Ultraviolet Tolerance Research Articles

Accumulated melanin in molds provides wavelength-dependent UV tolerance.

Fungal contamination poses a serious threat to public health and food safety because molds can grow under stressful conditions through melanin accumulation. Although ultraviolet (UV) irradiation is popular for inhibiting microorganisms, its effectiveness is limited by our insufficient knowledge about UV tolerance in melanin-accumulating molds. In this study, we first confirmed the protective effect of melanin by evaluating the UV sensitivity of young and mature spores. Additionally, we compared UV sensitivity between spores with accumulated melanin and spores prepared with melanin biosynthesis inhibitors. We found that mature spores were less UV-sensitive than young spores, and that reduced melanin accumulation by inhibitors led to reduced UV sensitivity. These results suggest that melanin protects cells against UV irradiation. To determine the most effective wavelength for inhibition, we evaluated the wavelength dependence of UV tolerance in a yeast (Rhodotorula mucilaginosa) and in molds (Aspergillus fumigatus, Cladosporium halotolerans, Cladosporium sphaerospermum, Aspergillus brasiliensis, Penicillium roqueforti, and Botrytis cinerea). We assessed UV tolerance using a UV-light emitting diode (LED) irradiation system with 13 wavelength-ranked LEDs between 250 and 365nm, a krypton chlorine (KrCl) excimer lamp device, and a low pressure (LP) Hg lamp device. The inhibition of fungi peaked at around 270nm, and most molds showed reduced UV sensitivity at shorter wavelengths as they accumulated pigment. Absorption spectra of the pigments showed greater absorption at shorter wavelengths, suggesting greater UV protection at these wavelengths. These results will assist in the development of fungal disinfection systems using UV, such as closed systems of air and water purification.

Recovery of insect-pathogenic fungi from solar UV damage: Molecular mechanisms and prospects.

Molecular mechanisms underlying insect-pathogenic fungal tolerance to solar ultraviolet (UV) damage have been increasingly understood. This chapter reviews the methodology established to quantify fungal response to solar UV radiation, which consists of UVB and UVA, and characterize a pattern of the solar UV dose (damage) accumulated from sunrise to sunset on sunny summer days. An emphasis is placed on anti-UV mechanisms of fungal insect pathogens in comparison to those well documented in model yeast. Principles are discussed for properly timing the application of a fungal pesticide to improve pest control during summer months. Fungal UV tolerance depends on either nucleotide excision repair (NER) or photorepair of UV-induced DNA lesions to recover UV-impaired cells in the darkness or the light. NER is a slow process independent of light and depends on a large family of anti-UV radiation (RAD) proteins studied intensively in model yeast but rarely in non-yeast fungi. Photorepair is a rapid process that had long been considered to depend on only one or two photolyases in filamentous fungi. However, recent studies have greatly expanded a genetic/molecular basis for photorepair-dependent photoreactivation that serves as a primary anti-UV mechanism in insect-pathogenic fungi, in which photolyase regulators required for photorepair and multiple RAD homologs have higher or much higher photoreactivation activities than do photolyases. The NER activities of those homologs in dark reactivation cannot recover the severe UV damage recovered by their activities in photoreactivation. Future studies are expected to further expand the genetic/molecular basis of photoreactivation and enrich principles for the recovery of insect-pathogenic fungi from solar UV damage.

Novel formulations improve the environmental tolerance of entomopathogenic nematodes

Entomopathogenic nematodes (EPN) are susceptible to environmental factors such as ultraviolet (UV), for which formulations of EPN infective juveniles (IJs) were explored. In the screening test for Steinernema carpocapsae, liquid starch displayed the greatest UV protection, followed by white kaolin clay (WKC), watergel and Barricade. In concentration tests, after 20-min UV S. carpocapsae IJs mixed in WKC caused 100% mortality of Galleria mellonella larvae at ≥5% concentration and had lower IJ mortality than aqueous IJs at 0.5%-10% WKC. Liquid starch showed increased UV tolerance with concentrations up to 30% and 50% for IJ viability and virulence, respectively. The WKC, liquid starch, Barricade and watergel were further compared to water using S. carpocapsae and Heterorhabditis floridensis. H. floridensis was more susceptible to UV radiation than S. carpocapsae. Liquid starch provided the strongest protection of both EPNs in IJ viability and virulence, with more prominent effects for S. carpocapsae than H. floridensis. The WKC and Barricade improved the viability of both EPNs after UV treatment, but only WKC enhanced S. carpocapsae virulence. The number of invading S. carpocapsae decreased as UV exposure time increased for WKC and only decreased at 20-min UV for liquid starch. Very few H. floridensis invaded insects after UV exposure regardless of treatments. In outdoor soil tests, liquid starch and WKC protected S. carpocapsae but were inferior to Barricade and watergel, probably associated with anti-desiccation. Overall, liquid starch and WKC improved the UV tolerance of EPNs, although the effects were less pronounced in soil involving other factors.

Arabidopsis RAD16 Homologues Are Involved in UV Tolerance and Growth.

In plants, prolonged exposure to ultraviolet (UV) radiation causes harmful DNA lesions. Nucleotide excision repair (NER) is an important DNA repair mechanism that operates via two pathways: transcription coupled repair (TC-NER) and global genomic repair (GG-NER). In plants and mammals, TC-NER is initiated by the Cockayne Syndrome A and B (CSA/CSB) complex, whereas GG-NER is initiated by the Damaged DNA Binding protein 1/2 (DDB1/2) complex. In the yeast Saccharomyces cerevisiae (S. cerevisiae), GG-NER is initiated by the Radiation Sensitive 7 and 16, (RAD7/16) complex. Arabidopsis thaliana has two homologues of yeast RAD16, At1g05120 and At1g02670, which we named AtRAD16 and AtRAD16b, respectively. In this study, we characterized the roles of AtRAD16 and AtRAD16b. Arabidopsis rad16 and rad16b null mutants exhibited increased UV sensitivity. Moreover, AtRAD16 overexpression increased plant UV tolerance. Thus, AtRAD16 and AtRAD16b contribute to plant UV tolerance and growth. Additionally, we found physical interaction between AtRAD16 and AtRAD7. Thus, the Arabidopsis RAD7/16 complex is functional in plant NER. Furthermore, AtRAD16 makes a significant contribution to Arabidopsis UV tolerance compared to the DDB1/2 and the CSB pathways. This is the first time the role and interaction of DDB1/2, RAD7/16, and CSA/CSB components in a single system have been studied.

Tolerance Increase in Escherichia coli O157:H7 and Methicillin-Resistant Staphylococcus Aureus USA300 Exposed to Low-Power Continuous Ultraviolet Radiation from Narrow-Wavelength Sources

Escherichia coli O157:H7 is a major cause of foodborne disease outbreaks throughout the world, while methicillin-resistant Staphylococcus aureus (MRSA) is responsible for many difficult-to-treat infections in humans. Ultraviolet (UV) irradiation is commonly used for disinfection in food processing, medical facilities, and water treatment to prevent the transmission of these pathogen. With increased use of UV disinfection technologies over the last few years because of COVID-19 and concerns about other communicable disease, it has become a concern that microbial species could develop tolerance to UV irradiation, especially when it is applied continuously. To elucidate the effect of continuous UV exposure at different wavelengths and power levels on the tolerance development of bacteria, Escherichia coli O157:H7 and MRSA)USA300 growths were investigated by continuously exposing inoculated agar plates to six different commercially available UV sources at wavelengths of 222 nm, 254 nm, 275 nm, and 405 nm. The agar plates in these experiments were partially covered by a thin acrylic sheet, which provided either complete protection from the UV to the cells directly under the sheet, no protection if significantly away from the sheet, or partial protection near the edges of the sheet due to shading or small amounts of UV reflection under the sheet at the edges. In these experiments, tolerant cells of E. coli and S. aureus were found from the 222 nm, the 405 nm, and one of the 254 nm sources. Upon examination of the power of each UV source, it was shown that the 275 nm and 254 nm sources that resulted in no tolerant cells had surface power densities [at 25 cm (10 in.)] that were more than 10–200 times greater than those that had tolerant cells. These results suggests that bacterial cells have a higher chance to develop UV tolerance under lower power UV sources (under the experimental conditions in our laboratory). Genome investigation of the tolerant colonies revealed that there are no significant differences between the cells that developed tolerance and the original organism, hinting at the need to explore the role of epigenetics mechanisms in the development of UV tolerance in these bacteria.

Genomic Analysis of Haloarchaea from Diverse Environments, including Permian Halite, Reveals Diversity of Ultraviolet Radiation Survival and DNA Photolyase Gene Variants

Ultraviolet (UV) radiation responses of extremophilic and archaeal microorganisms are of interest from evolutionary, physiological, and astrobiological perspectives. Previous studies determined that the halophilic archaeon, Halobacterium sp. NRC-1, which survives in multiple extremes, is highly tolerant of UV radiation. Here, Halobacterium sp. NRC-1 UV tolerance was compared to taxonomically diverse Haloarchaea isolated from high-elevation salt flats, surface warm and cold hypersaline lakes, and subsurface Permian halite deposits. Haloterrigena/Natrinema spp. from subsurface halite deposits were the least tolerant after exposure to photoreactivating light. This finding was attributed to deviation of amino acid residues in key positions in the DNA photolyase enzyme or to the complete absence of the photolyase gene. Several Halobacterium, Halorubrum and Salarchaeum species from surface environments exposed to high solar irradiance were found to be the most UV tolerant, and Halorubrum lacusprofundi from lake sediment was of intermediate character. These results indicate that high UV tolerance is not a uniform character trait of Haloarchaea and is likely reflective of UV exposure experienced in their environment. This is the first report correlating natural UV tolerance to photolyase gene functionality among Haloarchaea and provides insights into their survival in ancient halite deposits and potentially on the surface of Mars.

Tolerance of Steinernema carpocapsae infective juveniles in novel nanoparticle formulations to ultraviolet radiation

Entomopathogenic nematodes (EPNs) are susceptible to abiotic environmental factors including ultraviolet (UV) radiation, which affects the survival and efficacy. This study evaluated nanoparticle (NP) formulations for protecting Steinernema carpocapsae infective juveniles (IJs) from UV radiation. First, silica-NH2 NPs at oil-to-water ratios of 2:8, 3:7 and 4:6 were compared with Barricade Fire Gel (1 % and 2 %) and a water control (aqueous IJs) by exposing IJs to UV light (254 nm) for 0, 10 and 20 min. Barricade gel (especially 2 % Barricade) significantly improved IJs viability after UV treatment, while all three NPs had adverse effects on IJ viability after UV radiation. Subsequently, two silica (SiO2 basic and advanced) and one titania (TiO2) based formulations were tested with Barricade (1 % and 2 %) and a water control. The titania-NH2 NPs provided the highest UV protection, and IJ viability and virulence were not reduced even after 20-min UV. Except TiO2, only 2 % Barricade at 10-min UV and SiO2 basic at 20-min UV had lower IJ mortality than the water control. Only TiO2 formulated IJs caused higher insect mortality and infection levels than aqueous IJs after UV treatment. The UV tolerance of TiO2 was further examined by assessing the number of nematodes invading the hosts. Consistent with virulence tests, the number of invading nematodes in titania-NH2 NPs did not decrease after UV radiation for 10 or 20 min compared with the no-UV control. The anti-UV capability of titania-NH2 NPs has promise as a tool to enhance biocontrol efficacy of EPNs under field conditions.

Creation of an Industrial Bacillus thuringiensis Strain With High Melanin Production and UV Tolerance by Gene Editing.

Bacillus thuringiensis produces insecticidal crystal proteins (ICPs) which exhibit strong insecticidal toxicity. But when used in the field, ICPs would be destroyed by ultraviolet (UV) radiation in sunlight, thus decreasing the insecticidal activity and shortening the persistence. To improve the duration of B. thuringiensis preparations, we endowed a highly toxic industrial B. thuringiensis HD-1 with UV tolerance by making it produce melanin, a pigment that absorbs UV radiation. In B. thuringiensis, melanin is derived from homogentisate (HGA), an intermediate in the tyrosine pathway. And the absence of homogentisate-1,2-dioxygenase (HmgA) will lead to the formation of melanin. In this study, we used the CRISPR/Cas9 system to knock out the hmgA gene and obtained a melanin-producing mutant HD-1-ΔhmgA from strain HD-1. The melanin yield by mutant HD-1-ΔhmgA reached 3.60 mg/mL. And the anti-UV test showed that melanin serves as a protection to both the organism and the ICPs. After UV irradiation for 3 h, mutant HD-1-ΔhmgA still had an 80% insecticidal activity against the cotton bollworm, Helicoverpa armigera, while the control line only had about 20%. This study creates a light-stable biopesticide prototype based on a classic industrial strain that can be applied directly and takes the melanin-producing strain as a concept to improve the preparation validity.

Bryophyte ultraviolet-omics: from genes to the environment.

Ultraviolet (UV) radiation has contributed to the evolution of organisms since the origins of life. Bryophytes also have evolutionary importance as the first clearly identified lineage of land plants (embryophytes) colonizing the terrestrial environment, thus facing high UV and water scarcity, among other new challenges. Here we review bryophyte UV-omics, the discipline relating bryophytes and UV, with an integrative perspective from genes to the environment. We consider species and habitats investigated, methodology, response variables, protection mechanisms, environmental interactions, UV biomonitoring, molecular and evolutionary aspects, and applications. Bryophyte UV-omics shows convergences and divergences with the UV-omics of other photosynthetic organisms, from algae to tracheophytes. All these organisms converge in that UV damage may be limited under realistic UV levels, due to structural protection and/or physiological acclimation capacity. Nevertheless, bryophytes diverge because they have a unique combination of vegetative and reproductive characteristics to cope with high UV and other concomitant adverse processes, such as desiccation. This interaction has both evolutionary and ecological implications. In addition, UV effects on bryophytes depend on the species and the evolutionary lineage considered, with mosses more UV-tolerant than liverworts. Thus, bryophytes do not constitute a homogeneous functional type with respect to their UV tolerance.

Timing and duration of light exposure during conidia development determine tolerance to ultraviolet radiation.

Metarhizium is an important genus of soil-inhabiting fungi that are used for the biological control of insects. The efficiency of biocontrol is dependent on the maintenance of inoculum viability under adverse field conditions such as solar ultraviolet (UV) radiation. Therefore, increasing the tolerance of Metarhizium to UV radiation is necessary. It was previously established that, in mycelium, exposure to visible light increases tolerance to UV radiation. Similarly, growth under visible light for 14 days induces the production of tolerant conidia. However, a study evaluating if and how visible light affects conidia and their relationship with UV radiation was never performed. Here, we report that a relatively short and timed exposure to light around the time of conidiation is sufficient to induce the production of conidia with increased photoreactivating capacity and UV tolerance in Metarhizium acridum. Conidia produced by this method retain their characteristic higher tolerance even after many days of being transferred to the dark. Furthermore, we show that mature conidia of M. acridum and Metarhizium brunneum can still answer to light and regulate UV tolerance, suggesting that gene expression is possible even in dormant spores. Being able to respond to light in the dormant stages of development is certainly an advantage conferring improved environmental persistence to Metarhizium.

Spatially explicit depiction of a floral epiphytic bacterial community reveals role for environmental filtering within petals.

The microbiome of flowers (anthosphere) is an understudied compartment of the plant microbiome. Within the flower, petals represent a heterogeneous environment for microbes in terms of resources and environmental stress. Yet, little is known of drivers of structure and function of the epiphytic microbial community at the within‐petal scale. We characterized the petal microbiome in two co‐flowering plants that differ in the pattern of ultraviolet (UV) absorption along their petals. Bacterial communities were similar between plant hosts, with only rare phylogenetically distant species contributing to differences. The epiphyte community was highly culturable (75% of families) lending confidence in the spatially explicit isolation and characterization of bacteria. In one host, petals were heterogeneous in UV absorption along their length, and in these, there was a negative relationship between growth rate and position on the petal, as well as lower UV tolerance in strains isolated from the UV‐absorbing base than from UV reflecting tip. A similar pattern was not seen in microbes isolated from a second host whose petals had uniform patterning along their length. Across strains, the variation in carbon usage and chemical tolerance followed common phylogenetic patterns. This work highlights the value of petals for spatially explicit explorations of bacteria of the anthosphere.

TiO2 electron transport bilayer for all-inorganic perovskite photodetectors with remarkably improved UV stability toward imaging applications

High ultraviolet (UV) stability and low dark current (Idark) are necessary for high-quality perovskite photodetectors (PDs). TiO2 thin film is known as effective electron-transport-layer (ETL) for perovskite devices. However, common spin-coated TiO2 ETLs endow many surface defects and have strong UV photocatalytic effect to decompose perovskite materials, resulting in inferior stability of devices. In this work, TiO2 bilayer film (Bi-TiO2) has been fabricated by combining spin-coating and atomic-layer-deposition process and its positive effects on UV stability and Idark of Cs2AgBiBr6-based PDs have been revealed for the first time. It is demonstrated that Bi-TiO2 possesses fewer surface defects and smoother morphology with type Ⅱ band alignment, which is beneficial to suppress photocatalytic activity of TiO2 and reduce carrier recombination at the interface. After accelerated strong UV aging treatment, the PD with Bi-TiO2 maintains excellent performance, whereas the PD with spin-coated TiO2 film dramatically deteriorate with on-off ratio drops from ∼102 to ∼2. Besides, the Idark of PD remarkably decreases from ∼10−8 A to ∼10−10 A after bilayer optimization. Furthermore, we have integrated the corresponding PDs into a self-built imaging system adopting diffuse reflection mode. This work suggests a feasible approach to fabricate TiO2/Cs2AgBiBr6-based PDs with remarkable UV tolerance for imaging applications.

Nitrogen removal mechanism of marine anammox bacteria treating nitrogen-laden saline wastewater in response to ultraviolet (UV) irradiation: High UV tolerance and microbial community shift

Salt stress can be naturally overcome by marine anammox bacteria (MAB), while their low growth rate and sensitivity to operational conditions are still challenges for the application of anammox. To enhance the enrichment of MAB and decipher the effects of ultraviolet (UV) irradiation on MAB, UV was introduced in the nitrogen removal of MAB treating nitrogen-laden saline wastewater for the first time. The results indicated that MAB were resistant to a fairly high UV-C dose, 12000 mJ/cm2. Their relative abundance was enhanced by 1.2 folds under 12000 mJ/cm2 UV-C. However, the relative abundance of Actinobacteria, Acidobacteria, Chloroflexi and Marinicella were greatly dropped with enhanced UV-C dose. The tolerance mechanism was diversified, e.g. excessive extracellular polymeric substances, special structure of MAB and interspecific competition/cooperation. Although further study was still needed, the findings shed a light on MAB enrichment and exploited great potentials of MAB in nitrogen-laden saline wastewater treatment.

UVSSA, UBP12, and RDO2/TFIIS Contribute to Arabidopsis UV Tolerance

Plant DNA is damaged by exposure to solar radiation, which includes ultraviolet (UV) rays. UV damaged DNA is repaired either by photolyases, using visible light energy, or by nucleotide excision repair (NER), also known as dark repair. NER consists of two subpathways: global genomic repair (GGR), which repairs untranscribed DNA throughout the genome, and transcription-coupled repair (TCR), which repairs transcribed DNA. In mammals, CSA, CSB, UVSSA, USP7, and TFIIS have been implicated in TCR. Arabidopsis homologs of CSA (AtCSA-1/2) and CSB (CHR8) have previously been shown to contribute to UV tolerance. Here we examine the role of Arabidopsis homologs of UVSSA, USP7 (UBP12/13), and TFIIS (RDO2) in UV tolerance. We find that loss of function alleles of UVSSA, UBP12, and RDO2 exhibit increased UV sensitivity in both seedlings and adults. UV sensitivity in atcsa-1, uvssa, and ubp12 mutants is specific to dark conditions, consistent with a role in NER. Interestingly, chr8 mutants exhibit UV sensitivity in both light and dark conditions, suggesting that the Arabidopsis CSB homolog may play a role in both NER and light repair. Overall our results indicate a conserved role for UVSSA, USP7 (UBP12), and TFIIS (RDO2) in TCR.

Spores potentially dispersed to longer distances are more tolerant to ultraviolet radiation: A case study in the moss genus Orthotrichum.

Ultraviolet (UV) radiation influences the viability of algal spores and seed-plant pollen depending on the species, the dose, and the wavelength. In bryophytes, one of the dominant groups of plants in many habitats, UV radiation could determine their spore dispersal strategy, and such data are critical for reconstructing the ancestral state in plants and for determining the distribution range and persistence of bryophyte species. Spores of four bryophyte species of the moss genus Orthotrichum that were either hygrochastic or xerochastic (spores dispersed under wet or dry conditions, respectively) were exposed to realistic doses of UV radiation under laboratory conditions. Spore viability was evaluated through germination experiments and, for the first time in bryophytes, ultrastructural observations. Given that the UV-B doses used were relatively higher than the UV-A doses, the UV effect was probably due more to UV-B than UV-A wavelengths. All four species reduced their spore germination capacity in a UV dose-dependent manner, concomitantly increasing spore ultrastructural damage (cytoplasmic and plastid alterations). Most spores eventually died when exposed to the highest UV dose. Interestingly, spores of hygrochastic species were much more UV-sensitive than those of xerochastic species. UV tolerance determines moss spore viability, as indicated by germination capacity and ultrastructural damage, and differs between spores of species with different dispersal strategies. Specifically, the higher UV tolerance of xerochastic spores may enable them to be dispersed to longer distances than hygrochastic spores, thus extending more efficiently the distribution range of the corresponding species.

Correlated Molecular Structural Motions for Photoprotection after Deep-UV Irradiation.

Exposure to ultraviolet (UV) light could cause photodamage to biomolecular systems and degrade optoelectronic devices. To mitigate such detrimental effects from the bottom up, we strategically select a photosensitive molecule pyranine and implement femtosecond electronic and Raman spectroscopies to elucidate its ultrafast photoprotection mechanisms in solution. Our results show that pyranine undergoes excited-state proton transfer in water, while this process is blocked in methanol regardless of excitation wavelengths (267, 400 nm). After 267 nm irradiation, the molecule relaxes from a higher lying electronic state into a lower lying singlet state with a <300 fs time constant, followed by solvation events. Transient Raman marker bands exhibit different patterns of intensity dynamics and frequency shift that elucidate the real-time interplay among conformational motions, photochemical reaction, and vibrational cooling after excitation. More energetic photons are revealed to selectively enhance certain relaxation pathways. These mechanistic findings offer new guidelines to improve the UV tolerance and stability of the engineered functional molecules in materials and life sciences.

Responses of entomopathogenic fungi to the mutagen 4-nitroquinoline 1-oxide

Survival of entomopathogenic fungi under solar ultraviolet (UV) radiation is paramount to the success of biological control of insect pests and disease vectors. The mutagenic compound 4-nitroquinoline 1-oxide (4-NQO) is often used to mimic the biological effects of UV radiation on organisms. Therefore, we asked whether tolerance to 4-NQO could predict tolerance to UV radiation in thirty isolates of entomopathogenic fungi and one isolate of a xerophilic fungus. A dendrogram obtained from cluster analyses based on the 50 and 90 % inhibitory concentrations (IC50 and IC90, respectively) divided the fungal isolates into six clusters numbered consecutively based on their tolerance to 4-NQO. Cluster 6 contained species with highest tolerance to 4-NQO (IC50 > 4.7 μM), including Mariannaea pruinosa, Lecanicillium aphanocladii, and Torrubiella homopterorum. Cluster 1 contained species least tolerant to 4-NQO (IC50 < 0.2 μM), such as Metarhizium acridum (ARSEF 324), Tolypocladium geodes, and Metarhizium brunneum (ARSEF 7711). With few exceptions, the majority of Metarhizium species showed moderate to low tolerances (IC50 between 0.4 and 0.9 μM) and were placed in cluster 2. Cluster 3 included species with moderate tolerance (IC50 between 1.0 and 1.2 μM). In cluster 4 were species with moderate to high tolerance (IC50 between 1.3 and 1.6 μM). Cluster 5 contained the species with high tolerance (IC50 between 1.9 and 4.0 μM). The most UV tolerant isolate of M. acridum, ARSEF 324, was the least tolerant to 4-NQO. Also, L. aphanocladii, which is very susceptible to UV radiation, showed high tolerance to 4-NQO. Our results indicate that tolerance to 4-NQO does not correlate with tolerance to UV radiation. Therefore this chemical compound is not a predictor of UV tolerance in entomopathogenic fungi.

Cell compartmentation of ultraviolet‐absorbing compounds: An underexplored tool related to bryophyte ecology, phylogeny and evolution

Abstract Excessive exposure to ultraviolet (UV) radiation can be harmful to photosynthetic organisms, that most frequently respond with the accumulation of protective UV‐absorbing compounds (UVACs). UVACs location in different cell compartments can influence their preferential protective role as antioxidants and/or UV screens. However, the phylogenetic, ecological and evolutionary implications of UVACs compartmentation has been little studied, particularly in bryophytes. We analysed UVACs in the methanol‐soluble and ‐insoluble fractions (SUVACs and IUVACs respectively) in extracts of 87 bryophytes belonging to their three evolutionary lineages: 22 liverworts, 64 mosses and one hornwort. Assuming that the cell wall‐bound IUVACs are more effective UV screens than the mainly vacuolar SUVACs, thus conferring a higher UV tolerance, we evaluated whether UVACs levels and compartmentation were related to: (1) the bryophyte phylogeny down to the Order level; and (2) the bryophyte ecological attributes, including sun exposure and Ellenberg indicator values (numerical system classifying species' habitat along gradients of environmental factors). A similar phylogenetic and ecological analysis was conducted on the sclerophylly index (the ratio between the dry mass and the surface area of the bryophyte shoot). Mosses showed lower SUVACs but higher IUVACs and total UVACs, together with higher IUVAC/SUVAC ratios, than liverworts. Thus, mosses would better tolerate UV radiation than liverworts, which matches well with their general ecological preferences. As bryophytes were the earliest diverging land plants, we could infer that the different UVACs compartmentation between mosses and liverworts could have influenced their ecological segregation upon plant land colonization. UVACs compartmentation also differed between the two major moss lineages (acrocarpous and pleurocarpous), while the anatomically peculiar Sphagnales were the best characterized Order. There were not solid relationships between UVACs and the ecological attributes considered. Hence, UVACs might be mainly constitutive in bryophytes, depending more on the species phylogeny than on the habitat occupied in nature. Liverworts were less sclerophyllous than mosses, and, as in tracheophytes, water restrictions and high sun exposures increased sclerophylly. In conclusion, UVACs compartmentation represents an ecophysiological trait useful to understand the bryophyte ecophylogeny, because different compartmentation modalities seem to imply different UV adaptations and tolerance. A plain language summary is available for this article.

RAD4 and RAD23/HMR Contribute to Arabidopsis UV Tolerance.

In plants, exposure to solar ultraviolet (UV) light is unavoidable, resulting in DNA damage. Damaged DNA causes mutations, replication arrest, and cell death, thus efficient repair of the damaged DNA is essential. A light-independent DNA repair pathway called nucleotide excision repair (NER) is conserved throughout evolution. For example, the damaged DNA-binding protein Radiation sensitive 4 (Rad4) in Saccharomyces cerevisiae is homologous to the mammalian NER protein Xeroderma Pigmentosum complementation group C (XPC). In this study, we examined the role of the Arabidopsis thaliana Rad4/XPC homologue (AtRAD4) in plant UV tolerance by generating overexpression lines. AtRAD4 overexpression, both with and without an N-terminal yellow fluorescent protein (YFP) tag, resulted in increased UV tolerance. YFP-RAD4 localized to the nucleus, and UV treatment did not alter this localization. We also used yeast two-hybrid analysis to examine the interaction of AtRAD4 with Arabidopsis RAD23 and found that RAD4 interacted with RAD23B as well as with the structurally similar protein HEMERA (HMR). In addition, we found that hmr and rad23 mutants exhibited increased UV sensitivity. Thus, our analysis suggests a role for RAD4 and RAD23/HMR in plant UV tolerance.

Stimulation of RNA Polymerase II ubiquitination and degradation by yeast mRNA 3′-end processing factors is a conserved DNA damage response in eukaryotes

The quality and retrieval of genetic information is imperative to the survival and reproduction of all living cells. Ultraviolet (UV) light induces lesions that obstruct DNA access during transcription, replication, and repair. Failure to remove UV-induced lesions can abrogate gene expression and cell division, resulting in permanent DNA mutations. To defend against UV damage, cells utilize transcription-coupled nucleotide excision repair (TC-NER) to quickly target lesions within active genes. In cases of long-term genotoxic stress, a slower alternative pathway promotes degradation of RNA Polymerase II (Pol II) to allow for global genomic nucleotide excision repair (GG-NER). The crosstalk between TC-NER and GG-NER pathways and the extent of their coordination with other nuclear events has remained elusive. We aimed to identify functional links between the DNA damage response (DDR) and the mRNA 3′-end processing complex. Our labs have previously shown that UV-induced inhibition of mRNA processing is a conserved DDR between yeast and mammalian cells. Here we have identified mutations in the yeast mRNA 3′-end processing cleavage factor IA (CFIA) and cleavage and polyadenylation factor (CPF) that confer sensitivity to UV-type DNA damage. In the absence of TC-NER, CFIA and CPF mutants show reduced UV tolerance and an increased frequency of UV-induced genomic mutations, consistent with a role for RNA processing factors in an alternative DNA repair pathway. CFIA and CPF mutants impaired the ubiquitination and degradation of Pol II following DNA damage, but the co-transcriptional recruitment of Pol II degradation factors Elc1 and Def1 was undiminished. Overall these data are consistent with yeast 3′-end processing factors contributing to the removal of Pol II stalled at UV-type DNA lesions, a functional interaction that is conserved between homologous factors in yeast and human cells.