Evaluation of the immunogenicity of the P97R1 adhesin of Mycoplasma hyopneumoniae as a mucosal vaccine in mice.

The immunogenicity of P97 adhesin repeat region R1 (P97R1) of Mycoplasma hyopneumoniae, an important pathogenesis-associated region of P97, was evaluated in mice as a mucosal vaccine. Mice were immunized orally with attenuated Salmonella typhimurium aroA strain CS332 harbouring a eukaryotic or prokaryotic expression vector encoding IP97R1. Local and systemic immune responses were analysed by ELISA on mouse sera, lung washes and splenocyte supernatants following splenocyte stimulation with specific antigens in vitro. Although no P97R1-specific antibody responses were detected in serum and lung washes, significant gamma interferon was produced by P97R1-stimulated splenocytes from mice immunized orally with S. typhimurium aroA harbouring either expression system, indicating induction of a cell-mediated immune response. These results suggested that live bacterial vectors carrying DNA vaccines or expressing heterologous antigens preferentially induce a Th1 response. Surprisingly, however, mice immunized with the vaccine carrier S. typhimurium aroA CS332 induced serum IgG, but not mucosal IgA, against P97R1 or S. typhimurium aroA CS332 whole-cell lysate, emphasizing the importance of assessing the suitability of attenuated S. typhimurium antigen-carrier delivery vectors in the mouse model prior to their evaluation as potential vaccines in the target species, which in this instance was pigs.

Comparative immunogenicity of M. hyopneumoniae NrdF encoded in different expression systems delivered orally via attenuated S. typhimurium aroA in mice.

The Mycoplasma hyopneumoniae ribonucleotide reductase R2 subunit (NrdF) gene fragment was cloned into eukaryotic and prokaryotic expression vectors and its immunogenicity evaluated in mice immunized orally with attenuated Salmonella typhimurium aroA CS332 harboring either of the recombinant expression plasmids. We found that NrdF is highly conserved among M. hyopneumoniae strains. The immunogenicity of NrdF was examined by analyzing antibody responses in sera and lung washes, and the cell-mediated immune (CMI) response was assessed by determining the INF-[gamma] level produced by splenocytes upon in vitro stimulation with NrdF antigen. S. typhimurium expressing NrdF encoded by the prokaryotic expression plasmid (pTrcNrdF) failed to elicit an NrdF-specific serum or secretory antibody response, and IFN-[gamma] was not produced. Similarly, S. typhimurium carrying the eukaryotic recombinant plasmid encoding NrdF (pcNrdF) did not induce a serum or secretory antibody response, but did elicit significant NrdF-specific IFN-[gamma] production, indicating induction of a CMI response. However, analysis of immune responses against the live vector S. typhimurium aroA CS332 showed a serum IgG response but no mucosal IgA response in spite of its efficient invasiveness in vitro. In the present study we show that the DNA vaccine encoding the M. hyopneumoniae antigen delivered orally via a live attenuated S. typhimurium aroA can induce a cell-mediated immune response. We also indicate that different live bacterial vaccine carriers may have an influence on the type of the immune response induced.

Transgenic gene silencing strategies for virus control.

Co-suppression of transgenes and their homologous viral sequences by RNA silencing is a powerful strategy for achieving high-level virus resistance in plants. This review provides a brief overview of RNA silencing mechanisms in plants and discusses important transgene construct design features underpinning successful RNA silencing-mediated transgenic virus control. Application of those strategies to protect horticultural and field crops from virus infection and results of field tests are also provided. The effectiveness and stability of RNA-mediated transgenic resistance are assessed taking into account effects of viral, plant and environmental factors.

Transgenic rootstocks for production of GM-free fruit

Development of regeneration and transformation methods for genetic improvement of rootstocks for mango, avocado and citrus.

Improved Integrated Weed Management systems in transgenic farming landscapes

This project will develop and deliver improved integrated weed management strategies for weeds at risk of glyphosate resistance and species shift in transgenic farming landscapes. It will also facilitate the stewarship of glyphosate and transgenic technology, improving the sustainability of both the herbicide and the genes.

Population dynamics of house mice in Queensland grain-growing areas

Context. Irregular plagues of house mice cause high production losses in grain crops in Australia. If plagues can be forecast through broad-scale monitoring or model-based prediction, then mice can be proactively controlled by poison baiting. Aims. To predict mouse plagues in grain crops in Queensland and assess the value of broad-scale monitoring. Methods. Regular trapping of mice at the same sites on the Darling Downs in southern Queensland has been undertaken since 1974. This provides an index of abundance over time that can be related to rainfall, crop yield, winter temperature and past mouse abundance. Other sites have been trapped over a shorter time period elsewhere on the Darling Downs and in central Queensland, allowing a comparison of mouse population dynamics and cross-validation of models predicting mouse abundance. Key results. On the regularly trapped 32-km transect on the Darling Downs, damaging mouse densities occur in 50% of years and a plague in 25% of years, with no detectable increase in mean monthly mouse abundance over the past 35 years. High mouse abundance on this transect is not consistently matched by high abundance in the broader area. Annual maximum mouse abundance in autumn–winter can be predicted (R2 = 57%) from spring mouse abundance and autumn–winter rainfall in the previous year. In central Queensland, mouse dynamics contrast with those on the Darling Downs and lack the distinct annual cycle, with peak abundance occurring in any month outside early spring.Onaverage, damaging mouse densities occur in 1 in 3 years and a plague occurs in 1 in 7 years. The dynamics of mouse populations on two transects ~70 km apart were rarely synchronous. Autumn–winter rainfall can indicate mouse abundance in some seasons (R2 = ~52%). Conclusion. Early warning of mouse plague formation in Queensland grain crops from regional models should trigger farm-based monitoring. This can be incorporated with rainfall into a simple model predicting future abundance that will determine any need for mouse control. Implications. A model-based warning of a possible mouse plague can highlight the need for local monitoring of mouse activity, which in turn could trigger poison baiting to prevent further mouse build-up.

Field evaluation of transgenic pineapple (Ananas comosus (L.) Merr.) cv. ‘Smooth Cayenne’ for resistance to blackheart under subtropical conditions

A comparative analysis of transgenic pineapple lines transformed with a polyphenol oxidase (PPO) gene (ppo) and the untransformed cultivar ‘Smooth Cayenne’ was made from plants grown in a series of field trials under cool subtropical conditions in southeast Queensland. In the four field trials where blackheart was recorded, all of the control lines expressed blackheart on each occasion and exhibited the greatest incidence (50%) and severity (34%) of symptoms. Irrespective of the gene transfer method or the gene construct used, 38% of the lines produced were regarded as blackheart resistant, having no blackheart symptoms in two or more trials. Five blackheart resistant transgenic lines consistently performed as well as or better than control plants in terms of fruit characteristics and quality.

IPM in the transgenic era: a review of the challenges from emerging pests in Australian cotton systems

The Cotton Catchment Communities Cooperative Research Centre began during a period of rapid uptake of Bollgard II® cotton, which contains genes to express two Bt proteins that control the primary pests of cotton in Australia, Helicoverpa armigera and H. punctigera. The dramatic uptake of this technology presumably resulted in strong selection pressure for resistance in Helicoverpa spp. against the Bt proteins. The discovery of higher than expected levels of resistance in both species against one of the proteins in Bollgard II® cotton (Cry2Ab) led to significant re-evaluation of the resistance management plan developed for this technology, which was a core area of research for the Cotton CRC. The uptake of Bollgard II® cotton also led to a substantial decline in pesticide applications against Helicoverpa spp. (from 10–14 to 0–3 applications per season). The low spray environment allowed some pests not controlled by the Bt proteins to emerge as more significant pests, especially sucking species such as Creontiades dilutus and Nezara viridula. A range of other minor pests have also sporadically arisen as problems. Lack of knowledge and experience with these pests created uncertainty and encouraged insecticide use, which threatened to undermine the gains made with Bollgard II® cotton. Here we chronicle the achievements of the Cotton CRC in providing the industry with new knowledge and management strategies for these pests.