The use of ultraviolet-blocking films in insect pest management in the UK; effects on naturally occurring arthropod pest and natural enemy populations in a protected cucumber crop

Studies in polytunnels were conducted to investigate the effects of ultraviolet (UV)-blocking films on naturally occurring insect pests and their arthropod natural enemies on a cucumber crop. Within tunnels clad with Antibotrytis (blocks light < 400 nm) and UVI/EVA (UV transmitting), 5.8 and 23.4 times more aphids, respectively, were recorded on traps compared with those on traps within tunnels clad with XL 385 (blocks light < 385 nm). When all plants within the UVI/EVA tunnels had become heavily infested with aphids, half of the plants in XL 385 tunnels were uninfested. More Coleoptera and thrips (approximately two times) were recorded under the UVI/EVA film than under the UV-blocking films, but for other arthropod pests (e. g. whitefly, leafhoppers), clear conclusions could not be drawn as low numbers were recorded. Substantial numbers of chalcid parasitoids and syrphids were found under the UV-blocking films, but further research is needed to evaluate fully the effect of such films on biological control of aphids. Higher syrphid numbers and more aphid mummies were recorded under the UVI/EVA film, probably because of the higher numbers of aphids present in tunnels clad with this film. The potential that UV-blocking films have as an effective component of commercial Integrated Pest Management (IPM) systems, for protected horticultural crops, is discussed.

The effect of potato variety mixtures on epidemics of late blight, in relation to plot size and level of resistance

Potatoes of a number of varieties of contrasting levels of resistance were planted in pure or mixed stands in four experiments over 3 years. Three experiments compared the late blight severity and progress in mixtures with that in pure stands. Disease on susceptible or moderately resistant varieties typical of those in commercial use was similar in mixtures and pure stands. In 2 of 3 years, there were slight reductions on cv. Sante, which is moderately susceptible, in mixture with cv. Cara, which is moderately resistant. Cara was unaffected by this mixture. Mixtures of an immune or near-immune partner with Cara or Sante substantially reduced disease on the latter. The effect of the size of plots of individual varieties or mixtures on blight severity was compared in two experiments. Larger plots had a greater area under the disease progress curve, but the average rate of disease progress was greater in smaller plots; this may be because most disease progress took place later, under more favourable conditions, in the smaller plots. In one experiment, two planting densities were used. Density had no effect on disease and did not interact with mixture effects. The overall conclusion is that, while mixtures of potato varieties may be desirable for other reasons, they do not offer any improvement on the average of the disease resistance of the components.

Diversity and systematics of the Malacosporea (Myxozoa)

Myxozoans belonging to the recently described class Malacosporea parasitize freshwater bryozoans during at least part of their life cycle. There are at present only two species described in this class: Buddenbrockia plumatellae and Tetracapsuloides bryosalmonae. The former can exist as vermiform and sac-like stages in bryozoan hosts. The latter, in addition to forming sac-like stages in bryozoans, is the causative agent of salmonid proliferative kidney disease (PKD). We undertook molecular and ultrastructural investigations of new malacosporean material to further resolve malacosporean diversity and systematics. Phylogenetic analyses of 18S rDNA sequences provided evidence for two new putative species belonging to the genus Buddenbrockia, revealing a two-fold increase in the diversity of malacosporeans known to date. One new malacosporean is a vermiform parasite infecting the bryozoan Fredericella sultana and the other occurs as sac-like stages in the rare bryozoan, Lophopus crystallinus. Both bryozoans represent new hosts for the genus Buddenbrockia. Our results have established that the malacosporean which infected F. sultana was not a vermiform stage of T. bryosalmonae, although it was collected from a site endemic for PKD. Ultrastructural investigation of new material of B. plumatellae revealed the presence of numerous external tubes associated with developing polar capsules, confirming that the absence of external tubes should no longer be considered as a character of the class Malacosporea.

The effects of temperature and light integral on early vegetative growth and chlorophyll fluorescence of four contrasting genotypes of cacao (Theobroma cacao)

The effect of temperature on early vegetative growth, leaf chlorophyll fluorescence and chlorophyll content was examined on four genotypes of cacao (Amelonado, AMAZ 15–15, SCA 6 and SPEC 54/1). A controlled environment glasshouse was used to simulate the temperature conditions of three cacao-growing regions (Bahia, Brazil; Tafo, Ghana and Lower Perak, Malaysia) over the course of a year. Base temperatures calculated from increments in main stem growth varied from 18.6°C for AMAZ 15/15 to 20.8°C for SPEC 54/1. Temporal variation in Fv/Fm observed for two of the clones (SCA 6 and SPEC 54/1) in two of the compartments were correlated with temperature differences over time. Significant differences were also recorded between genotypes in leaf chlorophyll content. It was shown that variation over time in leaf chlorophyll content could be quantified accurately as a function of temperature and light integral. The results imply that genetic variability exists in cacao in response to temperature stress.

Acclimation of photosynthesis to elevated CO2 in onion (allium cepa) grown at a range of temperatures

Onion (Allium cepa) was grown in the field within temperature gradient tunnels (providing about -2.5 degrees C to +2.5 degrees C from outside temperatures) maintained at either 374 or 532 mumol mol (-1) CO2. Plant leaf area was determined non-destructively at 7 day intervals until the time of bulbing in 12 combinations of temperature and CO2 concentration. Gas exchange was measured in each plot at the time of bulbing, and the carbohydrate content of the leaf (source) and bulb (sink) was determined. Maximum rate of leaf area expansion increased with mean temperature. Leaf area duration and maximum rate of leaf area expansion were not significantly affected by CO2. The light-saturated rates of leaf photosynthesis (A(sat)) were greater in plants grown at normal than at elevated CO2 concentrations at the same measurement CO2 concentration. Acclimation of photosynthesis decreased with an increase in growth temperature, and with an increase in leaf nitrogen content at elevated CO2. The ratio of intercellular to atmospheric CO2 (C-i/C-a ratio) was 7.4% less for plants grown at elevated compared with normal CO2. A(sat) in plants grown at elevated CO2 was less than in plants grown at normal CO2 when compared at the same C-i Hence, acclimation of photosynthesis was due both to stomatal acclimation and to limitations to biochemical CO2 fixation. Carbohydrate content of the onion bulbs was greater at elevated than at normal CO2. In contrast, carbohydrate content was less at elevated compared with normal CO2 in the leaf sections in which CO2 exchange was measured at the same developmental stage. Therefore, acclimation of photosynthesis in fully expanded onion leaves was detected despite the absence of localised carbohydrate accumulation in these field-grown crops.

The assessment of enriched apoplastic extracts using proteomic approaches

In plant tissues the extracellular environment or apoplast, incorporating the cell wall, is a highly dynamic compartment with a role in many important plant processes including defence, development, signalling and assimilate partitioning. Soluble apoplast proteins from Arabidopsis thaliana, Triticum aestivum and Oryza sativa were separated by two-dimensional electrophoresis. The molecular weights and isoelectric points for the dominant proteins were established prior to excision, sequencing and identification by matrix-assisted laser-desorption ionisation time of flight mass spectrometry (MALDI - TOF MS). From the selected spots, 23 proteins from O. sativa and 25 proteins from A. thaliana were sequenced, of which nine identifications were made in O. sativa (39%) and 14 in A. thaliana (56%). This analysis revealed that: (i) patterns of proteins revealed by two-dimensional electrophoresis were different for each species indicating that speciation could occur at the level of the apoplast, (ii) of the proteins characterised many belonged to diverse families reflecting the multiple functions of the apoplast and (iii), a large number of the apoplast proteins could not be identified indicating that the majority of extracellular proteins are yet to be assigned. The principal proteins identified in the aqueous matrix of the apoplast were involved in defence, i.e. germin-like proteins or glucanases, and cell expansion, i.e. β-D-glucan glucohydrolases. This study has demonstrated that proteomic analysis can be used to resolve the apoplastic protein complement and to identify adaptive changes induced by environmental effectors.

Chlorophyll fluorescence technique as a rapid diagnostic test of the effects of the photosynthetic inhibitor chlorotoluron on two winter wheat cultivars

A modified chlorophyll fluorescence technique was evaluated as a rapid diagnostic test of the susceptibility of wheat cultivars to chlorotoluron. Two winter wheat cultivars (Maris Huntsman and Mercia) exhibited differential response to the herbicide. All of the parameters of chlorophyll fluorescence examined were strongly influenced by herbicide concentration. Additionally, the procedure adopted here for the examination of winter wheat cultivar sensitivity to herbicide indicated that the area above the fluorescence induction curve and the ratio F-V/F-M are appropriate chlorophyll fluorescence parameters for detection of differential herbicide response between wheat cultivars. The potential use of this technique as an alternative to traditional methods of screening new winter wheat cultivars for their response to photosynthetic inhibitor herbicide is demonstrated here.

Inter-simple sequence repeat and aggressiveness analyses revealed high genetic diversity, recombination and long-range dispersal in Fusarium culmorum

Inter-simple sequence repeat (ISSR) analysis and aggressiveness assays were used to investigate genetic variability within a global collection of Fusarium culmorum isolates. A set of four ISSR primers were tested, of which three primers amplified a total of 37 bands out of which 30 (81%) were polymorphic. The intraspecific diversity was high, ranging from four to 28 different ISSR genotypes for F. culmorum depending on the primer. The combined analysis of ISSR data revealed 59 different genotypes clustered into seven distinct clades amongst 75 isolates of F. culmorum examined. All the isolates were assayed to test their aggressiveness on a winter wheat cv. 'Armada'. A significant quantitative variation for aggressiveness was found among the isolates. The ISSR and aggressiveness variation existed on a macro- as well as micro-geographical scale. The data suggested a long-range dispersal of F. culmorum and indicated that this fungus may have been introduced into Canada from Europe. In addition to the high level of intraspecific diversity observed in F. culmorum, the index of multilocus association calculated using ISSR data indicated that reproduction in F. culmorum cannot be exclusively clonal and recombination is likely to occur.

Development and application of diazirines in biological and synthetic macromolecular systems

Many different reagents and methodologies have been utilised for the modification of synthetic and biological macromolecular systems. In addition, an area of intense research at present is the construction of hybrid biosynthetic polymers, comprised of biologically active species immobilised or complexed with synthetic polymers. One of the most useful and widely applicable techniques available for functionalisation of macromolecular systems involves indiscriminate carbene insertion processes. The highly reactive and non-specific nature of carbenes has enabled a multitude of macromolecular structures to be functionalised without the need for specialised reagents or additives. The use of diazirines as stable carbene precursors has increased dramatically over the past twenty years and these reagents are fast becoming the most popular photophors for photoaffinity labelling and biological applications in which covalent modification of macromolecular structures is the basis to understanding structure-activity relationships. This review reports the synthesis and application of a diverse range of diazirines in macromolecular systems.

Structure and rheology of aqueous micellar solutions and gels formed from an associative poly(oxybutylene)-poly(oxyethylene)-poly(oxybutylene) triblock copolymer

The structure and shear flow behaviour of aqueous micellar solutions and gels formed by an amphiphilic poly(oxybutylene)-poly(oxyethylene)-poly(oxybutylene) triblock copolymer with a lengthy hydrophilic poly(oxyethylene) block has been investigated by rheology, small angle neutron scattering (SANS) and small-angle X-ray scattering (SAXS). SANS revealed that bridging of chains between micelles introduces, in the micellar solution, an attractive long-range component which can be described through a potential of interaction corresponding to sticky soft spheres. The strength of the attractive interaction increases with increasing concentration. Rheology showed that the dependence of the storage modulus with temperature can be explained as a function of the micellar bridging, micellisation and phase morphology. SAXS studies showed that the orientation adopted by the system in the get phase under shear is similar to that previously observed by us for the gel phase of a poly(oxyethylene)-poly(oxybutylene) diblock copolymer with a long poly(oxyethylene) chain, suggesting that the micellar corona/core length ratio and not the architecture of the block copolymer influences the alignment of the gel phase under shear.

Nanoshells and nanotubes from block copolymers

Recent work exploring the use of block copolymer vesicles and tubules is reviewed. The stability and toughness of block copolymer vesicles are enhanced compared to those formed by low molar mass amphiphiles. Functionality can also readily be introduced through the polymer chemistry or by incorporating additional components (for example pore-forming membrane proteins). This design flexibility leads to numerous potential applications in encapsulation, in targeted drug delivery, templating of inorganic materials and many others.

Ordering on multiple lengthscales in a series of side group liquid crystal block copolymers containing a cholesteryl-based mesogen

Hierarchical ordering in a side group liquid crystal block copolymer is investigated by differential scanning calorimetry, polarized optical microscopy, small-angle X-ray and neutron scattering (SAXS and SANS) and transmission electron microscopy (TEM). A series of block copolymers with a range of compositions was prepared by atom transfer radical polymerization, comprising a polystyrene block and a poly(methyl methacrylate) block bearing chiral cholesteryl mesogens. Smectic ordering is observed as well as microphase separation of the block copolymer. Lamellar structures were observed for far larger volume fractions than for coil-coil copolymers (up to a volume fraction of liquid crystal block, f(LC) = 0.8). A sample with f(LC) = 0.86 exhibited a hexagonal-packed cylinder morphology, as confirmed by SAXS and TEM. The matrix comprised the liquid crystal block, with the mesogens forming smectic layers. For the liquid crystal homopolymer and samples with high f(LC), a smectic-smectic phase transition was observed below the clearing point. At low temperature, the smectic phase comprises coexisting domains with monolayer S-A,S-1 coexisting with interdigitated S-A,S-d domains. At high temperature a SA,1 phase is observed. This is the only structure observed for samples with lower f(LC). These unprecedented results point to the influence of block copolymer microphase separation on the smectic ordering.

Proteomic profiling of neuromas reveals alterations in protein composition and local protein synthesis in hyper-excitable nerves

Neuropathic pain may arise following peripheral nerve injury though the molecular mechanisms associated with this are unclear. We used proteomic profiling to examine changes in protein expression associated with the formation of hyper-excitable neuromas derived from rodent saphenous nerves. A two-dimensional difference gel electrophoresis ( 2D-DIGE) profiling strategy was employed to examine protein expression changes between developing neuromas and normal nerves in whole tissue lysates. We found around 200 proteins which displayed a > 1.75-fold change in expression between neuroma and normal nerve and identified 55 of these proteins using mass spectrometry. We also used immunoblotting to examine the expression of low-abundance ion channels Nav1.3, Nav1.8 and calcium channel alpha 2 delta-1 subunit in this model, since they have previously been implicated in neuronal hyperexcitability associated with neuropathic pain. Finally, S(35)methionine in vitro labelling of neuroma and control samples was used to demonstrate local protein synthesis of neuron-specific genes. A number of cytoskeletal proteins, enzymes and proteins associated with oxidative stress were up-regulated in neuromas, whilst overall levels of voltage-gated ion channel proteins were unaffected. We conclude that altered mRNA levels reported in the somata of damaged DRG neurons do not necessarily reflect levels of altered proteins in hyper-excitable damaged nerve endings. An altered repertoire of protein expression, local protein synthesis and topological re-arrangements of ion channels may all play important roles in neuroma hyper-excitability.

Nanostructure formation in poly(gamma-benzyl-L-glutamate)-poly(ethylene glycol)-poly(gamma-benzyl-L-glutamate) triblock copolymers in the solid state

The morphology in the solid state of a series of triblock copolymers comprising a poly(ethylene glycol) (PEG) midblock and symmetric poly(gamma-benzyl-L-glutamate) (PBLG) end blocks has been studied using X-ray scattering and microscopy techniques. Transmission electron microscopy (TEM) on samples selectively stained with uranyl acetate provided clear assignment of morphologies for as-cast and annealed samples. The thickness of both PEG and PBLG domains was in good agreement with calculations based on the conformations of the respective chains, allowing for the crystal or amorphous state of PEG and the a-helical or P-sheet structure of the PBLG. Atomic force microscopy provided complementary information on surface morphology for several samples that was in good agreement with the structure observed by TEM. A morphology diagram was constructed. Cylindrical structures were observed for ordered samples with low f(PBLG), whereas at higher f(PLBG) there was evidence for broken lamellar and "hockey puck" nanostructures. Regular lamellae were observed for intermediate compositions.

Quantitative model for the kinetics of lyotropic phase transitions involving changes in monolayer curvature

We present a kinetic model for transformations between different self-assembled lipid structures. The model shows how data on the rates of phase transitions between mesophases of different geometries can be used to provide information on the mechanisms of the transformations and the transition states involved. This can be used, for example, to gain an insight into intermediate structures in cell membrane fission or fusion. In cases where the monolayer curvature changes on going from the initial to the final mesophase, we consider the phase transition to be driven primarily by the change in the relaxed curvature with pressure or temperature, which alters the relative curvature elastic energies of the two mesophase structures. Using this model, we have analyzed previously published kinetic data on the inter-conversion of inverse bicontinuous cubic phases in the 1-monoolein-30 wt% water system. The data are for a transition between QII(G) and QII(D) phases, and our analysis indicates that the transition state more closely resembles the QII(D) than the QII(G) phase. Using estimated values for the monolayer mean curvatures of the QII(G) and QII(D) phases of -0.123 nm(-1) and -0.133 nm(-1), respectively, gives values for the monolayer mean curvature of the transition state of between -0.131 nm(-1) and -0.132 nm(-1). Furthermore, we estimate that several thousand molecules undergo the phase transition cooperatively within one "cooperative unit", equivalent to 1-2 unit cells of QII(G) or 4-10 unit cells of QII(D).