Boron and other elements in sporophores of ectomycorrhizal and saprotrophic fungi

Fungi are usually thought not to have a boron (B) requirement. It is not known if mycorrhizas take up B from low concentrations that are common in forest soils, as fungi might also immobilise B. Here, we studied the B concentrations in sporophores of 49 ectomycorrhizal and 10 saprotrophic fungi to assess whether B is translocated in mycelium or not. Additionally, P and metal concentrations were measured for comparison. Variability both within species and between species was very large, as the lowest measured B concentration was 0.01 mg kg⁻¹ in Amanita muscaria, and the highest was 280 mg kg⁻¹ in Paxillus involutus. There was no clear difference between saprotrophic and mycorrhizal fungi. The majority of species did not accumulate B at more than 0.01–3 mg kg⁻¹, but there were some species that consistently had median concentration values higher than 5–6 mg kg⁻¹ and much higher maximum values, particularly Paxillus involutus, Lactarius necator and several Russula species. Most species increased their B concentration in B fertilised plots, but there were exceptions, particularly Rozites caperatus and Lactarius camphoratus. Boron concentrations did not correlate with those of other elements. In conclusion, B is translocated in the mycelia of most of the studied species. The differences between species may be due to differences in their water use, or carbohydrates used in translocation. It remains to be studied, if B concentrations in mycorrhizas or mycelia in soil are in the same order of magnitude as the larger ones found here, and if this has any effects on the host plants.     

Fungal diversity and natural occurrence of fusaproliferin,beauvericin, deoxynivalenol and nivalenol in wheat cultivated in Santa Fe Province,Argentina

The Fusarium diversity and the mycobiota associated with moldy wheat kernels from Santa Fe province, Argentine, was assessed. The wheat cultivated area in Santa Fe province is divided according to agrometeorological conditions into two zones: Zone I (north-central) and Zone II (south). The natural occurrence of Fusarium toxins BEA, FUP, DON and NIV was also determined. Cladosporium was the most abundant of the 19 genera identified, followed by Fusarium, Phoma and Alternaria. Zone II shows a predominance of F. graminearum and F. culmorum. In Zone I, DON was present in 13/32 samples (range 0.43–3.60 mg kg⁻¹) and NIV in 6/32 samples (range 0.11–0.40 mg kg⁻¹). In zone II, DON was found in 11/21 samples (range 0.57–9.50 mg kg⁻¹) and NIV in 4/21 samples (range 0.10–0.60 mg kg⁻¹). BEA and FP were not detected in both zones.     

Major and Trace Elements in Zooplankton from the Northern Gulf of California During Summer

We report the distribution of major and trace element concentrations in epipelagic zooplankton collected in the Northern Gulf of California in August 2003. The Bray–Curtis index defined three element assemblages in zooplankton: (1) major metals, which included only two elements, Na (3.6–17.0%) and Ca (1.0–4.8%). Na had its highest concentrations in the shallow tidally mixed Upper Gulf, where high salinity, temperature, and zooplankton biomass (dominated by copepods) prevailed. Ca showed its highest concentrations south of Ballenas Channel, characterized by tidal mixing and convergence-induced upwelling, indicated by low sea-surface temperature, salinity, and zooplankton biomass; (2) Six biological essential elements, like Fe (80–9,100 mg kg⁻¹) and Zn (20–2,570 mg kg⁻¹), were detected in high concentrations in zooplankton collected near Guaymas Basin, which had high surface temperature and chlorophyll a concentrations. (3) Metals of terrigenous origin, such as Sc (0.01–1.4 mg kg⁻¹) and Th (0.03–2.3 mg kg⁻¹), and redox-sensitive metals, like Co (3–23.8 mg kg⁻¹); this was the assemblage with the largest number of elements (15). Both types of elements of assemblage 3 had maximum concentrations in the cyclonic eddy that dominates the summer circulation in the Northern region. We concluded that sediment resuspension by tidal mixing in the Upper Gulf, upwelling south of Ballenas Channel, and the cyclonic eddy were key oceanographic features that affected the element concentrations of epipelagic zooplankton in the Northern Gulf of California. Oceanographic mechanisms such as these may contribute to element incorporation in marine organisms in other seas.     

Sulphur protects mustard (<Emphasis Type="Italic">Brassica campestris</Emphasis> L.) from cadmium toxicity by improving leaf ascorbate and glutathione

In a pot-soil culture ameliorative effect of sulphur (S) (0 or 40 mg S kg⁻¹ soil) on cadmium (Cd) (0, 25, 50 and 100 mg Cd kg⁻¹ soil)-induced growth inhibition and oxidative stress in mustard (Brassica campestris L.) cultivar Pusa Gold was studied. Cadmium at 100 mg kg⁻¹ soil caused maximum increase in the contents of Cd and thiobarbituric acid reactive substances (TBARS) in leaves. Maximum reductions in growth (plant dry mass, leaf area), chlorophyll content, net photosynthetic rate (P_N) and the contents of ascorbate (AsA), glutathione (GSH) were observed with 100 mg Cd kg⁻¹ soil compared to control. The application of S helped in reducing Cd toxicity, which was greater for 25 and 50 mg Cd kg⁻¹ soil) compared to 100 mg Cd kg⁻¹ soil. Addition of S to Cd-treated plants showed decrease in Cd and TBARS content in leaves and restoration of growth and photosynthesis through increase in the contents of AsA and GSH. Net photosynthetic rate and plant dry mass were strongly and positively correlated with the contents of AsA and GSH. It is suggested that S may ameliorate Cd toxicity and protects growth and photosynthesis of mustard involving AsA and GSH.     

Biomass production and nutrient accumulation in <Emphasis Type="Italic">Sparganium emersum</Emphasis> Rehm. after sediment treatment with mineral and organic fertilisers in three mesocosm experiments

The response of the aquatic plant Sparganium emersum to different sediment nutrient levels was studied in three mesocosm experiments. The aim was to assess plant growth parameters and nutrient accumulation in the plant tissue under conditions relevant for habitats with sediments affected by anthropogenic nutrient enrichment. The experimental treatments were produced by fertilisation of the rooting medium (washed river sand) with differing doses of either NPK mineral fertiliser or digested sludge from solid pig slurry waste. Growth inhibition by high nutrient levels was not observed in any treatment (highest nutrient concentrations in the sediment with mineral fertiliser: N 250 mg kg⁻¹, P 50 mg kg⁻¹; organic fertiliser: N 6300 mg kg⁻¹, P 1800 mg kg⁻¹), which confirms the tolerance of S. emersum to high nutrient loads. The sediment nutrient concentration was best reflected in shoot dry mass. Nutrient contents in plant tissues were similar for most nutrient concentrations in the rooting media; only N increased significantly with N levels in the sediment in belowground parts. Nutrient standing stocks in plants, however, generally corresponded to the nutrient supply, and reached highest values (max. N 3.7 g m⁻², P 1.2 g m⁻²) in the richest treatments with organic fertiliser. The capability of S. emersum to use nutrients from high sediment concentrations and in organically polluted environments recommends this species for use in water quality management including tertiary wastewater treatment.     

Vanadium Uptake by Alfalfa Grown in V–Cd-Contaminated Soil by Pot Experiment

In order to characterize uptake of vanadium in alfalfa grown in vanadium–cadmium (Cd)-contaminated soil, 104 soil samples and 94 plant samples were collected from pot experiment. The results showed alfalfa had strong metal adaptability (up to 400 mg kg⁻¹) and high accumulation (up to 3,440.14 mg kg⁻¹) of vanadium. Root had higher contents and better absorption to vanadium than overground part. Moreover, both root and overground part had direct correlation with vanadium in soil, especially with the sum of first three fractions and reducible fraction. With the increasing of vanadium, higher concentration of Cd may inhibit the absorption of vanadium in alfalfa.     

Elicitor-induced accumulation of stilbenes in cell suspension cultures of Cayratia trifolia (L.) Domin

Cell cultures of Cayratia trifolia (Vitaceae), a tropical lianas, were maintained in Murashige and Skoog’s medium containing 0.25 mg l⁻¹ NAA, 0.2 mg l⁻¹ kinetin and casein hydrolysate 250 mg l⁻¹. Cell suspension cultures of C. trifolia accumulate stilbenes (piceid, resveratrol, viniferin, ampelopsin), which on elicitation by any of 500 μM salicylic acid, 100 μM methyl jasmonate, 500 μM ethrel and 500 mg l⁻¹ yeast extract, added on the 7th day, were enhanced by 3- to 6-fold (5–11 mg l⁻¹) by the 15th day.     

Production of monoterpenoids and aroma compounds from cell suspension cultures of <Emphasis Type="Italic">Camellia sinensis</Emphasis>

Cell suspension cultures of Camellia sinensis were established in 250 ml shake flasks. Flasks contained 50 ml liquid medium of either Murashige and Skoog (MS), N/5 MS or Heller medium containing different levels of 6-benzyladenine (BA) (0.05–2 mg l⁻¹), 2,4-dichlorophenoxyacetic acid (2,4-D) (1–10 mg l⁻¹), and sucrose (10–50 g l⁻¹). Moreover, the pH of the medium was varied from 5.2–6.2. In addition, cultures were subjected to light irradiation as well as to complete darkness. Following optimization of aroma and terpenoid extraction methods, cell cultures were analyzed for the volatile compounds using GC/MS. A total of 43 compounds were identified using the micro SDE apparatus. Among the major monoterpenoids obtained were α-terpineol and nerol. Moreover, other high aroma-value compounds, including 2-ethyl hexanol, benzyl alcohol, benzene acetaldehyde, nonanal and phenylethylalcohol were also detected. The highest levels of these compounds were obtained from cell suspension cultures grown in MS medium containing 5 mg l⁻¹ 2,4-D, 1 mg l⁻¹ BA and 30 g l⁻¹ sucrose at pH of 5.8 with incubation in complete darkness.     

Biogeochemical conditions and ice algal photosynthetic parameters in Weddell Sea ice during early spring

Physical, biogeochemical and photosynthetic parameters were measured in sea ice brine and ice core bottom samples in the north-western Weddell Sea during early spring 2006. Sea ice brines collected from sackholes were characterised by cold temperatures (range −7.4 to −3.8°C), high salinities (range 61.4–118.0), and partly elevated dissolved oxygen concentrations (range 159–413 μmol kg⁻¹) when compared to surface seawater. Nitrate (range 0.5–76.3 μmol kg⁻¹), dissolved inorganic phosphate (range 0.2–7.0 μmol kg⁻¹) and silicic acid (range 74–285 μmol kg⁻¹) concentrations in sea ice brines were depleted when compared to surface seawater. In contrast, NH₄ ⁺ (range 0.3–23.0 μmol kg⁻¹) and dissolved organic carbon (range 140–707 μmol kg⁻¹) were enriched in the sea ice brines. Ice core bottom samples exhibited moderate temperatures and brine salinities, but high algal biomass (4.9–435.5 μg Chl a l⁻¹ brine) and silicic acid depletion. Pulse amplitude modulated fluorometry was used for the determination of the photosynthetic parameters F _v/F _m, α, rETR_max and E _k. The maximum quantum yield of photosystem II, F _v/F _m, ranged from 0.101 to 0.500 (average 0.284 ± 0.132) and 0.235 to 0.595 (average 0.368 ± 0.127) in the sea ice internal and bottom communities, respectively. The fluorometric measurements indicated medium ice algal photosynthetic activity both in the internal and bottom communities of the sea ice. An observed lack of correlation between biogeochemical and photosynthetic parameters was most likely due to temporally and spatially decoupled physical and biological processes in the sea ice brine channel system, and was also influenced by the temporal and spatial resolution of applied sampling techniques.     

Transformation of carotenoid biosynthetic genes using a micro-cross section method in kiwifruit (Actinidia deliciosa cv. Hayward)

Genetic transformation using a micro-cross section (MCS) technique was conducted to improve the carotenoid content in kiwifruit (Actinidia deliciosa cv. Hayward). The introduced carotenoid biosynthetic genes include geranylgeranyl diphosphate synthase (GGPS), phytoene desaturase (PDS), ζ-carotene desaturase (ZDS), β-carotene hydroxylase (CHX), and phytoene synthase (PSY). The transformed explants were selected on half-strength MS medium containing 0.001 mg l⁻¹ of 2,4-D and 0.1 mg l⁻¹ of zeatin, either 5 mg l⁻¹ hygromycin or 25 mg l⁻¹ kanamycin, and 500 mg l⁻¹ cefotaxime. The genomic PCR, genomic Southern blot analysis, and RT-PCR were performed to confirm the integration and expression of the transgenes. The transformation efficiencies of either kanamycin- or hygromycin-resistant shoots ranged from 2.9 to 22.1% depending on the target genes, and from 2.9 to 24.2% depending on the reporter genes. The selection efficiencies ranged from 66.7 to 100% for the target genes and from 95.8 to 100% for the reporter genes. Changes of carotenoid content in the several PCR-positive plants were determined by UPLC analysis. As a result, transgenic plants expressing either GGPS or PSY increased about 1.2- to 1.3-fold in lutein or β-carotene content compared to non-transgenic plants. Our results suggest that the Agrobacterium-mediated transformation efficiency of kiwifruit can be greatly increased by this MCS method and that the carotenoid biosynthetic pathway can be modified in kiwifruit by genetic transformation. Our results further suggest that GGPS and PSY genes could be major target genes to increase carotenoid contents in kiwifruit.     

Permissible Value for Vanadium in Allitic Udic Ferrisols Based on Physiological Responses of Green Chinese Cabbage and Soil Microbes

Greenhouse experiments were conducted to study the permissible value of vanadium (V) based on the growth and physiological responses of green Chinese cabbage (Brassica chinensis L.), and effects of V on microbial biomass carbon (MBC) and enzyme activities in allitic udic ferrisols were also studied. The results showed that biomass of cabbage grown on soil treated with 133 mg V kg⁻¹ significantly decreased by 25.1% compared with the control (P < 0.05). Vanadium concentrations in leaves and roots increased with increasing soil V concentration. Contents of vitamin C (Vc) increased by 10.3%, while that of soluble sugar in leaves significantly decreased by 54.0% when soil V concentration was 133 mg kg⁻¹, respectively. The uptake of essential nutrient elements by cabbage was disturbed when soil V concentration exceeded 253 mg kg⁻¹. Soil MBC was significantly stimulated by 15.5%, while dehydrogenase activity significantly decreased by 62.8% and urease activity slightly changed at treatment of 133 mg V kg⁻¹ as compared with the control, respectively. Therefore, the permissible value of V in allitic udic ferrisols is proposed as 130 mg kg⁻¹.     

Tumor-inhibitory and liver-protective effects of <Emphasis Type="Italic">Phellinus igniarius</Emphasis> extracellular polysaccharides

The tumor-inhibitory and liver-protective effects of crude extracellular polysaccharides (EPS) extracted from the liquid mycelial culture of the mushroom Phellinus igniarius were studied in mice. The mice were injected with murine sarcoma S180 and murine hepatoma H22. Crude EPS at 100, 200, 400 mg kg⁻¹ body weight was administered to EPS groups each day in the twelve consecutive days. The result showed that EPS 200 mg kg⁻¹ body weight significantly inhibited S180 and H22 at 65.0 and 46.3%, respectively. Moreover, EPS could not only keep the numbers of WBC, RBC, PLT and the concentration of HGB in a normal range, but also normalize the activities of AST, ALT and ALP. For example, in EPS-treated mice, AST significantly reduced with the percentage of A/G reverse in S180 (P < 0.05) and H22 (P < 0.01) when the mice took EPS 200 mg kg⁻¹ body weight. In conclusion, it was remarkable that P. igniarius EPS exhibited antitumor activity related to dosage and protected liver function by sustaining the blood routine as well as keeping the blood biochemical indexes normal.     

Inhibitory effect of curcumin on liver injury in a murine model of endotoxemic shock

The effect of curcumin on lipopolysaccharide/d-galactosamine (LPS/GalN)-induced acute shock model of liver injury was examined in mice. The simultaneous administration of LPS (5–20 μg kg⁻¹, i.p.) and GalN (700 mg kg⁻¹, i.p.) markedly increased the serum tumor necrosis factor-α (TNF-α), glutamic oxaloacetic transaminase/glutamic pyruvic transaminase (GOT/GPT), and massive hepatic necrosis and inflammation, leading to 100% lethality. Pre-administration of curcumin (100 mg kg⁻¹, i.p.) 3 h before induction with LPS/GalN imparted a large extent of protection against acute elevation in serum TNF-α and serum GOT/GPT. Hepatic necrosis and lethality caused by LPS/GalN was also greatly reduced by curcumin treatment. The results demonstrated that curcumin could protect mice from LPS/GalN-induced hepatic injury and inflammation through blockading TNF-α production, eventually raising the survival rate of septic-shock-induced mice.     

Enhanced regeneration of haploid plantlets from microspores of <Emphasis Type="Italic">Brassica napus</Emphasis> L. using bleomycin,PCIB, and phytohormones

The effects of three periods of incubation (10, 20 and 30 min) at different levels of bleomycin (0, 0.1, 0.2, 0.3, 0.4 and 0.5 μg ml⁻¹), as well as three periods of exposure (12, 24 and 48 h) to different levels of the anti-auxin p-chlorophenoxyisobutyric acid (PCIB), including 1, 2, 3, 4 and 5 mg l⁻¹, on microspore embryogenesis of rapeseed cv. ‘Amica’ were investigated. Microspore embryogenesis was significantly enhanced following 20 min treatment with 0.2 μg ml⁻¹ bleomycin compared with untreated cultures. Highest embryo yield (163 embryos Petri dish⁻¹) was observed with 24 h treatment of 4 mg l⁻¹ PCIB. The highest percentage of secondary embryogenesis was observed on B₅ medium containing 0.15 mg l⁻¹ of gibberellic acid (GA₃) and 0.2 mg l⁻¹ 6-benzyladenine (BA) in 4–6 mm microspore-derived embryos (MDEs). Most callus formed on B₅ medium containing 0.15 mg l⁻¹ GA₃, 0.1 mg l⁻¹ BA and 0.1 mg l⁻¹ indole-3-acetic acid (IAA) when 4–6 mm embryos were used. Regeneration was highest on B₅ medium containing 0.05 mg l⁻¹ GA₃ or 0.1 mg l⁻¹ BA and 0.2 mg l⁻¹ IAA with 2–4 mm embryos. Microspore embryogenesis and plant regeneration could be improved by both bleomycin and PCIB when the appropriate MDE length and phytohormone level were selected.     

Aerobic biodegradation of nitrobenzene by a defined microbial consortium immobilized in polyurethane foam

An aerobic microbial consortium constructed by the combination of Rhodotorula mucilaginosa Z1, Streptomyces albidoflavus Z2 and Micrococcus luteus Z3 was immobilized in polyurethane foam and its ability to degrade nitrobenzene was investigated. Batch experimental results showed that polyurethane-foam-immobilized cells (PFIC) more efficiently degrade 200–400 mg l⁻¹ nitrobenzene than freely suspended cells (FSC). Kinetics of nitrobenzene degradation by PFIC was well described by the Andrews equation. Compared with FSC, PFIC exhibited better reusability (over 100 times) and tolerated higher shock-loadings of nitrobenzene (1,000 mg l⁻¹). Moreover, In the presence of salinity (≤5% NaCl, w/v), phenol (≤150 mg l⁻¹) and aniline (≤50 mg l⁻¹), respectively, degradation efficiency of nitrobenzene by PFIC reached over 95%. Even in the presence of both 100 mg l⁻¹ phenol and 50 mg l⁻¹ aniline, over 75% nitrobenzene was removed by PFIC in 36 h. Therefore, the immobilization of the defined consortium in polyurethane foam has application potential for removing nitrobenzene in industrial wastewater treatment system.     

The reserve of weatherable primary silicates impacts the accumulation of biogenic silicon in volcanic ash soils

Banana plantlets (Musa acuminata cv Grande Naine) cultivated in hydroponics take up silicon proportionally to the concentration of Si in the nutrient solution (0–1.66 mM Si). Here we study the Si status of banana plantlets grown under controlled greenhouse conditions on five soils developed from andesitic volcanic ash, but differing in weathering stage. The mineralogical composition of soils was inferred from X-ray diffraction, elemental analysis and selective chemical/mineralogical extractions. With increasing weathering, the content of weatherable primary minerals decreased. Conversely, clay content increased and stable secondary minerals were increasingly dominant: gibbsite, Fe oxides, allophane, halloysite and kaolinite. The contents of biogenic Si in plant and soil were governed by the reserve of weatherable primary minerals. The largest concentrations of biogenic Si in plant (6.9–7 g kg⁻¹) and soil (50–58 g kg⁻¹) occurred in the least weathered soils, where total Si content was above 225 g kg⁻¹. The lowest contents of biogenic Si in plant (2.8–4.3 g kg⁻¹) and soil (8–31 g kg⁻¹) occurred in the most weathered desilicated soils enriched with secondary oxides and clay minerals. Our data imply that soil weathering stage directly impacted the soil-to-plant transfer of silicon, and thereby the stock of biogenic Si in a soil–plant system involving a Si-accumulating plant. They further imply that soil type can influence the silicon soil–plant cycle and its hydrological output.     

Photosynthetic property and primary production of phytoplankton in sublittoral sand bank area in the Seto Inland Sea,Japan

We investigated the photosynthesis–light intensity (P–I) relationships of phytoplankton collected from a sublittoral sand bank in the Seto Inland Sea, Japan, under different temperature conditions. In spite of low chlorophyll a concentration (<3 mg m⁻³), phytoplankton had considerably high photosynthetic potential (>10 mg C (mg chl a)⁻¹ h⁻¹) in the study area. Based on the P–I relationships, we conducted numerical simulation of areal primary production using published data on water temperature, chlorophyll a concentration, and irradiance. The areal primary production ranged between 159 and 187 g C m⁻² year⁻¹. This production was within the range of typical values reported previously in deeper areas of the Seto Inland Sea. The productivity in the sand bank area was discussed in relation to water current, allochthonous resource input, and fisheries.     

Development of cell culture system from the giant freshwater prawn <Emphasis Type="Italic">Macrobrachium rosenbergii</Emphasis> (de Man)

A new cell culture system (MRH) was developed for the first time from 2 months old freshwater prawn, Macrobrachium rosenbergii. Primary cultures were developed from heart tissues by explant culture technique. Cell outgrowth was obtained from the heart explant after 14 days of explant culture. The culture medium used was Leibovitz-15 supplemented with 20% Fetal Bovine Serum along with 1% prawn hemolymph serum, 0.1% glucose, 0.5% NaCl and antibiotics (Penicillin 10,000 Units ml⁻¹, Streptomycin 10,000 μg ml⁻¹, Amphotericin B 500 mg ml⁻¹) with a final osmomolality of 470–550 mmol kg⁻¹. The pH of the growth medium found suitable for the growth of the cells was 7.20. The viability of cells was found to be 60% when revived after a month of storage in liquid nitrogen.     

High stilbenes accumulation in root cultures of <Emphasis Type="Italic">Cayratia trifolia</Emphasis> (L.) Domin grown in shake flasks

The Root cultures of Cayratia trifolia (Vitaceae) a tropical lianas, were maintained in liquid Murashige and Skoog’s medium containing 0.5 mg l⁻¹ NAA, 0.1 mg l⁻¹ kinetin with 3% sucrose. These root cultures when grown with 6% sucrose accumulated stilbenes (piceid, resveratrol, viniferin, ampelopsin) in high amounts, which on elicitation by 500 mg l⁻¹ yeast extract, 50 μM salicylic acid (SA), 50 μM methyl jasmonate (MeJa), 500 μM ethrel added at 25th day, increased up to ninefolds (7.1 mg l⁻¹). Addition of alar or phenylalanine along with the elicitors further enhanced the stilbenes content. In the present study, stilbenes accumulation up to 12 folds (9.2 mg l⁻¹) was obtained with SA and alar. The SA was the most effective in increasing the stilbenes contents while less than control values were recorded in the cells treated with MeJa. The roots could be grown up to 2 l flasks. The present work demonstrates that presence of precursor and sucrose during elicitation at an appropriate time combined with growth retardation significantly increased the production of stilbenes in C. trifolia cell cultures.