Roles of PprA,IrrE, and RecA in the resistance of <Emphasis Type="Italic">Deinococcus radiodurans</Emphasis> to germicidal and environmentally relevant UV radiation

To study the role of different DNA repair genes in the resistance of Deinococcus radiodurans to mono- and polychromatic UV radiation, wild-type strain and knockout mutants in RecA, PprA, and IrrE of D. radiodurans were irradiated with UV-C (254 nm), UV-(A + B) (280–400 nm) and UV-A (315–400 nm) radiation, and survival was monitored. The strain deficient in recA was highly sensitive to UV-C radiation compared to the wild-type, but showed no loss of resistance against irradiation with UV-(A + B) and UV-A, while pprA and irrE-deficient strains exhibited elevated sensitivity to UV-A and UV-(A + B) radiation. These results suggest that the repair of DNA double-strand breaks is essential after treatment with highly energetic UV-C radiation, whereas protection from oxidative stress may play a greater role in resistance to environmentally relevant UV radiation.     

Survival of Spacecraft-Associated Microorganisms under Simulated Martian UV Irradiation

Spore-forming microbes recovered from spacecraft surfaces and assembly facilities were exposed to simulated Martian UV irradiation. The effects of UVA (315 to 400 nm), UVA+B (280 to 400 nm), and the full UV spectrum (200 to 400 nm) on the survival of microorganisms were studied at UV intensities expected to strike the surfaces of Mars. Microbial species isolated from the surfaces of several spacecraft, including Mars Odyssey, X-2000 (avionics), and the International Space Station, and their assembly facilities were identified using 16S rRNA gene sequencing. Forty-three Bacillus spore lines were screened, and 19 isolates showed resistance to UVC irradiation (200 to 280 nm) after exposure to 1,000 J m⁻² of UVC irradiation at 254 nm using a low-pressure mercury lamp. Spores of Bacillus species isolated from spacecraft-associated surfaces were more resistant than a standard dosimetric strain, Bacillus subtilis 168. In addition, the exposure time required for UVA+B irradiation to reduce the viable spore numbers by 90% was 35-fold longer than the exposure time required for the full UV spectrum to do this, confirming that UVC is the primary biocidal bandwidth. Among the Bacillus species tested, spores of a Bacillus pumilus strain showed the greatest resistance to all three UV bandwidths, as well as the total spectrum. The resistance to simulated Mars UV irradiation was strain specific; B. pumilus SAFR-032 exhibited greater resistance than all other strains tested. The isolation of organisms like B. pumilus SAFR-032 and the greater survival of this organism (sixfold) than of the standard dosimetric strains should be considered when the sanitation capabilities of UV irradiation are determined.     

UV‐A/BLUE LIGHT–INDUCED REACTIVATION OF SPORE GERMINATION IN UV‐B IRRADIATED ULVA PERTUSA (CHLOROPHYTA)1

Recent reduction in the ozone shield due to manufactured chlorofluorocarbons raised considerable interest in the ecological and physiological consequences of UV‐B radiation (λ=280–315 nm) in macroalgae. However, early life stages of macroalgae have received little attention in regard to their UV‐B sensitivity and UV‐B defensive mechanisms. Germination of UV‐B irradiated spores of the intertidal green alga Ulva pertusa Kjellman was significantly lower than in unexposed controls, and the degree of reduction correlated with the UV doses. After exposure to moderate levels of UV‐B irradiation, subsequent exposure to visible light caused differential germination in an irradiance‐ and wavelength‐dependent manner. Significantly higher germination was found at higher photon irradiances and in blue light compared with white and red light. The action spectrum for photoreactivation of germination in UV‐B irradiated U. pertusa spores shows a major peak at 435 nm with a smaller but significant peak at 385 nm. When exposed to December sunlight, the germination percentage of U. pertusa spores exposed to 1 h of solar radiation reached 100% regardless of the irradiation treatment conditions. After a 2‐h exposure to sunlight, however, there was complete inhibition of germination in PAR+UV‐A+UV‐B in contrast to 100% germination in PAR or PAR+UV‐A. In addition to mat‐forming characteristics that would act as a selective UV‐B filter for settled spores under the parental canopy, light‐driven repair of germination after UV‐B exposure could explain successful continuation of U. pertusa spore germination in intertidal settings possibly affected by intense solar UV‐B radiation.     

Long-Term Survival of <Emphasis Type="Italic">Bacillus</Emphasis> Spores in Alcohol and Identification of 90% Ethanol as Relatively More Spori/Bactericidal

This study was taken up with a view to generate basic information on spore hardiness to ethanol in various Bacillus species and related genera, and to assess the effectiveness of different levels of ethanol as a bacterial disinfectant. Predominantly spore-bearing cultures of five Bacillus spp. (B. pumilus, B. subtilis, B. megaterium, B. fusiformis and B. flexus) that were isolated from the spent-alcohol used during plant tissue culture work were challenged with aqueous ethanol (25, 50, 60, 70, 80 and 90% v/v) in 1 ml volumes at 10¹⁰⁻¹¹ CFU ml⁻¹. Monitoring the spore endurance through spotting and plating revealed prolonged tolerance (>12 months) at different alcohol levels depending on the organism except in 90% where no survival was observed beyond 2–12 months. Spores of related genera like Paenibacillus and Lysinibacillus also showed long-term ethanol survival. Alcohol tolerance of spore-forming organisms depended on the extent of spores and spore hardiness, which in turn varied with the organism, strain, age of culture, growing conditions and other factors as authenticated with ATCC strains of B. pumilus and B. subtilis. Aqueous 90% ethanol caused instant inactivation of vegetative cells in different spore formers and twelve other non-sporulating Gram-positive and Gram-negative organisms tested. Taking into account both vegetative cells and spores, the appropriate concentration of ethanol as a disinfectant emerged to be 90% followed by absolute ethanol compared with the generally recommended 70–80% level.     

Comparative Sensitivity to UV-B Radiation of Two Bacillus thuringiensis Subspecies and Other Bacillus sp.

Susceptibility of Bacillus thuringiensis spores and toxins to the UV-B range (280–330 nm) of the solar spectrum reaching Earth's surface may be responsible for its inactivation and low persistence in nature. Spores of the mosquito larvicidal B. thuringiensis subsp. israelensis were significantly more resistant to UV-B than spores of the lepidopteran-active subsp. kurstaki. Spores of subsp. israelensis were as resistant to UV-B as spores of B. subtilis and more resistant than spores of the closely related B. cereus and another mosquito larvicidal species B. sphaericus. Sensitivity of B. thuringiensis subsp. israelensis spores to UV-B radiation depended upon their culture age; 24-h cultures, approaching maximal larvicidal activity, were still sensitive. Maximal resistance to UV-B was achieved only at 48 h. Received: 13 December 2000/Accepted: 19 January 2001     

Reconstruction of daily solar UV irradiation from 1893 to 2002 in Potsdam,Germany

Long-term records of solar UV radiation reaching the Earth’s surface are scarce. Radiative transfer calculations and statistical models are two options used to reconstruct decadal changes in solar UV radiation from long-term records of measured atmospheric parameters that contain information on the effect of clouds, atmospheric aerosols and ground albedo on UV radiation. Based on earlier studies, where the long-term variation of daily solar UV irradiation was derived from measured global and diffuse irradiation as well as atmospheric ozone by a non-linear regression method [Feister et al. (2002) Photochem Photobiol 76:281–293], we present another approach for the reconstruction of time series of solar UV radiation. An artificial neural network (ANN) was trained with measurements of solar UV irradiation taken at the Meteorological Observatory in Potsdam, Germany, as well as measured parameters with long-term records such as global and diffuse radiation, sunshine duration, horizontal visibility and column ozone. This study is focussed on the reconstruction of daily broad-band UV-B (280–315 nm), UV-A (315–400 nm) and erythemal UV irradiation (ER). Due to the rapid changes in cloudiness at mid-latitude sites, solar UV irradiance exhibits appreciable short-term variability. One of the main advantages of the statistical method is that it uses doses of highly variable input parameters calculated from individual spot measurements taken at short time intervals, which thus do represent the short-term variability of solar irradiance.     

Effect of radioprotective agents in sporulation medium on Bacillus subtilis spore resistance to hydrogen peroxide, wet heat and germicidal and environmentally relevant UV radiation

Aims: To determine the effects of cysteine, cystine, proline and thioproline as sporulation medium supplements on Bacillus subtilis spore resistance to hydrogen peroxide (H₂O₂), wet heat, and germicidal 254 nm and simulated environmental UV radiation. Methods and Results: Bacillus subtilis spores were prepared in a chemically defined liquid medium, with and without supplementation of cysteine, cystine, proline or thioproline. Spores produced with thioproline, cysteine or cystine were more resistant to environmentally relevant UV radiation at 280–400 and 320–400 nm, while proline supplementation had no effect. Spores prepared with cysteine, cystine or thioproline were also more resistant to H₂O₂ but not to wet heat or 254‐nm UV radiation. The increases in spore resistance attributed to the sporulation supplements were eliminated if spores were chemically decoated. Conclusions: Supplementation of sporulation medium with cysteine, cystine or thioproline increases spore resistance to solar UV radiation reaching the Earth’s surface and to H₂O₂. These effects were eliminated if the spores were decoated, indicating that alterations in coat proteins by different sporulation conditions can affect spore resistance to some agents. Significance and Impact of the Study: This study provides further evidence that the composition of the sporulation medium can have significant effects on B. subtilis spore resistance to UV radiation and H₂O₂. This knowledge provides further insight into factors influencing spore resistance and inactivation.     

Soluble polyamines in Salix myrsinifolia and S. myrsinites × S. myrsinifolia plantlets exposed to increased UV-B irradiation and decreased watering

Hybridisation between certain willow species is a common feature leading to novel genotypes varying in growth rate and stress tolerance. The objective of this 4-week study was to investigate the effects of decreased watering, enhanced ultraviolet-B irradiation (UV-B_BE, 280–315 nm, 7.2 kJ m⁻² day⁻¹) and combined decreased watering and enhanced UV-B irradiation on di- and polyamines in the leaves of Salix myrsinifolia and its hybrid with S. myrsinites. Control plantlets were well-watered and exposed to ambient UV-B irradiation (UV-B_BE, 3.6 kJ m⁻² day⁻¹). HPLC analyses showed that the constitutive concentrations of soluble di- and polyamines varied markedly between S. myrsinifolia and its hybrids. The degree of responses to treatments also varied: in S. myrsinifolia, concentrations of free putrescine were clearly increased by reduced watering, while in the hybrid willow, change in putrescine was less pronounced and not significant. Results also showed that the increase in putrescine in S. myrsinifolia by reduced watering was mitigated by concurrent enhancement of UV-B irradiation. There were no direct UV-B effects on the soluble polyamines.     

Effects of solar ultraviolet radiation on photochemical efficiency of <Emphasis Type="Italic">Chaetoceros curvisetus</Emphasis> (Bacillariophyceae)

To assess the short- and long-term impacts of Ultraviolet radiation (UVR, 280–400 nm) on the red tide alga Chaetoceros curvisetus, we exposed cells to three different solar radiation treatments–PAB:280–700 nm, PA:320–700 nm, and P:400–700 nm under 20°C incubated temperature. Short-term exposures were investigated: the photochemical efficiency (Φ_PSII) versus irradiance curves under six levels of solar radiation by covering the incubators with a variable number of neutral density screens (the irradiance thus varied from 100 to 3%) lasting 1 h, and long-term exposures were designed to assess how the cells acclimate to solar radiation (the growth, UV_abc and ratio of repair to damage rates of D1 protein were detected). A significant decrease in the photochemical efficiency (Φ_PSII) at high irradiance (100% of incident solar radiation, 261.6 Wm⁻²) was observed in short-term exposure (1 h). UVR-induced photoinhibition was reduced to 7% in 3% solar radiation (4.08 Wm⁻²), compared with 66% in 100% solar radiation (261.6 Wm⁻²). In long-term experiments (11 days) using batch cultures, cell densities during the first 6 days were relatively constant for treatment P, and decreased slightly under PA and PAB treaments, reflecting a change in the irradiance experienced in the laboratory to that of incident solar irradiance. Thereafter, cell density increased and UV-induced photoinhibition decreased with the following days, indicating acclimation to solar UV. At the end of experiment, cells were found to exhibit both higher ratios of repair to UV-related damage and increased concentrations of UV-absorbing compounds, whose maximum absorption was found to be at 329 nm. Our data indicate that C. curvisetus is sensitive to ultraviolet radiation, but was able to acclimate relatively rapidly (ca. 6 days) by synthesizing UV-absorbing compounds and by increasing the rates of repair processes of D1 protein in PSII.     

The short- and long-term response of <Emphasis Type="Italic">Scrippsiella trochoidea</Emphasis> (Pyrrophyta) to solar ultraviolet radiation

To assess the short- and long-term impacts of UV radiation (UVR, 280–400 nm) on the microalga Scrippsiella trochoidea, we exposed cells to three different radiation treatments (PAB: 280–700 nm, PA: 320–700 nm, and P: 400–700 nm). A significant decrease in the photochemical efficiency (Φ_PSII) at high irradiance (100% of incident solar radiation, 216.0 W m⁻²) was observed. Photoinhibition was reduced from 62.7 to 10.9% when the cells were placed in 12% solar radiation (26.1 W m⁻²). In long-term experiments (11 days) using batch cultures, cell densities during the first 5 days were decreased under treaments P, PA, and PAB, reflecting a change in the irradiance experienced in the laboratory to that of incident solar irradiance. Thereafter, specific growth rates increased and UV-induced photoinhibition decreased, indicating acclimation to solar UV. Cells were found to exhibit both higher ratios of repair to UV-related damage, shorter period for recovery and increased concentrations of UV-absorbing compounds (UV_abc), whose maximum absorption was found to be at 336 nm. Our data indicate that S. trochoidea is sensitive to ultraviolet radiation, but was able to acclimate relatively rapidly (ca. 6 days) by synthesizing UV_abc and by increasing the rates of repair processes of D1 protein in PSII.     

Enhancement in leghemoglobin content of root nodules by exclusion of solar UV-A and UV-B radiation in soybean

The impact of exclusion of solar UV-B (280–320 nm) and UV-A+B (280–400 nm) radiation on the root nodules was studied in soybean(Glycine max var. MACS 330). Soybean plants were grown in the tropical region of Indore (Latitude-22.4°N), India under field conditions in metal cages covered with polyester exclusion filters that specifically cut off UV-B (<320 nm) and UV-A+B (<400 nm) radiation; control plants were grown under ambient solar radiation. Leghemoglobin content was analyzed in the root nodules on the 50^th day after emergence of seedlings. Exclusion of UV radiations significantly enhanced the leghemoglobin content in the nodules on fresh weight basis; 25% and 45% higher amount of leghemoglobin were present in the nodules after the exclusion of UV-B and UV-A+B radiation respectively. Analysis by native and SDS-PAGE showed high intense bands of leghemoglobin after the exclusion of UV-A+B as compared to control. Exclusion of UV radiation also enhanced the growth of roots as well as aerial parts of the plants. UV Exclusion increased nodulation by increase in the number and size of nodules. The results are discussed in the light of advantage of exclusion for enhancing protein/nitrogen content in the plants.     

Killing bacterial spores by organic hydroperoxides

Killing of wild-type spores of Bacillus subtilis by t-butyl hydroperoxide, cumene hydroperoxide and peracetic acid was not through DNA damage, as shown by the absence of mutations in the survivors and the identical sensitivity of spores of strains with or without a recA mutation. In contrast, B. subtilis spores (termed α⁻β⁻) lacking the DNA protective α/β-type small, acid-soluble spore proteins (SASP) were more sensitive to t-butyl hydroperoxide and cumene hydroperoxide, and their killing was in large part through DNA damage, as shown by the high frequency of mutations in the survivors and the greater sensitivity of α⁻β⁻ recA spores. Analysis of t-butyl hydroperoxide-treated spores showed that generation of DNA damage in α⁻β⁻ spores was more rapid than in wild-type spores; α/β-type SASP also protected against DNA strand breakage in vitro caused by t-butyl hydroperoxide. α/β-Type SASP appeared to play no role in protection of spores from killing by peracetic acid; wild-type and α⁻β⁻ spores exhibited identical peracetic acid sensitivity and their killing by this agent appeared to be not through DNA damage. Received 17 December 1996/ Accepted in revised form 13 March 1997     

A Method for Extracting RNA from Dormant and Germinating Bacillus subtilis Strain 168 Endospores

RNA was extracted from dormant and germinating Bacillus subtilis 168 spores (intact spores and chemically decoated spores) by using rapid rupture followed by acid–phenol extraction. Spore germination progress was monitored by assaying colony forming ability before and after heat shock and by reading the optical density at 600 nm. The purity, yield, and composition of the extracted RNA were determined spectrophotometrically from the ratio of absorption at 260 nm to that at 280 nm; in a 2100 BioAnalyzer, giving the RNA yield/10⁸ spores or cells and the distribution pattern of rRNA components. The method reported here for the extraction of RNA from dormant spores, as well as during different phases of germination and outgrowth, has proven to be fast, efficient and simple to handle. RNA of a high purity was obtained from dormant spores and during all phases of germination and growth. There was a significant increase in RNA yield during the transition from dormant spores to germination and subsequent outgrowth. Chemically decoated spores were retarded in germination and outgrowth compared with intact spores, and less RNA was extracted; however, the differences were not significant. This method for RNA isolation of dormant, germinating, and outgrowing bacterial endospores is a valuable prerequisite for gene expression studies, especially in studies on the responses of spores to hostile environmental conditions.     

Survival of Akinetes (Resting-State Cells of Cyanobacteria) in Low Earth Orbit and Simulated Extraterrestrial Conditions

Cyanobacteria are photosynthetic organisms that have been considered for space applications, such as oxygen production in bioregenerative life support systems, and can be used as a model organism for understanding microbial survival in space. Akinetes are resting-state cells of cyanobacteria that are produced by certain genera of heterocystous cyanobacteria to survive extreme environmental conditions. Although they are similar in nature to endospores, there have been no investigations into the survival of akinetes in extraterrestrial environments. The aim of this work was to examine the survival of akinetes from Anabaena cylindrica in simulated extraterrestrial conditions and in Low Earth Orbit (LEO). Akinetes were dried onto limestone rocks and sent into LEO for 10 days on the ESA Biopan VI. In ground-based experiments, the rocks were exposed to periods of desiccation, vacuum (0.7 × 10⁻³ kPa), temperature extremes (−80 to 80°C), Mars conditions (−27°C, 0.8 kPa, CO₂) and UV radiation (325–400 nm). A proportion of the akinete population was able to survive a period of 10 days in LEO and 28 days in Mars simulated conditions, when the rocks were not subjected to UV radiation. Furthermore, the akinetes were able to survive 28 days of exposure to desiccation and low temperature with high viability remaining. Yet long periods of vacuum and high temperature were lethal to the akinetes. This work shows that akinetes are extreme-tolerating states of cyanobacteria that have a practical use in space applications and yield new insight into the survival of microbial resting-state cells in space conditions.     

UV-absorbing compounds in <Emphasis Type="Italic">Porphyra haitanensis</Emphasis> (Rhodophyta) with special reference to effects of desiccation

The intertidal red alga Porphyra haitanensis Chang et Zheng is episodically desiccated and exposed to high levels of solar radiation at low tide during emersion. However, little has been documented on the relationship between the stresses during desiccation and related chemical compounds. We found that P. haitanensis thalli, when desiccated under indoor (artificial radiation) or outdoor (solar radiation) conditions, with or without UV radiation (UVR: 280–400 nm), contained significantly higher concentrations of UV-absorbing compounds (peak at 336 nm) than those maintained submerged (without desiccation). Solar UVR had no effect on the content of UV-absorbing compounds. Even though the concentration of these compounds decreased with time in all treatments, a slower decrease was observed in the desiccated samples. The samples with higher levels of UV-absorbing compounds showed higher photochemical efficiency of photosystem II (PS II) during the exposure or subsequent recovering process than samples with low concentration of UV-absorbing compounds, reflecting their protective role. The concentration of these compounds varied in different parts of the thallus, with the middle and marginal parts containing 60–80% more UV-absorbing compounds than the basal parts in both female and male plants. In addition, the marginal parts of male thalli contained more UV-absorbing compounds than the corresponding parts of female thalli. Our data suggest that desiccation plays a key role in this alga to maintain high concentration of UV-absorbing compounds, and that this might provide a beneficial advantage to compete in the intertidal zone where the organism is normally exposed to high levels of UVR.     

Photosynthesis and yield in cotton (Gossypium hirsutum L.) Var. Vikram after exclusion of ambient solar UV-B/A

The impact of ambient solar UV was studied on the photosynthesis and yield of cotton (Gossypium hirsutum) var. Vikram in a field experiment by excluding either UV-B (<315 nm) or UV-B/A (<400 nm) components of solar spectrum. Cotton plants were grown in cages covered with polyester filters that could specifically cut off UV-B or UV-B/A part of the solar spectrum. The control plants were grown under a filter transmissible to UV. Exclusion of UV enhanced plant height, leaf area, total biomass, and the yield parameters (number and weight of bolls, length of fiber and number of seeds) of cotton. Enhancement in the vegetative growth and yield of the plants could be related to enhanced rate of photosynthesis in the leaves. Polyphasic chlorophyll a fluorescence (OJIP) transients from UV excluded plants gave a higher fluorescence yield at I–P phase. Fluorescence measurements indicated enhanced F _v/F _m ratio and reduction capacity after exclusion of solar UV. Exclusion also enhanced stomatal conductance and intercellular CO₂ concentration and reduced the stomatal resistance. Total soluble proteins were higher after UV exclusion, and in SDS–PAGE analysis, bands corresponding to smaller subunits (14 kDa) of Rubisco were more intensely stained. Experiments indicated suppressive action of ambient UV on carbon fixation and yield of cotton plants. Exclusion of solar UV proved to be beneficial in enhancing the yield of cotton plants.     

Comparative genomic study of <Emphasis Type="Italic">spo0E</Emphasis> family genes and elucidation of the role of Spo0E in <Emphasis Type="Italic">Bacillus anthracis</Emphasis>

The propensity of bacterium to sporulate or retain the vegetative form depends on the amount of phosphorylated Spo0A (Spo0A^-P), regulated by Spo0E multigene family of phosphatases (Spo0E, YisI and YnzD). Phylogenetic analysis revealed that Spo0E multigene family of phosphatases (SMFP) descends in two distinct clades of aerobic (Bacillus cluster) and anaerobic (Clostridia cluster) sporulating bacteria. High sequence conservation within species gives a notion that these members could have evolved through lineage and species-specific duplication event. Of the five genes in Bacillus cereus group, three are pathogen specific, and their synteny suggests that these paralogs could be involved in the regulation of amino acid metabolism and its transport. Overexpression of B. subtilis Spo0E, an ortholog of SMFP members in B. anthracis (BAS1251), resulted in sporulation deficient phenotype in B. anthracis. B. anthracis Spo0A^-P binds to a consensus DNA sequence 5′-TGNCGAA-3′ (‘0A-like box’) and loses its DNA binding ability following treatment with B. subtilis Spo0E. Thus, B. subtilis Spo0E acts on B. anthracis Spo0A^-P and, therefore could complement the function of BAS1251. Further, since ‘0A-like box’ are present in the promoter region of abrB gene, a known regulator of anthrax toxin gene expression, cross talk among SMFP members and Spo0A^-P–AbrB could regulate the expression of anthrax toxin genes. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Water- and temperature-dependence of DNA damage and repair in the fruticose lichen Cladonia arbuscula ssp. mitis exposed to UV-B radiation

The induction of cyclobutane pyrimidine dimers (CPDs) by ultraviolet‐B radiation (UV‐B, 280–315 nm) and repair mechanisms were studied in the lichen Cladonia arbuscula ssp. mitis exposed to different temperatures and water status conditions. In addition, the development and repair of CPDs were studied in relation to the different developmental stages of the lichen thallus podetial branches. Air‐dried lichen thalli exposed to UV‐B radiation combined with relatively high visible light (HL, 800 μmol m^?2 s^?1; 400–700 nm) for 7 days showed a progressive increase of CPDs with no substantial repair, although HL was present during and after irradiation with UV‐B. Fully hydrated lichen thalli, that had not been previously exposed to UV‐B radiation for 7 days, were given short‐term UV‐B radiation treatment at 25°C, and accumulated DNA lesions in the form of CPDs, with repair occurring when they were exposed to photoreactivating conditions (2 h of 300 μmol m^?2 s^?1, 400–700 nm). A different pattern was observed when fully hydrated thalli were exposed to short‐term UV‐B radiation at 2°C, in comparison with exposure at 25°C. High levels of CPDs were induced at 2°C under UV‐B irradiation, without significant repair under subsequent photoreactivating light. Likewise, when PAR (300 μmol m^?2 s^?1) and UV‐B radiation were given simultaneously, the CPD levels were not lowered. Throughout all experiments the youngest, less differentiated parts of the lichen thallus – namely ‘tips’, according to our arbitrary subdivision – were the parts showing the highest levels of CPD accumulation and the lowest levels of repair in comparison with the older thallus tissue (‘stems’). Thus the experiments showed that Cladonia arbuscula ssp. mitis is sensitive to UV‐B irradiation in the air‐dried state and is not able to completely repair the damage caused by the radiation. Furthermore, temperature plays a role in the DNA damage repairing capacity of this lichen, since even when fully hydrated, C. arbuscula ssp. mitis did not repair DNA damage at the low temperatures.     

In vitro and in vivo responses of fungal biocontrol agents to gradient doses of UV-B and UV-A irradiation

Conidia of Beauveria bassiana and Metarhizium spp. smeared on glass slides were assayed for their responses to irradiation with weighted 312-nm UV-B and 365-nm UV-A at gradient doses of 0.005–1.1 and 1.0–18.0 J cm⁻², respectively. All inverted, sigmoid dose–survival trends showed good fit to a survival model (r ² ≥ 0.97), yielding respective UV-B LD₅₀s of 0.23–0.59 and 0.05–0.65 J cm⁻² for 24 B. bassiana and 36 Metarhizium isolates, and UV-A LD₅₀s of 2.78–10.46 J cm⁻² for 24 Metarhizium isolates. Myzus persicae apterae on detached leaves were sprayed with a concentrated spore suspension of B. bassiana or M. anisopliae, followed by exposure to the UV-B doses to cause 10–90% viability losses. These doses caused aphid mortality reductions as expected but affected neither spray-to-death period nor fungal growth on cadavers. The results highlight the merits of using UV-tolerant candidates and photoprotection measures in fungal formulations for pest control.