LOW MOLECULAR WEIGHT CARBOHYDRATE PATTERNS IN THE BANGIOPHYCEAE (RHODOPHYTA)

The qualitative and quantitative occurrence of low molecular weight carbohydrates (LMWCs) in the Bangiophyceae is surveyed. Members of the orders Erythropeltidales ( Sahlingia and Erythrotrichia ), Compsopogonales ( Compsopogon ), and Bangiales ( Bangia and Porphyra ) all contain floridoside and, in some cases, D- and L-isofloridoside, making the group chemotaxonomically the most homogeneous; L-isofloridoside occurred only in the Bangiales. The Porphyridiales showed great variation in carbohydrate types: Chroodactylon contained only sorbitol, Porphyridium contained only floridoside, Dixoniella and Rhodella species exhibited only mannitol, and Rhodosorus showed digeneaside and sorbitol, whereas Stylonema contained floridoside, D-isofloridoside, digeneaside, and sorbitol. The extensive variation in LMWC composition within and between the different orders suggests that the enzyme systems for each might have developed repeatedly in different Bangiophyceae and Florideophyceae. Nonetheless, a phylogenetic tree based on small-subunit ribosomal RNA supports the LMWC patterns in the Bangiophyceae.     

SYNTHESIS OF MYCOSPORINE-LIKE AMINO ACIDS IN CHONDRUS CRISPUS (FLORIDEOPHYCEAE) AND THE CONSEQUENCES FOR SENSITIVITY TO ULTRAVIOLET B RADIATION

The induction and protective role of the UV-absorbing compounds known as mycosporine-like amino acids (MAAs) were examined in sublittoral Chondrus crispus Stackh. transplanted for 2 weeks in the spring and summer to shallow water under three irradiance conditions: PAR (photosynthetically active radiation; 400–700 nm), PAR + UVA (PAR + 320– 400 nm), PAR + UVA + UVB (PAR + UVA + 280– 320 nm). Sublittoral thalli collected around Helgoland, North Sea, Germany, from 6 m below the mean low water of spring tides contained less than 0.1 mg·g⁻¹ dry weight (DW) total MAAs, whereas eulittoral samples contained over 1 mg·g⁻¹ DW. Transplantation to shallow water led to the immediate synthesis of three MAAs in the following temporal order: shinorine (λ_max 334 nm), asterina (λ_max 330 nm), and palythine (λ_max 320 nm), with the shinorine content peaking and then declining after 2 days (exposure to 100 mol photons·m⁻²). Maximum total MAA content (2 mg·g⁻¹ DW) also occurred after 2 days of induction, exceeding the content normally found in eulittoral samples. Furthermore, the relative proportion of the different MAAs at this time was different than that in eulittoral samples. After 2 days the total content declined to the eulittoral value, with palythine as the principal MAA. Similar data were obtained for all treatments, indicating that MAA synthesis in C. crispus was induced by PAR and not especially stimulated by UV radiation. The ability of photosystem II (PSII) to resist damage by UVB was tested periodically during the acclimation period by exposing samples to a defined UVB dose in the lab. Changes in chlorophyll fluorescence (F_v/F_m and effective quantum yield, φ_II) indicated that PSII function was inhibited during the initial stage of acclimation but gradually improved with time. No difference among screening treatments was detected except in spring for the samples acclimating to PAR + UVA + UVB. In this treatment F_v/F_m and φ_II were significantly lower than in the other treatments. During the first week of each experiment, growth rates were also significantly reduced by UVB. The reductions occurred despite maximum MAA content, indicating an incomplete protection of photosynthetic and growth-related processes.     

The effects of ultraviolet radiation on photosynthetic performance,growth and sunscreen compounds in aeroterrestrial biofilm algae isolated from building facades

The effects of artificial ultraviolet radiation [UVR; 8 W m⁻² ultraviolet-A (UVA), 0.4 W m⁻² ultraviolet-B (UVB)] on photosynthetic performance, growth and the capability to synthesise mycosporine-like amino acids (MAAs) was investigated in the aeroterrestrial green algae Stichococcus sp. and Chlorella luteoviridis forming biofilms on building facades, and compared with the responses of two green algae, from soil (Myrmecia incisa) and brackish water (Desmodesmus subspicatus). All species exhibited decreasing quantum efficiency (F _v/F _m) after 1–3 days exposure to UVR. After 8–12 days treatment, however, all aeroterrestrial isolates exhibited full recovery under UVA and UVA/B. In contrast, D. subspicatus showed only 80% recovery after treatment with UVB. While Stichococcus sp. and C. luteoviridis exhibited a broad tolerance in growth under all radiation conditions tested, M. incisa showed a significant decrease in growth rate after exposure to UVA and UVA/B. Similarly D. subspicatus grew with a reduced rate under UVA, but UVA/B led to full inhibition. Using HPLC, an UV-absorbing MAA (324 nm-MAA) was identified in Stichococcus sp. and C. luteoviridis. While M. incisa contained a specific 322 nm-MAA, D. subspicatus lacked any trace of such compounds. UV-exposure experiments indicated that all MAA-containing species are capable of synthesizing and accumulating these compounds, thus supporting their function as an UV-sunscreen. All data well explain the conspicuous ecological success of aeroterrestrial green algae in biofilms on facades. Biosynthesis and accumulation of MAAs under UVR seem to result in a reduced UV-sensitivity of growth and photosynthesis, which consequently may enhance survival in the environmentally harsh habitat.     

Assessment of oxidative stress in the planktonic diatom Thalassiosira pseudonana in response to UVA and UVB radiation

Induction of oxidative stress by UVA and UVB in the diatom Thalassiosirapseudonana was experimentally studied. Cells, pre-grown in alight-limited continuous culture, were incubated for 4 h at175 µmol m^-2s^-1photosynthetically active radiation, withoptional supplementary UVA at an unweighted dose rate of 0.70W m^-2, or 2.79 W m^-2UVA plus 0.45 W m^-2UVB (unweighted). A fluorescence-basedmeasure of photosynthetic efficiency (F_v/F_m) decreased from0.69 to 0.58 in the presence of UVB, whereas UVA caused a minordecrease of F_v/F_m. Quantitative analysis of confocal imagesshowed a minor increase of active oxygen production associatedwith supplemental UVA alone, and a 100% increase with additionalUVB. Cellular malondialdehyde, an indicator of lipid peroxidationby active oxygen, almost doubled under UVA and increased three-foldwith additional UVB. Activities of superoxide dismutase (scavengingactive oxygen) and glutathione reductase (reducing GSSG to GSH)increased in response to UVB exposure, whereas ascorbate peroxidaseactivities did not. UVB caused a minor decrease in the glutathioneratio GSH : (GSH + 0.5GSSG), which indicates a moderate oxidativestress.     

Effects of temperature and UV radiation increases on the photosynthetic efficiency in four scleractinian coral species

Experiments were performed on coral species containing clade A (Stylophora pistillata, Montipora aequituberculata) or clade C (Acropora sp., Pavona cactus) zooxanthellae. The photosynthetic efficiency (F(v)/F(m)) of the corals was first assessed during a short-term increase in temperature (from 27 degrees C to 29 degrees C, 32 degrees C, and 34 degrees C) and acute exposure to UV radiation (20.5 W m(-2) UVA and 1.2 W m(-2) UVB) alone or in combination. Increasing temperature to 34 degrees C significantly decreased the F(v)/F(m) in S. pistillata and M. aequituberculata. Increased UV radiation alone significantly decreased the F(v)/F(m) of all coral species, even at 27 degrees C. There was a combined effect of temperature and UV radiation, which reduced F(v)/F(m) in all corals by 25% to 40%. During a long-term exposure to UV radiation (17 days) the F(v)/F(m) was significantly reduced after 3 days' exposure in all species, which did not recover their initial values, even after 17 days. By this time, all corals had synthesized mycosporine-like amino acids (MAAs). The concentration and diversity of MAAs differed among species, being higher for corals containing clade A zooxanthellae. Prolonged exposure to UV radiation at the nonstressful temperature of 27 degrees C conferred protection against independent, thermally induced photoinhibition in all four species.     

Floridoside,L-Isofloridoside,and D-Isofloridoside in the Red Alga Porphyra columbina (Seasonal and Osmotic Effects)

The quantitative heteroside distribution in Porphyra columbina Montagne and Bangia atropurpurea (Roth) C. Agardh (Bangiales, Rhodophyta) has been measured using 13C-nuclear magnetic resonance spectroscopy and gas-liquid chromatography. In P. columbina, floridoside and both D- and L-isofloridoside were recorded, with concentrations of L-isofloridoside exceeding those of floridoside. All three compounds were also measured in B. atropurpurea. Marked changes in the relative amounts of the heterosides were recorded throughout the season. The role of L-isofloridoside in the osmotic acclimation of P. columbina has been demonstrated.     

Mycosporine-like amino acid (MAAs) production by Heterocapasa sp. (Dinophyceae) in indoor cultures

The possibility of using mycosporine-like amino acids (MAAs), with an apparent sunscreen function in nature, as ultraviolet radiation (UVR) blockers to prevent skin injury has been raised by diverse authors. Production of MAAs by the dinoflagellate Heterocapsa sp. (Dinophyceae) is shown here. Three major peaks with absorption maxima at 330.8, 332.0 and 333.2 nm were detected by high performance liquid chromatography (HPLC) analysis of methanolic extracts in all tested conditions. Analysis of crude extract by mass spectroscopy with electrospray ionization (MS-EI) showed a set of molecular ions ([M+H](+)) with main peaks being at m/z 242.4, 288.4, 303.3 and 333.3 u.m.a. According to these data, along with retention times, the MAA profile of Heterocapsa sp. is assumed to be composed of shinorine (lambda(max)=334 nm), mycosporine-2-glycine (lambda(max)=331 nm) and palythinol (lambda(max)=332 nm). A constitutive MAA content of about 4 microg (10(6) cells)(-1) was measured under exposure to PAR only. A maximal accumulation of MAA per culture volume of 1.1 mg l(-1) was obtained after 72 h of exposure to PAR+UVA, while the highest production rate (0.025 mg l(-1) h(-1)) was computed after 24 h of exposure to PAR+UVA+UVB.     

VARIATIONS IN FLORIDOSIDE CONTENT AND FLORIDOSIDE PHOSPHATE SYNTHASE ACTIVITY IN PORPHYRA PERFORATA (RHODOPHYTA)1

The diurnal and seasonal variations in floridoside content and floridoside phosphate synthase (FPS) activity were measured in samples of Porphyra perforata J. Ag. growing in the field. Floridoside content generally increased about fourfold from early morning to the middle of the day and then dropped gradually in the afternoon and the evening. On a monthly basis, there was a steady increase in floridoside content from February to May. A similar trend in monthly increase in FPS activity was also observed from February to April. The level of FPS activity varied with the time of day. The highest activity was observed early in the morning shortly after dawn; subsequently, it decreased, reaching about 50% of the peak value late in the afternoon/evening. On a daily as well as seasonal basis, a change in FPS activity correlated with a change in floridoside content. Among environmental factors, floridoside content and FPS activity showed a positive correlation with daylength and temperature on a seasonal basis. However, on a daily basis, salinity and water temperature did not seem to affect the floridoside content or the FPS activity.     

Reduction of Solar UV Radiation Due to Urban High-Rise Buildings – A Coupled Modelling Study

Solar UV radiation has both adverse and beneficial effects to human health. Using models (a radiative transfer model coupled to a building shading model), together with satellite and surface measurements, we studied the un-obstructed and obstructed UV environments in a sub-tropical urban environment featured with relatively high pollution (aerosol) loadings and high-rise buildings. Seasonal patterns of the erythemal UV exposure rates were governed by solar zenith angles, seasonal variations of aerosol loadings and cloud effects. The radiative transfer modelling results agreed with measurements of erythemal UV exposure rates and spectral irradiances in UVA and UVB ranges. High-rise buildings and narrow road width (height to width, H/W, ratios up to 15) reduced the modelled total UV (UVA+UVB) radiation and leave 10% of the un-obstructed exposure rate at ground-level at noon. No more than 80% of the un-obstructed exposure rate was received in the open area surrounded by 20-storey buildings. Our modelled reduction of UVB radiation in the urban environment was consistent with similar measurements obtained for Australia. However, our results in more extreme environments (higher H/W ratios) were for the first time reported, with 18% of the un-obstructed exposure rate remained at the ground-level center of the street canyon.     

Sources of mycosporine-like amino acids in planktonic Chlorella-bearing ciliates (Ciliophora)

1. Mycosporine‐like amino acids (MAAs) are a family of secondary metabolites known to protect organisms exposed to solar UV radiation. We tested their distribution among several planktonic ciliates bearing Chlorella isolated from an oligo‐mesotrophic lake in Tyrol, Austria. In order to test the origin of these compounds, the MAAs were assessed by high performance liquid chromatography in both the ciliates and their symbiotic algae. 2. Considering all Chlorella‐bearing ciliates, we found: (i) seven different MAAs (mycosporine‐glycine, palythine, asterina‐330, shinorine, porphyra‐334, usujirene, palythene); (ii) one to several MAAs per species and (iii) qualitative and quantitative seasonal changes in the MAAs (e.g. in Pelagodileptus trachelioides). In all species tested, concentrations of MAAs were always <1% of ciliate dry weight. 3. Several MAAs were also identified in the Chlorella isolated from the ciliates, thus providing initial evidence for their symbiotic origin. In Uroleptus sp., however, we found evidence for a dietary source of MAAs. 4. Our results suggest that accumulation of MAAs in Chlorella‐bearing ciliates represents an additional benefit of this symbiosis and an adaptation for survival in sunlit, UV‐exposed waters.     

MAA Synthesis and Accumulation in Polar Macroalgae are Controlled by Abiotic Factors

Mycosporine‐like amino acids (MAAs) are regarded as powerful sunscreens protecting the algae against harmful UV radiation. The MAA protection efficiency was tested in algal samples by measuring the optimum quantum yield of photosynthesis using photosystem II fluorescence. It could be demonstrated that the recovery of photosynthesis after exposure to enhanced UV radiation is faster in individuals with high MAA content. MAAs can be synthesized in several polar macroalgae in response to different radiation conditions. Although MAA induction patterns are very species‐specific, some similarities can be found. Field studies indicate that plants from different growth habitats providing distinct radiation climate can be grouped into three physiological categories depending on their MAA content. The first group (I) includes mainly deep‐water species, typically lacking MAAs. The second group (II), algal species found in a broad range of water depths (eu‐ and sublittoral), which are able to flexibly synthesize and accumulate MAAs. The third group (III) includes supra‐ and eulittoral taxa, which always contain high MAA concentrations. In laboratory studies, we showed that taxa of group II and III responded in three different ways based on MAA accumulation when exposed to different radiation conditions (PAR, PAR + UVA, PAR + UVA + UVB). Either they: (a) exhibit highest total MAA concentration under the full artificial spectrum; (b) increase their MAA concentration after exposure to PAR and PAR + UVA or (c) MAA concentration declines after exposure to the full spectrum. Our studies have indicated that when coupled with UVR, exposure to temperature fluctuations ranging from 0 to 10 °C also affect MAA biosynthesis.     

AMMONIUM AND UV RADIATION STIMULATE THE ACCUMULATION OF MYCOSPORINE‐LIKE AMINO ACIDS IN PORPHYRA COLUMBINA (RHODOPHYTA) FROM PATAGONIA,ARGENTINA1

The combined effects of ammonium concentration and UV radiation on the red alga Porphyra columbina (Montagne) from the Patagonian coast (Chubut, Argentina) was determined using short‐term (less than a week) experimentation. Discs of P. columbina were incubated with three ammonium concentrations (0, 50, and 300 μM NH₄Cl) in anilluminated chamber (PAR=300 μmol photons·m^?2·s^?1, UVA=15 W·m^?2, UVB=0.7 W·m^?2) at 15°C. Algae incubated at 300 μM ammonium showed a significant increase (P<0.05) in the concentration of mycosporine‐like amino acids (MAAs) compared with the initial value or with the other ammonium treatments. The increase of MAAs was, however, a function of the quality of irradiance received, with a higher increase in samples exposed to UVA compared with UVB (29% and 5% increase, respectively). However, UVB radiation was more effective in inducing MAA synthesis per unit energy received by the algae. Samples exposed to PAR only had an intermediate increase in MAA concentration of 16%. Chl a concentration decreased through the incubation with the greatest decrease at high ammonium concentration. Phycobiliprotein (BP) decreased through time with the smallest decrease occurring at high ammonium concentration. Photoinhibition (as a decrease of optimal quantum yield) was significantly greater under nitrogen‐deprived conditions than that under replete ammonium levels. Maximal gross photosynthesis (GP_max), as oxygen evolution, and maximal electron transport rate (ETR_max), as chl fluorescence, increased with the ammonium concentration. Positive relationships between maximal GP or ETR and pigment ratios (BP/chl a and MAAs/chl a) and negative relationships with chl a concentration were found.     

Metabolic response to maximal exercise of 800 and 2,000 m in the thoroughbred horse

To define the metabolic response to maximal exercise in the thoroughbred horse under field conditions, muscle biopsies and venous blood samples were taken from five horses after a single 800-m gallop and from four horses after a single 2,000-m gallop. Muscle and blood samples were also collected during 60 min of recovery. After exercise muscle ATP contents were decreased by 30 +/- 7 (SD) and 47 +/- 3% after the 800- and 2,000-m gallops, respectively. As indicators of purine catabolism, ammonia and uric acid increased in plasma, the accumulation being greater after the 2,000-m gallop. Blood ammonia peaked immediately after exercise and uric acid after 40-60 min of recovery. Muscle glycogen utilization over the 800- and 2,000-m gallops averaged 2.68 +/- 0.90 and 1.06 +/- 0.12 mmol glucosyl units.kg dry muscle-1.s-1, respectively, and the total used amounted to 27.3 +/- 6.6 and 32.5 +/- 8.8% of the initial store. Muscle lactate accumulation averaged 123.5 +/- 49.7 and 167.3 +/- 20.7 mmol/kg dry muscle, respectively, and declined during recovery with half times of 22.9 +/- 4.2 and 18.9 +/- 6.6 min. Blood lactate peaked 5-10 min after exercise. Exercise resulted in only a small increase in muscle glycerol content, but this continued to rise during recovery reaching 9-12 mmol/kg dry muscle after 20 min. During this time the increase in muscle glycerol content exactly matched the decline in glycerol 3-phosphate.     

Raman spectroscopic analysis of the effect of ultraviolet irradiation on calf thymus DNA

Raman spectroscopy was used for the first time to detect the effect of independent UVA (ultraviolet-A: 320-400nm) and UVB (ultraviolet-B: 280-320 nm) irradiation on the calf thymus DNA in aqueous solution. After both UVA and UVB irradiation for 1h or 3h, the damage to the conformation of DNA was moderate, but the reduction of the B-form DNA component was obvious. Both UVA and UVB caused significant damage to the deoxyribose moiety and bases, among which the pyrimidine base pairs were more seriously affected. There appeared to be preferential damaging sites on DNA molecules caused by UVA and UVB irradiation. UVA irradiation caused more damage to the deoxyribose than UVB irradiation, while UVB irradiation caused more significant damage to the pyrimidine moiety than UVA irradiation. After UVB irradiation for 3h, unstacking of the AT base pairs and the cytosine ring took place, severe damage to the thymine moiety occurred, and some base pairs were modified. Moreover, with either UVA or UVB irradiation for 3h,the photoreactivation of DNA occurred. The damage to the DNA caused by UVB was immediate, while the damage caused by UVA was proportional to the irradiation duration. The experimental results partly indicate the formation of some cyclobutane pyrimidine dimers and (6-4) photoproducts.     

Separation of floridoside and isofloridosides by HPLC and complete (1)H and (13)C NMR spectral assignments for D-isofloridoside

Isofloridosides (1-O-alpha-D-galactopyranosylglycerol) and floridoside (2-O-alpha-D-galactopyranosylglycerol) were extracted from the red alga Porphyra umbilicalis (Linné) Kützing (Bangiales, Rhodophyta). Their separation was achieved by HPLC (NH(2) P50 column) after successive purification of the crude extract by ion-exchange chromatography and HPLC (Sugar-Pak TM1 column). 1D and 2D NMR spectroscopy experiments allowed to completely assign the (1)H and (13)C spectra of D-isofloridoside.     

Ultraviolet-B Wavelengths Regulate Changes in UV Absorption of Cleaner Fish Labroides dimidiatus Mucus

High-energy wavelengths in the ultraviolet-B (UVB, 280-315 nm) and the UVA (315-400-nm) portion of the spectrum are harmful to terrestrial and aquatic organisms. Interestingly, UVA is also involved in the repair of UV induced damage. Organisms living in shallow coral reef environments possess UV absorbing compounds, such as mycosporine-like amino acids, to protect them from UV radiation. While it has been demonstrated that exposure to UV (280-400 nm) affects the UV absorbance of fish mucus, whether the effects of UV exposure vary between UVB and UVA wavelengths is not known. Therefore, we investigated whether the UVB, UVA, or photosynthetically active radiation (PAR, 400-700 nm) portions of the spectrum affected the UV absorbance of epithelial mucus and Fulton’s body condition index of the cleaner fish Labroides dimidiatus. We also compared field-measured UV absorbance with laboratory based high-performance liquid chromatography measurements of mycosporine-like amino acid concentrations. After 1 week, we found that the UV absorbance of epithelial mucus was higher in the UVB+UVA+PAR treatment compared with the UVA+PAR and PAR only treatments; after 2 and 3 weeks, however, differences between treatments were not detected. After 3 weeks, Fulton’s body condition index was lower for fish in the UVB+UVA+PAR compared with PAR and UVA+PAR treatments; furthermore, all experimentally treated fish had a lower Fulton’s body condition index than did freshly caught fish. Finally, we found a decrease with depth in the UV absorbance of mucus of wild-caught fish. This study suggests that the increase in UV absorbance of fish mucus in response to increased overall UV levels is a function of the UVB portion of the spectrum. This has important implications for the ability of cleaner fish and other fishes to adjust their mucus UV protection in response to variations in environmental UV exposure.     

UV irradiance as a major influence on growth, development and secondary products of commercial importance in Lollo Rosso lettuce ‘Revolution’ grown under polyethylene films

The growth and production of anthocyanin, flavonoid and phenolic compounds were evaluated in Lollo Rosso lettuce ‘Revolution’ grown continuously under films varying in their ability to transmit UV radiation (completely transparent to UV, transparent above 320, 350, 370 and 380 nm and completely opaque to UV radiation). Plants were grown from seed under UV transparent and UV blocking films and destructively harvested 3–4 weeks after transplanting. Plants under a complete UV blocking film (UV400) produced up to 2.2 times more total above ground dry weight than plants under the UV transparent film. In contrast, anthocyanin content in plants under the UV blocking film was approximately eight times lower than in plants under a UV transparent film. Furthermore, there was a curvilinear relationship between the anthocyanin content and UV wavelength cutoff such that above 370 nm there was no further reduction in anthocyanin content. Fluorescence measurements indicated that photosynthetic performance index was 15% higher under the presence of UVB and UVA (UV280) than under the presence of UVA (UV320) and 53% higher than in the absence of UV radiation suggesting protection of the photosynthetic apparatus possibly by phenolic compounds. These findings are of particular importance as the potential of UV transmitting films to increase secondary compounds may offer the opportunity to produce plants commercially with increased health benefits compared to those grown under conventional films.     

Effect of solar ultraviolet radiation on survival of krill larvae and copepods in Antarctic Ocean

The effect of solar ultraviolet radiation on the survival rate of Antarctic zooplankton was examined in February–March in 2002. We investigated survival rate of calyptopis larvae of Euphausia superba and late copepodite stages (IV and V) of large dominant calanoid species, Calanoides acutus and Calanus propinquus reared in quartz jars with three different radiation regimes (total radiation, exclusion of UVB, exclusion of UVA and UVB) and a dark control. The survival rates of the krill larvae decreased after 3 days from start of the experiment, being below 50% at 4 days in the treatments with total radiation and exclusion of UVB, although most individuals could survive until the end of the experiments in the treatments with exclusion of both UVA and UVB and dark control. The calanoid juveniles showed almost same pattern of survival curves as the krill larvae did, but survived slightly longer. Although >10% of surface UVA radiation at 340 and 380 nm penetrated down to 30 m, both C. acutus and C. propinquus were mostly distributed above 20 m. Surface swarm of the krill larvae can be often recognized in the previous studies. These results suggest that not only solar UVB but also UVA radiation potentially lower the survival rate of Antarctic zooplankton at depth less than 20 m.     

Seasonal ectomycorrhizal fungal biomass development on loblolly pine (Pinus taeda L.) seedlings

Ergosterol, a membrane sterol found in fungi but not in plants, was used to estimate live mycelial biomass in ectomycorrhizae. Loblolly pine (Pinus taeda L.) seeds were sown in April 1993 and grown with standard nursery culture practices. Correlations between total seedling ergosterol and visual assessment of mycorrhizal colonization were high during July and August but low as ectomycorrhizal development continued into the growing season. Percentages of mycelial dry weight over lateral roots decreased from 9% in July to 2.5% in November because seedling lateral root dry weight accumulated faster than mycelial dry weight. Total ergosterol per seedling increased from July through February. As lateral root dry weight ceased to increase during winter months, ectomycorrhizal mycelia became the major carbohydrate sink of pine seedlings. No distinctive seasonal pattern of soil ergosterol content was observed. The impact of ectomycorrhizal fungi on plant carbohydrate source-sink dynamics can be quantitatively estimated with ergosterol analysis but not with conventional visual determination.     

Bipyrimidine photoproducts rather than oxidative lesions are the main type of DNA damage involved in the genotoxic effect of solar UVA radiation

Exposure to solar UV radiation gives rise to mutations that may lead to skin cancer. UVA (320-340 nm) constitutes the large majority of solar UV radiation but is less effective than UVB (290-320 nm) at damaging DNA. Although UVA has been implicated in photocarcinogenesis, its contribution to sunlight mutagenesis has not been elucidated, and DNA damage produced by UVA remains poorly characterized. We employed HPLC-MS/MS and alkaline agarose gel electrophoresis in conjunction with the use of specific DNA repair proteins to determine the distribution of the various classes and types of DNA lesions, including bipyrimidine photoproducts, in Chinese hamster ovary cells exposed to pure UVA radiation, as well as UVB and simulated sunlight (lambda > 295 nm) for comparison. At UVA doses compatible with human exposure, oxidative DNA lesions are not the major type of damage induced by UVA. Indeed, single-strand breaks, oxidized pyrimidines, oxidized purines (essentially 8-oxo-7,8-dihydroguanine), and cyclobutane pyrimidine dimers (CPDs) are formed in a 1:1:3:10 ratio. In addition, we demonstrate that, in contrast to UVB and sunlight, UVA generates CPDs with a large predominance of TT CPDs, which strongly suggests that they are formed via a photosensitized triplet energy transfer. Moreover, UVA induces neither (6-4) photoproducts nor their Dewar isomers via direct absorption. We also show that UVA photons contained in sunlight, rather than UVB, are implicated in the photoisomerization of (6-4) photoproducts, a quickly repaired damage, into poorly repaired and highly mutagenic Dewar photoproducts. Altogether, our data shed new light on the deleterious effect of UVA.