Desiccation-time limits of photosynthetic recovery in Equisetum hyemale (Equisetaceae) spores

The chlorophyllous spores of Equisetum survive desiccation, yet cannot tolerate this quiescent state for more than ~2 wk. The hypothesis that spore viability of Equisetum hyemale L. is limited by inhibition of photosynthetic recovery was tested using chlorophyll a fluorescence and oxygen-exchange analyses. Experimental spores were desiccated at 2% relative humidity and 25C for time periods of 24 h, 1 wk, and 2 wk, and then rehydrated at 200 mmol photons/m2s (PAR) and 25C for up to 24 h. Spores desiccated for 24 h recovered photosynthetic competence very rapidly during rehydration, reaching the O2 compensation point in 6.3 ~ 0.3 (mean +/- SE) min. Recovery of photosynthetic performance of spores desiccated for 1 wk was slower, as judged by significantly slower increases of (1) photochemical efficiency of photosystem (PS) II, (2) PS II quinoneB-reducing center concentration, (3) quinoneB concentration, (4) water-oxidation activity, (5) rate of light-induced O2 evolution, and (6) apparent quantum yield of net O2 exchange. Photosystem-II and whole-spore photosynthetic competence of 2-wk desiccated spores was increasingly impaired, and did not recover during rehydration. Origin fluorescence yield and dark respiration were not affected by desiccation time following rehydration. The results suggest that the extremely short viability of disseminated spores of Equisetum hyemale is due to the inability to recover losses of water oxidation and photosystem II-core function following 2 wk of desiccation.     

Activation of photosynthesis and resistance to photoinhibition in cyanobacteria within biological desert crust

Filamentous cyanobacteria are the main primary producers in biological desert sand crusts. The cells are exposed to extreme environmental conditions including temperature, light, and diurnal desiccation/rehydration cycles. We have studied the kinetics of activation of photosynthesis during rehydration of the cyanobacteria, primarily Microcoleus sp., within crust samples collected in the Negev desert, Israel. We also investigated their susceptibility to photoinhibition. Activation of the photosynthetic apparatus, measured by fluorescence kinetics, thermoluminescence, and low temperature fluorescence emission spectra, did not require de novo protein synthesis. Over 50% of the photosystem II (PSII) activity, assembled phycobilisomes, and photosystem I (PSI) antennae were detected within less than 5 min of rehydration. Energy transfer to PSII and PSI by the respective antennae was fully established within 10 to 20 min of rehydration. The activation of a fraction of PSII population (about 20%-30%) was light and temperature-dependent but did not require electron flow to plastoquinone [was not inhibited by 3-(3,4-dichlorophenyl)-1,1-dimethylurea]. The cyanobacteria within the crusts are remarkably resistant to photoinhibition even in the absence of protein synthesis. The rate of PSII repair increased with light intensity and with time of exposure. Consequently, the extent of photoinhibition in high-light-exposed crusts reached a constant, relatively low, level. This is in contrast to model organisms such as Synechocystis sp. strain PCC 6803 where PSII activity declined continuously over the entire exposure to high illumination. Ability of the crust's organisms to rapidly activate photosynthesis upon rehydration and withstand photoinhibition under high light intensity may partly explain their ability to survive in this ecosystem.     

Resurrection kinetics of photosynthesis in desiccation‐tolerant terrestrial green algae (Chlorophyta) on tree bark

The rough bark of orchard trees (Malus) around Darmstadt is predominantly covered in red to purple‐brown layers (biofilms) of epiphytic terrestrial alga of Trentepohlia umbrina. The smooth bark of forest trees (Fagus sylvatica L. and Acer sp.) in the same area is covered by bright green biofilms composed of the green algae Desmococcus, Apatococcus and Trebouxia, with a few cells of Coccomyxa and ‘Chlorella’ trebouxioides between them. These algae are desiccation tolerant. After samples of bark with the biofilms were kept in dry air in darkness for various periods of time, potential quantum yield of PSII, F_v/F_m, recovered during rehydration upon rewetting. The kinetics and degree of recovery depended on the length of time that the algae were kept in dry air in the desiccated state. Recovery was better for green biofilm samples, i.e. quite good even after 80 days of desiccation (F_v/F_m = ca. 50% of initial value), than the red samples, where recovery was only adequate up to ca. 30–40 days of desiccation (F_v/F_m = ca. 20–55% of initial value). It is concluded that the different bark types constitute different ecophysiological niches that can be occupied by the algae and that can be distinguished by their capacity to recover from desiccation after different times in the dry state.     

PSI-driven cyclic electron flow allows intertidal macro-algae Ulva sp. (Chlorophyta) to survive in desiccated conditions

Ulva sp. (Chlorophyta) is a representative species of the intertidal macro-algae responsible for the green tides that occurred along the shores of Qingdao in 2008 and had detrimental effects on the preparation for the 2008 Beijing Olympic Games sailing competition. In view of its significance, we have investigated the photosynthetic performance of the photosystems and the changes in photosynthetic electron transport that occur during desiccation and rehydration of Ulva sp. The PSII activity in Ulva sp. declined gradually during the course of desiccation, which was reflected by the decreased maximum quantum yield and effective quantum yield, whereas the PSI activity fluctuated significantly. In contrast, the electron transport rates of PSII approached zero at severe levels of desiccation, but the electron transport of PSI, which still operated, could be suppressed effectively by a specific inhibitor. Furthermore, the electron transport of PSI during rehydration of desiccated thalli was recovered faster than that of PSII. All these results implied that the linear electron flow was abolished in desiccated Ulva sp., whereas the cyclic PSI activity was significantly elevated, was still active at severe levels of desiccation and could be restored faster than PSII activity. Based on these results, we concluded the PSI-driven cyclic electron flow might provide desiccation tolerance and additional flexibility for the cell physiology of Ulva sp. under desiccation conditions, which might be one of the most important factors that make Ulva sp. well suited to experience daily cycles of desiccation at low tide and rehydration at high tide.     

Dehydration,temperature, and light tolerance in members of the aeroterrestrial green algal genus Interfilum (Streptophyta) from biogeographically different temperate soils

Unicellular green algae of the genus Interfilum (Klebsormidiales, Streptophyta) are typical components of biological soil crusts. Four different aeroterrestrial Interfilum strains that have previously been molecular‐taxonomically characterized and isolated from temperate soils in Belgium, Czech Republic, New Zealand, and Ukraine were investigated. Photosynthetic performance was evaluated under different controlled abiotic conditions, including dehydration, as well as under a light and temperature gradient. For standardized desiccation experiments, a new methodological approach with silica gel filled polystyrol boxes and effective quantum yield measurements from the outside were successfully applied. All Interfilum isolates showed a decrease and inhibition of the effective quantum yield under this treatment, however with different kinetics. While the single cell strains exhibited relatively fast inhibition, the cell packet forming isolates dried slower. Most strains fully recovered effective quantum yield after rehydration. All Interfilum isolates exhibited optimum photosynthesis at low photon fluence rates, but with no indication of photoinhibition under high light conditions suggesting flexible acclimation mechanisms of the photosynthetic machinery. Photosynthesis under lower temperatures was generally more active than respiration, while the opposite was true for higher temperatures. The presented data provide an explanation for the regular occurrence of Interfilum species in soil habitats where environmental factors can be particularly harsh.     

APPARENT LIGHT REQUIREMENT FOR ACTIVATION OF PHOTOSYNTHESIS UPON REHYDRATION OF DESICCATED BEACHROCK MICROBIAL MATS1

Photosynthetic electron transport of beachrock microbial mats growing in the intertidal zone of Heron Island (Great Barrier Reef, Australia) was investigated with a pulse amplitude modulation chl fluorometer providing four different excitation wavelengths for preferential excitation of the major algal groups (cyanobacteria, green algae, diatoms/dinoflagellates). A new type of fiberoptic emitter‐detector unit (PHYTO‐EDF) was used to measure chl fluorescence at the sample surface. Fluorescence signals mainly originated from cyanobacteria, which could be almost selectively assessed by 640‐nm excitation. Even after desiccation for long time periods under full sunlight, beachrock showed rapid recovery of photosynthesis after rehydration in the light (t_1/2～ 15 min). However, when rehydrated in the dark, the quantum yield of energy conversion of PSII remained zero over extended periods of time. Parallel measurements of O₂ concentration with an oxygen microoptode revealed zero oxygen concentration in the surface layer of rehydrated beachrock in the dark. Upon illumination, O₂ concentration increased in parallel with PSII quantum yield and decreased again to zero in the dark. It is proposed that oxygen is required for preventing complete dark reduction of the PSII acceptor pools via the NADPH‐dehydrogenase/chlororespiration pathway. This hypothesis is supported by the observation that PSII quantum yield could be partially induced in the dark by flushing with molecular oxygen. Abbreviations: EDF, emitter‐detector unit; F_o, fluor‐escence yield of dark‐adapted sample; F_m, maximal fluorescence yield measured during saturation pulse; F_v, variable fluorescence yield; LED, light‐emitting diode; PAM, pulse amplitude modulation; PQ, plastoquinone     

Chlororespiration in Green Algae Isolated From Desert Crusts

Photosynthetic organisms enduring extreme temperatures, low water availability, or high light require photoprotective mechanisms to prevent sustained damage to photosynthetic machinery. Green microalgae living in desert crust communities of the south‐western US experience all these environmental stresses, yet photophysiological studies of green algae in the literature have focused on only a handful of common aquatic and marine species. We are examining the variation in green algal photoprotective mechanisms that is the result of natural selection acting independently in multiple lineages of highly diverse desert green algae (Chlorophyta) within the classes Chlorophyceae and Trebouxiophyceae. We have found that unusually extensive dark reduction of the plastoquinone pool is a prominent photophysiological feature among these desert algae; this reduction may be linked with enhanced chlororespiration. Recently, chlororespiration in higher plants has been linked through mutant analysis to control of the carotenoid synthetic pathway, heat stress, and starch metabolism among other pathways, though the function of chlororespiration remains controversial. Given that green algae and higher plants are monophyletic, analysis of potential chlororespiration in desert green algae may help decipher the evolution of the chlororespiratory process as well as its potential role in photoprotection in desert habitats.     

Dehydration rate and time of desiccation affect recovery of the lichenic algae <Emphasis Type="Italic">Trebouxia erici</Emphasis>: alternative and classical protective mechanisms

The mechanisms involved in desiccation tolerance of lichens and their photobionts are still poorly understood. To better understand these mechanisms we have studied dehydration rate and desiccation time in Trebouxia, the most abundant chlorophytic photobiont in lichen. Our findings indicate that the drying rate has a profound effect on the recovery of photosynthetic activity of algae after rehydration, greater than the effects of desiccation duration. The basal fluorescence (F′_o) values in desiccated algae were significantly higher after rapid dehydration, than after slow dehydration, suggesting higher levels of light energy dissipation in slow-dried algae. Higher values of PSII electron transport were recovered after rehydration of slow-dried Trebouxia erici compared to rapid-dried algae. The main component of non-photochemical quenching after slow dehydration was energy dependent (q _E), whereas after fast dehydration it was photoinhibition (q _I). Although q _E seems to play a role during desiccation recovery, no significant variations were detected in the xanthophyll cycle components. Desiccation did not affect PSI functionality. Classical antioxidant activities like superoxide dismutase or peroxidase decreased during desiccation and early recovery. Dehydrins were detected in the lichen-forming algae T. erici and were constitutively expressed. There is probably a minimal period required to develop strategies which will facilitate transition to the desiccated state in this algae. In this process, the xanthophyll cycle and classical antioxidant mechanisms play a very limited role, if any. However, our results indicate that there is an alternative mechanism of light energy dissipation during desiccation, where activation is dependent on a sufficiently slow dehydration rate.     

Widespread desert affiliation of trebouxiophycean algae (Trebouxiophyceae,Chlorophyta) including discovery of three new desert genera

Unlike most other green algae, trebouxiophyceans are predominantly aerophytic and contain many symbiotic representatives. In recent years, a number of new terrestrial trebouxiophycean taxa were described from soils, tree bark, and lichens. The present phylogenetic study reveals three new lineages of free‐living trebouxiophyceans found in North American desert soil crusts and proposes new generic names to accommodate them: Desertella, Eremochloris, and Xerochlorella. This survey of desert isolates also led to discovery of representatives of seven existing genera of trebouxiophyceans. Two of these genera have never been reported to contain desert representatives and one was known previously only from aquatic habitats. Furthermore, we expand the known geographic range of the recently described genus Chloropyrula, heretofore only known from the Ural Mountains. We demonstrate that the diversity of trebouxiophyceans is still underestimated and poorly understood, and that most major trebouxiophycean lineages contain desert‐dwelling taxa.     

Efficiency of light utilization of Chlamydomonas reinhardtii under medium-duration light/dark cycles

The light regime inside a photobioreactor is characterized by a light gradient with full (sun)light at the light-exposed surface and darkness in the interior of the bioreactor. Consequently, depending on the mixing characteristics, algae will be exposed to certain light/dark cycles. In this study the green alga Chlamydomonas reinhardtii was cultivated under five different light regimes: (1) continuous illumination; (2) a square-wave light/dark cycle with a light fraction (epsilon) of 0.5 and a duration (t(c)) of 6.1 s; (3) epsilon=0.5, t(c)=14.5 s; (4) epsilon=0.5, t(c)=24.3 s and (5) epsilon=0.8, t(c)=15.2 s. The biomass yield on light energy, protein per photons, decreased under light/dark cycles (epsilon=0. 5) in comparison to continuous light (CL), from 0.207 (CL) to 0.117-0.153 g mol(-1) (epsilon=0.5). Concomitantly, the maximal specific photosynthetic activity, oxygen production per protein, decreased from 0.94 (CL) to 0.64-0.66 g g(-1) h(-1) (epsilon=0.5). Also the quantum yield of photochemistry, yield of the conversion of light energy into chemical energy, decreased from 0.47 (CL) to 0. 23 (epsilon=0.5, t(c)=24.3 s). Apparently, C. reinhardtii is not able to maintain a high photosynthetic capacity under medium-duration light/dark cycles and since specific light absorption did not change, light utilization efficiency decreased in comparison to continuous illumination.     

Desiccation and rehydration dynamics in the epiphytic resurrection fern Pleopeltis polypodioides

The epiphytic resurrection—or desiccation-tolerant (DT)—fern Pleopeltis polypodioides can survive extreme desiccation and recover physiological activity within hours of rehydration. Yet, how epiphytic DT ferns coordinate between deterioration and recovery of their hydraulic and photosynthetic systems remains poorly understood. We examined the functional status of the leaf vascular system, chlorophyll fluorescence, and photosynthetic rate during desiccation and rehydration of P. polypodioides. Xylem tracheids in the stipe embolized within 3–4 h during dehydration. When the leaf and rhizome received water, tracheids refilled after ∼24 h, which occurred along with dramatic structural changes in the stele. Photosynthetic rate and chlorophyll fluorescence recovered to predesiccation values within 12 h of rehydration, regardless of whether fronds were connected to their rhizome. Our data show that the epiphytic DT fern P. polypodioides can utilize foliar water uptake to rehydrate the leaf mesophyll and recover photosynthesis despite a broken hydraulic connection to the rhizome.     

Ecophysiological responses of desiccation-tolerant cryptobiotic crusts

In our present studies, the recovery of photosynthetic activity after rehydration was demonstrated. We measured chlorophyll fluorescence, CO₂ gas exchange and the pigment composition in the previously long-term air-dried cryptogamic inselberg crusts collected from two tropical areas. The cryptobiotic crusts were collected from different localities on similar ecological and climatic conditions from extreme habitats of inselbergs (outcrops). These inselbergs are characterized by a dry microclimate and are covered by scarce soil. We found that the ecophysiological responses of both cryptogamic inselberg crusts showed an extremely high degree of desiccation-tolerance due to the fast and full recovery during rehydration. The photosynthetic activity of the cryptobiotic crusts were restored and regained within 15 and 40 min, respectively, after rehydration. Photosynthetic activity of the crusts was retained at all applied light intensities when enough water was available, however the degree of the recovery was different between the crusts. Photosynthetic pigment contents were strongly and positively correlated with water content. Our results indicated that tropical desiccation-tolerant cryptogamic crusts found on inselberg rock surfaces have CO₂ fixation ability in the range of cyanobacteria and lichens, suggesting that at a global scale they can assimilate CO₂ in a significant amount.     

Unearthing the molecular phylodiversity of desert soil green algae (Chlorophyta)

Deserts are not usually considered biodiversity hotspots, but desert microbiotic crust communities exhibit a rich diversity of both eukaryotic and prokaryotic life forms. Like many communities dominated by microscopic organisms, they defy characterization by traditional species-counting approaches to assessing biodiversity. Here we use exclusive molecular phylodiversity (E) to quantify the amount of evolutionary divergence unique to desert-dwelling green algae (Chlorophyta) in microbiotic crust communities. Given a phylogenetic tree with branch lengths expressed in units of expected substitutions per site, E is the total length of all tree segments representing exclusively desert lineages. Using MCMC to integrate over tree topologies and branch lengths provides 95% Bayesian credible intervals for phylodiversity measures. We found substantial exclusive molecular phylodiversity based on 18S rDNA data, showing that desert lineages are distantly related to their nearest aquatic relatives. Our results challenge conventional wisdom, which holds that there was a single origin of terrestrial green plants and that green algae are merely incidental visitors rather than indigenous components of desert communities. We identify examples of lineage diversification within deserts and at least 12 separate transitions from aquatic to terrestrial life apart from the most celebrated transition leading to the embryophyte land plants. [Bayesian phylogenetics; biodiversity; exclusive molecular phylodiversity; microbiotic crusts.].     

EFFECTS OF SALINITY AND LIGHT INTENSITY ON THE RESUMPTION OF PHOTOSYNTHESIS IN REHYDRATED CYANOBACTERIAL MATS FROM BAJA CALIFORNIA SUR,MEXICO1

Lyngbya mats in the intertidal of the Laguna Ojo de Liebre are metabolically active for only a few days every 2 weeks during spring tides, with environmental conditions varying greatly during these periods of hydration. Pulse amplitude modulated fluorometry (PAM) and oxygen measurements were used to measure photosynthetic activity during the first few hours after rehydration under various light intensities and salinities. Upon rehydration, a transitory maximum in respiratory activity (10–30 min) occurred before the resumption of photosynthesis, which could recover in about 2 h. Salinities outside the mats' natural range (35–50 psu) were detrimental to photosynthetic recovery. Both high (100 psu) and low (0–10 psu) salinities slowed recovery as well as lowered the overall photosynthetic yield. Photosynthesis was initiated earlier and recovered more rapidly with increasing light intensity. In addition, the positive effect of light on rates of recovery was disproportionately greater at lower salinities (10–25 psu) where high light (500 W·m^?2) counteracted the negative effects of low‐salt stress early in recovery. However, higher light intensities became photoinhibitory later in recovery (>2 h). Photosynthesis did not recover uniformly within the mat. Filaments deeper in the mat most likely recovered later than those near the surface due to high light attenuation. The ability of the mats to tolerate desiccation and take advantage of hydration periods even when conditions are suboptimal enables these mats to predominate in the intertidal environment.     

ECOPHYSIOLOGICAL PERFORMANCE OF THE AEROTERRESTRIAL GREEN ALGA KLEBSORMIDIUM CRENULATUM (CHAROPHYCEAE,STREPTOPHYTA) ISOLATED FROM AN ALPINE SOIL CRUST WITH AN EMPHASIS ON DESICCATION STRESS1

Aeroterrestrial filamentous green algae of the genus Klebsormidium (Klebsormidiales, Streptophyta) are typical components of biological soil crusts, which occur worldwide in arid and semiarid habitats including alpine regions. In the present study, Klebsormidium crenulatum (Kütz.) Lokhorst was isolated from an alpine soil crust above the timberline of the Austrian Alps. Growth responses, photosynthetic performance, and desiccation tolerance were measured under controlled laboratory conditions. K. crenulatum exhibited optimal growth and the highest photosynthetic efficiency under relatively low photon fluence densities (30 and 21.9 μmol photons · m^?2 · s^?1, respectively), indicating low‐light requirements. It grew in a narrow range of salinities between 1.2 and 15 practical salinity units (psu), pointing to a pronounced stenohaline response pattern. Increasing temperatures from 5°C to 40°C led to different effects on photosynthetic oxygen evolution and respiratory oxygen consumption in K. crenulatum. While at low temperatures (5°C–10°C) photosynthesis was relatively high, respiration was not detectable or was at a very low level. Conversely, at the highest temperature of 40°C, photosynthesis was inhibited, and respiration unaffected, indicating strong differences in temperature sensitivity between both physiological processes. K. crenulatum was capable of photosynthesizing efficiently for up to 2.5 h under desiccation, followed by a decrease to 15% of the initial value after 3 h. Complete recovery took place within 2 h after rehydration. All ecophysiological data explain the widespread abundance of K. crenulatum in soil crusts of the alpine regions of the European Alps.     

Photoinhibition in common atlantic macroalgae measured on site in Gran Canaria

The photosynthetic quantum yield was analysed in four common atlantic macroalgae, the Rhodophytes Gelidium arbuscula and Halopithys incurvus and the Phaeophytes Halopteris scoparia and Lobophora variegata in Gran Canaria, Canary Islands at their growth site. The fluorescence parameters were measured using a portable pulse amplitude modulated (PAM) fluorometer (PAM 2000) instrument and a diving PAM under water without removing the thalli from their growth sites. Solar radiation was monitored continuously above and under water during the whole experimental period using two three-channel dosimeters (European light dosimeter network; ELDONET) (Real Time Computer, M?hrendorf, Germany). These instruments measure solar radiation in three wavelength ranges, ultraviolet (UV)-A, UV-B and photosynthetic active radiation (PAR). In all four algae the effective photosynthetic quantum yield decreased significantly from the optimal values measured after dark adaptation due to exposure to 15 min solar radiation, but at least partially recovered subsequently in the shade within several hours. Increasing the exposure period to 30 min intensified the photoinhibition. In some algae no recovery was observed after this treatment and in others no significant recovery could be detected. Exposure to unfiltered solar radiation caused a significantly higher photoinhibition than PAR-only radiation or PAR plus UV-A. A substantial inhibition was found in all algae at their growth sites in the water column when the sun was at high angles, as measured with the diving PAM. Received in revised form: 15 May 2000 Electronic Publication     

Quantifying the responses of calcareous periphyton crusts to rehydration: A microcosm study (Florida Everglades)

We examined the high-resolution temporal dynamics of recovery of dried periphyton crusts following rapid rehydration in a phosphorus (P)-limited short hydroperiod Everglades wetland. Crusts were incubated in a greenhouse in tubs containing water with no P or exogenous algae to mimic the onset of the wet season in the natural marsh when heavy downpours containing very low P flood the dry wetland. Algal and bacterial productivity were tracked for 20 days and related to compositional changes and P dynamics in the water. A portion of original crusts was also used to determine how much TP could be released if no biotic recovery occurred. Composition was volumetrically dominated by cyanobacteria (90%) containing morphotypes typical of xeric environments. Algal and bacterial production recovered immediately upon rehydration but there was a net TP loss from the crusts to the water in the first 2 days. By day 5, however, cyanobacteria and other bacteria had re-absorbed 90% of the released P. Then, water TP concentration reached a steady-state level of 6.6 μg TP/L despite water TP concentration through evaporation. Phosphomonoesterase (PMEase) activity was very high during the first day after rehydration due to the release of a large pre-existing pool of extracellular PMEase. Thereafter, the activity dropped by 90% and increased gradually from this low level. The fast recovery of desiccated crusts upon rehydration required no exogenous P or allogenous algae/bacteria additions and periphyton largely controlled P concentration in the water.     

Energy Transfer from Phycobilisomes to Photosystems of Nostoc flagelliforme Born. et Flah. During the Rewetting Course and Its Physiological Significance

During the non-frost season, the condensation of dew makes Nostocflagelliforme Born. et Flah., a highly drought-tolerant terrestrial cyanobacterium, frequently undergo rehydration-dehydration.Rehydration begins in the dark at night. After rewetting in the dark, photochemical activity and the structure of photosystem (PS)II were not recovered at all; the structure of PSI, energy transfer in phycobilisomes, and energy transfer from phycobilisomes to PSI were recovered within 5 min, as in the light. The recovery of energy transfer from phycobilisomes to PSII was light dependent and energy transfer from phycobilisomes to PSII was only partially recovered in the dark. These results suggest that the two-trigger control (water and light) of photosynthetic recovery may make IV, flagelliforme avoid unnecessary energy consumption and, at the same time, the partial recovery of energy transfer from phycobilisomes to PSII in the dark could help N. flagelliforme accumulate more photosynthetic products during the transient period of rehydration-dehydration.     

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.     

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.