Biosynthesis of Depsipeptide Mycotoxins in Fusarium

The cyclic hexadepsipeptide enniatin is known as a phytopathogenic compound from Fusaria causing necrosis and wilt. The molecule consists of three alternating residues each of a branched chain amino acid and D-hydroxyisovaleric acid (D-Hiv). Enniatins are synthesized by a 347kDa multienzyme (enniatin synthetase) via a thiol template mechanism. The corresponding gene esyn1 has an open reading frame of 9393 nucleotides and harbours two modules, one responsible for D-hydroxy acid activation and one for L-amino acid activation with an integrated N-methyltransferase domain. Such methyltransferases build an homologous group among N-methyl peptide synthetases. Enniatins are synthesized by step-wise condensation of dipeptidol building blocks in an iterative manner resembling fatty acid synthesis. A key enzyme in enniatin biosynthesis is the NADPH-dependent D-2-hydroxyisovalerate dehydrogenase, that supplies enniatin synthetase with D-Hiv. Enniatins contribute to the wilt toxic character of Fusaria. Virulence was significantly reduced in F. avenaceum after disruption of the esyn1 gene.     

Evaluation of a combined transcutaneous carbon dioxide pressure and pulse oximetry sensor in adult sheep and dogs

OBJECTIVE: To evaluate a combined transcutaneous carbon dioxide pressure (tcPCO(2)) and pulse oximetry sensor in sheep and dogs. ANIMALS: 13 adult sheep and 11 adult dogs. PROCEDURES: During inhalation anesthesia, for the first 10 minutes following sensor placement, arterial blood gas was analyzed and tcPCO(2) was recorded every 2 minutes. Subsequently, the animals were hyper-, normo-, and hypoventilated. The simultaneously obtained tcPCO(2) and PaCO(2) values were analyzed by use of Bland-Altman statistical analysis. RESULTS: Mean +/- SD overall difference between tcPCO(2) and PaCO(2) 10 minutes after sensor application was 13.3 +/- 8.4 mm Hg in sheep and 8.9 +/- 12 mm Hg in dogs. During hyper-, normo-, and hypoventilation, mean difference (bias) and precision (limits of agreement [bias +/- 2 SD]) between tcPCO(2) and PaCO(2) values were 13.2 +/- 10.4 mm Hg (limits of agreement, -7.1 and 33.5 mm Hg) in sheep and 10.6 +/- 10.5 mm Hg (limits of agreement, -9.9 and 31.2 mm Hg) in dogs, respectively. Changes in PaCO(2) induced by different ventilation settings were detected by the tcPCO(2) sensor with a lag (response) time of 4.9 +/- 3.5 minutes for sheep and 6.2 +/- 3.6 minutes for dogs. CONCLUSIONS AND CLINICAL RELEVANCE: The tcPCO(2) sensor overestimated PaCO(2) in sheep and dogs and followed changes in PaCO(2) with a considerable lag time. The tcPCO(2) sensor might be useful for noninvasive monitoring of changes but cannot be used as a surrogate measure for PaCO(2).     

Stability and Biological Activity of Cyfluthrin Isomers

A GC-MS method capable of completely separating the four pairs of diastereoisomers of cyfluthrin is presented and the method used to show that isomerisation of the cyfluthrin enantiomers occurs in methanol. This methanol-induced isomerisation could also be demonstrated by bioassays using water fleas. The biological activities of the various cyfluthrin isomers contained in the commercial products cyfluthrin and beta-cyfluthrin were assayed using several strains of lepidopteran larvae including Plutella xylostella, Heliothis virescens and Spodoptera frugiperda. With the susceptible strains, the efficiencies of the isomers mixtures of cyfluthrin and beta-cyfluthrin were shown to obey the rules of additivity. However, in tests with a resistant strain of P. xylostella originating from Thailand, the ‘inactive’ isomer III acted synergistically with the active isomer IV. Resistance factors in strains of H. virescens and P. xylostella were found to be higher with cis than with trans isomers. This probably contributes to the superior action of cyfluthrin and beta-cyfluthrin against various pests of agricultural importance since the commercial products contain a high content of trans isomers (‘high trans pyrethroids’).     

Biochemical and toxicological differences in the modes of action of the benzoylureas

The two insecticidal benzoylurea compounds, diflubenzuron and chlorfluazuron, show large differences in their toxicity against the larvae of insects like the tobacco budworm, Heliothis virescens, or the Egyptian cotton leafworm, Spodoptera littoralis, chlorfluazuron being about 100 times more toxic. This difference is due mainly to a much faster metabolism of diflubenzuron. Its half-life within the larvae is about 5 h, compared to about 50 h for chlorfluazuron. Chlorfluazuron is also the much better ovicide of the two, following injection of the compounds into the females of H. virescens. Again the difference in the rate of metabolism is the main cause. The rate of excretion of the parent benzoylureas is relatively low, but their metabolites are excreted very quickly.     

Age effects on Norway spruce (Picea abies) susceptibility to ozone uptake: a novel approach relating stress avoidance to defense

Cumulative ozone (O3) uptake and O3 flux were related to physiological, morphological and biochemical characteristics of Norway spruce (Picea abies (L.) Karst.) trees of different ages. Under ambient CO2 conditions, photosynthetic capacity (Amax) declined in mature trees when cumulative O3 uptake into needles, which provides a measure of effective O3 dose, exceeded 21 mmol m-2 of total needle surface area. A comparable decline in Amax of seedlings occurred when cumulative O(3) uptake was only 4.5 mmol m-2. The threshold O3 flux causing a significant decline in Amax ranged between 2.14 and 2.45 nmol m-2 s-1 in mature trees and seedlings subjected to exposure periods of > or = 70 and > or = 23 days, respectively. The greater O3 sensitivity of young trees compared with mature trees was associated with needle morphology. Biomass of a 100-needle sample increased significantly with tree age, whereas a negative correlation was found for specific leaf area, these changes parallel those observed during differentiation from shade-type to sun-type needles with tree ontogeny. Age-dependent changes in leaf morphology were related to changes in detoxification capacity, with area-based concentrations of ascorbate increasing during tree ontogeny. These findings indicate that the extent of O3-induced injury is related to the ratio of potentially available antioxidants to O3 influx. Because this ratio, when calculated for ascorbate, increased with tree age, we conclude that the ratio may serve as an empirical basis for characterizing age-related differences in tree responses to O3.     

Estimating “autotrophic” belowground respiration in spruce and beech forests: decreases following girdling

Seasonal fluxes of CO₂ from soil and the contribution of autotrophic (root + mycorrhizal) to total soil respiration (SR) were estimated for a mixed stand of European beech (Fagus sylvatica) and Norway spruce (Picea abies) in Central Europe. Mature trees of each species were girdled in August 2002 to eliminate carbohydrate allocation to roots. SR was measured at distances of 0.5, 1.0, and 1.5/2.0 m from the bole of each tree at 1–2 weeks intervals throughout the fall of 2002 and monthly during the spring and summer of 2003. The contribution of roots and mycorrhizae to total SR was estimated by the decrease in SR compared to ungirdled control trees to account for seasonal patterns evident in controls. SR decreased with soil temperature in the fall 2002 and increased again in 2003 as soil warmed. During most of the study period, SR was strongly related to soil temperature. During the dry summer of 2003, however, SR appeared to be uncoupled from temperature and was strongly related to soil water content (SWC). Mean rates of SR in beech and spruce control plots as well as root densities did not show a clear pattern with distance from the bole. SR decreased to levels below controls in beech within a few days after girdling, whereas spruce did not show a significant decrease until October 2002, 6 weeks after girdling. In both beech and spruce, decreased SR in response to girdling was greatest closest to the bole, possibly reflecting increased mycorrhizal activity close to the bole. Autotrophic respiration was estimated in beech to be as much as 50% of the total SR in the stand. The contribution of autotrophic respiration was less certain for spruce, although close to the bole, the autotrophic fraction may contribute to total SR as much as in beech. The large fraction of autotrophic respiration in total SR requires better understanding of tree level stresses that affect carbon allocation below ground.     

Nitrogen turnover and N2O production in the forest floor of beech stands as influenced by forest management

Effects of forest management (thinning) on gross ammonification, net ammonification, net nitrification, microbial biomass, and N₂O production were studied in the forest floor of adjacent untreated control (“C”) and thinned (“T”) plots in three beech (Fagus sylvatica L.) stands in the Swabian Jura (Southern Germany) during three intensive field campaigns in the year 2004. The investigated sites are located less than 1 km apart on the slopes of a narrow valley. Due to different exposure (southwest, northeast, northwest), the three sites are characterized by warm‐dry microclimate (southwest site, SW) and cool‐moist microclimate (northeast site, NE; and northwest site, NW). Measurements at the NW site covered the second year (13 to 20 months) after thinning, and measurements at the SW and NE sites covered the sixth year (61 to 68 months) after thinning. Mean gross ammonification varied insignificantly across the six plots (range: 37.5 ? 31.2 to 51.0 ? 10.5 mg N (kg dry soil)^–1 d^–1). The SW site was characterized by very low net nitrification and nitrate (NO ) concentrations that were not significantly different between control and thinned plot. In contrast, for the thinned plot at the NE site (NET), significantly increased mean net nitrification (2.3 ? 1.2 mg N (kg dry soil)^–1 d^–1 at the NET plot vs. 0.4 ? 0.2 mg N (kg dry soil)^–1 d^–1 at the NEC plot) and mean extractable NO concentrations (43.9 ? 22.8 mg N (kg dry soil)^–1 at the NET plot vs. 4.1 ? 0.8 mg N (kg dry soil)^–1 at the NEC plot) were observed. The differences in net nitrification and NO concentrations across the research plots were related to differences in the forest‐floor C : N ratios: net nitrification increased exponentially below a threshold C : N value of about 25. The results of this study indicate that the forest floor of the warm‐dry SW site is very resistant to N loss triggered by thinning due to high C : N ratios around 30. Under the cool‐moist microclimate of the NE site, a significantly lower C : N ratio of 22.1 at the thinned plot (control plot: 26.7) coincided with significantly increased net nitrification. Thus, different responses of net nitrification to thinning under different microclimate appear to be triggered by different C : N ratios. Nitrous oxide production was mainly governed by forest‐floor water content, and since differences in water content at adjacent control and thinned plots were low, N₂O production was not significantly different between adjacent control and thinned plots.