Lichen Xanthoria Elegans Research Articles

Resistance of Primary Photosynthesis to Photoinhibition in Antarctic Lichen Xanthoria elegans: Photoprotective Mechanisms Activated during a Short Period of High Light Stress.

The Antarctic lichen, Xanthoria elegans, in its hydrated state has several physiological mechanisms to cope with high light effects on the photosynthetic processes of its photobionts. We aim to investigate the changes in primary photochemical processes of photosystem II in response to a short-term photoinhibitory treatment. Several chlorophyll a fluorescence techniques: (1) slow Kautsky kinetics supplemented with quenching mechanism analysis; (2) light response curves of photosynthetic electron transport (ETR); and (3) response curves of non-photochemical quenching (NPQ) were used in order to evaluate the phenomenon of photoinhibition of photosynthesis and its consequent recovery. Our findings suggest that X. elegans copes well with short-term high light (HL) stress due to effective photoprotective mechanisms that are activated during the photoinhibitory treatment. The investigations of quenching mechanisms revealed that photoinhibitory quenching (qIt) was a major non-photochemical quenching in HL-treated X. elegans; qIt relaxed rapidly and returned to pre-photoinhibition levels after a 120 min recovery. We conclude that the Antarctic lichen species X. elegans exhibits a high degree of photoinhibition resistance and effective non-photochemical quenching mechanisms. This photoprotective mechanism may help it survive even repeated periods of high light during the early austral summer season, when lichens are moist and physiologically active.

UV-B absorbing and bioactive secondary compounds in lichens Xanthoria elegans and Xanthoria parietina: A review

Secondary metabolites are the bioactive compounds of plants which are synthesized during primary metabolism, have no role in the development process but are needed for defense and other special purposes. These secondary metabolites, such as flavonoids, terpenes, alkaloids, anthraquinones and carotenoids, are found in Xanthoria genus lichens. These lichens are known as lichenized fungi in the family Teloschistaceae, which grows on rock and produce bioactive compounds. A lot of secondary compounds in plants are induced by UV (100-400 nm) spectra. The present review showcases the present identified bioactive compounds in Xanthoria elegans and Xanthoria parietina lichens, which are stimulated by different amounts of UV-B light (280-320 nm), as well as the biochemistry of the UV-B absorbingcompounds.

Special issue in honour of Prof. Reto J. Strasser - Analysis of K- and L-band appearance in OJIPs in Antarctic lichens in low and high temperature

In this study, we evaluated the effect of temperature on the fast chlorophyll fluorescence (ChlF) transient (OJIP) and OJIP-derived parameters in Antarctic lichens Xanthoria elegans, Usnea antarctica, and Dermatocarpon polyphyllizum. Samples were exposed to a range of temperatures (-5 to +45 degrees C) and measured after 15-min equilibration. High temperature (+45 degrees C) caused a decrease of ChlF, an increased J-step, and shortened time to reach peak ChlF (FP). Temperature below +5 degrees C caused the increase of ChlF and J-step. The K-band was identified in X. elegans (above +20 degrees C), U. antarctica (+35 degrees C), and D. polyphyllizum (+45 degrees C). L-band was well distinguishable in X. elegans (+45 degrees C). As indicated by the OJIP-derived parameters, high temperature inhibited photosystem II function. The inhibition was apparent as less effective energetic connectivity. The OJIP transients and auxiliary measurement of ChlF temperature curves suggested that X. elegans had the lowest termostability among the experimental species.

The evaluation of time‐resolved Raman spectroscopy for the suppression of background fluorescence from space‐relevant samples

AbstractOne of the primary goals in space research is the search for signs of extant or extinct extraterrestrial life, and Raman spectroscopy can play a role in this field. Raman spectrometers are planned for future missions to Mars and possibly the Moon to identify the mineralogical surface composition and potentially existing organic compounds (especially on Mars). However, a major challenge in Raman spectroscopy, especially in the visible range, is the strong fluorescence background. Time‐resolved Raman spectroscopy (TRRS) can provide selective detection of Raman signals over the generally longer living fluorescence. This study investigates the potential of a TRRS system, using 3‐ps, 440‐nm laser pulses and time‐gated detection with an intensified charge‐coupled device (CCD) camera. Test samples were the lichen Xanthoria elegans as an extraterrestrial life analogue, and a lunar regolith analogue material (LRS) as a planetary surface analogue. The TRRS technique is evaluated by comparing gated to nongated Raman spectroscopy using different detectors but with otherwise the same instrument and identical measurement conditions. The gated spectra of X. elegans showed significant signal‐to‐noise ratio (SNR) improvements compared to the nongated spectra. The visible Raman lines could be assigned to the photoprotective pigment parietin. For the LRS sample, measurement spots with a good SNR in the nongated spectrum were not significantly improved by measuring in gated mode. However, spots dominated by fluorescence showed significant improvement in gated mode because of fluorescence suppression. Minerals such as plagioclase, diopside, olivine, apatite, and a carbonate mineral were detected. In most cases, TRRS provided better results compared to nongated measurements, demonstrating the suitability for future space‐exploration missions.

The Effect of High-Dose Ionizing Radiation on the Astrobiological Model Lichen Circinaria gyrosa.

The lichen Circinaria gyrosa is an astrobiological model defined by its high capacity of resistance to space conditions and to a simulated martian environment. Therefore, it became part of the currently operated BIOMEX experiment on board the International Space Station and the recent STARLIFE campaign to study the effects of four types of space-relevant ionizing radiation. The samples were irradiated with helium and iron ions at doses up to 2 kGy, with X-rays at doses up to 5 kGy and with γ rays at doses from 6 to 113 kGy. Results on C. gyrosa's resistance to simulated space ionizing radiation and its post-irradiation viability were obtained by (i) chlorophyll a fluorescence of photosystem II (PSII), (ii) epifluorescence microscopy, (iii) confocal laser scanning microscopy (CLSM), and (iv) field emission scanning electron microscopy (FESEM). Results of photosynthetic activity and epifluorescence show no significant changes up to a dose of 1 kGy (helium ions), 2 kGy (iron ions), 5 kGy (X-rays)-the maximum doses applied for those radiation qualities-as well as a dose of 6 kGy of γ irradiation, which was the lowest dose applied for this low linear energy transfer (LET) radiation. Significant damage in a dose-related manner was observed only at much higher doses of γ irradiation (up to 113 kGy). These data corroborate the findings of the parallel STARLIFE studies on the effects of ionizing radiation on the lichen Circinaria gyrosa, its isolated photobiont, and the lichen Xanthoria elegans. Key Words: Simulated space ionizing radiation-Gamma rays-Extremotolerance-Lichens-Circinaria gyrosa-Photosynthetic activity. Astrobiology 17, 145-153.

Simulated Space Radiation: Impact of Four Different Types of High-Dose Ionizing Radiation on the Lichen Xanthoria elegans.

This study addresses the viability of the lichen Xanthoria elegans after high-dose ionizing irradiation in the frame of the STARLIFE campaign. The first set of experiments was intended to resemble several types of galactic cosmic radiation (GCR) as present beyond the magnetic shield of Earth. In the second set of experiments, γ radiation up to 113 kGy was applied to test the limit of lichen resistance to ionizing radiation. Entire thalli of Xanthoria elegans were irradiated in the anhydrobiotic state. After STARLIFE 1, the metabolic activity of both symbionts was quantified by live/dead staining with confocal laser scanning microscopy. The photosynthetic activity was measured after the respective irradiation to assess the ability of the symbiotic green algae to restore photosynthesis after irradiation. The STARLIFE campaign complements the results of the LIFE experiments at the EXPOSE-E facility on the International Space Station by testing the model organism Xanthoria elegans on its resistance to hazardous radiation that might be accumulated during long-term space exposure. In addition, the photosynthetic activity of metabolically active lichen was investigated after X-ray irradiation up to 100 Gy (3.3 Gy/min). Since previous astrobiological experiments were mostly performed with anhydrobiotic lichen, these experiments will broaden our knowledge on the correlation of physiological state and astrobiological stressors. Key Words: Astrobiology-Extremotolerance-Gamma rays-Ionizing radiation-Lichens-Viability. Astrobiology 17, 136-144.

Characterisation of Growth and Ultrastructural Effects of the Xanthoria elegans Photobiont After 1.5Years of Space Exposure on the International Space Station.

The lichen Xanthoria elegans has been exposed to space and simulated Mars-analogue environment in the Lichen and Fungi Experiment (LIFE) on the EXPOSE-E facility at the International Space Station (ISS). This long-term exposure of 559 days tested the ability of various organisms to cope with either low earth orbit (LEO) or Mars-analogue conditions, such as vacuum, Mars-analogue atmosphere, rapid temperature cycling, cosmic radiation of up to 215 ± 16 mGy, and insolation of accumulated doses up to 4.87 GJm(-2), including up to 0.314 GJm(-2) of UV irradiation. In a previous study, X. elegans demonstrated considerable resistance towards these conditions by means of photosynthetic activity as well as by post-exposure metabolic activity of 50-80% in the algal and 60-90% in the fungal symbiont (Brandt et al. Int J Astrobiol 14(3):411-425, 2015). The two objectives of the present study were complementary: First, to verify the high post-exposure viability by using a qualitative cultivation assay. Second, to characterise the cellular damages by transmission electron microscopy (TEM) which were caused by the space and Mars-analogue exposure conditions of LIFE. Since the algal symbiont of lichens is considered as the more susceptible partner (de Vera and Ott 2010), the analyses focused on the photobiont. The study demonstrated growth and proliferation of the isolated photobiont after all exposure conditions of LIFE. The ultrastructural analysis of the algal cells provided an insight to cellular damages caused by long-term exposure and highlighted that desiccation-induced breakdown of cellular integrity is more pronounced under the more severe space vacuum than under Mars-analogue atmospheric conditions. In conclusion, desiccation-induced damages were identified as a major threat to the photobiont of X. elegans. Nonetheless, a fraction of the photobiont cells remained cultivable after all exposure conditions tested in LIFE.

Humidity interaction of lichens under astrobiological aspects: the impact of UVC exposure on their water retention properties

AbstractWe quantitatively studied the hydration and dehydration behaviour of the three astrobiological model lichens Xanthoria elegans, Buellia frigida and Circinaria gyrosa by thermoanalysis and gravimetric isotherm measurements under close-to-Martian environmental conditions in terms of low temperature and low pressure. Additionally, the impact of UVC exposure on the isolated symbionts of B. frigida and X. elegans was studied by thermoanalysis and mass spectrometry as well as by gravimetric isotherm measurements. The thermal analysis revealed whewellite as a component of C. gyrosa which was not found in B. frigida and X. elegans. Neither the water retention nor the thermal behaviour of symbionts changed when irradiated with UVC under dry conditions. On the other hand, UVC irradiation of the wet mycobiont of B. frigida had a distinct impact on the hydration/dehydration ability which was not observed for the mycobiont of X. elegans. Possibly the melanin of B. frigida's mycobiont, that is not present in X. elegans, or a specifically damaged acetamido group of the chitin of B. frigida may be the sources of additional UVC-induced sorption sites for water associated with the UVC exposure.

Production of UV-B screens and changes in photosynthetic efficiency in Antarctic Nostoc commune colonies and a lichen Xanthoria elegans depend on a dose and duration of UV-B stress

The survival of non-vascular autotrophs in the extreme polar conditions and the principles of their high tolerance to extreme physical factors have been intriguing scientists in last decades. Therefore, this study focuses on the capacity of production of UV-B screening pigments in two model Antarctic species, one algal lichen, and colony of a cyanobacterium. Dose-dependent activation of protective mechanisms of Antarctic cyanobacterium (Nostoc commune) and algal lichen (Xanthoria elegans), synthesis of UV-B screening compounds in particular, were studied together with the changes in photosynthetic efficiency induced by a background photosynthetically active radiation (PAR) supplemented with UV-B radiation. The samples were exposed to different doses of UV-B (280–320 nm), low (0.7 W m-2), medium (1.5 W m-2) and high (3.0 W m-2) for 5 days. Untreated samples (control) were shielded from UV-B radiation during experiment. Chlorophyll fluorescence parameters and secondary UV-B protective metabolites were analysed in the intervals of 24 h, 48 h and 120 h. Amount of UV-B screening pigments was measured spectrophotometrically using several specific wavelengths in UV-B absorption range. Results showed that if exposed to a low dose of UV-B radiation or a short-term treatment, both species exhibited an increase in UV-B screening pigments to protect the lichen photobiont against UV-B damage. However, if exposed to a high dose of UV-B radiation or a long-term treatment, a decrease of UV-B screening compounds occured. This implies that Antarctic lichen and cyanobacterium can protect themselves against an increase of stress factors ranging within physiological limits, like e.g. increased synthesis of UV-B screening compounds thanks to a thinning of the ozone layer and consequent increase in UV radiation doses incident on Antarctic terrestrial ecosystems. Nevertheless, the likely increased UV-B radiation due to more intense depletion of stratospheric ozone layer may lead to alterations in UV-B tolerance in Antarctic lichens in future.

Survival Potential and Photosynthetic Activity of Lichens Under Mars-Like Conditions: A Laboratory Study

Lichens were repetitively exposed over 22 days to thermophysical Mars-like conditions at low-and mid-latitudes. The simulated parameters and the experimental setup are described. Natural samples of the lichen Xanthoria elegans were used to investigate their ability to survive the applied Mars-like conditions. The effects of atmospheric pressure, CO(2) concentration, low temperature, water availability, and light on Mars were also studied. The results of these experiments indicate that no significant decrease in the vitality of the lichen occurred after exposure to simulated martian conditions, which was demonstrated by confocal laser scanning microscopy analysis, and a 95% CO(2) atmosphere with 100% humidity, low pressure (partial pressure of CO(2) was 600 Pa), and low temperature has a balancing effect on photosynthetic activity as a function of temperature. This means a starting low photosynthetic activity at high CO(2) concentrations with Earth-like pressure has a reduction of 60%. But, if the simulated atmospheric pressure is reduced to Mars-like conditions with the maintenance of the same Mars-like 95% CO(2) concentration, the photosynthetic activity increases and again reaches similar values as those exhibited under terrestrial atmospheric pressure and concentration. Based on these results, we presume that, in any region on Mars where liquid water might be available, even for short periods of time, a eukaryotic symbiotic organism would have the ability to survive, at least over weeks, and to temporarily photosynthesize.

Life at the limits: capacities of isolated and cultured lichen symbionts to resist extreme environmental stresses.

Lichens are described as a symbiosis formed by a myco- and photobiont, capable of colonizing habitats where their separate symbionts would not be able to survive. Space simulation studies on the separated symbionts of the lichen Xanthoria elegans have been performed to test their capacity to resist the most extreme conditions. The isolated cultured symbiont cells were exposed to different doses of the UV spectrum, and to vacuum. Cultures of both symbionts were analysed by specific vitality tests (LIVE/DEAD-staining detected by Confocal Laser Scanning Microscopy). Growth capacity of symbiont cultures on different media was analysed after exposure to extreme environmental stresses. The data obtained support the hypothesis that the symbiotic state considerably enhances the ability of the respective symbionts to survive exposure to extreme conditions, including the conditions of space simulation. Species such as X. elegans may, therefore, be suitable for use as model organisms in exobiological studies.

Microbial Rock Inhabitants Survive Hypervelocity Impacts on Mars-Like Host Planets: First Phase of Lithopanspermia Experimentally Tested

The scenario of lithopanspermia describes the viable transport of microorganisms via meteorites. To test the first step of lithopanspermia, i.e., the impact ejection from a planet, systematic shock recovery experiments within a pressure range observed in martian meteorites (5-50 GPa) were performed with dry layers of microorganisms (spores of Bacillus subtilis, cells of the endolithic cyanobacterium Chroococcidiopsis, and thalli and ascocarps of the lichen Xanthoria elegans) sandwiched between gabbro discs (martian analogue rock). Actual shock pressures were determined by refractive index measurements and Raman spectroscopy, and shock temperature profiles were calculated. Pressure-effect curves were constructed for survival of B. subtilis spores and Chroococcidiopsis cells from the number of colony-forming units, and for vitality of the photobiont and mycobiont of Xanthoria elegans from confocal laser scanning microscopy after live/dead staining (FUN-I). A vital launch window for the transport of rock-colonizing microorganisms from a Mars-like planet was inferred, which encompasses shock pressures in the range of 5 to about 40 GPa for the bacterial endospores and the lichens, and a more limited shock pressure range for the cyanobacterium (from 5-10 GPa). The results support concepts of viable impact ejections from Mars-like planets and the possibility of reseeding early Earth after asteroid cataclysms.

Metabolic processes sustaining the reviviscence of lichen Xanthoria elegans (Link) in high mountain environments.

To survive in high mountain environments lichens must adapt themselves to alternating periods of desiccation and hydration. Respiration and photosynthesis of the foliaceous lichen, Xanthoria elegans, in the dehydrated state were below the threshold of CO2-detection by infrared gas analysis. Following hydration, respiration totally recovered within seconds and photosynthesis within minutes. In order to identify metabolic processes that may contribute to the quick and efficient reactivation of lichen physiological processes, we analysed the metabolite profile of lichen thalli step by step during hydration/dehydration cycles, using 31P- and 13C-NMR. It appeared that the recovery of respiration was prepared during dehydration by the accumulation of a reserve of gluconate 6-P (glcn-6-P) and by the preservation of nucleotide pools, whereas glycolytic and photosynthetic intermediates like glucose 6-P and ribulose 1,5-diphosphate were absent. The large pools of polyols present in both X. elegans photo- and mycobiont are likely to contribute to the protection of cell constituents like nucleotides, proteins, and membrane lipids, and to preserve the integrity of intracellular structures during desiccation. Our data indicate that glcn-6-P accumulated due to activation of the oxidative pentose phosphate pathway, in response to a need for reducing power (NADPH) during the dehydration-triggered down-regulation of cell metabolism. On the contrary, glcn-6-P was metabolised immediately after hydration, supplying respiration with substrates during the replenishment of pools of glycolytic and photosynthetic intermediates. Finally, the high net photosynthetic activity of wet X. elegans thalli at low temperature may help this alpine lichen to take advantage of brief hydration opportunities such as ice melting, thus favouring its growth in harsh high mountain climates.

Protective pigmentation in UVB‐screened Antarctic lichens studied by Fourier transform Raman spectroscopy: an extremophile bioresponse to radiation stress

AbstractFT‐Raman spectra were obtained for two Antarctic extremophiles, the epilithic lichens Xanthoria elegans and Caloplaca sublobulata from the maritime ecological long‐term research site on Leonie Island. Twelve specimens from cloches designed for the filtering out and transmission of UVB radiation over a 2 year period and two specimens from the natural habitat outside the cloches were analysed in terms of their characteristic Raman bands from the two photoprotective pigments parietin and β‐carotene. Following chemometric analysis, the specimens inside the UVB‐protective cloches exhibited a lower parietin:β‐carotene ratio than specimens from the same habitat that did not have UVB protection. The relative roles of parietin, a passive UVB photoprotectant, and β‐carotene are discussed and a possible duality of biological function is suggested for these pigments. Copyright © 2004 John Wiley & Sons, Ltd.

Phototolerance of lichens, mosses and higher plants in an alpine environment: analysis of photoreactions.

Adaptation to excessive light is one of the requirements of survival in an alpine environment particularly for poikilohydric organisms which in contrast to the leaves of higher plants tolerate full dehydration. Changes in modulated chlorophyll fluorescence and 820-nm absorption were investigated in the lichens Xanthoria elegans (Link) Th. Fr. and Rhizocarpon geographicum (L.) DC, in the moss Grimmia alpestris Limpr. and the higher plants Geum montanum L., Gentiana lutea L. and Pisum sativum L., all collected at altitudes higher than 2000 m above sea level. In the dehydrated state, chlorophyll fluorescence was very low in the lichens and the moss, but high in the higher plants. It increased on rehydration in the lichens and the moss, but decreased in the higher plants. Light-induced charge separation in photosystem II was indicated by pulse-induced fluorescence increases only in dried leaves, not in the dry moss and dry lichens. Strong illumination caused photodamage in the dried leaves, but not in the dry moss and dry lichens. Light-dependent increases in 820-nm absorption revealed formation of potential quenchers of chlorophyll fluorescence in all dehydrated plants, but energy transfer to quenchers decreased chlorophyll fluorescence only in the moss and the lichens, not in the higher plants. In hydrated systems, coupled cyclic electron transport is suggested to occur concurrently with linear electron transport under strong actinic illumination particularly in the lichens because far more electrons became available after actinic illumination for the reduction of photo-oxidized P700 than were available in the pool of electron carriers between photosystems II and I. In the moss Grimmia, but not in the lichens or in leaves, light-dependent quenching of chlorophyll fluorescence was extensive even under nitrogen, indicating anaerobic thylakoid acidification by persistent cyclic electron transport. In the absence of actinic illumination, acidification by ca. 8% CO2 in air quenched the initial chlorophyll fluorescence yield Fo only in the hydrated moss and the lichens, not in leaves of the higher plants. Under the same conditions, 8% CO2 reduced the maximal fluorescence yield Fm strongly in the poikilohydric organisms, but only weakly or not at all in leaves. The data indicate the existence of deactivation pathways which enable poikilohydric organisms to avoid photodamage not only in the hydrated but also in the dehydrated state. In the hydrated state, strong nonphotochemical quenching of chlorophyll fluorescence indicated highly sensitive responses to excess light which facilitated the harmless dissipation of absorbed excitation energy into heat. Protonation-dependent fluorescence quenching by cyclic electron transport, P700 oxidation and, possibly, excitation transfer between the photosystems were effectively combined to produce phototolerance.

DNA isolation from lichen ascomata

Analysis of DNA from the fungal component of lichens requires selective protocols to isolate its DNA from that of its symbiotic partner. In the present study, we describe a method for extraction of DNA from fungal ascomata, a source of algal-free mycelium. This method, which includes a DNA precipitation onto glassmilk (= ground SiO2), is particularly useful for limited amounts of starting material, as exemplified by the isolation of DNA from ascomata of Arthonia molendoi growing parasitically on the lichen Xanthoria elegans. The protocol is effective for the isolation of high-quality DNA from cultured fungi, herbarium specimens and lichens high in polysaccharide content. This new protocol makes possible the examination of many fungi until now thought intractable to DNA methods.