heavy metals, which induce free radicals. Nitrogen monoxide (NO) regulates important cellular functions, but the research on lichen NO is still very scarce. In Ramalina farinacea thalli, NO seems to be involved in the peroxidative damage caused by air pollution, antioxidant defence and regulation of lipid peroxidation and photosynthesis. Our hypothesis is that NO also has a critical role during the rehydration and in the responses to lead of its isolated phycobionts (Trebouxia sp. TR9 and Trebouxia jamesii). Therefore, we studied the intracellular reactive oxygen species (ROS) production, lipid peroxidation and chlorophyll autofluorescence during rehydration of thalli and isolated microalgae in the presence of a NO scavenger and Pb(NO3)2. During rehydration, NO scavenging modulates free radical release and chlorophyll autofluorescence but not lipid peroxidation in both thalli and phycobionts. Pb(NO3)2 reduced free radical release (hormetic effect) both in the whole thallus and in microalgae. However, only in TR9, the ROS production, chlorophyll autofluorescence and lipid peroxidation were dependent on NO. In conclusion, Pb hormetic effect seems to depend on NO solely in TR9, while is doubtful for T. jamesii and the whole thalli.


Heavy metals Lichen Microalgae Nitric oxide Rehydration Free radicals 



arbitrary units


2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (carboxy-PTIO)


2,7-dichlorodihydrofluorescein diacetate


Heavy Metals




Plant growth-promoting rhizobacteria


Reactive oxygen species


Reactive nitrogen species


2-thiobarbituric acid

T. jamesii

Trebouxia jamesii


Trebouxia sp. TR9



We want to acknowledge the technical assistance of Álvarez R. and Del Hoyo A. who cultured and dehydrated the samples of phycobionts, Díaz C. who made the analysis of free radicals, chlorophyll autofluorescence and lipid peroxidation in Ramalina farinacea and Mejuto I. who helped in microscopy. We want to thank del Campo. E for the revision of the draft.

Authors’ Contributions

The study was designed by Barreno E., Catalá M. and Casano L. M. Catalá M. performed the measurements of free radical production kinetics and chlorophyll autofluorescence. Coello A.J. did the study of lipid peroxidation in phycobionts. Casano L.M. provided biological material. Expósito J.R. analysed the data, made the figures and wrote the draft under the supervision of Catalá M. All the authors have revised and approved the final draft.

Funding Information

This study was funded by the Spanish Ministry of Economy and Competitiveness (MINECO CGL2016-79158-P and CGL2016-40058-P), Comunidad de Madrid - European Commission (Youth Employment Intiative, Spain) (PEJ-2017-AI/AMB-6337), FEDER and the Generalitat Valenciana (PROMETEOIII/2017/039 GVA).


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Authors and Affiliations

  1. 1.Departamento de Biología y Geología, Física y Química InorgánicaUniversidad Rey Juan Carlos, ESCETMóstoles, MadridSpain
  2. 2.Departamento de Biodiversidad y ConservaciónReal Jardín Botánico (RJB-CSIC)MadridSpain
  3. 3.Departamento de Botánica, Instituto Cavanilles de Biodiversidad y Biología Evolutiva, Facultad de Ciencias BiológicasUniversitat de ValènciaBurjassot, ValenciaSpain
  4. 4.Departamento de Ciencias de la VidaUniversidad de AlcaláAlcalá de Henares, MadridSpain

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