Dissecting the T-cell response to hordeins in coeliac disease can develop barley with reduced immunotoxicity.

BACKGROUND: Wheat, rye and barley prolamins are toxic to patients with coeliac disease. Barley is diploid with pure inbred cultivars available, and is attractive for genetic approaches to detoxification. AIM: To identify barley hordein fractions which activated the interferon-γ (IFN-γ) secreting peripheral blood T-cells from coeliac volunteers, and compare immunotoxicity of hordeins from experimental barley lines. METHODS: To reactivate a T-cell response to hordein, volunteers underwent a 3-day oral barley challenge. Peripheral blood mononuclear cells (PBMC) were isolated from twenty-one HLA DQ2(+) patients with confirmed coeliac disease. IFN-γ ELISpot assays enumerated T-cells activated by purified prolamins and positive controls. RESULTS: Hordein-specific T-cells were induced by oral barley challenge. All prolamin fractions were immunotoxic, but D- and C-hordeins were most active. Barley lines lacking B- and C-hordeins had a 5-fold reduced hordein-content, and immunotoxicity of hordein extracts were reduced 20-fold compared with wild-type barley. CONCLUSIONS: In vivo oral barley challenge offers a convenient and rapid approach to test the immunotoxicity of small amounts of purified hordeins using fresh T-cells from patients in high throughput overnight assays. Barley lines that did not accumulate B- and C-hordeins were viable, yet had substantially reduced immunotoxicity. Creation of hordein-free barley may therefore be possible.

Impact of the biological control agent Mesoclanis polana (Tephritidae) on bitou bush (Chrysanthemoides monilifera subsp. rotundata) in eastern Australia.

The seed fly Mesoclanis polana (Diptera: Tephritidae) was released in Australia in 1996. Its impact on seed production of bitou bush (Chrysanthemoides monilifera subsp. rotundata) was monitored at eight sites along the New South Wales coast from 1996 to 2004. Peak flowerhead production occurred in autumn (March to May); therefore, samples collected in May of each year were used to compare abundance and impact of M. polana across sites and between years. Latitude had a significant effect on abundance and impact of M. polana. By May 2004, 99.6% of flowerheads at the five most northern sites contained at least one egg, while 64% of flowerheads from the two most southern sites contained at least one egg. In May 2004, mean numbers of M. polana eggs per flowerhead were between 13 and 17 at four of the six northern sites and below two at the two southern sites. In May 2004, average seed destruction by M. polana was 58% at the four most northern sites and 11% at the two most southern sites. The highest level of seed destruction recorded was 86% at Kingscliff in May 2003. Percentage seed destruction in May provided a reasonable estimate of seed destruction for the whole year. Parasitoids were found throughout the range of M. polana in Australia. Their attack rate on M. polana was less than 10% at all sites, except at the two most northern sites where parasitism of up to 27% was recorded. Parasitism results are compared with predictions made following an earlier study. The release of Mesoclanis magnipalpis, a species suited to cooler environments, is recommended for the southern range of C. monilifera subsp. rotundata.

Research Note: The five families of sucrose-phosphate synthase genes in Saccharum spp. are differentially expressed in leaves and stem.

Sucrose-phosphate synthase (SPS) is a key enzyme in the pathway of sucrose synthesis. Five different gene families encoding SPS have been reported in the Poaceae [Castleden CK, Aoki N, Gillespie VJ, MacRae EA, Quick WP, Buchner P, Foyer CH, Furbank RT, Lunn JE (2004) Evolution and function of the sucrose-phosphate synthase gene families in wheat and other grasses. Plant Physiology 135, 1753-1764]. Expression of the five families in leaf and stem tissues of Saccharum spp. at different stages of development was determined by quantitative real-time PCR. The type B and C families of SPS genes were predominantly expressed in both immature and mature leaves, whereas the two subfamilies making up the type D family were expressed at similar levels in all tissues examined. In the type A family, expression was lowest in leaves and increased from the meristem region down to internode 7 of the stem.

Nuclear DNA regulates the level of ribulose 1,5-bisphosphate carboxylase oxygenase in Medicago sativa L.

The response to selection for leaf proteins was studied during three selection cycles. Selection for high total nitrogen content showed 75% heritability, and the levels of both ribulose 1,5-bisphosphate carboxylase oxygenase (Rubisco) and cytoplasmic protein were strongly under nuclear DNA control. High and low protein content were correlated with chloroplast area. Although the amounts of nuclear DNA were similar, the ratio of Rubisco/DNA and chlorophyll/DNA changed during the selection process. It can be concluded that the levels of Rubisco achieved in mature plants of M. sativa are under nuclear DNA control. The possible involvement of small subunit (SSU) genes in controlling these levels is discussed.

Effects of certain cations on the formation and infectivity of Phytophthora zoospores. 1. Effects of calcium, magnesium, potassium, and iron ions.

The effects of four cations, Ca2+, Mg2+, K+, and Fe3+, upon the production of zoosporangia by isolates of four species of Phytophthora in axenic culture have been investigated. A response surface design was used to examine main effects and possible interactions. Responses to Ca2+ and Fe3+ were strongly quadratic with the higher concentrations inhibiting sporangial production in P. cinnamomi and P. drechsleri isolates. Responses to Mg2+ and K+ were weaker and, in the case of magnesium, were linear rather than quadratic. There was no significant interaction between cations except in the case of P. cambivora where the interaction between Mg2+ and K+ was significant at the 5% level. For each cation, the optimum concentration for zoosporangial production was calculated. The processes of zoospore release and infection of plant material by P. drechsleri showed a strong linear and weak quadratic response to Ca2+, higher concentrations of Ca2+ favouring these processes. A strong quadratic response was obtained to Mg2+, K+, and Fe3+. Interaction between Ca2+ and Mg2+ was evident for all three host materials used; Mg2+ and K+ showed an interaction when Pinus radiata was used as the host material. The implications and limitations of these results are discussed.