Responses of antioxidants in the lichen Ramalina lacera may serve as an early-warning bioindicator system for the detection of air pollution stress.

The aim of this study was to identify, in the lichen Ramalina lacera, antioxidants that could provide indications of air pollution stress, and respond earlier than traditionally used structural/physiological parameters. The pollution-sensitive lichen R. lacera was transplanted from its relatively unpolluted natural habitat to two air-polluted sites for a period of up to 6 months. The superoxide dismutase and catalase activities, total water- and lipid-soluble low-molecular-weight antioxidant capacities and chlorophyll b/chlorophyll a ratios were assessed every 6 weeks. The earliest signs of oxidative stress were detected in the activities of fungal copper/zinc-superoxide dismutase, algal iron-superoxide dismutase and water-soluble low-molecular-weight antioxidants, which increased significantly as early as 42 days after exposure to pollution. Catalase activity increased in lichens transplanted to the polluted sites after 90 days. All activities decreased towards the end of the experiment. The impact of air pollution on R. lacera, using the traditionally employed parameter of chlorophyll b/chlorophyll a ratio, was only detected after 6 months of exposure to air pollution. Our results indicate that antioxidant parameters may serve as improved early-warning indicators of air pollution stress in lichens.

Characterization of enzymatic antioxidants in the lichen Ramalina lacera and their response to rehydration.

Lichens are slow-growing associations of fungi and green algae or cyanobacteria. This symbiotic association forms a common thallus that does not possess roots or a waxy cuticle and depends mainly on atmospheric input of mineral nutrients. The lifestyle of most lichens is composed of alternating periods of desiccation with low metabolic activity and hydration that induces increase in their metabolism. We have previously shown that rehydration of the naturally desiccated lichen Ramalina lacera resulted in a rapid increase in photosynthesis and was accompanied by a burst of intracellular production of reactive oxygen species and nitric oxide, as well as a transient decrease in water-soluble antioxidant capacity. We report here on enzymatic antioxidants of R. lacera and their response to rehydration. Native gel electrophoresis of crude extracts of R. lacera stained for superoxide dismutase (SOD) activity revealed four Fe-SOD and four Mn-SOD electromorphs that are synthesized by the alga, a Cu/Zn-SOD and a Mn-SOD that are the product of the fungus, and two catalases synthesized one by the fungus and the other by the algae. In addition, we detected glutathione reductase and glucose-6-phosphate dehydrogenase activities in crude extracts of R. lacera. Rehydration of the thalli resulted in a decrease in SOD activity of all forms, and a transient decrease in total catalase activity, as well as a decrease in the antioxidant auxiliary enzymes glutathione reductase and glucose-6-phosphate dehydrogenase.

Rehydration of the lichen Ramalina lacera results in production of reactive oxygen species and nitric oxide and a decrease in antioxidants.

Lichens are slow-growing associations of fungi and unicellular green algae or cyanobacteria. They are poikilohydric organisms whose lifestyle in many cases consists of alternating periods of desiccation, with low metabolic activity, and hydration, which induces increase in their metabolism. Lichens have apparently adapted to such extreme transitions between desiccation and rehydration, but the mechanisms that govern these adaptations are still poorly understood. In this study, the effect of rehydration on the production of reactive oxygen species and nitric oxide as well as low-molecular-weight antioxidants was investigated with the lichen Ramalina lacera. Rehydration of R. lacera resulted in the initiation of and a rapid increase in photosynthetic activity. Recovery of photosynthesis was accompanied by bursts of intracellular production of reactive oxygen species and nitric oxide. Laser-scanning confocal microscopy using dichlorofluorescein fluorescence revealed that formation of reactive oxygen species following rehydration was associated with both symbiotic partners of the lichen. The rate and extent of reactive oxygen species production were similar in the light and in the dark, suggesting a minor contribution of photosynthesis. Diaminofluorescein fluorescence, indicating nitric oxide formation, was detected only in fungal hyphae. Activities associated with rehydration did not have a deleterious effect on membrane integrity as assessed by measurement of electrolyte leakage, but water-soluble low-molecular-weight antioxidants decreased significantly.

Lichens as biomonitors around a coal-fired power station in Israel.

In the present study epiphytic lichens were applied as biomonitors of air pollution to determine the environmental impact of a coal-fired power station. Thalli of the lichen Ramalina lacera (With.) J.R. Laund. growing on carob twigs (Ceratonia siliqua L.) were collected with their substrate in July 2000 in a relatively unpolluted forest near HaZorea, Ramoth Menashe, Northeast Israel, and transplanted to 10 biomonitoring sites in the vicinity of the coal-fired power station Oroth Rabin near the town of Hadera. The lichens were retrieved in January 2001. We examined the following parameters of lichen vitality: (a) potential quantum yield of photosynthesis expressed as fluorescence ratio F(v)/F(m), (b) stress-ethylene production, and (c) electric conductivity expressing integrity of cell membranes. Following an exposure of 7 months, the lichens were retrieved and physiological parameters and data of elemental content were analyzed comparatively. Electric conductivity values correlated positively with B, Fe, Mg, Mn, Na, Pb, S, Sn, and Ti content. Concentrations of stress-ethylene correlated positively with Al, Ba, Pb, S, and V content and negatively with Cu and Sn. F(v)/F(m) ratios correlated negatively with S content. Some of the heavy metals reached lower levels than those reported in the relevant literature despite a wind regime that should have blown pollutants toward the biomonitoring sites.

Biomonitoring air pollution with the desert lichen Ramalina maciformis.

To investigate the environmental impact of anthropogenic activity in the Negev Desert, we monitored the following in order to determine the physiological integrity of the epilithic fruticose lichen Ramalina maciformis: (1) production of ethylene; (2) potential quantum yield of photosystem II expressed as chlorophyll fluorescence ratio Fv/Fm; and (3) electrolyte leakage of cell membranes in terms of electric conductivity. The degree of pollution was assessed by measuring the amounts of B, Cd, Co, Cu, Fe, K, Mg, Mn, Na, Ni, P, Pb, Sr and Zn in the lichen thallus. Some of the lichen-carrying stones collected in the relatively unpolluted control site were relocated on the same hill, to test the possible impact of relocation. An additional amount of lichen-carrying stones was marked as controls. The greater part of the lichen-carrying stones was transferred to 10 biomonitoring sites in and around Ramat Hovav, an industrial area in the Negev Desert, Israel. Transplanted lichen in polluted sites contained higher amounts of mineral elements than lichens in the control site after an exposure period of 7 months. Statistical analysis revealed positive correlation of ethylene production and Ni content, negative correlation of Fv/Fm ratios and B, Cu, Mn, Na, Ni, Sr and Zn, and positive correlation of electric conductivity and B, Mn, Ni and Sr. Both elemental content and physiological alterations in transferred samples of R. maciformis point to a high degree of contamination in Ramat Hovav. The present study suggests that the lichen R. maciformis should be classified as a sensitive species endangered by anthropogenic activity in the desert. Furthermore, this lichen species is likely to be suitable as a bioindicator of pollution in other parts of the Negev Desert. Ethylene production and electric conductivity as parameters of lichen-vitality, indicated stress in 5 of the 12 biomonitoring sites (42%) and may therefore be considered as more sensitive than the Fv/Fm ratio, which indicated stress in 3 of the 12 sites (25%).

Changes in the potential quantum yield of photosystem II and the integrity of cell membranes relative to the elemental content of the epilithic desert lichen Ramalina maciformis.

The present study used the epilithic fruticose lichen Ramalina maciformis to investigate the occurrence of mineral elements, including heavy metals, at a distance of up to 50 km from the industrial region in Ramat Hovav in the Negev Desert, Israel. The major objective of this study was an analytical comparison of elemental content and physiological parameters of lichen vitality, apart from a test of the applicability of this specific lichen in investigations of air pollution. The Ca, Cr, Cu, K, Mg, Na, Pb, S, Sr, and Zn content of thalli from the unpolluted Tellalim site collected in August 1997, transferred to 24 biomonitoring sites, and retrieved in April 1998 was analyzed in comparison with the following parameters: The potential quantum yield of photosystem 11 (PSII), and the integrity of cell membranes. Transplanted thalli in several sites at Ramat Hovav accumulated large amounts of most of the elements. The K content of the transplants located in the polluted sites indicated a leakage of this element, because this content was lower than that of thalli in unpolluted sites. Calcium, Cu, Mn, and Na showed an inverse correlation with the K content of the lichen. Calcium, Cu, and Sr showed an inverse correlation with the Fv/Fm ratio expressing the potential quantum yield of PSII. Calcium, Cr, Cu, Mg, Na, S, and Sr showed a positive correlation with the electrical conductivity corresponding with cell-membrane disintegration. The present study demonstrated a meaningful connection between enlarged concentrations of certain elements and physiological phenomena. The capability of the lichen to detect air pollution was found to be satisfactory. The dispersion of airborne heavy metals was found, however, to be local and limited to a few hundred meters from the source of pollution.