Tommaso Pisani

The integrity of lichen cell membrane as a suitable parameter for monitoring biological effects of acute nitrogen pollution

This study aimed at testing the suitability of cell membrane integrity in the lichen Evernia prunastri (L.) Ach. as sensitive indicator of nitrogen (N) stress, to set up a rapid and effective method for monitoring biological effects of acute N pollution. Lichen samples were incubated in solutions of potassium nitrate, ammonium nitrate and ammonium sulphate at different concentrations, and cell membrane damage, expressed in terms of increased electrolyte leakage, was measured after 0, 24, 48 and 96 h. Cell membrane damage was observed in E. prunastri in the presence of high or very high N concentrations, irrespective of the compound supplied. Since the mycobiont represents the large majority of a lichen biomass, it is reasonable to assume that ion leakage mainly occurred from fungal cells. Although in biomonitoring studies the photobiont is usually regarded as the most sensitive partner of the lichen symbiosis, our findings suggest that the mycobiont is most affected in the case of N-excess, and that this feature can be used as suitable indicator of acute N stress episodes.

Effects of ammonia from livestock farming on lichen photosynthesis

This study investigated if atmospheric ammonia (NH3) pollution around a sheep farm influences the photosynthetic performance of the lichens Evernia prunastri and Pseudevernia furfuracea. Thalli of both species were transplanted for up to 30 days in a semi-arid region (Crete, Greece), at sites with concentrations of atmospheric ammonia of ca. 60 microg/m3 (at a sheep farm), ca. 15 microg/m3 (60 m from the sheep farm) and ca. 2 microg/m3 (a remote area 5 km away). Lichen photosynthesis was analysed by the chlorophyll a fluorescence emission to identify targets of ammonia pollution. The results indicated that the photosystem II of the two lichens exposed to NH(3) is susceptible to this pollutant in the gas-phase. The parameter PI(ABS), a global index of photosynthetic performance that combines in a single expression the three functional steps of the photosynthetic activity (light absorption, excitation energy trapping, and conversion of excitation energy to electron transport) was much more sensitive to NH3 than the FV/FM ratio, one of the most commonly used stress indicators.

Antiproliferative activity of lichen extracts on murine myeloma cells

In the present study we report some preliminary results concerning the evaluation of antiproliferative activity on murine myeloma cells (P3X63-Ag8.653) of crude extracts of two common lichen species, Evernia prunastri and Xanthoria parietina.The results were evaluated by means of the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] test, which is commonly used to assess the activity of living cells through mitochondrial dehydrogenases. They indicated that extracts of E. prunastri had no effect, while those of X. parietina significantly affected murine myeloma cell proliferation, with a reduction down to 75% for methanolic extracts. This opens perspectives for deeper investigations extended also to other mammalian cell lines.

Physiological effects of arsenic in the lichen Xanthoria parietina (L.) Th. Fr.

The aim of this study was to test in a short term laboratory experiment the accumulation and physiological effects of As in the epiphytic lichen Xanthoria parietina. Arsenic content in treated samples increased progressively with increasing concentration in treatment solutions. Treatment of X. parietina thalli with 0.1, 1, 10 ppm As solutions caused significant decrease of viability, measured as intensity of respiratory activity, and damages to cell membranes, assessed by increase of electric conductivity of rinsing water and lipid peroxidation products. Soluble proteins content decreased and H₂O₂ content increased already at the lowest As concentration tested (0.01 ppm). Photosynthetic efficiency, measured in terms of F(V)/F(M) ratio, decreased significantly only at the highest As concentration (10 ppm). It was concluded that As exposure causes physiological stress both on the mycobiont and the photobiont and that cell membrane damage, expressed in terms of electric conductivity of rinsing water, is the parameter most affected by As treatment.

Bioacumulation and ultrastructural effects of Cd, Cu, Pb and Zn in the moss Scorpiurum circinatum (Brid.) Fleisch. & Loeske

This paper tested if culturing the moss Scorpiurum circinatum (Brid.) Fleisch. & Loeske with metal solutions (Cd, Cu, Pb and Zn) for 30 days causes metal bioaccumulation and ultrastructural changes. The results showed that despite the high heavy metal concentrations in treatment solutions, treated samples did not show severe ultrastructural changes and cells were still alive and generally well preserved. Bioaccumulation highlighted that moss cells survived to heavy metal toxicity by immobilizing most toxic ions extracellularly, likely in binding sites of the cell wall, which is the main site of metal detoxification.

Physiological and chemical response of lichens transplanted in and around an industrial area of south Italy: relationship with the lichen diversity.

The lichen Evernia prunastri (L.) Ach. has been exposed for 3 months in and around an industrial area of Mediterranean Italy for monitoring physiological (photosynthetic efficiency, membrane lipids peroxidation and cell membrane integrity) and chemical (bioaccumulation of the heavy metals Cr, Ni, Pb, V and Zn) effects and investigate the consistency with the environmental quality status depicted by the diversity of epiphytic lichens (index of lichen diversity (ILD)). The results showed that thalli transplanted close to the industrial area exhibited early stress symptoms, as revealed by the increase in electrical conductivity indicating a damage endured by lichen cell membranes. The electrical conductivity was inversely correlated with the diversity of epiphytic lichens recorded at the same sites. The ILD negatively correlated also with membrane lipid peroxidation and the rate of accumulation of Pb, V and Zn. Reciprocal correlations found among trace elements pinpointed vehicular traffic and metal processing in the industrial area as main sources. The damage endured by cell membranes was the best physiological indicator consistent with the air quality status depicted by the diversity of epiphytic lichens.

Effects of high temperature on epiphytic lichens: Issues for consideration in a changing climate scenario

Abstract The effect of high temperatures on the lichen Evernia prunastri was investigated by analysing the content of photosynthetic pigments, chlorophyll degradation, and the integrity of cell membranes. Results indicate that E. prunastri can tolerate a temperature of 40°C for 24 h, but for longer periods a reduction in chlorophyll b is apparent, suggesting that chlorophyll b could be used as a valuable parameter to detect changes caused by prolonged exposure to such temperatures. Exposure of E. prunastri to a temperature of 80°C had strongly negative effects on the content of photosynthetic pigments, causing both inhibition of chlorophyll synthesis and chlorophyll degradation to phaeophytin; slow acclimation was not capable of preventing such damage. Damage to cell membranes, as indicated by changes in electrical conductivity of the medium, proved to be a useful and simple tool for measuring damage caused by high temperatures. Besides the effects of temperature, results also indicate that exposure to a prolonged dark period has an important negative influence on the chlorophyll a/b ratio, suggesting that lichens could be profitably used not only to monitor climate changes, but also to monitor the effects of global dimming.

Time- and dose-dependency of the effects of nitrogen pollution on lichens

The present work aims at testing if exposure time and dose play a role in the response of lichen species to nitrogen (N) pollution. To this purpose, samples of the N-sensitive Evernia prunastri and the N-tolerant Xanthoria parietina were treated for 5 weeks either with solutions of NH(4)NO(3) 0.05 and 1 M, or (NH(4))(2)SO(4) 0.025 and 0.5 M. Photosynthetic efficiency was measured as an indicator of sample vitality. The results showed that the lowest concentrations were ineffective at the beginning, but after several supplies both compounds inhibited photosynthetic activity of E. prunastri. The highest concentrations had a deleterious effect, but with a temporal trend. For X. parietina no effect was found for the lowest concentrations, while the same trend shown by E. prunastri was instead observed following treatments with the highest concentrations. It was concluded that the response of lichens to N supply is not only species-specific, but also time- and dose-dependent. The results give a clue on field studies on the relationships between lichens and N pollution.

Physiological effects of mercury in the lichens Cladonia arbuscula subsp. mitis (Sandst.) Ruoss and Peltigera rufescens (Weiss) Humb.

This study aimed at investigating the cellular distribution of Hg in the lichens Cladonia arbuscula subsp. mitis and Peltigera rufescens treated with Hg²(+) and at testing if Hg treatment affects selected physiological parameters. In both species, increasing Hg accumulation under increasing Hg supply in the treatment solutions was found. P. rufescens showed a higher intracellular accumulation. Photosynthetic parameters were negatively affected in both species, as indicated by the decrease in photosynthetic pigments content, photosynthetic efficiency and chlorophyll integrity. Cell membranes of both species endured damage as indicated by the increase in the concentration of products of lipid peroxidation and decrease in ergosterol content. Nevertheless, differences between the two species were found, suggesting a differential sensitivity to Hg.

Influence of sun irradiance and water availability on lichen photosynthetic pigments during a Mediterranean summer

This study was carried out to investigate changes in lichen photosynthetic pigments induced by different combinations of light irradiance and water availability during a Mediterranean summer. To this purpose, thalli of three epiphytic lichens with a markedly different ecology concerning photo-hygrophytism, namely Evernia prunastri (hygro-mesophytic), Flavoparmelia caperata (mesophytic) and Xanthoria parietina (xero-mesophytic), were transplanted for 30 days to N- and S-facing cardinal exposures in central Italy. To investigate the effect of thallus hydration, at each cardinal exposure 50% of thalli were hydrated daily with deionised water. The results showed that changes in the concentrations of photosynthetic pigments are species-specific and consist in a general depression of photosynthetic pigments only in the hygro-mesophytic species E. prunastri. The pattern of photosynthetic pigments was also investigated in spontaneous samples along contrasting aspects. In this case, X. parietina from S-facing slopes, adapted to direct solar influx, showed higher pigment content than N-facing thalli; F. caperata and E. prunastri avoid direct extreme solar radiation and assimilation pigments were influenced more by the shadowing of tree canopies than by the cardinal exposure. The influence of drought on lichen photosynthetic pigments in the Mediterranean area is discussed.