[show abstract][hide abstract] ABSTRACT: Host use by herbivores is largely determined by host properties such as nutrient content and chemical defence against foragers. The impacts of these attributes on a herbivore may largely depend on its life cycle stage. Lichen species are known to differ in nutritional quality and level of chemical defence and, consequently, vary as fodder for herbivores. The aim of this study was to explore the impact of several lichen species and the presence of their secondary metabolites on their use as hosts by a specialist lichen-feeder, Cleorodes lichenaria. This study also addressed, for the first time, how a specialist lichen-feeder deals with different lichen secondary metabolites. In the beginning of their development, larvae grew better on Xanthoria parietina than on the other host lichens, whereas older larvae grew best on Ramalina fraxinea. Lichen secondary chemicals in R. fraxinea and Parmelia sulcata hindered larval growth in the beginning but after 75 days lichen secondary chemicals had no impact on the mass of larvae. Physodic acids in Hypogymnia physodes were lethal to larvae. In general, larvae metabolized 70-95% of ingested lichen secondary chemicals and the rest of these were excreted in frass. Lichen secondary metabolites in P. sulcata restrict and in H. physodes prevent their use as a host for C. lichenaria larvae. Our main finding, the ability of larvae to metabolize several lichen secondary metabolites, indicates digestive adaptation to these chemicals. No signs of sequestration of these chemicals were found.
[show abstract][hide abstract] ABSTRACT: Lecanoric acid (1), orsellinic acid methyl ester (2), orcinol (3), and usnic acid (4) were isolated from the lichen Parmelia subrudecta, collected on Palma of the Canary Islands, Spain. Compounds 1, 2, 3, and 4 were purified by solvent extraction, silica gel column chromatography, and preparative high-performance liquid chromatography (HPLC) consecutively. The structures of the four compounds were elucidated by one- and two-dimensional nuclear magnetic resonance (NMR) experiments and mass spectrometric investigations. These compounds showed activity against important gram-positive and gram-negative pathogens like mycobacteria and multiresistant staphylococci. This activity is combined with antiproliferative activity and cytotoxicity.
[show abstract][hide abstract] ABSTRACT: The photobiont is considered as the more sensitive partner of lichen symbiosis in metal pollution. For this reason the presence of a metal tolerant photobiont in lichens may be a key factor of ecological success of lichens growing on metal polluted substrata. The photobiont inventory was examined for terricolous lichen community growing in Cu mine-spoil heaps derived by historical mining. Sequences of internal transcribed spacer (ITS) were phylogenetically analyzed using maximum likelihood analyses. A total of 50 ITS algal sequences were obtained from 22 selected lichen taxa collected at three Cu mine-spoil heaps and two control localities. Algae associated with Cladonia and Stereocaulon were identified as members of several Asterochloris lineages, photobionts of cetrarioid lichens clustered with Trebouxia hypogymniae ined. We did not find close relationship between heavy metal content (in localities as well as lichen thalli) and photobiont diversity. Presence of multiple algal genotypes in single lichen thallus has been confirmed.
Ecotoxicology and Environmental Safety 05/2010; 73(4):603-12. · 2.20 Impact Factor
[show abstract][hide abstract] ABSTRACT: The phytotoxic effects of the lichen secondary metabolite-usnic acid on cultures of free living alga-Scenedesmus quadricauda (UTEX 76) and aposymbiotically grown lichen photobiont Trebouxia erici (UTEX 911) were assessed. We found a relatively strong inhibition effect of usnic acid on the growth of alga Scenedesmus, accompanied by an increase of cell size, an alteration of assimilation pigment composition, followed by strong degradation of chlorophyll a, a decrease of chlorophyll a fluorescence, and an increase of reactive oxygen species in the cells. The content of soluble proteins remained a stable parameter. Phytotoxicity of usnic acid on cultures of Trebouxia photobiont was significantly lower. Usnic acid in lichens may act as an allochemical that controls the division of photobiont cells, thereby regulating the balance between the photobiont and mycobiont forming thallus. Higher tolerance to usnic acid in Trebouxia cultures may be an adaptation resulting from the long term co-evolution of these algae with fungi that produce secondary metabolites.
Journal of Chemical Ecology 03/2010; 36(4):405-11. · 2.46 Impact Factor
[show abstract][hide abstract] ABSTRACT: Despite intensive research focused on the effects of UV-B, deeper metabolic responses to UV-A and UV-C are still scarce. Besides, especially microalgal species had to develop efficient protective features in comparison with tissue structure of vascular plants. We exposed axenic cultures of Scenedesmus quadricauda (Chlorophyceae) to UV-A (366 nm) and UV-C (254 nm) light over 1 h. Both wavelengths stimulated increase in soluble proteins, superoxide radical and hydrogen peroxide, but had a nonsignificant effect on cell viability. Within 17 detected free amino acids, five (including proline) increased in response to UV-A while only aspartic acid and histidine increased in UV-C treatment. Total soluble phenols and flavonoids were influenced neither by UV-A nor by UV-C while selected flavonols (quercetin and kaempferol) decreased in UV-A and were not detected in UV-C treatment. Benzoic acid derivatives increased preferentially after UV-A illumination (vanillic acid and vanillin) while cinnamic derivatives (caffeic, chlorogenic and p-coumaric acids) decreased in both UV-A and UV-C. It is concluded that UV light stimulated oxidative stress while exposure time was not sufficient to stimulate larger changes in phenolic metabolites. Present findings in the context of available data and with emphasis on phenolics in algae are discussed.
Photochemistry and Photobiology 02/2010; 86(3):612-6. · 2.29 Impact Factor
[show abstract][hide abstract] ABSTRACT: The results of a study aimed at investigating the effects of boron excess on a set of ecophysiological parameters in the lichen Xanthoria parietina, to set up a monitoring system to trace early biological effects of boron pollution in geothermal areas, are reported. To this purpose, lichen thalli have been incubated for 24 h in solutions at boron concentrations of 0.1, 1, 10 and 100 ppm, which were within the range in bulk deposition and geothermal fluids. The results showed a general trend of decreasing sample viability and increasing cell membrane damage and membrane lipid peroxidation under increasing boron concentrations, while photosynthetic efficiency, chlorophyll degradation and the contents of H(2)O(2) and water-soluble proteins were not affected. It was argued that the fungal partner, that represents the large majority of the lichen biomass, is more sensitive to boron excess than the algal partner.
[show abstract][hide abstract] ABSTRACT: We examined accumulation of phenolic acids, total soluble phenolics and flavonoids, and activities of phenolic metabolism-related enzymes (shikimate dehydrogenase (SKDH), phenylalanine ammonia-lyase (PAL), cinnamyl alcohol dehydrogenase (CAD), polyphenol oxidase (PPO)) in Matricaria chamomilla plants exposed to 3, 60 and 120 microM of nickel (Ni) for 10 days. Ni showed low toxicity as indicated by unaltered content of total soluble phenolics in the leaf rosettes. In the roots, the effects of Ni were more visible, including increased total phenolics and PAL activity, but a decrease in PPO activity was observed. CAD activity was not affected by any of the Ni concentrations. Cinnamic acid derivatives were affected more than benzoic acid derivatives. Accumulation of chlorogenic acid, an important antioxidant compound, was enhanced by Ni treatment (ca. 4-fold in 120 microM Ni). Accumulation of protocatechuic acid, a phenol with high chelating strength, even decreased in the leaf rosettes. These observations are discussed in connection to antioxidative properties of phenolic metabolites and previously tested metals (cadmium and copper).
Journal of plant physiology 05/2009; 166(13):1460-4. · 2.50 Impact Factor
[show abstract][hide abstract] ABSTRACT: Influence of 100 mM NaCl and 50 microM salicylic acid (SA) and their combination on the metabolism of chamomile (Matricaria chamomilla) during 7 days was studied. NaCl reduced growth and selected physiological parameters and SA in combined treatment (NaCl + SA) reversed majority of these symptoms. Application of SA reduced NaCl-induced increase of Na+ in the rosettes, but not in the roots. Accumulation of total amino acids was stimulated in NaCl-treated roots, especially due to exceptional increase of proline (4.4-fold). Among phenolic acids, accumulation of protocatechuic acid was the most enhanced in NaCl-exposed leaf rosettes (ca. 3-fold) while chlorogenic and caffeic acids in the roots (2.4- and 2.8-fold, respectively). Total soluble phenols increased after NaCl and SA treatments, but root lignin content was not affected. Activity of phenylalanine ammonia-lyase and shikimate dehydrogenase increased in response to NaCl, but cinnamyl alcohol dehydrogenase was not affected and polyphenol oxidase decreased. Stress parameters were elevated by NaCl treatment (superoxide radical and malondialdehyde content, activities of catalase, ascorbate- and guaiacol-peroxidase) and substantially prevented by SA, while accumulation of hydrogen peroxide decreased. Overall, SA showed strong beneficial properties against NaCl-induced negative symptoms. Protective effect of SA was the most visible at the level of guaiacol-peroxidase and through amelioration of stress parameters and mineral nutrient contents.
[show abstract][hide abstract] ABSTRACT: The effect of Atropa belladonna L. (AB) aqueous extract on skin wound healing was studied in male Sprague-Dawley rats subjected to two parallel full-thickness skin incisions on the back. Specimens for histological evaluation were collected on days 2 and 5 whereas for biomechanical testing, they were collected on day 5. In the in vitro study, a different concentration of AB extract was used to test the differentiation of keratinocytes using a panel of selected antibodies, proliferation, and cell survival of 3T3 fibroblasts and human umbilical vein endothelial cells using the MTT-assay. Results of the in vivo experiments showed in AB-treated wounds a shortened process of inflammation and accelerated collagen formation, as well as significantly increased wound stiffness as compared with control tissues. The in vitro examination showed that control keratinocytes were cytokeratin 19 free, while samples exposed to the highest AB extract concentration expressed CK19. Moreover, all concentrations were stimulatory to human umbilical vein endothelial cell proliferation. In addition, only the AB extract at the lowest tested concentration increased fibroblast growth, but higher concentrations decreased cell survival. In conclusion, our results indicate that the AB water extract positively affects early phases of skin wound healing in rats. However, the in vitro results on the inverse relation between the concentration of the AB extract and its effects on cell proliferation may be important for future research.
Wound Repair and Regeneration 01/2009; 17(3):378-86. · 2.76 Impact Factor
[show abstract][hide abstract] ABSTRACT: The influence of salicylic acid (SA) doses of 50 and 250 microM, for a period of up to 7 days, on selected physiological aspects and the phenolic metabolism of Matricaria chamomilla plants was studied. SA exhibited both growth-promoting (50 microM) and growth-inhibiting (250 microM) properties, the latter being correlated with decrease of chlorophylls, water content and soluble proteins. In terms of phenolic metabolism, it seems that the higher SA dose has a toxic effect, based on the sharp increase in phenylalanine ammonia-lyase (PAL) activity (24 h after application), which is followed by an increase in total soluble phenolics, lignin accumulation and the majority of the 11 detected phenolic acids. Guaiacol-peroxidase activity was elevated throughout the experiment in 250 microM SA-treated plants. In turn, some responses can be explained by mechanisms associated with oxidative stress tolerance; these mitigate acute SA stress (which is indicated by an increase in malondialdehyde content). However, PAL activity decreased with prolonged exposure to SA, indicating its inhibition. Accumulation of coumarin-related compounds (umbelliferone and herniarin) was not affected by SA treatments, while (Z)- and (E)-2-beta-D: -glucopyranosyloxy-4-methoxycinnamic acids increased in the 250 microM SA-treated rosettes. Free SA content in the rosettes increased significantly only in the 250 microM SA treatment, with levels tending to decrease towards the end of the experiment and the opposite trend was observed in the roots.
[show abstract][hide abstract] ABSTRACT: Cadmium (Cd) and copper (Cu) uptake by the plants of Matricaria chamomilla and relation to activities of guaiacol peroxidase (GPX, EC 184.108.40.206), catalase (CAT, EC 220.127.116.11) and glutathione reductase (GR, EC 18.104.22.168) up to 7 days of exposure to 3, 60 and 120 microM Cd or Cu was studied. Cd content in rosettes was ca. 10-fold higher in comparison to Cu while Cu was preferentially accumulated in the roots. In line with this observation, increase of CAT and GPX activity was similar in rosettes of Cd and Cu-treated plants, indicating non-redox active properties of Cd and low Cu accumulation. In the roots, Cu showed strong pro-oxidant effect, as judged from extreme stimulation of CAT and GPX, followed by increase of hydrogen peroxide and malondialdehyde. However, GPX seemed to be more important for alleviation of oxidative stress (ca. 93-250-fold higher activity in 120 microM Cu-treated roots). Cd had substantially lower influences and stimulated GR activity more than Cu. Activities of hydrogen peroxide-scavenging enzymes in relation to its accumulation are also discussed.
[show abstract][hide abstract] ABSTRACT: Comparisons were made between the content of Al, Cd, Cu, Fe, Mn, Pb, Sb and Zn in thalli of the lichen Xanthoria parietina and selected physiological processes. Assimilation pigments, activity of photosystem II, parietin, reactive substances produced using thiobarbituric acid (TBA) and CO(2) gas exchange were measured in order to assess tolerance to atmospheric sources of pollution. As expected, lichen thalli accumulated high amounts of the measured elements in relation to distance from pollution sources in Kosice, Slovak Republic (US Steel factory and vehicular traffic in the city center). However, except for TBA reactive substances production and to some extent CO(2) gas exchange at the most polluted station, none of the tested physiological parameters showed a clear correlation between accumulation of elements and physiological damage. This reflected a high degree of pollution tolerance in this lichen and corresponded with its high abundance in Kosice.
Ecotoxicology and Environmental Safety 07/2008; 70(2):319-26. · 2.20 Impact Factor
[show abstract][hide abstract] ABSTRACT: Physiological responses of Matricaria chamomilla plants exposed to cadmium (Cd) and copper (Cu) excess (3, 60, and 120 microM for 7 days) with special emphasis on phenolic metabolism were studied. Cu at 120 microM reduced chamomile growth, especially in the roots where it was more abundant than Cd. Notwithstanding the low leaf Cu amount (37.5 microg g(-1) DW) in comparison with Cd (237.8 microg g(-1) DW) at 120 microM, it caused reduction of biomass accumulation, F(v)/F(m) ratio and soluble proteins. In combination with high accumulation of phenolics, strong reduction of proteins and high GPX activity in the roots, this supports severe redox Cu properties. In terms of leaf phenylalanine ammonia-lyase (PAL) activity, it seems that Cd had a stimulatory effect during the course of the experiment, whereas Cu was found to stimulate it after 7-day exposure. The opposite trend was visible in the roots, where Cd had a stimulatory effect at high doses but Cu mainly at the highest dose. This supports the assumption of different PAL time dynamics under Cd and Cu excess. A dose of 60 and 120 microM Cu led to 2- and 3-times higher root lignin accumulation while the same Cd doses increased it by 33 and 68%, respectively. A Cu dose of 120 microM can be considered as limiting for chamomile growth under conditions of present research, while resistance to high Cd doses was confirmed. However, PAL and phenolics seemed to play an important role in detoxification of Cd- and Cu-induced oxidative stress.
[show abstract][hide abstract] ABSTRACT: The mycobiont of lichens usually determines the morphology of the symbiotic organism and is also dominates in terms of biomass. However, its role for sensitivity or tolerance of lichens to heavy metals is almost unknown. In the present study, the influence of copper (Cu) on the aposymbiotically-grown mycobiont of Cladonia cristatella was assessed. Intracellular Cu uptake was correlated with increasing Cu concentrations over a 24-h exposure time. Viability, measured as the degree of reduction of triphenyltetrazolium chloride to triphenyl formazan, as well as to ergosterol levels, decreased with growing Cu concentrations tested. Reduced glutathione (GSH) was found to be the most abundant low-molecular-weight thiol in the hyphae of C. cristatella and its intracellular content increased at concentrations of 10mum Cu. Higher Cu concentrations caused a significant decrease in GSH, possibly due to heavy metal-induced oxidation of GSH to glutathione disulphide (GSSG). Free cysteine levels were relatively constant. As expected, we did not observe the production of phytochelatins in the mycobiont, contrary to what is found in intact lichens and axenic cultures of their photobionts.
Mycological Research 09/2006; 110(Pt 8):994-9. · 2.81 Impact Factor
[show abstract][hide abstract] ABSTRACT: The influence of copper was assessed on wild and tolerant strains of the lichen photobiont Trebouxia erici and shown to have multiple toxic effects. Addition of 4 mM copper chloride into Trebouxia media reduced growth rates of the wild type to less than 1% of control levels. It also injured cell membranes, inhibited dehydrogenase activity, altered pigment composition, and reduced activity of photosystem II. Membrane damage was assessed through measuring electrical conductivity and potassium content, dehydrogenase activity by degree of conversion of 2,3,5-triphenyl tetrazolium chloride to red triphenylformazan, and functioning of PS II by chlorophyll a fluorescence. In respect to most parameters the tolerant strain was usually less affected by copper than the wild strain and, in some cases, not at all. We demonstrated by inductively coupled plasma atomic emission spectrometry and energy-dispersive X-ray microanalysis that enriched copper in the medium resulted in increased uptake by both wild and tolerant photobiont cells, but extracellular concentrations were significantly higher in the tolerant strain. This suggested that, at least in vitro, extracellular deposition was a mechanism of copper tolerance.
Archives of Environmental Contamination and Toxicology 09/2003; 45(2):159-67. · 2.01 Impact Factor