A CPD photolyase gene PnPHR1 from Antarctic moss Pohlia nutans is involved in the resistance to UV-B radiation and salinity stress.

In Antarctic continent, the organisms are exposed to high ultraviolet (UV) radiation because of damaged stratospheric ozone. UV causes DNA lesions due to the accumulation of photoproducts. Photolyase can repair UV-damaged DNA in a light-dependent process by electron transfer mechanism. Here, we isolated a CPD photolyase gene PnPHR1 from Antarctic moss Pohlia nutans, which encodes a protein of theoretical molecular weight of 69.1 KDa. The expression level of PnPHR1 was increased by UV-B irradiation. Enzyme activity assay in vitro showed that PnPHR1 exhibited photoreactivation activity, which can repair CPD photoproducts in a light-dependent manner. The complementation assay of repair-deficient E. coli strain SY2 demonstrated that PnPHR1 gene enhanced the survival rate of SY2 strain after UV-B radiation. Additionally, overexpression of PnPHR1 enhanced the Arabidopsis resistance to UV-B radiation and salinity stress, which also conferred plant tolerance to oxidative stress by decreasing ROS production and increasing ROS clearance. Our work shows that PnPHR1 encodes an active CPD photolyase, which may participate in the adaptation of P. nutans to polar environments.

Transcriptional profiling and physiological analysis reveal the critical roles of ROS-scavenging system in the Antarctic moss Pohlia nutans under Ultraviolet-B radiation.

Organisms suffer more harmful ultraviolet radiation in the Antarctica due to the ozone layer destruction. Bryophytes are the dominant flora in the Antarctic continent. However, the molecular mechanism of Antarctic moss adaptation to UV-B radiation remains unclear. In the research, the transcriptional profiling of the Antarctic moss Pohlia nutans under UV-B radiation was conducted by Illumina HiSeq2500 platform. Totally, 72,922 unigenes with N50 length of 1434 bp were generated. Differential expression analysis demonstrated that 581 unigenes were markedly up-regulated and 249 unigenes were significantly down-regulated. The gene clustering analysis showed that these differentially expressed genes (DEGs) includes several transcription factors, photolyases, antioxidant enzymes, and flavonoid biosynthesis-related genes. Further analyses suggested that the content of malondialdehyde (MDA), the activities of several antioxidant enzymes (i.e., catalase, peroxidase, and glutathione reductase) were significantly enhanced upon UV-B treatment. Furthermore, the content of flavonoids and the gene expression levels of their synthesis-related enzymes were also markedly increased when plants were exposed to UV-B light. Therefore, these results suggested that the pathways of antioxidant enzymes, flavonoid synthesis and photolyases were the main defense systems that contributed to the adaption of Pohlia nutans to the enhanced UV-B radiation in Antarctica.

Phenolic compounds from different bryophyte species and cell compartments respond specifically to ultraviolet radiation, but not particularly quickly.

To study the potential quick responses to ultraviolet (UV) radiation of bryophyte phenolic compounds, we cultivated two thalloid liverworts, two leafy liverworts, and two mosses under three moderate realistic UV levels in the laboratory for 22 days. At the end of the daylight period on the first and last culture days, we measured the bulk levels and individual contents of phenolic UV-absorbing compounds (UVACs) of each species, differentiating in both cases the UVACs located in the methanol-soluble (mainly vacuolar) and -insoluble (cell wall-bound) fractions (SUVACs and IUVACs, respectively). The bulk levels of SUVACs and IUVACs mostly showed linear or hyperbolic relationships with the UV dose applied. Thirteen flavones (apigenin and luteolin derivatives) and two hydroxycinnamic acids (p-coumaric and ferulic acids) were identified in the soluble and insoluble fractions, respectively. Only two compounds (p-coumaric and ferulic acids) from the insoluble fraction of the leafy liverwort Plagiochila asplenioides showed a significant quick accumulation in response to UV radiation in the first day of culture, whereas six UVACs (mainly soluble apigenin and luteolin derivatives) from different species (mainly liverworts) were significantly accumulated at the end of the culture. In conclusion, the responses of bryophyte UVACs to UV radiation were influenced by the specific compound considered, the fraction in which each UVAC was located, the global or individual way of UVACs quantification, the bryophyte species and evolutionary lineage, and the experimental conditions used. Particularly, SUVACs were more UV-responsive than IUVACs and liverworts than mosses, and responses were not especially quick.

Photoprotection enhanced by red cell wall pigments in three East Antarctic mosses

BACKGROUND: Antarctic bryophytes (mosses and liverworts) are resilient to physiologically extreme environmental conditions including elevated levels of ultraviolet (UV) radiation due to depletion of stratospheric ozone. Many Antarctic bryophytes synthesise UV-B-absorbing compounds (UVAC) that are localised in their cells and cell walls, a location that is rarely investigated for UVAC in plants. This study compares the concentrations and localisation of intracellular and cell wall UVAC in Antarctic Ceratodon purpureus, Bryum pseudotriquetrum and Schistidium antarctici from the Windmill Islands, East Antarctica. RESULTS: Multiple stresses, including desiccation and naturally high UV and visible light, seemed to enhance the incorporation of total UVAC including red pigments in the cell walls of all three Antarctic species analysed. The red growth form of C. purpureus had significantly higher levels of cell wall bound and lower intracellular UVAC concentrations than its nearby green form. Microscopic and spectroscopic analyses showed that the red colouration in this species was associated with the cell wall and that these red cell walls contained less pectin and phenolic esters than the green form. All three moss species showed a natural increase in cell wall UVAC content during the growing season and a decline in these compounds in new tissue grown under less stressful conditions in the laboratory. CONCLUSIONS: UVAC and red pigments are tightly bound to the cell wall and likely have a long-term protective role in Antarctic bryophytes. Although the identity of these red pigments remains unknown, our study demonstrates the importance of investigating cell wall UVAC in plants and contributes to our current understanding of UV-protective strategies employed by particular Antarctic bryophytes. Studies such as these provide clues to how these plants survive in such extreme habitats and are helpful in predicting future survival of the species studied.

Bryophyte gas-exchange dynamics along varying hydration status reveal a significant carbonyl sulphide (COS) sink in the dark and COS source in the light.

Carbonyl sulphide (COS) is a potential tracer of gross primary productivity (GPP), assuming a unidirectional COS flux into the vegetation that scales with GPP. However, carbonic anhydrase (CA), the enzyme that hydrolyses COS, is expected to be light independent, and thus plants without stomata should continue to take up COS in the dark. We measured net CO2 (AC ) and COS (AS ) uptake rates from two astomatous bryophytes at different relative water contents (RWCs), COS concentrations, temperatures and light intensities. We found large AS in the dark, indicating that CA activity continues without photosynthesis. More surprisingly, we found a nonzero COS compensation point in light and dark conditions, indicating a temperature-driven COS source with a Q10 (fractional change for a 10°C temperature increase) of 3.7. This resulted in greater AS in the dark than in the light at similar RWC. The processes underlying such COS emissions remain unknown. Our results suggest that ecosystems dominated by bryophytes might be strong atmospheric sinks of COS at night and weaker sinks or even sources of COS during daytime. Biotic COS production in bryophytes could result from symbiotic fungal and bacterial partners that could also be found on vascular plants.

Effect of UV-B radiation on UV absorbing compounds and pigments of moss and lichen of Schirmacher oasis region, East Antarctica.

The survival of Antarctic flora under ozone depletion depends on their ability to acclimate against increasing UV—B radiation by employing photo protective mechanisms either by avoiding or repairing UV—B damage. A fifteen days experiment was designed to study moss (Bryum argenteum) and lichen (Umbilicaria aprina) under natural UV—B exposure and under UV filter frames at the Maitri region of Schirmacher oasis, East Antarctica. Changes in UV absorbing compounds, phenolics, carotenoids and chlorophyll content were studied for continuous fifteen days and significant changes were observed in the UV exposed plants of B. argenteum and U. aprina. The change in the UV absorbing compounds was more significant in B. argenteum (P<0.0001) than U. aprina (P<0.0002). The change in phenolic contents and total carotenoid content was significant (P<0.0001) in both B. argenteum and lichen U. aprina indicating that the increase in UV absorbing compounds, phenolic contents and total carotenoid content act as a protective mechanism against the deleterious effect of UV—B radiations.

Impacts of long-term enhanced UV-B radiation on bryophytes in two sub-Arctic heathland sites of contrasting water availability.

BACKGROUND AND AIMS: Anthropogenic depletion of stratospheric ozone in Arctic latitudes has resulted in an increase of ultraviolet-B radiation (UV-B) reaching the biosphere. UV-B exposure is known to reduce above-ground biomass and plant height, to increase DNA damage and cause accumulation of UV-absorbing compounds in polar plants. However, many studies on Arctic mosses tended to be inconclusive. The importance of different water availability in influencing UV-B impacts on lower plants in the Arctic has been poorly explored and might partially explain the observed wide variation of responses, given the importance of water in controlling bryophyte physiology. This study aimed to assess the long-term responses of three common sub-Arctic bryophytes to enhanced UV-B radiation (+UV-B) and to elucidate the influence of water supply on those responses. METHODS: Responses of three sub-Arctic bryophytes (the mosses Hylocomium splendens and Polytrichum commune and the liverwort Barbilophozia lycopodioides) to +UV-B for 15 and 13 years were studied in two field experiments using lamps for UV-B enhancement with identical design and located in neighbouring areas with contrasting water availability (naturally mesic and drier sites). Responses evaluated included bryophyte abundance, growth, sporophyte production and sclerophylly; cellular protection by accumulation of UV-absorbing compounds, ß-carotene, xanthophylls and development of non-photochemical quenching (NPQ); and impacts on photosynthesis performance by maximum quantum yield (F(v) /F(m)) and electron transport rate (ETR) through photosystem II (PSII) and chlorophyll concentrations. RESULTS: Responses were species specific: H. splendens responded most to +UV-B, with reduction in both annual growth (-22 %) and sporophyte production (-44 %), together with increased ß-carotene, violaxanthin, total chlorophyll and NPQ, and decreased zeaxanthin and de-epoxidation of the xanthophyll cycle pool (DES). Barbilophozia lycopodioides responded less to +UV-B, showing increased ß-carotene and sclerophylly and decreased UV-absorbing compounds. Polytrichum commune only showed small morphogenetic changes. No effect of UV-B on bryophyte cover was observed. Water availability had profound effects on bryophyte ecophysiology, and plants showed, in general, lower growth and ETR, together with a higher photoprotection in the drier site. Water availability also influenced bryophyte responses to +UV-B and, in particular, responses were less detectable in the drier site. CONCLUSIONS: Impacts of UV-B exposure on Arctic bryophytes were significant, in contrast to modest or absent UV-B effects measured in previous studies. The impacts were more easily detectable in species with high plasticity such as H. splendens and less obvious, or more subtle, under drier conditions. Species biology and water supply greatly influences the impact of UV-B on at least some Arctic bryophytes and could contribute to the wide variation of responses observed previously.

Seasonal acclimation of the moss Polytrichum juniperinum Hedw. to natural and enhanced ultraviolet radiation.

Short- and long-term changes in the methanol-extractable UV-absorbing compounds and biomass of the pioneer moss Polytrichum juniperinum in response to natural and enhanced UV radiation were studied. Under natural conditions, the compounds were found to fluctuate seasonally. In summer these compounds correlated negatively with irradiation. The concentration was low in July after a period of simultaneous heat, drought and high irradiation. Transient positive correlation between daily concentration and UV was seen in June. The concentration increased towards autumn and was relatively high under snow. Two enhanced UV experiments were performed. Seasonality in the compounds was again observed, with negative correlations with irradiation. During the first weeks, a transient inhibition of compound production was observed after the daily UV-B treatment. After six years of modulated UV-treatment in situ, photosynthesizing biomass decreased under UV-B and increased under UV-A. A larger variation in the UV-absorbing compounds was observed under UV-B treatment.

[Effects of enhanced UV-B radiation on the growth of five bryophytes in Changbai Mountains].

Five bryophytes (Rhytidium rugosum, Rhytidiadelphus triquetrus, Hylocomium splendens, Hylocomium pyrenaicum, and Polytrichum alpinum) were exposed to 0.2 kJ x m(-2) x d(-1) (visible light under native condition, CK), 3.0 kJ x m(-2) x d(-1) (simulated dose of UV-B irradiance at the tundra in Changbai Mountains, medium dose of UV-B irradiance, T1), and 6.0 kJ x m(-2) x d(-1) (high dose of UV-B irradiance, T2) to investigate the effects of enhanced UV-B radiation on plant height, biomass, and chlorophyll content. The results indicated that medium and high UV-B radiations decreased the plant height, biomass, and chlorophyll content of R. triquetrus and H. splendens by 32.3%, 62.4%, and 81.3%, and 21.4%, 59.4%, and 62.8%, respectively, and the relative growth rates were negative. Enhanced UV-B radiations had less effect on P. alpinum chlorophyll content but doubled its below-ground biomass, and slightly increased the biomass of R. rugosum. P. alpinum and R. rugosum had higher tolerance against UV-B radiation, while R. triquetrus and H. splendens were more sensitive to UV-B radiation.