Ultraviolet radiation and Vitamin D3 in amphibian health, behaviour, diet and conservation

Amphibians are currently suffering a period of mass extinction with approximately 20% of species under severe threat and more than 120 species already extinct. In light of this crisis there is an urgency to establish viable ex situ populations and also find the causes of in situ declines. The role of ultraviolet radiation and Vitamin D₃ in amphibian health directly influences both ex situ and in situ populations. Vitamin D₃ can be photosynthesised endogenously via UV-B radiation (UV-B), or acquired through the diet, and then metabolised to calcitriol the biologically active hormonal form. Although, there is a lack of literature concerning Vitamin D₃ requirements and calcitriol synthesis in amphibians, amphibians are likely to have similar Vitamin D₃ requirements and metabolic processes as other vertebrates due to the phylogenetically conservative nature of calcitriol biosynthesis. Deficiencies in calcitriol in amphibians result in nutritional metabolic bone disease (NMBD) and could compromise reproduction and immunity. However, excess biologically active UV radiation has also proven detrimental across all three amphibian life stages and therefore could impact both in situ and ex situ populations. Here we review the role and necessity of UV-B and calcitriol in amphibians and the potential for negative impacts due to excessive exposure to UV radiation. We also identify priorities for research that could provide critical information for maintaining healthy in ex situ and in situ populations of amphibians.     

Exposure of red-legged frog embryos to ambient UV-B radiation in the field negatively affects larval growth and development

Exposure to ultraviolet-B radiation (UV-B; 280-320 nm) has a wide array of effects on aquatic organisms, including amphibians, and has been implicated as a possible factor contributing to global declines and range reductions in amphibian populations. Both lethal and sublethal effects of UV-B exposure have been documented for many amphibian species at various life-history stages. Some species, such as red legged frogs, Rana aurora, appear to be resistant to current ambient levels of UV-B, at least at the embryonic and larval stages, despite the fact that they have experienced range reductions in the Willamette Valley of Oregon, USA. However, UV-B is lethal to embryonic and larval R. aurora at levels slightly above those currently experienced during development. Therefore, we predicted that exposure of embryos to ambient UV-B radiation would result in sublethal effects on larval growth and development. We tested this by exposing R. aurora embryos to ambient UV-B in the field and then raising individuals in the laboratory for 1 month after hatching. Larvae that were exposed to UV-B as embryos were smaller and less developed than the non-exposed individuals 1 month post-hatching. These types of sublethal effects of UV-B exposure indicate that current levels of UV-B could already be influencing amphibian development.     

Survival of three species of anuran metamorphs exposed to UV-B radiation and the pathogenic fungus Batrachochytrium dendrobatidis

When exploring the possible factors contributing to population declines, it is necessary to consider multiple, interacting environmental stressors. Here, we investigate the impact of 2 factors, ultraviolet radiation and disease, on the survival of anuran amphibians. Exposure to ultraviolet-B (UV-B) radiation increases mortality and results in various sub-lethal effects for many amphibian species. Infectious diseases can also negatively impact amphibian populations. In this study, we exposed metamorphic individuals (metamorphs) to both UV-B and Batrachochytrium dendrobatidis (BD), a fungal pathogen and cause of the disease chytridiomycosis, and monitored survival for 3 wk. We tested for possible interactions between UV-B and BD in 3 species: the Cascades frog Rana cascadae; the Western toad Bufo boreas; and the Pacific treefrog Hyla regilla. We found strong interspecific differences in susceptibility to BD. For example, R. cascadae suffered a large increase in mortality when exposed to BD; B. boreas also experienced mortality, but this effect was small relative to the R. cascadae response. H. regilla did not show any decrease in survival when exposed to either factor. No synergistic interactions between UV-B and BD were found for any of the test species. A previous study investigating the impact of BD on larval amphibians showed different species responses (Blaustein et al. 2005a). Our results highlight the importance of studying multiple life history stages when determining the impact of environmental stressors. The contrast between these 2 studies emphasizes how vulnerability to a pathogen can vary between life history stages within a single species.     

Association of amphibians with attenuation of ultraviolet-b radiation in montane ponds

Ambient ultraviolet-b (UV-B) radiation (280-320 nm) has increased at north-temperate latitudes in the last two decades. UV-B can be detrimental to amphibians, and amphibians have shown declines in some areas during this same period. We documented the distribution of amphibians and salmonids in 42 remote, subalpine and alpine ponds in Olympic National Park, Washington, United States. We inferred relative exposure of amphibian habitats to UV-B by estimating the transmission of 305- and 320-nm radiation in pond water. We found breeding Ambystoma gracile, A. macrodactylum and Rana cascadae at 33%, 31%, and 45% of the study sites, respectively. Most R. cascadae bred in fishless shallow ponds with relatively low transmission of UV-B. The relationship with UV-B exposure remained marginally significant even after the presence of fish was included in the model. At 50 cm water depth, there was a 55% reduction in incident 305-nm radiation at sites where breeding populations of R. cascadae were detected compared to other sites. We did not detect associations between UV-B transmission and A. gracile or A. macrodactylum. Our field surveys do not provide evidence for decline of R. cascadae in Olympic National Park as has been documented in Northern California, but are consistent with the hypothesis that the spatial distribution of R. cascadae breeding sites is influenced by exposure to UV-B. Substrate or pond depth could also be related to the distribution of R. cascadae in Olympic National Park.     

Linking species performance to community structure as affected by UV-B radiation: an attenuation experiment

Aims UV-B radiation is known to affect plant physiology and growth rate in ways that can influence community species composition and structure. Nevertheless, comparatively little is known about how UV-B radiation induced changes in the performance of individual species cascades to affect overall community properties. Because foliage leaves are primarily responsible for photosynthesis and carbon gain and are the major organ that senses and responds to UV-B radiation, we hypothesized that, under reduced UV-B radiation, species with larger leaf areas per plant would manifest higher growth rates and hence tend to improve their community status compared to species with smaller leaf areas per plant in herbaceous plant communities.Methods We tested this hypothesis by examining plant traits (leaf area per plant and plant height), plant growth rate (aboveground biomass per plant and plant biomass per area) and community status (species within-community relative biomass) for 19 common species in a two-year field experiment in an alpine meadow on Tibetan Plateau.Important findings Aboveground biomass per plant, as well as per area, progressively increased in a 39% reduced (relative to ambient) UV-B treatment during the experimental period. At the second year, 11 out of 19 species significantly or marginally significantly increased their plant height, leaf area per plant and aboveground biomass per plant. No species was negatively affected by reducing UV-B. As hypothesized, the increase in aboveground biomass per plant increased with increasing leaf area per plant, as indicated by cross-species regression analysis. Moreover, the change in species within-community status increased with increasing leaf area per plant. Our study demonstrates that UV-B radiation has differential effects on plant growth rate across species and hence significantly affects species composition and plant community structure. We suggest that UV-B radiation is an ecological factor structuring plant communities particularly in alpine and polar areas.     

Global increases in ultraviolet B radiation: potential impacts on amphibian development and metamorphosis

Levels of ultraviolet B radiation (UVBR) reaching the Earth's surface have increased since the 1970s as a result of stratospheric ozone depletion caused by the emission of ozone-depleting substances (ODSs) such as chlorofluorocarbons. Despite international agreements to phase out harmful ODSs, these substances are persistent, and even under the most optimistic scenarios, stratospheric ozone levels will not return to pre-1980 levels for several decades. Furthermore, climate change may enhance chemical stratospheric ozone depletion. Global phenomena such as climate change, ozone depletion, and acidification of aquatic ecosystems interact to modify dissolved organic carbon levels in aquatic systems, thereby increasing the penetration of UVBR. Since amphibians inhabit both aquatic and terrestrial habitats and have unshelled eggs and permeable skin, they are vulnerable to changes in environmental conditions and habitat quality. Increased exposure of amphibians to UVBR can produce lethal and sublethal effects, especially in individuals that do not possess adequate defense mechanisms to protect themselves. In this article, we discuss worldwide increases in UVBR and the adverse effects of UVBR exposure on amphibians. Specifically, studies on the effects of UVBR on amphibian development and metamorphosis are summarized, and possible mechanisms of thyroid system disruption caused by UVBR exposure are considered.     

Ultraviolet radiation, toxic chemicals and amphibian population declines

Abstract. As part of an overall ‘biodiversity crisis’, many amphibian populations are in decline throughout the world. Numerous factors have contributed to these declines, including habitat destruction, pathogens, increasing ultraviolet (UV) radiation, introduced non‐native species and contaminants. In this paper we review the contribution of increasing UV radiation and environmental contamination to the global decline of amphibian populations. Both UV radiation and environmental contaminants can affect amphibians at all life stages. Exposure to UV radiation and to certain contaminants can kill amphibians and induce sublethal affects in embryos, larvae and adults. Moreover, UV radiation and contaminants may interact with one another synergistically. Synergistic interactions of UV radiation with contaminants can enhance the detrimental effects of the contaminant and UV radiation.     

Integration and scaling of UV-B radiation effects on plants: the relative sensitivity of growth forms and interspecies interactions

Aims The relative plant type sensitivity and selected community interactions under increased UV-B radiation where examined. Specifically, we investigated: (i) if there are differences among growth forms in regard to their sensitivity to UV-B radiation, (ii) if increased UV-B radiation influences the plant competitive balance in plant communities and (iii) the response mechanisms of the UV-B radiation-sensitive species that might increase their fitness.Methods To answer our research questions, we used a mechanistic model that, for the first time, integrated the effects of increased UV-B radiation from molecular level processes, whole plant growth and development, and community interactions.Important findings In the model simulations, species types exhibited different levels of sensitivity to increased UV-B radiation. Summer C₃ and C₄ annuals showed similar growth inhibition rates, while biennials and winter C₃ annuals were the most sensitive. Perennials exhibited inhibitions in growth only if increased UV-B radiation results in increases in metabolic rates. In communities, species sensitive to UV-B radiation may have a competitive disadvantage compared to resistant plant species. But, sensitive species may have a wide array of responses that can increase their fitness and reproductive success in the community, such as, increased secondary metabolites production, changes in timing of emergence and reproduction, and changes in seed size. While individual plants may exhibit significant inhibitions in growth and development, in communities, these inhibitions can be mitigated by small morphological and physiological adaptations. Infrequent or occasional increased UV-B radiation events should not have any lasting effect on the structure of the community, unless other environmental factors are perturbing the dynamic equilibrium.     

A small increase in UV-B increases the susceptibility of tadpoles to predation

Increased ultraviolet-B (UV-B) radiation as a consequence of ozone depletion is one of the many potential drivers of ongoing global amphibian declines. Both alone and in combination with other environmental stressors, UV-B is known to have detrimental effects on the early life stages of amphibians, but our understanding of the fitness consequences of these effects remains superficial. We examined the independent and interactive effects of UV-B and predatory chemical cues (PCC) on a suite of traits of Limnodynastes peronii embryos and tadpoles, and assessed tadpole survival time in a predator environment to evaluate the potential fitness consequences. Exposure to a 3 to 6 per cent increase in UV-B, which is comparable to changes in terrestrial UV-B associated with ozone depletion, had no effect on any of the traits measured, except survival time in a predator environment, which was reduced by 22 to 28 per cent. Exposure to PCC caused tadpoles to hatch earlier, have reduced hatching success, have improved locomotor performance and survive for longer in a predator environment, but had no effect on tadpole survival, behaviour or morphology. Simultaneous exposure to UV-B and PCC resulted in no interactive effects. These findings demonstrate that increased UV-B has the potential to reduce tadpole fitness, while exposure to PCCs improves their fitness.     

Distribution Patterns of Lentic-Breeding Amphibians in Relation to Ultraviolet Radiation Exposure in Western North America

An increase in ultraviolet-B (UV-B) radiation has been posited to be a potential factor in the decline of some amphibian population. This hypothesis has received support from laboratory and field experiments showing that current levels of UV-B can cause embryo mortality in some species, but little research has addressed whether UV-B is influencing the distribution of amphibian populations. We compared patterns of amphibian presence to site-specific estimates of UV-B dose at 683 ponds and lakes in Glacier, Olympic, and Sequoia–Kings Canyon National Parks. All three parks are located in western North America, a region with a concentration of documented amphibian declines. Site-specific daily UV-B dose was estimated using modeled and field-collected data to incorporate the effects of elevation, landscape, and water-column dissolved organic carbon. Of the eight species we examined (Ambystoma gracile, Ambystoma macrodactylum, Bufo boreas, Pseudacris regilla, Rana cascadae, Rana leuteiventris, Rana muscosa, Taricha granulosa), two species (T. granulosa and A. macrodactylum) had quadratic relationships with UV-B that could have resulted from negative UV-B effects. Both species were most likely to occur at moderate UV-B levels. Ambystoma macrodactylum showed this pattern only in Glacier National Park. Occurrence of A. macrodactylum increased as UV-B increased in Olympic National Park despite UV-B levels similar to those recorded in Glacier. We also found marginal support for a negative association with UV-B for P. regilla in one of the two parks where it occurred. We did not find evidence of a negative UV-B effect for any other species. Much more work is still needed to determine whether UV-B, either alone or in concert with other factors, is causing widespread population losses in amphibians.     

Effects of UV-B Radiation on Anti-predator Behavior in Three Species of Amphibians

Ultraviolet radiation has been suggested as a possible contributing cause of amphibian declines around the world. Both laboratory and field studies have demonstrated that exposure to ultraviolet radiation can lead to increased mortality of developing amphibians. Virtually no studies have examined the sub‐lethal effects of ultraviolet on amphibian behavior. In this study, we examine the anti‐predator behavior of three species of amphibians after short‐term exposure to ultraviolet‐B radiation. Toad (Bufo boreas) juveniles that had been exposed to ultraviolet radiation did not respond to chemical extracts from conspecifics and heterospecifics as much as juveniles that had not been exposed. Both newt larvae (Taricha granulosa) that had been exposed to ultraviolet radiation and those that had not been exposed responded to chemical cues from conspecific predators by increasing the amount of time spent in shelter. Frog tadpoles (Rana cascadae) that had been exposed to ultraviolet radiation did not reduce their movement in response to chemical cues from predators as much as tadpoles that had not been exposed. These results indicate that ultraviolet exposure may have important sub‐lethal effects in amphibians that could adversely effect their fitness.     

Elevational differences in trait response to UV-B radiation by long-toed salamander populations

Amphibian species capable of optimizing trait response to environmental stressors may develop complex strategies for defending against rapid environmental change. Trait responses may differ between populations, particularly if stressor strength varies across spatial or temporal gradients. Ultraviolet-B (UV-B) radiation is one such stressor that poses a significant threat to amphibian species. We examined the ability of long-toed salamanders (Ambystoma macrodactylum) at high- and low-elevation breeding sites to cooperatively employ behavioral and physiological trait responses to mediate UV-B damage. We performed a microhabitat survey to examine differences in oviposition behavior and UV-B conditions among breeding populations at high- (n = 3; >1,500 m) and low-elevation (n = 3; <100 m) sites. We found significant differences in oviposition behavior across populations, with females at high-elevation sites selecting oviposition substrates in UV-B protected microhabitats. We also collected eggs (n = 633) from each of the breeding sites for analysis of photolyase activity, a photoreactivating enzyme that repairs UV-B damage to the DNA, using a photoproduct immunoassay. Our results revealed no significant differences in photolyase activity between long-toed salamander populations at high and low elevations. For high-elevation salamander populations, relatively low physiological repair capabilities in embryos appear to be buffered by extensive behavioral modifications to reduce UV-B exposure and standardize developmental temperatures. This study provides valuable insight into environmental stress responses via the assessment of multiple traits in allowing sensitive species to persist in rapidly changing landscapes.     

Composition of symbiotic bacteria predicts survival in Panamanian golden frogs infected with a lethal fungus

Symbiotic microbes can dramatically impact host health and fitness, and recent research in a diversity of systems suggests that different symbiont community structures may result in distinct outcomes for the host. In amphibians, some symbiotic skin bacteria produce metabolites that inhibit the growth of Batrachochytrium dendrobatidis (Bd), a cutaneous fungal pathogen that has caused many amphibian population declines and extinctions. Treatment with beneficial bacteria (probiotics) prevents Bd infection in some amphibian species and creates optimism for conservation of species that are highly susceptible to chytridiomycosis, the disease caused by Bd. In a laboratory experiment, we used Bd-inhibitory bacteria from Bd-tolerant Panamanian amphibians in a probiotic development trial with Panamanian golden frogs, Atelopus zeteki, a species currently surviving only in captive assurance colonies. Approximately 30% of infected golden frogs survived Bd exposure by either clearing infection or maintaining low Bd loads, but this was not associated with probiotic treatment. Survival was instead related to initial composition of the skin bacterial community and metabolites present on the skin. These results suggest a strong link between the structure of these symbiotic microbial communities and amphibian host health in the face of Bd exposure and also suggest a new approach for developing amphibian probiotics.     

Interacting Symbionts and Immunity in the Amphibian Skin Mucosome Predict Disease Risk and Probiotic Effectiveness

Pathogenesis is strongly dependent on microbial context, but development of probiotic therapies has neglected the impact of ecological interactions. Dynamics among microbial communities, host immune responses, and environmental conditions may alter the effect of probiotics in human and veterinary medicine, agriculture and aquaculture, and the proposed treatment of emerging wildlife and zoonotic diseases such as those occurring on amphibians or vectored by mosquitoes. Here we use a holistic measure of amphibian mucosal defenses to test the effects of probiotic treatments and to assess disease risk under different ecological contexts. We developed a non-invasive assay for antifungal function of the skin mucosal ecosystem (mucosome function) integrating host immune factors and the microbial community as an alternative to pathogen exposure experiments. From approximately 8500 amphibians sampled across Europe, we compared field infection prevalence with mucosome function against the emerging fungal pathogen Batrachochytrium dendrobatidis. Four species were tested with laboratory exposure experiments, and a highly susceptible species, Alytes obstetricans, was treated with a variety of temperature and microbial conditions to test the effects of probiotic therapies and environmental conditions on mucosome function. We found that antifungal function of the amphibian skin mucosome predicts the prevalence of infection with the fungal pathogen in natural populations, and is linked to survival in laboratory exposure experiments. When altered by probiotic therapy, the mucosome increased antifungal capacity, while previous exposure to the pathogen was suppressive. In culture, antifungal properties of probiotics depended strongly on immunological and environmental context including temperature, competition, and pathogen presence. Functional changes in microbiota with shifts in temperature provide an alternative mechanistic explanation for patterns of disease susceptibility related to climate beyond direct impact on host or pathogen. This nonlethal management tool can be used to optimize and quickly assess the relative benefits of probiotic therapies under different climatic, microbial, or host conditions.     

Microbial community dynamics and effect of environmental microbial reservoirs on red-backed salamanders (Plethodon cinereus)

Beneficial cutaneous bacteria on amphibians can protect against the lethal disease chytridiomycosis, which has devastated many amphibian species and is caused by the fungus Batrachochytrium dendrobatidis. We describe the diversity of bacteria on red-backed salamanders (Plethodon cinereus) in the wild and the stability of these communities through time in captivity using culture-independent Illumina 16S rRNA gene sequencing. After field sampling, salamanders were housed with soil from the field or sterile media. The captive conditions led to different trajectories of bacterial communities. Eight OTUs present on >90% of salamanders in the field, through time, and in both treatments were defined as the core community, suggesting that some bacteria are closely associated with the host and are independent of an environmental reservoir. One of these taxa, a Pseudomonas sp., was previously cultured from amphibians and found to be antifungal. As all host-associated bacteria were found in the soil reservoir, environmental microbes strongly influence host–microbial diversity and likely regulate the core community. Using PICRUSt, an exploratory bioinformatics tool to predict gene functions, we found that core skin bacteria provided similar gene functions to the entire community. We suggest that future experiments focus on testing whether core bacteria on salamander skin contribute to the observed resistance to chytridiomycosis in this species even under hygenic captive conditions. For disease-susceptible hosts, providing an environmental reservoir with defensive bacteria in captive-rearing programs may improve outcomes by increasing bacterial diversity on threatened amphibians or increasing the likelihood that defensive bacteria are available for colonization.     

Impact of UV-B exposure on amphibian embryos: linking species physiology and oviposition behaviour

Increasing ultraviolet-B radiation (UV-B) has recently captured the attention of ecologists as a key environmental stressor. Certain species may be particularly vulnerable as a result of either high natural exposure to UV-B or limited physiological capacity to withstand it. UV-B sensitivity has been examined at the cellular and individual level for a wide variety of taxa, but estimates of exposure to UV-B in natural systems are lacking and predictions of large-scale impacts are therefore limited. Here, we combine data on the physiological sensitivity to UV-B and patterns of field exposure across sites for embryos of several well-studied US Pacific Northwest amphibian species. We find substantial differences among species' physiological abilities to withstand UV-B and in the level of UV-B exposure of embryos in the field. More specifically, we find that species with the highest physiological sensitivity to UV-B are those with the lowest field exposures as a function of the location of embryos and the UV-B attenuation properties of water at each site. These results also suggest that conclusions made about species' vulnerability to UV-B in the absence of information on field exposures may often be misleading.     

The Effects of Ultraviolet Radiation on the Biology of Amphibians

SYNOPSIS. Potential causes for the global decline of amphibiansinclude habitat loss, disease, environmental contaminants andclimate changes. The diminishing ozone layer and consequentincrease of ultraviolet-B radiation reaching the earth's surfacehas been hypothesized to be a mortality factor, especially inhabitats otherwise undisturbed. We discuss the fundamental physicsof UV and types of biological damage after exposure. A deleteriouschange in DNA, especially the production of pyrimidine dimers,is a main effect of UVB exposure. Damaged DNA can be repairedby enzymes such as photolyase when organisms are irradiatedwith UVA or visible light. We review studies including laboratoryand field approaches on damage to amphibians from UVB exposure.Field studies in which embryos were exposed to natural sunlightor sunlight with UVB removed have shown conflicting results:some show increased embryonic mortality after UVB exposure,whereas others show that current levels of UVB are not detrimentalto amphibian embryos. The abiotic factors such as water depth,water color, and dissolved organic content of aquatic ovipositionsites effectively reduces UVB penetration through water andreduces exposure to UVB of all life history stages. Biotic factorssuch as jelly capsules around eggs, melanin pigmentation ofeggs, and color of larvae and metamorphosed forms further reduceeffectiveness of UVB penetration. We suggest areas of futureresearch to test the hypothesis of the causal connection betweencurrent UVB levels and amphibian decline.     

Native predators living in invaded areas: responses of terrestrial amphibian species to an Argentine ant invasion

Predator–prey interactions play a key role in the success and impacts of invasive species. However, the effects of invasive preys on native predators have been poorly studied. Here, we first reviewed hypotheses describing potential relationships between native predators and invasive preys. Second, we examined how an invasive prey, the Argentine ant (Linepithema humile), affected a native terrestrial amphibian community. In the field, we looked at the structure of the amphibian community in invaded versus uninvaded areas and characterized amphibian trophic ecology. The amphibian community sampled seemed to show a species-dependent response in abundance to invasion: adults of the natterjack toad (Bufo calamita), the species demonstrating the highest degree of ant specialization, were less abundant in invaded areas. Although available ant biomass was significantly greater in invaded than in uninvaded areas (only Argentine ants occurred in the former), amphibians consumed relatively fewer ants in invaded areas. In the lab, we quantified amphibian consumption of Argentine ants versus native ants and assessed whether consumption patterns could have been influenced by prior exposure to the invader. The lab experiments corroborated the field results: amphibians preferred native ants over Argentine ants, and prior exposure did not influence consumption. Differences in preference explained why amphibians consumed fewer Argentine ants in spite of their greater relative availability; they might also explain why the most ant-specialized amphibians seemed to avoid invaded areas. Our results suggest the importance to account for predator feeding capacities and dietary ranges to understand the effects of invasive species at higher trophic levels.     

Effects of UV-B radiation on photosynthesis and growth of terrestrial plants

The photosynthetic apparatus of some plant species appears to be well-protected from direct damage from UV-B radiation. Leaf optical properties of these species apparently minimizes exposure of sensitive targets to UV-B radiation. However, damage by UV-B radiation to Photosystem II and Rubisco has also been reported. Secondary effects of this damage may include reductions in photosynthetic capacity, RuBP regeneration and quantum yield. Furthermore, UV-B radiation may decrease the penetration of PAR, reduce photosynthetic and accessory pigments, impair stomatal function and alter canopy morphology, and thus indirectly retard photosynthetic carbon assimilation. Subsequently, UV-B radiation may limit productivity in many plant species. In addition to variability in sensitivity to UV-B radiation, the effects of UV-B radiation are further confounded by other environmental factors such as CO₂, temperature, light and water or nutrient availability. Therefore, we need a better understanding of the mechanisms of tolerance to UV-B radiation and of the interaction between UV-B and other environmental factors in order to adequately assess the probable consequences of a change in solar radiation.Abbreviations A_max light and CO₂ saturated rate of oxygen evolution - Ci internal CO₂ concentration - F_v/F_m ratio of variable to total fluorescence yield - PAR photosynthetically active radiation (400–700 nm) - PS II Photosystem II - _app apparent quantum yield of photosynthesis - SLW specific leaf weight - UV-B ultraviolet-B radiation between 290–320 nm