Bulk Carbohydrate Grain Filling of Barley β‐Glucan Mutants Studied by 1H HR MAS NMR

Temporal and genotypic differences in bulk carbohydrate accumulation in three barley genotypes differing in the content of mixed linkage β‐(1→3),(1→4)‐D‐glucan (β‐glucan) and starch were investigated using proton high‐resolution, magic angle spinning, nuclear magnetic resonance (¹H HR MAS NMR) during grain filling. For the first time, ¹H HR MAS NMR spectra of flour from immature barley seeds are analyzed. Spectral assignments are made using two‐dimensional (2D) NMR methods. Both α‐ and β‐glucan biosynthesis were characterized by inspection of the spectra as well as by calibration to the reference methods for starch and β‐glucan content. Starch was quantified with very good calibrations to the α‐(1→4) peak (5.29–5.40 ppm) and the region 3.67–3.83 ppm covering starch glycopyranosidic protons from H5 and H6. In contrast, the spectral inspection of the β‐anomeric region 4.45–4.85 ppm showed unexpected lack of intensity in the high β‐glucan mutant lys5f at seed maturity, resulting in poor calibration to reference β‐glucan content. We hypothesize that the lack of β‐glucan signal in lys5f indicates partial immobilization of the β‐glucan that appears to be either genotypic dependent or water/β‐glucan ratio dependent.     

Susceptibility of canine and feline bacterial pathogens to pradofloxacin and comparison with other fluoroquinolones approved for companion animals

In this study, 908 bacterial pathogens from defined infections of dogs and cats were tested for their susceptibility to the novel fluoroquinolone pradofloxacin, which was approved in 2011 for use in cats and dogs. Most of the bacteria tested (Staphylococcus aureus, Staphylococcus pseudintermedius, Escherichia coli, β-haemolytic streptococci, Pasteurella multocida and Bordetella bronchiseptica) exhibited low pradofloxacin MIC₉₀ values of ≤0.25 μg/ml. Solely Proteus spp. and Pseudomonas aeruginosa had higher MIC₉₀ values of ≥4 μg/ml. Only six (3.4%) of 177 S. pseudintermedius and 12 (5.3%) of 227 E. coli isolates showed pradofloxacin MICs of ≥2 μg/ml. Analysis of the quinolone resistance determining regions of the target genes identified double mutations in GyrA that resulted in amino acid exchanges S83L + D87N or S83L + D87Y and single or double mutations in ParC that resulted in amino acid exchanges S80I or S80I + E84G in all 12 E. coli isolates. The six S. pseudintermedius isolates exhibited amino acid exchanges S84L or E88K in GyrA and S80I in GrlA. Comparative analysis of the MICs of pradofloxacin and the MICs determined for enrofloxacin and its main metabolite ciprofloxacin, but also marbofloxacin, orbifloxacin, difloxacin and ibafloxacin was conducted for the target pathogens S. pseudintermedius, E. coli and P. multocida. This comparison confirmed that pradofloxacin MICs were significantly lower than those of the other tested fluoroquinolones.     

Towards the primary target of chloroacetamides –new findings pave the way

This review reports on research of the last ten years to find the primary target enzyme for chloroacetamides. As could be shown first with the green alga Scenedesmus, the formation of very‐long‐chain fatty acids (VLCFAs) is severely impaired. Subsequently, in short‐term experiments, labelled malonate or stearate could be incorporated into leaf discs of cucumber, barley or leek seedlings. While the formation of ‘normal’ long‐chain fatty acids (up to C18) was not influenced, phytotoxic chloroacetamides strongly inhibited the synthesis of VLCFAs of C20, 22 and 24, with I₅₀ values of 10–100 nM . Inhibition depends on the amide structure and on stereospecificity. Also cafenstrole or recently developed tetrazolinones and phosphosulfonates were found active to inhibit fatty‐acid elongation. Subsequently, a cell‐free elongase assay was developed using a microsomal preparation from leek seedlings (Allium porrum L), [¹⁴C]malonyl‐CoA and C18, 20, or C22 acyl‐CoA primer substrates. All elongation steps were strongly affected by those phytotoxic herbicides which were also active in vivo. The inhibitors form a tight‐binding complex with the condensing elongase enzyme system which develops with time and lowers the I₅₀ values markedly. Apparently, a nucleophilic attack of the inhibitor takes place at the specific target enzyme. Acyl‐CoA elongation inhibition is correlated with growth inhibition of the intact cell. Due to the low I₅₀ values and the specific inhibition, we assume that impaired VLCFA‐formation is the primary phytotoxic impact of chloroacetamides and functionally related structures. © 2000 Society of Chemical Industry     

Determination of the minimum alveolar concentration (MAC) and cardiopulmonary effects of sevoflurane in sheep

Objective

To determine sevoflurane’s minimum alveolar concentration (MAC_SEVO) and its cardiopulmonary effects in sheep.

A sensitivity analysis for assessing the relevance of fine‐root turnover for P and K uptake

Plant fine roots are subject to continual turnover, i.e., old roots die during the plant life cycle and are quickly replaced by new roots. New roots grow partly into undepleted soil areas and can take up nutrients at a higher rate than old roots. This is one possible advantage of root turnover. It has been shown that root turnover of several plant species increases when P and/or K supply is limited, indicating an efficiency mechanism. The objective of this study was to assess the maximum benefit for nutrient uptake by root turnover and to determine which soil or plant properties influence this process. Based on a data set of field‐grown faba beans, a sensitivity analysis with a transport and uptake model was performed, i.e., several input parameters were systematically varied to assess their importance for nutrient uptake of a root system with and without fine‐root turnover. The calculations were based on the assumptions that all new roots grow into undepleted soil areas and that no inter‐root competition occurs. Model calculations indicated that a root system with a high but realistic turnover rate can take up twice the amount of P or K compared to a stable root system without any turnover. This benefit on uptake is higher at low concentrations of these nutrients in soil solution, low soil water content, or high maximum inflow. However, measured uptake under poor conditions of nutrient supply is often higher than calculated uptake, even when root turnover is taken into account. This indicates that root turnover might be an efficiency mechanism, but not the only one.     

Land Application of Wastewater in Brazil - A scientific challenge: Chemical Characterization of Soil at Populina, S?o Paulo State (9 pp)

Background, Aims and Scope   The rapid increase of wastewater worldwide and the consequently urgently needed treatment measures towards disposal purposes has led to an expansion of alternative treatment methods. Besides conventional and cost intensive systems, domestic wastewater treatment by overland flow process represents an effective and low-cost alternative. However, in addition to beneficial elements in wastewaters, there is a release of other elements of the Periodic Table into the environment, which can be considered as potentially toxic. This requires a reliable chemical characterization of the wastewater element composition as well as the soils onto which it will be applied. This paper aims at presenting the chemical fingerprint methodology to characterize concentrations of a large set of elements in a tropical ferralsol in order to provide information concerning chemical modifications after long-term application of wastewater by overland flow process. The results are also used to estimate environmental implications according to guiding values for soils in São Paulo State.Methods   The wastewater treatment in Populina has been operating for 18 years. Composite soil samples were taken in a treated plot and a control plot. Wastewater was chemically characterized by using AAS. Total C and total N of soil samples were determined by Delta Plus Mass Spectrometer. The remaining elements (32) were analyzed using ICP-OES. Grain size analysis was carried out after oxidation by H2O2, and soil dispersion with NaOH and Na4P2O7 treatments. pH was measured in water. Quality control of all measurements was verified by corresponding reference materials. Fingerprint graphs were generated by standardizing the values of the control plot and by graphically displaying positive/negative percentage deviations of the element values at the treated plot from values of the control site. Results and Discussion   Chemical analyses of the wastewater used showed high BOD, COD, TOC and N values, while the majority of metals were below the detection limit that is attributed to the limitations in the instrumental techniques (AAS) utilized. The comparison of the two selected sites revealed substantial changes of the element contents associated with long-term application of domestic wastewater. Fingerprint graphs of the treated field indicated higher values for the vast majority of macro and micro-nutrients (C, N, P, K, Ca, Mg, Mo, Zn, Cu), as well as for other elements, e.g. Ba, Sr, Na, Li, or La. Enrichments of ecotoxic elements (e.g. As or Pb) occurred in the surface horizon despite the low contents of those elements in the wastewater. Other elements analyzed, i.e. Ag, Be, Bi, Cd, Sb, Sn, W or Sc, that are partially considered to be toxic, did not show considerable differences. Due to limitations in the analytical techniques applied, these results do not imply that there is no existence or no release of elements from wastewater at trace or ultra-trace levels. It is expected that more sophisticated techniques will be available in the future. Typical soil elements (Ti, Al, Fe, V, Zr) show higher values at the control site suggesting higher contents of mineralogical components. The highly positive deviation of C-contents linked with higher values of many other elements in the treated soil suggest element enrichments associated with organic matter. Concerning environmental implications, the comparison with guiding values for an enhanced monitoring showed partly an excess of acceptable metal concentration levels. Proposed intervention values were not exceeded, except for with Ba. Comparisons with phytotoxic element concentrations in soils indicate critical values for As, Cu and Cr. Conclusion   Using multi-element analysis, the study represents a first approach to access wastewater application by overland flow on the basis of a rather large range of chemical elements in Brazil. The comparison of the two selected areas revealed distinctly higher values for the majority of elements in the treated plot. The study also highlighted the importance of organic matter for element retention. The environmental implications of domestic wastewater application to soil surfaces can be grave. Since also toxic elements are enriched in these treated soils, the areas can partly be classified as polluted, and require long-term monitoring and detailed investigations.Recommendation and Outlook   The analytical data obtained may only allow for some conclusions concerning long-term feasibility and geochemical preconditions of wastewater irrigation. Therefore, not only the total concentrations of metals are of interest, but also element speciation must be analyzed in order to predict, for instance, how long the ion uptake capacity of the soil will last. Aside from the analytical procedures used in speciation, theoretical chemical (thermodynamic) concepts such as the principles of Pourbaix diagrams should also be used to predict solubility, immobilization and bioavailability of different element species.     

Non-homoeologous wheat-rye chromosomal translocations conferring resistance to greenbug

Summary C-banding andin situ hybridization were used to determine the chromosomal constitution of the greenbug-resistant germplasm GRS 1204. The results showed that this line had the radiation-induced non-homoeologous wheat-rye translocation chromosomes T2AS-1RS·1RL and T2AL·2AS-1RS. C-banding analysis further revealed the presence of a wheat-Agropyron elongatum translocation chromosome T1BL·1BS-3Ae#1L in line GRS 1204, that was derived from Teewon. The greenbug resistance of line GRS 1204 is similar to that of line GRS 1201 that was earlier shown to have the greenbug resistance geneGb6 located on the 1RS arm of the wheat-rye translocation chromosome T1AL·1RS. BecauseGb6 in line GRS 1204 is present on one of the non-homoeologous translocation chromosomes, agronomically line GRS 1201 should be the better adapted source ofGb6 resistance and be used in cultivar improvement.