Basking in the sun: how mosses photosynthesise and survive in Antarctica.

The Antarctic environment is extremely cold, windy and dry. Ozone depletion has resulted in increasing ultraviolet-B radiation, and increasing greenhouse gases and decreasing stratospheric ozone have altered Antarctica's climate. How do mosses thrive photosynthetically in this harsh environment? Antarctic mosses take advantage of microclimates where the combination of protection from wind, sufficient melt water, nutrients from seabirds and optimal sunlight provides both photosynthetic energy and sufficient warmth for efficient metabolism. The amount of sunlight presents a challenge: more light creates warmer canopies which are optimal for photosynthetic enzymes but can contain excess light energy that could damage the photochemical apparatus. Antarctic mosses thus exhibit strong photoprotective potential in the form of xanthophyll cycle pigments. Conversion to zeaxanthin is high when conditions are most extreme, especially when water content is low. Antarctic mosses also produce UV screening compounds which are maintained in cell walls in some species and appear to protect from DNA damage under elevated UV-B radiation. These plants thus survive in one of the harshest places on Earth by taking advantage of the best real estate to optimise their metabolism. But survival is precarious and it remains to be seen if these strategies will still work as the Antarctic climate changes.

Incorporating pressure-volume traits into the leaf economics spectrum.

In recent years, attempts have been made in linking pressure-volume parameters and the leaf economics spectrum to expand our knowledge of the interrelationships among leaf traits. We provide theoretical and empirical evidence for the coordination of the turgor loss point and associated traits with net CO2 assimilation (An ) and leaf mass per area (LMA). We measured gas exchange, pressure-volume curves and leaf structure in 45 ferns and angiosperms, and explored the anatomical and chemical basis of the key traits. We propose that the coordination observed between mass-based An , capacitance and the turgor loss point (πtlp ) emerges from their shared link with leaf density (one of the components of LMA) and, specially, leaf saturated water content (LSWC), which in turn relates to cell size and nitrogen and carbon content. Thus, considering the components of LMA and LSWC in ecophysiological studies can provide a broader perspective on leaf structure and function.

The ratio of electron transport to assimilation (ETR/AN): underutilized but essential for assessing both equipment's proper performance and plant status.

MAIN CONCLUSION: ETR/AN ratios should be in the range 7.5-10.5 for non-stressed C3 plants. Ratios extremely out of this range can be reflecting both uncontrolled plant status and technical mistakes during measurements. We urge users to explicitly refer to this ratio in future studies as a proof for internal data quality control. For the last few decades, the use of infra-red gas-exchange analysers (IRGAs) coupled with chlorophyll fluorometers that allow for measurements of net CO2 assimilation rate and estimates of electron transport rate over the same leaf area has been popularized. The evaluation of data from both instruments in an integrative manner can result in additional valuable information, such as the estimation of the light respiration, mesophyll conductance and the partitioning of the flux of electrons into carboxylation, oxygenation and alternative processes, among others. In this review, an additional and more 'straight' use of the combination of chlorophyll fluorescence and gas exchange-derived parameters is presented, namely using the direct ratio between two fully independently estimated parameters, electron transport rate (ETR)-determined by the fluorometer-and net CO2 assimilation rate (AN)-determined by the IRGA, i.e., the ETR/AN ratio, as a tool for fast detection of incongruencies in the data and potential technical problems associated with them, while checking for the study plant's status. To illustrate this application, a compilation of 75 studies that reported both parameters for a total of 178 species under varying physiological status is presented. Values of ETR/AN between 7.5 and 10.5 were most frequently found for non-stressed C3 plants. C4 species showed an average ETR/AN ratio of 4.7. The observed ratios were larger for species with high leaf mass per area and for plants subjected to stressful factors like drought or nutritional deficit. Knowing the expected ETR/AN ratio projects this ratio as a routinary and rapid check point for guaranteeing both the correct performance of equipment and the optimal/stress status of studied plants. All known errors associated with the under- or overestimation of ETR or AN are summarized in a checklist that aims to be routinely used by any IRGA/fluorometer user to strength the validity of their data.

Limitations to photosynthesis in bryophytes: certainties and uncertainties regarding methodology.

Bryophytes are the group of land plants with the lowest photosynthetic rates, which was considered to be a consequence of their higher anatomical CO2 diffusional limitation compared with tracheophytes. However, the most recent studies assessing limitations due to biochemistry and mesophyll conductance in bryophytes reveal discrepancies based on the methodology used. In this study, we compared data calculated from two different methodologies for estimating mesophyll conductance: variable J and the curve-fitting method. Although correlated, mesophyll conductance estimated by the curve-fitting method was on average 4-fold higher than the conductance obtained by the variable J method; a large enough difference to account for the scale of differences previously shown between the biochemical and diffusional limitations to photosynthesis. Biochemical limitations were predominant when the curve-fitting method was used. We also demonstrated that variations in bryophyte relative water content during measurements can also introduce errors in the estimation of mesophyll conductance, especially for samples which are overly desiccated. Furthermore, total chlorophyll concentration and soluble proteins were significantly lower in bryophytes than in tracheophytes, and the percentage of proteins quantified as Rubisco was also significantly lower in bryophytes (<6.3% in all studied species) than in angiosperms (>16% in all non-stressed cases). Photosynthetic rates normalized by Rubisco were not significantly different between bryophytes and angiosperms. Our data suggest that the biochemical limitation to photosynthesis in bryophytes is more relevant than so far assumed.

When time is not of the essence: constraints to the carbon balance of bryophytes.

The data available so far indicate that the photosynthetic and relative growth rates of bryophytes are 10% of those reported for tracheophytes. By examining the existing literature and reanalysing data published in over 100 studies, this review examines the ecophysiological, biochemical, and structural reasons behind this phenomenon. The limiting Rubisco content and surface for gas exchange are the internal factors that can explain the low photosynthetic and growth rates of bryophytes. The role of the thicker cell walls of bryophytes in limiting CO2 diffusion is unclear, due to the current uncertainties regarding their porosity and permeability to CO2. From this review, it is also evident that, despite bryophytes having low photosynthetic rates, their positive carbon balance is tightly related to their capacity to deal with extreme conditions. Contributing factors include their capacity to deal with large daily temperature oscillations, and their capacity to delay the cessation of photosynthesis under water deficit (or to tolerate desiccation in extreme situations). Although further studies on bryophytes are needed before more solid conclusions can be drawn, it seems that their success relies on their remarkable tolerance to a highly variable environment, possibly at the expense of their maximum photosynthetic rate.

Desiccation tolerance in bryophytes relates to elasticity but is independent of cell wall thickness and photosynthesis.

Mosses have been found outliers of the trade-off between photosynthesis and bulk elastic modulus described for vascular plants. Hence, potential trade-offs among physical features of cell walls and desiccation tolerance, water relations, and photosynthesis were assessed in bryophytes and other poikilohydric species. Long-term desiccation tolerance was quantified after variable periods of desiccation/rehydration cycles. Water relations were analyzed by pressure-volume curves. Mesophyll conductance was estimated using both CO2 curve-fitting and anatomical methods. Cell wall elasticity was the parameter that better correlated with the desiccation tolerance index for desiccation tolerant species and was antagonistic to higher absolute values of osmotic potential. Although high values of cell wall effective porosity were estimated compared with the values assumed for vascular plants, the desiccation tolerance index negatively correlated with the porosity in desiccation tolerant bryophytes. Neither cell wall thickness nor photosynthetic capacity were correlated with the desiccation tolerance index of the studied species. The existence of a potential evolutionary trade-off between cell wall thickness and desiccation tolerance is rejected. The photosynthetic capacity reported for bryophytes is independent of elasticity and desiccation tolerance. Furthermore, the role of cell wall thickness in limiting CO2 conductance would be overestimated under a scenario of high cell wall porosity for most bryophytes.

Differences in biochemical, gas exchange and hydraulic response to water stress in desiccation tolerant and sensitive fronds of the fern Anemia caffrorum.

Desiccation tolerant plants can survive extreme water loss in their vegetative tissues. The fern Anemia caffrorum produces desiccation tolerant (DT) fronds in the dry season and desiccation sensitive (DS) fronds in the wet season, providing a unique opportunity to explore the physiological mechanisms associated with desiccation tolerance. Anemia caffrorum plants with either DT or DS fronds were acclimated in growth chambers. Photosynthesis, frond structure and anatomy, water relations and minimum conductance to water vapour were measured under well-watered conditions. Photosynthesis, hydraulics, frond pigments, antioxidants and abscisic acid contents were monitored under water deficit. A comparison between DT and DS fronds under well-watered conditions showed that the former presented higher leaf mass per area, minimum conductance, tissue elasticity and lower CO2 assimilation. Water deficit resulted in a similar induction of abscisic acid in both frond types, but DT fronds maintained higher stomatal conductance and upregulated more prominently lipophilic antioxidants. The seasonal alternation in production of DT and DS fronds in A. caffrorum represents a mechanism by which carbon gain can be maximized during the rainy season, and a greater investment in protective mechanisms occurs during the hot dry season, enabling the exploitation of episodic water availability.

Cell wall thickness and composition are involved in photosynthetic limitation.

The key role of cell walls in setting mesophyll conductance to CO2 (gm) and, consequently, photosynthesis is reviewed. First, the theoretical properties of cell walls that can affect gm are presented. Then, we focus on cell wall thickness (Tcw) reviewing empirical evidence showing that Tcw varies strongly among species and phylogenetic groups in a way that correlates with gm and photosynthesis; that is, the thicker the mesophyll cell walls, the lower the gm and photosynthesis. Potential interplays of gm, Tcw, dehydration tolerance, and hydraulic properties of leaves are also discussed. Dynamic variations of Tcw in response to the environment and their implications in the regulation of photosynthesis are discussed, and recent evidence suggesting an influence of cell wall composition on gm is presented. We then propose a hypothetical mechanism for the influence of cell walls on photosynthesis, combining the effects of thickness and composition, particularly pectins. Finally, we discuss the prospects for using biotechnology for enhancing photosynthesis by altering cell wall-related genes.

Resurrection plants optimize photosynthesis despite very thick cell walls by means of chloroplast distribution.

Resurrection plants are vascular species able to sustain extreme desiccation in their vegetative tissues. Despite its potential interest, the role of leaf anatomy in CO2 diffusion and photosynthesis under non-stressed conditions has not been explored in these species. Net CO2 assimilation (An) and its underlying diffusive, biochemical, and anatomical determinants were assessed in 10 resurrection species from diverse locations, including ferns, and homoiochlorophyllous and poikilochlorophyllous angiosperms. Data obtained were compared with previously published results in desiccation-sensitive ferns and angiosperms. An in resurrection plants was mostly driven by mesophyll conductance to CO2 (gm) and limited by CO2 diffusion. Resurrection species had a greater cell wall thickness (Tcw) than desiccation-sensitive plants, a feature associated with limited CO2 diffusion in the mesophyll, but also greater chloroplast exposure to intercellular spaces (Sc), which usually leads to higher gm. This combination enabled a higher An per Tcw compared with desiccation-sensitive species. Resurrection species possess unusual anatomical features that could confer stress tolerance (thick cell walls) without compromising the photosynthetic capacity (high chloroplast exposure). This mechanism is particularly successful in resurrection ferns, which display higher photosynthesis than their desiccation-sensitive counterparts.

It Is Hot in the Sun: Antarctic Mosses Have High Temperature Optima for Photosynthesis Despite Cold Climate.

The terrestrial flora of Antarctica's frozen continent is restricted to sparse ice-free areas and dominated by lichens and bryophytes. These plants frequently battle sub-zero temperatures, extreme winds and reduced water availability; all influencing their ability to survive and grow. Antarctic mosses, however, can have canopy temperatures well above air temperature. At midday, canopy temperatures can exceed 15°C, depending on moss turf water content. In this study, the optimum temperature of photosynthesis was determined for six Antarctic moss species: Bryum pseudotriquetrum, Ceratodon purpureus, Chorisodontium aciphyllum, Polytrichastrum alpinum, Sanionia uncinata, and Schistidium antarctici collected from King George Island (maritime Antarctica) and/or the Windmill Islands, East Antarctica. Both chlorophyll fluorescence and gas exchange showed maximum values of electron transport rate occurred at canopy temperatures higher than 20°C. The optimum temperature for both net assimilation of CO2 and photoprotective heat dissipation of three East Antarctic species was 20-30°C and at temperatures below 10°C, mesophyll conductance did not significantly differ from 0. Maximum mitochondrial respiration rates occurred at temperatures higher than 35°C and were lower by around 80% at 5°C. Despite the extreme cold conditions that Antarctic mosses face over winter, the photosynthetic apparatus appears optimised to warm temperatures. Our estimation of the total carbon balance suggests that survival in this cold environment may rely on a capacity to maximize photosynthesis for brief periods during summer and minimize respiratory carbon losses in cold conditions.

What drives photosynthesis during desiccation? Mosses and other outliers from the photosynthesis-elasticity trade-off.

In vascular plants, more rigid leaves have been linked to lower photosynthetic capacity, associated with low CO2 diffusion across the mesophyll, indirectly resulting in a trade-off between photosynthetic capacity (An) and bulk modulus of elasticity (ε). However, we evaluated mosses, liverworts, and Chara sp., plus some lycophytes and ferns, and found that they behaved as clear outliers of the An-ε relationship. Despite this finding, when vascular and non-vascular plants were plotted together, ε still linearly determined the cessation of net photosynthesis during desiccation both in species with stomata (either actively or hydro-passively regulated) and in species lacking stomata, and regardless of their leaf structure. The latter result challenges our current view of photosynthetic responses to desiccation and/or water stress. Structural features and hydric strategy are discussed as possible explanations for the deviation of these species from the An-ε trade-off, as well as for the general linear dependency between ε and the full cessation of An during desiccation.

Symbiosis at its limits: ecophysiological consequences of lichenization in the genus Prasiola in Antarctica.

BACKGROUND AND AIMS: Lichens represent a symbiotic relationship between at least one fungal and one photosynthetic partner. The association between the lichen-forming fungus Mastodia tessellata (Verrucariaceae) and different species of Prasiola (Trebouxiophyceae) has an amphipolar distribution and represents a unique case study for the understanding of lichen symbiosis because of the macroalgal nature of the photobiont, the flexibility of the symbiotic interaction and the co-existence of free-living and lichenized forms in the same microenvironment. In this context, we aimed to (1) characterize the photosynthetic performance of co-occurring populations of free-living and lichenized Prasiola and (2) assess the effect of the symbiosis on water relations in Prasiola, including its tolerance of desiccation and its survival and performance under sub-zero temperatures. METHODS: Photochemical responses to irradiance, desiccation and freezing temperature and pressure-volume curves of co-existing free-living and lichenized Prasiola thalli were measured in situ in Livingston Island (Maritime Antarctica). Analyses of photosynthetic pigment, glass transition and ice nucleation temperatures, surface hydrophobicity extent and molecular analyses were conducted in the laboratory. KEY RESULTS: Free-living and lichenized forms of Prasiola were identified as two different species: P. crispa and Prasiola sp., respectively. While lichenization appears to have no effect on the photochemical performance of the alga or its tolerance of desiccation (in the short term), the symbiotic lifestyle involves (1) changes in water relations, (2) a considerable decrease in the net carbon balance and (3) enhanced freezing tolerance. CONCLUSIONS: Our results support improved tolerance of sub-zero temperature as the main benefit of lichenization for the photobiont, but highlight that lichenization represents a delicate equilibrium between a mutualistic and a less reciprocal relationship. In a warmer climate scenario, the spread of the free-living Prasiola to the detriment of the lichen form would be likely, with unknown consequences for Maritime Antarctic ecosystems.

Recent advances in understanding and improving photosynthesis.

Since 1893, when the word "photosynthesis" was first coined by Charles Reid Barnes and Conway MacMillan, our understanding of the elements and regulation of this complex process is far from being entirely understood. We aim to review the most relevant advances in photosynthesis research from the last few years and to provide a perspective on the forthcoming research in this field. Recent discoveries related to light sensing, harvesting, and dissipation; kinetics of CO2 fixation; components and regulators of CO2 diffusion through stomata and mesophyll; and genetic engineering for improving photosynthetic and production capacities of crops are addressed.

Anatomical constraints to nonstomatal diffusion conductance and photosynthesis in lycophytes and bryophytes.

Photosynthesis in bryophytes and lycophytes has received less attention than terrestrial plant groups. In particular, few studies have addressed the nonstomatal diffusion conductance to CO2 gnsd of these plant groups. Their lower photosynthetic rate per leaf mass area at any given nitrogen concentration compared with vascular plants suggested a stronger limitation by CO2 diffusion. We hypothesized that bryophyte and lycophyte photosynthesis is largely limited by low gnsd . Here, we studied CO2 diffusion inside the photosynthetic tissues and its relationships with photosynthesis and anatomical parameters in bryophyte and lycophyte species in Antarctica, Australia, Estonia, Hawaii and Spain. On average, lycophytes and, specially, bryophytes had the lowest photosynthetic rates and nonstomatal diffusion conductance reported for terrestrial plants. These low values are related to their very thick cell walls and their low exposure of chloroplasts to cell perimeter. We conclude that the reason why bryophytes lie at the lower end of the leaf economics spectrum is their strong nonstomatal diffusion conductance limitation to photosynthesis, which is driven by their specific anatomical characteristics.

A field portable method for the semi-quantitative estimation of dehydration tolerance of photosynthetic tissues across distantly related land plants.

Desiccation tolerant (DT) plants withstand complete cellular dehydration, reaching relative water contents (RWC) below 30% in their photosynthetic tissues. Desiccation sensitive (DS) plants exhibit different degrees of dehydration tolerance (DHT), never surviving water loss >70%. To date, no procedure for the quantitative evaluation of DHT extent exists that is able to discriminate DS species with differing degrees of DHT from truly DT plants. We developed a simple, feasible and portable protocol to differentiate between DT and different degrees of DHT in the photosynthetic tissues of seed plants and between fast desiccation (< 24 h) tolerant (FDT) and sensitive (FDS) bryophytes. The protocol is based on (1) controlled desiccation inside Falcon tubes equilibrated at three different relative humidities that, consequently, induce three different speeds and extents of dehydration and (2) an evaluation of the average percentage of maximal photochemical efficiency of PSII (Fv /fm) recovery after rehydration. Applying the method to 10 bryophytes and 28 tracheophytes from various locations, we found that (1) imbibition of absorbent material with concentrated salt-solutions inside the tubes provides stable relative humidity and avoids direct contact with samples; (2) for 50 ml capacity tubes, the optimal plant amount is 50-200 mg fresh weight; (3) the method is useful in remote locations due to minimal instrumental requirements; and (4) a threshold of 30% recovery of the initial Fv /fm upon reaching RWC ≤ 30% correctly categorises DT species, with three exceptions: two poikilochlorophyllous species and one gymnosperm. The protocol provides a semi-quantitative expression of DHT that facilitates comparisons of species with different morpho-physiological traits and/or ecological attributes.