Characterization of a Wheat Breeders’ Array suitable for high‐throughput SNP genotyping of global accessions of hexaploid bread wheat (Triticum aestivum)

Targeted selection and inbreeding have resulted in a lack of genetic diversity in elite hexaploid bread wheat accessions. Reduced diversity can be a limiting factor in the breeding of high yielding varieties and crucially can mean reduced resilience in the face of changing climate and resource pressures. Recent technological advances have enabled the development of molecular markers for use in the assessment and utilization of genetic diversity in hexaploid wheat. Starting with a large collection of 819 571 previously characterized wheat markers, here we describe the identification of 35 143 single nucleotide polymorphism‐based markers, which are highly suited to the genotyping of elite hexaploid wheat accessions. To assess their suitability, the markers have been validated using a commercial high‐density Affymetrix Axiom^® genotyping array (the Wheat Breeders’ Array), in a high‐throughput 384 microplate configuration, to characterize a diverse global collection of wheat accessions including landraces and elite lines derived from commercial breeding communities. We demonstrate that the Wheat Breeders’ Array is also suitable for generating high‐density genetic maps of previously uncharacterized populations and for characterizing novel genetic diversity produced by mutagenesis. To facilitate the use of the array by the wheat community, the markers, the associated sequence and the genotype information have been made available through the interactive web site ‘CerealsDB’.     

The application of genetic approaches for investigations of mycorrhizal symbioses

Genetic analyses of mycorrhizal symbioses have been far less common to date than molecular biological investigations. This review aims to address the problem that genetic research approaches are some of the least familiar to non specialists by providing some detailed explanations of the requirements and processes involved, including concepts of genetic variation and genetic mapping. Each section includes examples of research progress which is restricted to studies of arbuscular mycorrhizal (AM) and ectomycorrhizal (EcM) symbioses. Most such research has focussed on AM hosts or EcM fungi. For AM hosts, some early work on natural genetic variation has not been exploited yet, but new research with barley and clover will enable genetic mapping of mycorrhizal associated QTLs for the first time. EcM fungal studies have shown a genetic basis for mycorrhizal capacity and quantitative genetic differences in mycorrhizal capacity. Some recent work with EcM hosts has begun genetic mapping of QTLs associated with mycorrhizal status. Most AM genetic research has focussed on analysis of nodulation-defective mutants for their AM host status. Map-based cloning and characterisation of the first genes shown by these analyses to be essential for establishment of both nodulation and mycorrhizal symbioses are anticipated shortly. Comparisons with molecular and genetic research on plant disease resistance genes and signalling pathways may prove useful as those studies are more advanced and underlying biochemical and evolutionary relationships are likely to exist.     

The effect of fiber-type heterogeneity on optimized work and power output of hindlimb muscles of the salamander Ambystoma tigrinum

Most vertebrate muscles are composed of a mixture of fiber types. However, studies of muscle mechanics have concentrated on homogeneous bundles of fibers. Hindlimb muscles of the tiger salamander, Ambystoma tigrinum, present an excellent system to explore the consequences of fiber heterogeneity. Isometric twitches and work loops were obtained in vitro from two muscles, the m. iliotibialis pars posterior (heterogeneous, containing types I, IIa and IIb fibers) and the m. iliofibularis (nearly homogeneous for type IIa fibers). Maximal isometric twitch and tetanic stresses in m. iliotibialis posterior were significantly greater than in iliofibularis. Work loops were obtained over a range of frequencies (0.5-3.0 Hz) and strains (2-6% muscle length) that encompassed the observed ranges in vivo. Work per cycle from the homogeneous iliofibularis declined from 1.5-3.0 Hz, while that from the heterogeneous m. iliotibialis posterior increased from 0.5 Hz to 2.5 Hz and declined at 3.0 Hz. Power output from the iliofibularis rose with frequency to at least 3 Hz; power from the iliotibialis posterior rose with frequency to 2.5 Hz and declined thereafter. Mass-specific work per cycle and power output were higher in iliofibularis than iliotibialis posterior over most frequencies and strains tested.     

Site-Specific Management of Nematodes Pitfalls and Practicalities

The greatest constraint to potato production in the United Kingdom (UK) is damage by the potato cyst nematodes (PCN) Globodera pallida and G. rostochiensis. Management of PCN depends heavily on nematicides, which are costly. Of all the inputs in UK agriculture, nematicides offer the largest potential cost savings from spatially variable application, and these savings would be accompanied by environmental benefits. We mapped PCN infestations in potato fields and monitored the changes in population density and distribution that occurred when susceptible potato crops were grown. The inverse relationship between population density before planting and multiplication rate of PCN makes it difficult to devise reliable spatial nematicide application procedures, especially when the pre-planting population density is just less than the detection threshold. Also, the spatial dependence found suggests that the coarse sampling grids used commercially are likely to produce misleading distribution maps.     

Development of an edible subunit vaccine in corn against enterotoxigenic strains of escherichia coli

Summary Advances in the development of subunit vaccines and in the production of foreign proteins in plants together offer the prospect of stable and inexpensive vaccine delivery systems. Various bacterial and viral proteins stably produced in plants have been shown to elicit immune responses in feeding trials. We have extended this approach by using Zea mays as the plant production system. Corn has several advantages as a vaccine delivery vehicle, most notably established technologies to generate transgenic plants, to optimize traits through breeding and to process the seed into a palatable form. Here we report on the production in corn seed of the GM₁ receptor binding (B) subunit of the heat-labile toxin (Lt) from enterotoxigenic strains of Escherichia coli. Versions of the Lt-B gene were synthesized to give optimum codon usage for corn and to target the protein to either the cell surface or the cytoplasm. These synthetic genes were fused to a strong promoter and transformed into corn. Lt-B was highly expressed in corn seed at up to 1.8% of the total soluble protein and this was further increased approximately five-fold through plant breeding. As in E. coli. Lt-B produced in corn forms a functional pentamer that can bind to the GM₁ receptor. Furthermore, Lt-B pentamer stored in corn seed is much more resistant to heat than is the pure protein, allowing the transgenic corn to be readily processed into an edible form. This work demonstrates the potential of using products derived from transgenic corn seed as delivery vehicles for subunit vaccines.     

Construction of a linkage map of the Rennell Island Tall coconut type (Cocos nucifera L.) and QTL analysis for yield characters.

AFLP and SSR DNA markers were used to construct a linkage map in the coconut (Cocos nucifera L.; 2n = 32) type Rennell Island Tall (RIT). A total of 227 markers were arranged into 16 linkage groups. The total genome length corresponded to 1971 cM for the RIT map, with 5-23 markers per linkage group. QTL analysis for yield characters in two consecutive sampling periods identified nine loci. Three and two QTLs were detected for number of bunches and one and three QTLs for number of nuts. The correlation of trait values between characters and evaluation periods is partially reflected in identical QTLs. The QTLs represent characters that are important in coconut breeding. The cosegregation of markers with these QTLs provides an opportunity for marker-assisted selection in coconut breeding programmes.     

Effects of strobilurin containing fungicides on the deoxynivalenol content in winter wheat

The effects of the strobilurin fungicides Juwel, Juwel Top and Amistar on the deoxynivalenol contamination of winter wheat was studied in field experiments. In general, the application of strobilurins during stem elongation and inflorescence emergence of wheat resulted in increased deoxynivalenol contents in kernels as compared with the untreated control. This stimulating effect can be reversed by a following azole fungicide applied within a time period of one week during the stages of flowering.     

Effects of defoliation intensity on soil food-web properties in an experimental grassland community

We established a greenhouse experiment based on replicated mini‐ecosystems to evaluate the effects of defoliation intensity on soil food‐web properties in grasslands. Plant communities, composed of white clover (Trifolium repens), perennial ryegrass (Lolium perenne) and plantain (Plantago lanceolata) with well‐established root and shoot systems, were subjected to five defoliation intensity treatments: no trimming (defoliation intensity 0, or DI 0), and trimming of all plant material to 35 cm (DI 1), 25 cm (DI 2), 15 cm (DI 3) and 10 cm (DI 4) above soil surface every second week for 14 weeks. Intensification of defoliation reduced shoot production and standing shoot and root mass of plant communities but increased their root to shoot ratio. Soil microbial activity and biomass decreased with intensification of defoliation. Concentrations of NO₃–N in soil steadily increased with intensifying defoliation, whereas NH₄–N concentrations did not vary between treatments. Numbers of microbi‐detritivorous enchytraeids, bacterial‐feeding rotifers and bacterial‐feeding nematodes steadily increased with intensifying defoliation, while the abundance of fungal‐feeding nematodes was significantly enhanced only in DI 3 and DI 4 relative to DI 0. The abundance of herbivorous nematodes per unit soil mass was lower in DI 3 and DI 4 than in DI 0, DI 1 and DI 2, but when calculated per unit root mass, their abundance tended to increase with defoliation intensity. The abundance of omnivorous and predatory nematodes appeared to be highest in the most intensely defoliated systems. The ratio of abundance of fungal‐feeding nematodes to that of bacterial‐feeding nematodes was not significantly affected by defoliation intensity. The results infer that defoliation intensity may significantly alter the structure of soil food webs in grasslands, and that defoliation per se is able to induce patterns observed in grazing studies in the field. The results did not support hypotheses that defoliation per se would cause a shift between the bacterial‐based and fungal‐based energy channels in the decomposer food web, or that herbivore and detritivore densities in soil would be highest under intermediate defoliation. Furthermore, our data for microbes and microbial feeders implies that the effects of defoliation intensity on soil food‐web structure may depend on the duration of defoliation and are therefore likely to be dynamic rather than constant in nature.     

The Mycobacterium marinum G13 promoter is a strong sigma 70-like promoter that is expressed in Escherichia coli and mycobacteria species

A Mycobacterium marinum promoter, designated G13, was isolated from a promoter-trap library as a constitutive producer of the mutant green fluorescent protein. Sequence analysis, primer extension analysis, and computer promoter prediction analysis indicate that the G13 promoter is very similar to Escherichia coli consensus sigma 70 promoters. Expression of the green fluorescent protein from the G13 promoter in M. marinum is, however, up to 40 times higher than that seen from the mycobacterial hsp60 promoter during exponential growth. Further, expression from this promoter does not appear to affect the growth of the organism in culture media or in macrophages. The strong expression of the G13 promoter allows it to be developed as a useful molecular tool for high level expression of markers in vitro.