40 resultados para Crossing experiments, Baculoviridae, Yeast Two-Hybrid System, Resistance management, sex-linkage


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Cost effective methods are now available to identify physiological resistance in wild populations of Norway rat and House mice that are proving difficult to control. The new molecular methodology is a significant development for resistance management.

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A two-phase system composed by a leach bed and a methanogenic reactor was modified for the first time to improve volumetric substrate degradation and methane yields from a complex substrate (maize; Zea mays). The system, which was operated for consecutive feed cycles of different durations for 120 days, was highly flexible and its performance improved by altering operational conditions. Daily substrate degradation was higher the shorter the feed cycle, reaching 8.5 g TSdestroyed d�1 (7-day feed cycle) but the overall substrate degradation was higher by up to 55% when longer feed cycles (14 and 28 days) were applied. The same occurred with volumetric methane yields, reaching 0.839 m3 (m3)�1 d�1. The system performed better than others on specific methane yields, reaching 0.434 m3 kg�1 TSadded, in the 14-day and 28-day systems. The UASB and AF designs performed similarly as second stage reactors on methane yields, SCOD and VFA removal efficiencies.

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Interactions between different convection modes can be investigated using an energy–cycle description under a framework of mass–flux parameterization. The present paper systematically investigates this system by taking a limit of two modes: shallow and deep convection. Shallow convection destabilizes itself as well as the other convective modes by moistening and cooling the environment, whereas deep convection stabilizes itself as well as the other modes by drying and warming the environment. As a result, shallow convection leads to a runaway growth process in its stand–alone mode, whereas deep convection simply damps out. Interaction between these two convective modes becomes a rich problem, even when it is limited to the case with no large–scale forcing, because of these opposing tendencies. Only if the two modes are coupled at a proper level can a self–sustaining system arise, exhibiting a periodic cycle. The present study establishes the conditions for self–sustaining periodic solutions. It carefully documents the behaviour of the two mode system in order to facilitate the interpretation of global model behaviours when this energy–cycle is implemented as a closure into a convection parameterization in future.

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The Richards equation has been widely used for simulating soil water movement. However, the take-up of agro-hydrological models using the basic theory of soil water flow for optimizing irrigation, fertilizer and pesticide practices is still low. This is partly due to the difficulties in obtaining accurate values for soil hydraulic properties at a field scale. Here, we use an inverse technique to deduce the effective soil hydraulic properties, based on measuring the changes in the distribution of soil water with depth in a fallow field over a long period, subject to natural rainfall and evaporation using a robust micro Genetic Algorithm. A new optimized function was constructed from the soil water contents at different depths, and the soil water at field capacity. The deduced soil water retention curve was approximately parallel but higher than that derived from published pedo-tranfer functions for a given soil pressure head. The water contents calculated from the deduced soil hydraulic properties were in good agreement with the measured values. The reliability of the deduced soil hydraulic properties was tested in reproducing data measured from an independent experiment on the same soil cropped with leek. The calculation of root water uptake took account for both soil water potential and root density distribution. Results show that the predictions of soil water contents at various depths agree fairly well with the measurements, indicating that the inverse analysis is an effective and reliable approach to estimate soil hydraulic properties, and thus permits the simulation of soil water dynamics in both cropped and fallow soils in the field accurately. (C) 2009 Elsevier B.V. All rights reserved.

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BACKGROUND Methyl benzimidazole carbamate (MBC) fungicides are used to control the oilseed rape pathogen Pyrenopeziza brassicae. Resistance to MBCs has been reported in P. brassicae, but the molecular mechanism(s) associated with reductions in sensitivity have not been verified in this species. Elucidation of the genetic changes responsible for resistance, hypothesised to be target-site mutations in β-tubulin, will enable resistance diagnostics and thereby inform resistance management strategies. RESULTS P. brassicae isolates were classified as sensitive, moderately resistant or resistant to MBCs. Crossing P. brassicae isolates of different MBC sensitivities indicated that resistance was conferred by a single gene. The MBC-target encoding gene β-tubulin was cloned and sequenced. Reduced MBC sensitivity of field isolates correlated with β-tubulin amino acid substitutions L240F and E198A. The highest level of MBC resistance was measured for isolates carrying E198A. Negative cross-resistance between MBCs and the fungicides diethofencarb and zoxamide was only measured in E198A isolates. PCR-RFLP was used to screen isolates for the presence of L240F and E198A. The substitutions E198G and F200Y were also detected in DNA samples from P. brassicae populations after cloning and sequencing of PCR products. The frequencies of L240F and E198A in different P. brassicae populations were quantified by pyrosequencing. There were no differences in the frequencies of these alleles between P. brassicae populations sampled from different locations or after fungicide treatment regimes. CONCLUSIONS The molecular mechanisms affecting sensitivity to MBCs in P. brassicae have been identified. Pyrosequencing assays are a powerful tool for quantifying fungicide-resistant alleles in pathogen populations.

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Epigenetic modification of the genome via cytosine methylation is a dynamic process that responds to changes in the growing environment. This modification can also be heritable. The combination of both properties means that there is the potential for the life experiences of the parental generation to modify the methylation profiles of their offspring and so potentially to ‘pre-condition’ them to better accommodate abiotic conditions encountered by their parents. We recently identified high vapor pressure deficit (vpd)-induced DNA methylation at two gene loci in the stomatal development pathway and an associated reduction in leaf stomatal frequency.1 Here, we test whether this epigenetic modification pre-conditioned parents and their offspring to the more severe water stress of periodic drought. We found that three generations of high vpd-grown plants were better able to withstand periodic drought stress over two generations. This resistance was not directly associated with de novo methylation of the target stomata genes, but was associated with the cmt3 mutant’s inability to maintain asymmetric sequence context methylation. If our finding applies widely, it could have significant implications for evolutionary biology and breeding for stressful environments.

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We present a first overview of flows in the high latitude ionosphere observed at 15 s resolution using the U.K.-Polar EISCAT experiment. Data are described from experiments conducted on two days, 27 October 1984 and 29 August 1985, which together span the local times between about 0200 and 2130MLT and cover five different regions of ionospheric flow. With increasing local time, these are: the dawn auroral zone flow cell, the dayside region of low background flows equatorward of the flow cells, the dusk auroral zone flow cell, the boundary region between the dusk auroral zone and the polar cap, and the evening polar cap. Flows in both the equatorward and poleward portions of the auroral zone cells appear to be relatively smooth, while in the central region of high speed flow considerable variations are generally present. These have the form of irregular fluctuations on a wide range of time scales in the early morning dawn cell, and impulsive wave-like variations with periods of a few minutes in the afternoon dusk cell. In the dayside region between the flow cells, the ionosphere is often essentially stagnant for long intervals, but low amplitude ULF waves with a period of about 5 min can also occur and persist for many cycles. These conditions are punctuated at one to two hour intervals by sudden ‘flow burst’ events with impulsively generated damped wave trains. Initial burst flows are generally directed poleward and can peak at line-of-sight speeds in excess of 1 km s^{−1} after perhaps 45 s. Flows in the polar cap are reasonably smooth on time scales of a few minutes and show no evidence for the presence of ULF waves. Under most, but not all, of the above conditions, the beam-swinging algorithm used to determine background vector flows should produce meaningful results. Comparison of these flow data with simultaneous plasma and magnetic field measurements in the solar wind, made by the AMPTE IRM and UKS spacecraft, emphasizes the strong control exerted on high latitude flows by the north-south component of the IMF.

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There is growing recognition that hybrid organizations can play a critical role in tackling intractable global sustainable development challenges. At the same time, acute social, environmental, and economic challenges are opening up “opportunity” spaces for hybrids. Different institutional contexts are also leading to variable hybrid forms linked to the focus of their mission and their profit-oriented status. This article presents a process for identifying, mapping, and building impact indicators based on a study of 20 hybrid organizations in Sub-Saharan Africa.

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We present a novel algorithm for concurrent model state and parameter estimation in nonlinear dynamical systems. The new scheme uses ideas from three dimensional variational data assimilation (3D-Var) and the extended Kalman filter (EKF) together with the technique of state augmentation to estimate uncertain model parameters alongside the model state variables in a sequential filtering system. The method is relatively simple to implement and computationally inexpensive to run for large systems with relatively few parameters. We demonstrate the efficacy of the method via a series of identical twin experiments with three simple dynamical system models. The scheme is able to recover the parameter values to a good level of accuracy, even when observational data are noisy. We expect this new technique to be easily transferable to much larger models.

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There are over 500 candidate secreted effector proteins (CSEPs) or Blumeria effector candidates (BECs) specific to the barley powdery mildew pathogen Blumeria graminis f.sp. hordei. The CSEP/BEC proteins are expressed and predicted to be secreted by biotrophic feeding structures called haustoria. Eight BECs are required for the formation of functional haustoria. These include the RNase-like effector BEC1054 (synonym CSEP0064). In order to identify host proteins targeted by BEC1054, recombinant BEC1054 was expressed in E. coli, solubilized, and used in pull-down assays from barley protein extracts. Many putative interactors were identified by LC-MS/MS after subtraction of unspecific binders in negative controls. Therefore, a directed yeast-2-hybrid assay, developed to measure the effectiveness of the interactions in yeast, was used to validate putative interactors. We conclude that BEC1054 may target several host proteins, including a glutathione-S-transferase, a malate dehydrogenase, and a pathogen-related-5 protein isoform, indicating a possible role for BEC1054 in compromising well-known key players of defense and response to pathogens. In addition, BEC1054 interacts with an elongation factor 1 gamma. This study already suggests that BEC1054 plays a central role in barley powdery mildew virulence by acting at several levels.