Influence of humic, fulvic and hydrophilic acids on the growth, photosynthesis and respiration of the dinoflagellate Prorocentrum minimum (Pavillard) Schiller

Blooms of the dinoflagellate Prorocentrum minimum often occur in coastal regions characterized by variable salinity and elevated concentrations of terrestrially derived dissolved organic carbon (DOC). Humic, fulvic and hydrophilic acid fractions of DOC were isolated from runoff entering lower Narragansett Bay immediately after a rainfall event and the influence of these fractions upon P. minimum growth, cell yield, photosynthesis and respiration was examined. All organic fractions stimulated growth rates and cell yields compared with controls (no organic additions), but the extent of stimulation varied with the fraction and its molecular weight. Greatest stimulations were observed with humic and fulvic acids additions; cell yields were more than 2.5 and 3.5 times higher than with hydrophilic acid additions while growth rates were 21 and 44% higher, respectively. Responses to additions of different molecular weight fractions of each DOC fraction suggest that growth rate effects were attributable to specific molecular weight fractions: the >10,000 fraction of humic acids, both the >10,000 and <500 fractions of fulvic acids and the <10,000 fraction of hydrophilic acids. The form and concentration of nitrogen (as NO₃⁻ or NH₄⁺) present also influenced P. minimum response to DOC; 10–20 μg ml⁻¹ additions of fulvic acid had no effect upon growth rates in the presence of NH₄⁺ but significantly increased growth rates in the presence of NO₃⁻, a relationship probably related to fulvic acid effects upon trace metal bioavailability and subsequent regulation of the biosynthesis of enzymes required for NO₃⁻ assimilation. The influence of DOC additions on P. minimum respiration and production rates also varied with the organic fraction and its concentration. Production rates ranged from 1.1 to 3.4 pg O₂ cell⁻¹ h⁻¹, with highest rates observed upon exposure to fulvic and hydrophilic acid concentrations of >10 μm ml⁻¹. Low concentrations (5–10 μg ml⁻¹) of humic acid had no statistically significant effect upon production, but exposure to concentrations >25 μg ml⁻¹ resulted in a 30% decrease in O₂ evolution, probably due to light attenuation by the highly colored humic acid fraction. Respiration rates ranged from 1.2 to 2.7 pg O₂ cell⁻¹ h⁻¹ and were elevated upon exposure to both fulvic and hydrophilic acids, but not to humic acid. These results demonstrate that terrestrially derived DOC fractions play an active role in stimulation of P. minimum growth via direct effects upon growth, yield and photosynthesis as well as via indirect influences such as interactions with nitrogen and effects upon light attenuation.     

A divalent-ion binding site on the 16-kDa proton channel from Nephrops norvegicus—revealed by EPR spectroscopy

As purified from the hepatopancreas of Nephrops norvegicus, the 16-kDa proton channel proteolipid is found to contain an endogenous divalent ion binding site that is occupied by Cu²⁺. The EPR spectrum has g-values and hyperfine splittings that are characteristic of type 2 Cu²⁺. The copper may be removed by extensive washing with EDTA. Titration with Ni²⁺ then induces spin-spin interactions with nitroxyl spin labels that are attached either to the unique Cys⁵⁴, or to fatty acids intercalated in the membrane. Paramagnetic relaxation enhancement by the fast-relaxing Ni²⁺ is used to characterise the binding and to estimate distances from the dipolar interactions. The Ni²⁺-binding site on the protein is situated around 14-18 Å from the spin label on Cys⁵⁴, and is at a similar distance from a lipid chain spin-labelled on the 5 C-atom, but is more remote from the C-9 and C-14 positions of the lipid chains.     

氮磷限制对锥状斯氏藻孢囊形成的作用

在实验室研究了锥状斯氏藻(Scrippsiellatrochoidea)在N、P单因子营养限制(N:500μg.L-1,P:74～0.74μg.L-1和P:74μg.L-1,N:500～5μg.L-1)条件下的生长和孢囊形成。结果显示N、P限制不利于锥状斯氏藻的快速生长,其中低P对细胞生长的限制作用更显著。其孢囊形成率在15～99%之间,中度N限制能促进孢囊的形成,形成率几乎可达100%。孢囊一般在对数生长期结束、细胞数量达到最大值时开始形成。但由于接种后营养盐浓度的急剧降低,营养极度限制组孢囊可在接种后第1d就开始形成。结果显示稳定生长期孢囊的大量形成大大降低了锥状斯氏藻营养细胞数量,能在一定程度上促进其赤潮的消亡。     

N、P营养盐对塔玛亚历山大藻（Alexandrium tamarense）生长的影响

模拟自然海水营养盐浓度状况,在N、P浓度分别为10-500μg L-1 N和0.74-74μg L-1 P时,研究N、P双因子限制(N、P浓度同时降低,N:P固定为15:1)及单因子限制(保持N或P为最高浓度,只降低一种营养盐浓度)对有毒赤潮藻塔玛亚历山大藻(Alexandrium tamarense)生长的影响。结果表明,塔玛亚历山大藻细胞能较快进入对数生长期,但N、P双因子限制能明显影响其生长,在N、P浓度分别低于100μg L-1 N和15μg L-1 P时,细胞密度无明显增长;而N或P分别受限时,生长态势明显优于N、P同时受到限制的试验组,而且N、P单因子中度限制对生长影响较小。结果说明塔玛亚历山大藻对单因子营养元素限制较强的适应能力,可使其在常常出现单营养因子限制的自然水体中维持一定生长速率和细胞密度,并有助于滤食该藻的贝类体内麻痹性贝类毒素的积累。     

Simultaneous removal of carbon and nitrogen from municipal-type synthetic wastewater using net-like rotating biological contactor (NRBC)

The treatment of municipal-type synthetic wastewater was carried out using a three stages net-like rotating biological contactor (NRBC). The results indicated that, compared with conventional rotating biological contactor (RBC), NRBC have several advantages, such as quick start-up, high biomass concentration and can handle high organic loading rates. The COD and total nitrogen removal rates achieved were 78.8–89.7% and 40.2–61.4%, respectively, in aerobic treatment of low COD municipal-type wastewater at hydraulic retention times (HRT) from 5 to 9 h. The COD removal rate achieved 80–95% when organic loading varied between 16 and 40 gCOD/m² d. A large amount of nematodes were found in the NRBC system, which made the NRBC system produce relatively low amounts of waste sludge, due to their grazing.     

Contrasted Responses of Two Understorey Species to Direct and Indirect Effects of a Canopy Gap

Positive associations between adult trees and understorey species have been explained either by direct or indirect facilitation. We tested both models by comparing the performance of two understorey species with contrasted stress-tolerance abilities Galium odoratum and Deschampsia flexuosa. Individuals of both species were transplanted in the four combinations of two treatments (gap and removal of an herbaceous competitor, Molinia caerulea). Our experiment demonstrated that direct facilitation of adult trees may explain the restricted occurrence of the shade-demanding Galium within closed forest communities. In contrast, the shade-tolerant Deschampsia was subjected to additional competition within the forest, likely because adult trees had a higher negative effect on light availability and a similar negative effect on nitrogen availability within the forest than did Molinia in the gaps.     

Root responses to nutrient patches in grassland and forest

Differences between growth forms in root responses to experimentally created heterogeneity have been documented in many greenhouse and plot studies, but not in natural vegetation. Here we examined the response of roots to experimental nutrient patches in undisturbed grassland and forest at the northern edge of the North American Great Plains. Forest vegetation increases the spatial heterogeneity of soil resources, and we tested for differences between forest and grassland roots in response to patches. Ten minirhizotrons (clear tubes, 5 cm diameter, 180 cm long) were installed in both grassland and forest 3 years before the experiment. Minirhizotrons ran horizontally 10 cm beneath the soil surface. Patches of available nitrogen (N) were created over the tubes, using three concentrations (0, 3, 15 g N m⁻² yr⁻¹) and two patch sizes (1␣and 10 cm²). Root images were collected beneath patches over the course of a growing season. Root length was significantly greater in grassland than forest at the start and end of the growing season, but did not respond to N patches. Root production was also significantly greater in grassland than forest, and was significantly greater (about 20%) in high-N patches than in unfertilized patches. This increase, however, did not differ between vegetation types. Turnover did not vary with any treatment, and patch size had no effect on any response variable. Overall, differences caused by experimental patches were much smaller than differences between habitats, and did not vary between habitats. Realistic levels of experimentally imposed hetereogeneity in established vegetation may not be much greater than background levels, and field vegetation has extant root systems which respond to patches via uptake instead of growth. Both mechanisms should contribute to less root proliferation in field experiments than in greenhouse experiments.     

Grimmia homodictyon Dixon

Abstract

We examined photosynthetic responses of two dominant pleurocarpous mosses, Actinothuidium hookeri (Mitt.) Broth. and Hylocomium splendens (Hedw.) Schimp. to low-level nitrogen (N) addition. The study was conducted in an old-growth fir forest on the eastern Tibetan Plateau. The added N, 1 g N/m², was mainly absorbed by the new-growth. The concentrations of chlorophyll a and b both increased 8 days after N addition. The quantum yield of Photosystem II (Φ_PSII) also increased. However, no significant changes were found in terms of gas exchange parameters. The mass-based CO₂ assimilation rate, chlorophyll a content, and chlorophyll a/b ratio (which is related to antenna size of the photosystem), of H. splendens were all higher than those of A. hookeri. Shoot mass per area (SMA) of H. splendens was lower than that of A. hookeri. We conclude that the photosynthetic rate was less sensitive to low-level N addition than chlorophyll contents and chlorophyll fluorescence parameters, suggesting other limiting factors in the photosynthetic process. Additionally, the faster growing H. splendens has a higher photosynthetic capacity than A. hookeri, allocating fewer resources to structural tissue.     

Draft genome sequence of Sinorhizobium meliloti RU11/001, a model organism for flagellum structure,motility and chemotaxis

Sinorhizobium meliloti of the order Rhizobiales is a symbiotic nitrogen-fixing bacterium nodulating plants of the genera Medicago, Trigonella and Melilotus and hence is of great agricultural importance. In its free-living state it is motile and capable of modulating its movement patterns in response to chemical attractants. Here, the draft genome consisting of a circular chromosome, the megaplasmids pSymA and pSymB and three accessory plasmids of Sinorhizobium meliloti RU11/001, a model organism for flagellum structure, motility and chemotaxis, is reported.     

Modified immunoslotblot assay to detect hemi and sulfur mustard DNA adducts

Sulfur mustard (SM) is an old chemical warfare agent causing blisters (vesicant). Skin toxicity is thought to be partly caused by SM induced DNA damage. SM and the hemi mustard 2-chloroethyl ethyl sulfide (CEES) are bi- and monofunctional DNA alkylating agents, respectively. Both chemicals react especially with N₇ guanine. The most abundant adducts are 7-hydroxyethylthioethylguanine for SM (61%) and 7-ethyl thioethylguanine for CEES. Thus, DNA alkylation should serve as a biomarker of SM exposure. A specific monoclonal antibody (2F8) was previously developed to detect SM and CEES adducts at N₇ position by means of immunoslotblot (ISB) technique (van der Schans et al. (2004) [16]). Nitrogen mustards (HN-1, HN-2, HN-3) are alkylating agents with structural similarities, which can form DNA adducts with N₇ guanine. The aim of the presented work was to modify the van der Schans protocol for use in a field laboratory and to test the cross reactivity of the 2F8 antibody against nitrogen mustards. Briefly, human keratinocytes were exposed to SM and CEES (0–300 μM, 60 min) or HN-1, HN-2, HN-3 (120 min). After exposure, cells were scraped and DNA was isolated and normalized. 1 μg DNA was transferred to a nitrocellulose membrane using a slotblot technique. After incubation with 2F8 antibody, the DNA adducts were visualized with chromogen staining (3,3′-diaminobenzidine (DAB), SeramunGrün). Blots were photographed and signal intensity was quantified. In general, DAB was superior to SeramunGrün stain. A staining was seen from 30 nM to 300 μM of SM or CEES, respectively. However, statistically significant DNA adducts were detected after CEES and SM exposure above 30 μM which is below the vesicant threshold. No signal was observed after HN-1, HN-2, HN-3 exposure. The total hands-on time to complete the assay was about 36 h. Further studies are necessary to validate SM or CEES exposure in blister roofs of exposed patients.