Genetic mapping of a new flowering time gene on chromosome 3B of wheat

The single chromosome substitution lines of chromosome 3B of the Czech alternative wheat variety Česká Přesívka (CP 3B) into two spring varieties Zlatka and Sandra, revealed clear differences in flowering time compared to the recipient varieties. To map this gene(s), recombinant substitution lines for chromosome 3B were produced from crosses of the substitution lines with their recipient parents and genetic maps developed using SSR markers. Two populations were mapped, Sandra//Sandra 3B/Sandra (CP 3B) and Zlatka//Zlatka/Zlatka (CP 3B). Combining the genotype data with phenotype data on flowering time in five independent experiments under natural long day or controlled short day conditions revealed a single flowering time QTL. This gene had an additive effect of 1–6 days, depending on environment and genetic background, and was mapped in both populations to a position in the region of marker Xbarc164 near the centromere on the long arm of 3B. Comparisons of the genetic maps with other 3B maps developed by the authors indicated that the QTL may be homologous to a QTL segregating in UK germplasm.     

In vitro heparinization of canine whole blood with low molecular weight heparin (dalteparin) significantly and dose‐dependently prolongs heparinase‐modified tissue factor‐activated thromboelastography parameters and prothrombinase‐induced clotting time

Background: Low‐molecular‐weight heparin (LMWH) is being used increasingly in veterinary medicine for both treatment and prophylaxis of thromboembolic disease, but no predictable patient‐side method exists to monitor its effect. Objectives: The aim of this study was to evaluate thromboelastography (TEG) and prothombinase‐induced clotting time (PiCT) assays for detecting hemostatic alterations following in vitro heparinization of canine whole blood with dalteparin (Fragmin). Methods: Citrated whole‐blood samples were collected from 7 clinically healthy dogs. Dalteparin was added at concentrations of 0, 0.156, 0.625, 1.25, and 2.5 U/mL of whole blood. TEG was performed using heparinase cups with tissue factor (TF, 1:50,000) and kaolin as activators. Reaction time (R), clotting time (K), angle (α), and maximum amplitude (MA) were recorded. PiCT and anti‐FXa activity were measured in plasma. Results: With TF, increasing concentrations of dalteparin significantly prolonged R and K and significantly decreased α and MA. K, α, and MA ratios were significantly different from baseline at all dalteparin concentrations and R was significantly different from baseline at concentrations of 0.625, 1.25, and 2.5 U/mL. With kaolin, only R was significantly different from baseline at dalteparin concentrations of 0.625 and 2.5 U/mL. PiCT detected dalteparin concentrations ≤ 0.625 U/mL, with a good linear correlation (r²=.96, P<.0001). Conclusion: These results suggest that TF‐activated TEG and PiCT assays should be further evaluated as promising new methods for evaluating the effect of LMWH, using doses in the recommended clinical range and prospective clinical studies.     

Sampling plan and test protocol for the semiquantitative detection of genetically modified canola (Brassica napus) seed in bulk canola seed

Using a statistical approach, sampling plans for the semiquantitative detection of genetically modified (GM) canola within a bulk seed sample can be developed and tailored to meet different GM thresholds, costs, and confidence limits. This is achieved by changing the number of subsamples analyzed, the number of seeds per subsample, and the percentage of positive results allowed. These sampling plans must be devised carefully, taking into account the detection capability of the analytical assay. This is particularly important in the case of InVigor (a registered trademark of Bayer CropScience) canola, for which expression levels of the introduced protein in seed are very low. Lateral flow assays and enzyme-linked immunosorbent assays (ELISA) were both investigated for their suitability as a qualitative assay using a subsampling approach. On the basis of an ELISA, several sampling plans have been devised and validated to provide at least 99% confidence that bulk seed samples containing at least 0.9% (w/w) InVigor canola will be detected. Although the term "seed" is used throughout this paper to refer to the canola, the term "seed" is to be taken to include both seed and the canola seed (grain) that is harvested by the farmer/grower.     

Predicting Areas of 137Cs Loss and Accumulation in Upland Catchments

Anthropogenic radioisotopes with long physicalhalf-lives derived from atmospheric fallout remain inthe environment for decades after deposition. Process- and field-based studies within uplandcatchments show that radiocaesium is transported insolution as well as in particulate form. Catchmenthydrology is therefore an important control onradiocaesium transport and natural processes of soildevelopment. The topographic index, from thehydrological model TOPMODEL, has been used as a basisfor the development of a simple model for predictingradiocaesium redistribution in temperate uplandcatchments. The model is particularly suited topredicting ¹³⁷Cs redistribution within uplandenvironments as it is based on topography, which isreadily calculated from digital terrain models withingeographical information systems. A conceptual modelof radiocaesium losses on hillslopes and re-depositionon the valley floor was calibrated with atmosphericweapons testing ¹³⁷Cs inventories from soil coredata for the Raithburn catchment, Renfrewshire, U.K. The model fitted the observations well and showed thatin this catchment a topographic index value of about5.0, for 10 m grid cells, forms the threshold between¹³⁷Cs loss and accumulation. The resultsindicated that about 20% of the total atmosphericweapons testing ¹³⁷Cs deposited in the catchmenthad been transported out of the catchment over theca. 30 yr period since deposition.     

Is the decline of soil microbial biomass in late winter coupled to changes in the physical state of cold soils?

During winter when the active layer of Arctic and alpine soils is below 0 °C, soil microbes are alive but metabolizing slowly, presumably in contact with unfrozen water. This unfrozen water is at the same negative chemical potential as the ice. While both the hydrostatic and the osmotic components of the chemical potential will contribute to this negative value, we argue that the osmotic component (osmotic potential) is the significant contributor. Hence, the soil microorganisms need to be at least halotolerant and psychrotolerant to survive in seasonally frozen soils. The low osmotic potential of unfrozen soil water will lead to the withdrawal of cell water, unless balanced by accumulation of compatible solutes. Many microbes appear to survive this dehydration, since microbial biomass in some situations is high, and rising, in winter. In late winter however, before the soil temperature rises above zero, there can be a considerable decline in soil microbial biomass due to the loss of compatible solutes from viable cells or to cell rupture. This decline may be caused by changes in the physical state of the system, specifically by sudden fluxes of melt water down channels in frozen soil, rapidly raising the chemical potential. The dehydrated cells may be unable to accommodate a rapid rise in osmotic potential so that cell membranes rupture and cells lyse. The exhaustion of soluble substrates released from senescing plant and microbial tissues in autumn and winter may also limit microbial growth, while in addition the rising temperatures may terminate a winter bloom of psychrophiles.Climate change is predicted to cause a decline in plant production in these northern soils, due to summer drought and to an increase in freeze-thaw cycles. Both of these may be expected to reduce soil microbial biomass in late winter. After lysis of microbial cells this biomass provides nutrients for plant growth in early spring. These feedbacks, in turn, could affect herbivory and production at higher trophic levels.     

Environmental and spatial characterisation of bacterial community composition in soil to inform sampling strategies

Soil physicochemical properties and microbial communities are highly heterogeneous and vary widely over spatial scales, necessitating careful consideration of sampling strategies to provide representative and reproducible soil samples across field sites. To achieve this, the study aimed to establish appropriate sampling methodology and to determine links between the variability of parameters, utilising two sampling strategies. The first (design 1) involved extracting 25 cores from random locations throughout the field and pooling them into five sets of five cores. The second (design 2) involved a further 25 cores within five 1 m² sub-plots. Sub-samples from each sub-plot were pooled in order to determine between and within sub-plot variability. All samples were analysed independently and as pooled sub-samples. Results indicate that pooling spatially separated samples significantly reduced the variability in pH, compared to individual samples. Pooling samples from a small area resulted in lower within sub-plot variability than between sub-plots for pH and bacterial community composition assessed by terminal-restriction fragment length polymorphism analysis. Following multivariate statistical analysis, a large amount of variation in community composition was explained by soil pH, which is remarkable given the relatively small size of the sampling area and minor differences in pH. Moisture content was also important in determining bacterial communities in the random design (design 1). In the 1 m² sub-plot design (design 2), the spatial location of the plots explained a large degree of the variation in bacterial community composition between plots, which was due to spatial autocorrelation of pH and possible additional environmental parameters. This study emphasises the importance of sampling design for obtaining representative samples from soil.     

Investigation of isomeric transformations of chlorogenic acid in buffers and biological matrixes by ultraperformance liquid chromatography coupled with hybrid quadrupole/ion mobility/orthogonal acceleration time-of-flight mass spectrometry

Ultraperformance liquid chromatography coupled with hybrid quadrupole/ion mobility/orthogonal acceleration time-of-flight (oa-TOF) mass spectrometry (UPLC-IM-MS) was used to study the isomeric transformations of trans-5-caffeoylquinic acid, an extremely active compound present in multiple vegetables, fruits, and beverages. The UPLC/oa-TOF MS results proved that in phosphate buffer (pH 7.4), plasma, or urine sample, trans-5-caffeoylquinic acid first isomerizes to trans-4-caffeoylquinic acid and then to trans-3-caffeoylquinic acid by intramolecular acyl migration. When exposed to UV light, trans-3-, -4-, and -5-caffeoylquinic acids undergo cis/trans isomerization to form cis isomers. The isomerization was solely dependent on the pH of the matrix, as well as the incubation temperature, and was independent of metabolic enzymes. UPLC-IM-MS results revealed that a reversible cis/trans isomerization of caffeoylquinic acids could also be induced by the electric field in an electrospray source. Thus, understanding the possible role of electric field-induced isomerization of caffeoylquinic acids may help lessen the confusion between gas phase phenomena and liquid state chemistry when applying IM-MS analysis. The comprehensive understanding of caffeoylquinic acid isomerization transformations is crucial for the appropriate handling of samples and interpretation of experimental data.     

Microbial community composition and carbon cycling within soil microenvironments of conventional, low-input, and organic cropping systems

This study coupled stable isotope probing with phospholipid fatty acid analysis (¹³C-PLFA) to describe the role of microbial community composition in the short-term processing (i.e., C incorporation into microbial biomass and/or deposition or respiration of C) of root- versus residue-C and, ultimately, in long-term C sequestration in conventional (annual synthetic fertilizer applications), low-input (synthetic fertilizer and cover crop applied in alternating years), and organic (annual composted manure and cover crop additions) maize-tomato (Zea mays - Lycopersicum esculentum) cropping systems. During the maize growing season, we traced ¹³C-labeled hairy vetch (Vicia dasycarpa) roots and residues into PLFAs extracted from soil microaggregates (53-250 μm) and silt-and-clay (<53 μm) particles. Total PLFA biomass was greatest in the organic (41.4 nmol g⁻¹ soil) and similar between the conventional and low-input systems (31.0 and 30.1 nmol g⁻¹ soil, respectively), with Gram-positive bacterial PLFA dominating the microbial communities in all systems. Although total PLFA-C derived from roots was over four times greater than from residues, relative distributions (mol%) of root- and residue-derived C into the microbial communities were not different among the three cropping systems. Additionally, neither the PLFA profiles nor the amount of root- and residue-C incorporation into the PLFAs of the microaggregates were consistently different when compared with the silt-and-clay particles. More fungal PLFA-C was measured, however, in microaggregates compared with silt-and-clay. The lack of differences between the mol% within the microbial communities of the cropping systems and between the PLFA-C in the microaggregates and the silt-and-clay may have been due to (i) insufficient differences in quality between roots and residues and/or (ii) the high N availability in these N-fertilized cropping systems that augmented the abilities of the microbial communities to process a wide range of substrate qualities. The main implications of this study are that (i) the greater short-term microbial processing of root- than residue-C can be a mechanistic explanation for the higher relative retention of root- over residue-C, but microbial community composition did not influence long-term C sequestration trends in the three cropping systems and (ii) in spite of the similarity between the microbial community profiles of the microaggregates and the silt-and-clay, more C was processed in the microaggregates by fungi, suggesting that the microaggregate is a relatively unique microenvironment for fungal activity.     

Increased N availability in grassland soils modifies their microbial communities and decreases the abundance of arbuscular mycorrhizal fungi

Two complementary studies were performed to examine (1) the effect of 18 years of nitrogen (N) fertilization, and (2) the effects of N fertilization during one growing season on soil microbial community composition and soil resource availability in a grassland ecosystem. N was added at three different rates: 0, 5.44, and 27.2 g N m⁻² y⁻¹. In both studies, Schizachyrium scoparium was the dominant plant species before N treatments were applied. Soil microbial communities from each experiment were characterized using fatty acid methyl ester (FAME) analysis. Discriminant analysis of the FAMEs separated the three N fertilizer treatments in both experiments, indicating shifts in the composition of the microbial communities. In general, plots that received N fertilizer at low or high application rates for 18 years showed increased proportions of bacterial FAMEs and decreased fungal FAMEs. In particular, control plots contained a significantly higher proportion of fungal FAMEs C18:1(cis9) and C18:2(cis9,12) and of the arbuscular mycorrhizal fungal (AMF) FAME, C16:1(cis11), than both of the N addition treatment plots. A significant negative effect of N fertilization on the AMF FAME, C16:1(cis11), was measured in the short-term experiment. Our results indicate that high rates of anthropogenic N deposition can lead to significant changes in the composition of soil microbial communities over short periods and can even disrupt the relationship between AMF and plants.