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Lichen: from genome to ecosystem in a changing world 3I-O (3I-05) Submission ID: IAL0009-00008 ALLELOPATHIC EFFECTS OF LICHEN SECONDARY METABOLITES AGAINST POTENTIAL COMPETITORS FOR ROCK SURFACES Favero-Longo S. E. 1 , Gazzano C. 1 , Piervittori R. 1 1 Dip. Biologia Vegetale, Università di Torino, Torino, Italy Lichen secondary metabolites (LSM) are known to determine allelopathic effects on soil and mycorrhizal fungi, and on the spore/seed germination and early growth stages of bryophytes and vascular plants, thus likely supporting the competition of terricolous lichens for soil surfaces. In the current study, laboratory and field assays were performed to evaluate the potential allelopathic effects of LSM against potential competitors of saxicolous lichens for rock surfaces. Usnic acid, norstictic acid and parietin, solubilized in water, water-acetone 90:10 mixture and pure acetone, were tested against microcolonial fungi (MCF: Coniosporium apollinis, C. perforans, C. uncinatum, Phaeococcomyces cfr. chersonesus), black yeasts (BY: Sarcinomyces petricola), green algae (GA: Apatococcus lobatus, Scenedesmus ecornis) and cyanobacteria (CY: Pleurocapsa minor, Lyngbya sp.). The common biocide benzalkonium chloride (1%) was used as positive control. Fungal, algal and cyanobacterial colonies, cultured on standard media (MEA, Trebouxia-Medium, BG11), were poured with 50 μl of each solution and their areal growth was monitored for one month using image analysis (WinCAM software). Field assays on black patinas on the walls of the Roman Theatre of Aosta (NW-Italy) were also performed, using epifluorescence microscopy to check the algal and cyanobacterial sensibility. The different MCF species showed a slightly different sensibility to the different LSM solutions, but in all the cases the three metabolites in the water-acetone mixture (ca. 0.05 mM) and usnic acid in water (0.02 mM) determined the highest growth inhibition, displaying the same effects of benzalkonium chloride. On the other hand, all the LSM solutions did not significantly affect or even increased the areal growth of algal and cyanobacterial colonies. Epifluorescence observations of the treated photosynthetic microorganisms in laboratory confirmed that most of cells were still emitting a strong red fluorescence. However, in the field patinas, algal and cyanobacterial cells showed a higher sensibility to LSM, more often displaying an epifluorescence decrease in the treated areas with respect to negative controls. In conclusion, LSM may play a significant role in lichen competition for rock surfaces. LSM may be a potential resource for the MCF control on rocks, avoiding the use of human-toxic biocides. 42
The 7 th International Association for Lichenology Symposium 2012 (3I-06) Submission ID: IAL0226-00001 ROLE OF NITRIC OXIDE IN THE RESPONSE OF RAMALINA FARINACEA TO LEAD Barreno E. 1 , Diaz-Rodriguez C. 2 , Catala M. 2 1 Botany, Inst. Cavanilles of Biodiversity and Evolutionary Biology, Valencia, Burjassot, Spain 2 Biology and Geology, Rey Juan Carlos University, Mostoles, Spain Nitric oxide is a small multifaceted molecule with a plethora of biological functions. The roles of NO in biotic and abiotic stress are especially relevant and an involvement of NO in the establishment of symbiotic relationships (i.e. mycorrhizae) has been described. Despite its relevance, NO production in lichens has only been described recently and its roles are unknown. Both pro-oxidant and antioxidant functions have been described for NO and a recent study made in our laboratories suggests that during lichen rehydration NO seems to be related with contention of oxidative damage and chlorophyll stabilization. Preliminary studies have also showed that NO is important in the regulation of oxidative damage exerted by the air pollutant cumene hydroperoxide during rehydration. The toxic mechanisms of heavy metals, such as Pb, involve both the inhibition of enzymes and the production of free radicals. The aim of the present work is to study the effect of Pb in the intracellular oxidative burst occurred during rehydration and the role of NO in the lichen Ramalina farinacea (L.) Ach. Intracellular ROS specific fluorescent probe, dichlorodihydrofluorescein diacetate (DCFH2-DA) has been used. ROS kinetics and chlorophyll autofluorescence have been recorded during the first minutes after rehydration. Lipid peroxidation and NO-endproducts have been quantified at different time points. NO specific inhibitor c-PTIO has been used in order to elucidate NO functions. The results show that Pb induces decreases in intracellular ROS production and lipid peroxidation during rehydration; although a decrease in chlorophyll autofluorescence has also been observed. NO inhibition during Pb-conditioned rehydration does not affect the studied parameters. We conclude that NO is not involved in lichen response to Pb. Lichen tolerance to moderate doses of Pb may be related to the induction of a compensatory response known as hormesis. [MCINN (CGL2009-13429-C02-01/02), AECID (PCI_A_l024755/09) and Generalitat Valenciana (PROMETEO 174/2008 GVA)] 43 3I-O
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