Effect of solar UV-B radiation on a phyllosphere bacterial community.

The effect of solar UV-B radiation on the population dynamics and composition of the culturable bacterial community from peanut (Arachis hypogeae L.) was examined in field studies using plants grown under UV-B-transmitting (UV-B+) or UV-B-excluding (UV-B-) plastic filters. Our data demonstrate that solar UV-B selection alters phyllosphere bacterial community composition and that UV tolerance is a prevalent phenotype late in the season. The total bacterial population size was not affected by either UV-B treatment. However, isolates from the UV-B+ plots (n = 368) were significantly more UV tolerant than those from the UV-B- (n = 363) plots. UV sensitivity was determined as the minimal inhibitory dose of UV that resulted in an inhibition of growth compared to the growth of a nonirradiated control. The difference in minimal inhibitory doses among bacterial isolates from UV-B+ and UV-B- treatments was mainly partitioned among nonpigmented isolates, with pigmented isolates as a group being characterized as UV tolerant. A large increase in UV tolerance was observed within isolate groups collected late (89 and 96 days after planting) in the season. Identification of 200 late-season isolates indicated that the predominant UV-tolerant members of this group were Bacillus coagulans, Clavibacter michiganensis, and Curtobacterium flaccumfaciens. We selected C. michiganensis as a model UV-tolerant epiphyte to study if cell survival on UV-irradiated peanut leaves was increased relative to UV survival in vitro. The results showed an enhancement in the survival of C. michiganensis G7.1, especially following high UV-C doses (300 and 375 J m(-2)), that was evident between 24 and 96 h after inoculation. A dramatic increase in the in planta/in vitro survival ratio was observed over the entire 96-h experiment period for C. michiganensis T5.1.     

Comparative approach to capture bacterial diversity of coastal waters

Despite the revolutionary advancements in DNA sequencing technology and cultivation techniques, few studies have been done to directly compare these methods. In this study, a 16S rRNA gene-based, integrative approach combining culture-independent techniques with culture-dependent methods was taken to investigate the bacterial community structure of coastal seawater collected from the Yellow Sea, Korea. For culture-independent studies, we used the latest model pyrosequencer, Roche/454 Genome Sequencer FLX Titanium. Pyrosequencing captured a total of 52 phyla including 27 candidate divisions from the water column, whereas the traditional cloning approach captured only 15 phyla including 2 candidate divisions. In addition, of 878 genera retrieved, 92.1 % of the sequences were unique to pyrosequencing. For culture-dependent analysis, plate culturing, plate washing, enrichment, and high-throughput culturing (HTC) methods were applied. Phylogenetic analysis showed that the plate-washing clones formed a cluster devoid of any previously cultured representatives within the family Rhodobacteraceae. One HTC isolate (SF293) fell into the OM182 clade, which was not recovered by other culturing methods described here. By directly comparing the sequences obtained from cultures with those from culture-independent work, we found that only 33% of the culture sequences were identical to those from clone libraries and pyrosequences. This study presents a detailed comparison of common molecular and cultivation techniques available in microbial ecology. As different methods yielded different coverage, we suggest choosing the approach after carefully examining the scientific questions being asked.     

Mixotrophic haptophytes are key bacterial grazers in oligotrophic coastal waters

Grazing rate estimates indicate that approximately half of the bacterivory in oligotrophic oceans is due to mixotrophic flagellates (MFs). However, most estimations have considered algae as a single group. Here we aimed at opening the black-box of the phytoflagellates (PFs) <20 μm. Haptophytes, chlorophytes, cryptophytes and pigmented dinoflagellates were identified using fluorescent in situ hybridization or by standard 4′,6-diamidino-2-phenylindole staining. Their fluctuations in abundance, cell size, biomass and bacterivory rates were measured through an annual cycle in an oligotrophic coastal system. On average, we were able to assign to these groups: 37% of the total pico-PFs and 65% of the nano-PFs composition. Chlorophytes were mostly picoplanktonic and they never ingested fluorescently labeled bacteria. About 50% of the PF <20 μm biomass was represented by mixotrophic algae. Pigmented dinoflagellates were the least abundant group with little impact on bacterioplankton. Cryptophytes were quantitatively important during the coldest periods and explained about 4% of total bacterivory. Haptophytes were the most important mixotrophic group: (i) they were mostly represented by cells 3–5 μm in size present year-round; (ii) cell-specific grazing rates were comparable to those of other bacterivorous non-photosynthetic organisms, regardless of the in situ nutrient availability conditions; (iii) these organisms could acquire a significant portion of their carbon by ingesting bacteria; and (iv) haptophytes explained on average 40% of the bacterivory exerted by MFs and were responsible for 9–27% of total bacterivory at this site. Our results, when considered alongside the widespread distribution of haptophytes in the ocean, indicate that they have a key role as bacterivores in marine ecosystems.     

Nitrogen metabolism by heterotrophic bacterial assemblages in Antarctic coastal waters

Field studies to examine the in situ assimilation and production of ammonium (NH₄ ⁺) by bacterial assemblages were conducted in the northern Gerlache Strait region of the Antarctic Peninsula. Short term incubations of surface waters containing ¹⁵N-NH₄ ⁺ as a tracer showed the bacterial population taking up 0.041–0.128 g-atoms Nl^–1d^–1, which was 8–25% of total NH₄ ⁺ uptake rates. The large bacterial uptake of NH₄ ⁺ occurred even at low bacterial abundance during a rich phytoplankton bloom. Estimates of bacterial production using ³H-leucine and -adenine were l.0gCl^–1 d^–1 before the bloom and 16.2 g Cl^–1 d^–1 at the bloom peak. After converting bacterial carbon production to an estimate of nitrogen demand, NH₄ ⁺ was found to supply 35–60% of bacterial nitrogen requirements. Bacterial nitrogen demand was also supported by dissolved organic nitrogen, generally in the form of amino acids. It was estimated, however, that 20–50% of the total amino acids taken up were mineralized to NH₄ ⁺. Bacterial production of NH₄ ⁺ was occurring simultaneously to its uptake and contributed 27–55% of total regenerated NH₄ ⁺ in surface waters. Using a variety of ¹⁵N-labelled amino acids it was found that the bacteria metabolized each amino acid differently. With their large mineralization of amino acids and their relatively low sinking rates, bacteria appear to be responsible for a large portion of organic matter recycling in the upper surface waters of the coastal Antarctic ecosystem.     

Effect of natural sunlight on bacterial activity and differential sensitivity of natural bacterioplankton groups in northwestern Mediterranean coastal waters

We studied the effects of natural sunlight on heterotrophic marine bacterioplankton in short-term experiments. We used a single-cell level approach involving flow cytometry combined with physiological probes and microautoradiography to determine sunlight effects on the activity and integrity of the cells. After 4 h of sunlight exposure, most bacterial cells maintained membrane integrity and viability as assessed by the simultaneous staining with propidium iodide and SYBR green I. In contrast, a significant inhibition of heterotrophic bacterial activity was detected, measured by 5-cyano-2,3 ditolyl tetrazolium chloride reduction and leucine incorporation. We applied microautoradiography combined with catalyzed reporter deposition-fluorescence in situ hybridization to test the sensitivity of the different bacterial groups naturally occurring in the Northwestern Mediterranean to sunlight. Members of the Gammaproteobacteria and Bacteroidetes groups appeared to be highly resistant to solar radiation, with small changes in activity after exposure. On the contrary, Alphaproteobacteria bacteria were more sensitive to radiation as measured by the cell-specific incorporation of labeled amino acids, leucine, and ATP. Within Alphaproteobacteria, bacteria belonging to the Roseobacter group showed higher resistance than members of the SAR11 cluster. The activity of Roseobacter was stimulated by exposure to photosynthetic available radiation compared to the dark treatment. Our results suggest that UV radiation can significantly affect the in situ single-cell activity of bacterioplankton and that naturally dominating phylogenetic bacterial groups have different sensitivity to natural levels of incident solar radiation.     

Increased antioxidant activity in Cassia seedlings under UV-B radiation

Cassia auriculata L. seedlings were irradiated with ultraviolet B (UV-B) in an environment-control chamber. The two doses assayed (7.5 and 15.0 kJ m⁻²) induced oxidative damage with an increase in lipid peroxidation and hydrogen peroxide and a decrease in chlorophyll and total phenol contents. The ascorbate and dehydroascorbate content as well as the reduced glutathione/oxidized glutathione content and ratio were significantly increased. The UV-B stress led to significant increases of the activity of superoxide dismutase, catalase, peroxidase and polyphenol oxidase. It is suggested that Cassia seedlings try to counteract high concentrations of oxygen species produced under UV-B stress through a co-ordinated increase in the contents and activities of antioxidants involved in their detoxification.     

增强UV-B辐射对小麦叶肉细胞原生质体微丝骨架的影响

以增强UV-B（10.08 kJ/m2.d）辐射后的小麦幼苗叶肉细胞原生质体为材料,异硫氰酸荧光素标记的鬼笔环肽（FITC-Ph）为探针,利用激光共聚焦扫描显微镜,观察分析小麦叶肉细胞原生质体中微丝骨架的分布及形态变化。结果表明对照组中,叶肉细胞原生质体微丝呈现网状或平行状随机排列,形态上表现为纤维状结构。增强UV-B辐射处理后,原生质体中微丝的密集分布遭到破坏,纤丝状微丝消失,聚集成束,或呈点状、碎片状分布。     

Influence of supplemental UV-B radiation on photosynthetic characteristics of rice plants

In a field experiment with rice (Oryza sativa L. cv. Saket 4) grown under ambient and supplemental ultraviolet-B (UV-B) radiation at 20 % ozone depletion, differences in gas exchange, concentrations of photosynthetic pigments, anthocyanins and flavonoids, biomass accumulation, catalase and peroxidase activities, and contents of ascorbic acid and phenol were determined. Decline in photosynthesis was associated with reductions in stomatal conductance and concentrations of photosynthetic pigments. Enhanced UV-B radiation (eUV-B) increased the contents of flavonoid and phenolic compounds in leaves. Peroxidase activity increased and catalase activity was always lower at eUV-B. The total plant biomass decreased at eUV-B.     

Influence of supplemental UV-B radiation on photosynthetic characteristics of rice plants

In a field experiment with rice (Oryza sativa L. cv. Saket 4) grown under ambient and supplemental ultraviolet-B (UV-B) radiation at 20 % ozone depletion, differences in gas exchange, concentrations of photosynthetic pigments, anthocyanins and flavonoids, biomass accumulation, catalase and peroxidase activities, and contents of ascorbic acid and phenol were determined. Decline in photosynthesis was associated with reductions in stomatal conductance and concentrations of photosynthetic pigments. Enhanced UV-B radiation (eUV-B) increased the contents of flavonoid and phenolic compounds in leaves. Peroxidase activity increased and catalase activity was always lower at eUV-B. The total plant biomass decreased at eUV-B.     

增强UV-B辐射对小麦叶肉细胞核内F-actin的影响

以增强紫外线B(UV-B)处理的小麦幼苗叶肉细胞核为材料,以异硫氰酸荧光素标记鬼笔环肽(FITC-Ph)荧光标记后,利用流式细胞仪以及激光共聚焦显微镜,对核内肌动蛋白丝(F-actin)进行研究。结果表明:小麦细胞核内确实存在F-actin; 以FITC-Ph标记后用流式细胞仪检测显示,以10.08 kJ·m^-2·d^-1增强的UV-B处理后的小麦叶片细胞核内F-actin的含量升高; 激光共聚焦技术显示细胞核内F-actin的分布,发现小麦叶肉细胞核核仁处F-actin含量较高。推测可能是增强UV-B辐射导致细胞质内F-actin骨架断裂进入细胞核。究其具体作用还待进一步研究,该研究推测其与“分束分裂”有关。     

Inhibition of photosynthetic activity by UV-B radiation in radish seedlings

Inhibition of primary photosynthetic reactions by UV-B radiation (280 nm-320 nm) was demonstrated in radish leaves ( Raphanus sativus cv. Saxa Treib). Detached radish cotyledons from 10-day-old seedlings were irradiated with continuous white light and increasing UV-B irradiances using cut-off filters with increasing transmission for shorter wavelengths (WG 360, WG 345, WG 320, WG 305, WG 295, WG 280). Photosynthetic activity measured in terms of chlorophyll fluorescence induction (Kautsky effect) after 2, 4, 6, 8 and 24 h irradiation decreased in a wavelength dependent way with increasing UV-B irradiance and irradiation time.
Radish seedlings grown for 10 days from the time of germination under the same UV-B irradiation conditions exhibited similar reductions of the variable fluorescence as detached cotyledons irradiated for short time periods. They additionally had lower initial fluorescence at high UV-B radiation levels, although the chlorophyll content per leaf area increased. In contrast to short term experiments, the plastoquinone and flavonoid content increased with increasing UV-B irradiance when based on leaf area.     

Kinetic modulation of bacterial hydrolases by microbial community structure in coastal waters

In this study, we hypothesized that shifts in the kinetic parameters of extracellular hydrolytic enzymes may occur as a consequence of seasonal environmental disturbances and would reflect the level of adaptation of the bacterial community to the organic matter of the ecosystem. We measured the activities of enzymes that play a key role in the bacterial growth (leucine aminopeptidase, β- and α-glucosidases) in surface coastal waters of the Eastern Cantabrian Sea and determined their kinetic parameters by computing kinetic models of distinct complexity. Our results revealed the existence of two clearly distinct enzymatic systems operating at different substrate concentrations: a high-affinity system prevailing at low substrate concentrations and a low-affinity system characteristic of high substrate concentrations. These findings could be the result of distinct functional bacterial assemblages growing concurrently under sharp gradients of high-molecular-weight compounds. We constructed an ecological network based on contemporaneous and time-delayed correlations to explore the associations between the kinetic parameters and the environmental variables. The analysis revealed that the recurring phytoplankton blooms registered throughout the seasonal cycle trigger the wax and wane of those members of the bacterial community able to synthesize and secrete specific enzymes.     

Influence of UV-B radiation on young triticale plants with different wax cover

Ultraviolet-B radiation (biologically effective dose 2.6 kJ m⁻² d⁻¹) had negative influence on morphology and physiology of the young triticale plants. Plants exposed to UV-B were of lower height than control plants, their leaves were narrow, and the rate of net photosynthesis was decreased. The line RAH 336, which wax cover is lesser than that of traditional cultivar Magnat, was more susceptible to UV-B radiation, considering primary photosynthesis reactions, recorded by chlorophyll a fluorescence. An activation of protective mechanisms was observed: plants responded to UV-B by an increase of the content of UV-B absorbing compounds, and changes of antioxidant enzyme activities.     

The bacterial pollution of coastal waters of the southern suburbs of Tunis

The authors present in this paper a study of the bacterian pollution of inshored waters of southern suburbs of Tunis, on the basis of 180 samples collected in 15 different stations, 15 monthly (one for each station). Two districts have been considered: Ez-Zahra and Hammam-Lif. The two districts are submitted to an important bacterian pollution, mainly due to human factor but also to geographical and geomorphological data, to physico-chemical parameters and to some biological characters.     

Oil-oxidizing activity of bacteria isolated from south Sakhalin coastal waters

Ability of bacteria isolated from the southern coastal waters of the Sakhalin Island to degrade various hydrocarbons was studied. The population of marine microorganisms grown on oil was heterogeneous in terms of hydrocarbon degradation. The rate of bacterial degradation of oil hydrocarbons was shown to correlate with their growth rate on the model medium. The degradation rates were higher for aromatic hydrocarbons than for alkanes. Based on our data, the studied bacteria were conditionally assigned to three groups: active, intermediately active, and passive degraders. Ability to oxidize oil was previously not reported for members of the genus Cobetia.     

Influence of microwave radiation on bacterial community structure in biofilm

Organic matter exposed to microwave radiation triggers standard thermal effects as well as a range of so called non-thermal effects. The present work examined the non-thermal effects of microwave radiation on the biofilm in bioreactors with immobilised biomass. Microwave-exposed and conventionally heated reactors were used and both groups of reactors operated on analogous technological parameters. The analysis of the treated sewage demonstrated a significant increase in nitrification and denitrification efficiency in the bioreactors treated with microwave radiation. An analysis of bacteria diversity based on DGGE method showed significantly different bacterial communities developed in the reactors exposed to the microwave radiation in comparison to the control reactors. Moreover, bacterial richness measured by Shannon index was significantly higher in the microwave treated samples (Mann–Whitney's test, p < 0.05). These findings indicate that microwave radiation can affect the structure and function of bacterial communities independent of thermal effects.     

Diurnal variation of cell proliferation in three bacterial taxa from coastal North Sea waters

Pulse-labeling with bromodeoxyuridine (BrdU) in combination with fluorescence in situ hybridization was applied to quantify the percentage of proliferating cells in coastal North Sea waters. In order to assess diurnal variability, we sampled eight or nine times, respectively, within 3 consecutive days at two seasons. Bacteria affiliated with the Roseobacter, SAR86, and NOR5 lineages constituted on average 19% +/- 3%, 8% +/- 2%, and 6% +/- 1% of all cells in May 2002 and 17% +/- 3%, 10% +/- 2%, and 11% +/- 3% in August. The relative abundances of the three populations either remained stable, or they changed very gradually during the observation periods. On average, 38 and 39% of all Bacteria exhibited DNA de novo synthesis in May and August, respectively. The fractions of proliferating cells in bacteria of the SAR86 (May, 59%; August, 72%) and the Roseobacter (48 and 53%) lineages were significantly above the community average. A substantial cell proliferation of population NOR5 (34%) was only encountered in August, concomitant with a dinoflagellate bloom. Significant short-term fluctuations of DNA-synthesizing cells were observed in Roseobacter during May and in NOR5 during August, hinting at a pronounced (temporal or spatial) mesoscale patchiness of growth rates in these populations. Since the BrdU proliferation assay is susceptible to misinterpretation, we also modeled the expected number of labeled cells at increasing BrdU incubation times in a slowly growing bacterial population. We suggest that the absence of visible DNA synthesis in marine bacterioplankton cells after DNA pulse-labeling must not be interpreted as an indication of cell "inactivity."