Developing and communicating strategies for controlling virus diseases in vegetable cucurbit crops

Virus diseases cause serious yield and quality losses in field grown cucurbit crops worldwide. In Australia, the main viruses of cucurbits are Papaya ringspot virus (PRSV), Squash mosaic virus (SqMV), Watermelon mosaic virus (WMV) and Zucchini yellow mosaic virus (ZYMV). Plants infected early have severely distorted fruit. High infection incidences, of ZYMV and PRSV in crops cause losses of marketable fruit of up to 100% and infected crops are often abandoned. Two new alternative hosts of ZYMV were identified, the native cucurbit Cucumis maderaspatanus and wild legume Rhyncosia minima. No new alternative hosts of PRSV, SqMV or WMV were found in Western Australia or Queensland. Seed transmission of ZYMV (0.7%) was found in seedlings grown from ZYMV-infected fruit of zucchini but not of pumpkin. None was detected with PRSV or SqMV in zucchini or pumpkin seedlings, respectively. ZYMV spread to pumpkins by aphids was greater downwind than upwind of a virus source. Delaying sowing by 2 weeks decreased ZYMV spread. Millet non-host barriers between pumpkin plantings slowed ZYMV infection. Host resistance gene (zym) in cucumber cultivars was effective against ZYMV. Pumpkin cultivars with resistance gene (Zym) became infected under high virus pressure but leaf symptoms were milder and infected plants higher yielding with more market-acceptable fruit than those without Zym. Most zucchini cultivars with Zym developed severe leaf and fruit symptoms. ZYMV, PRSV, WMV and SqMV spread readily from infected to healthy cucurbit plants by direct leaf contact. ZYMV survives and remains infective on diverse surfaces for up to 6 hours but can be inactivated by some disinfectants. Phylogenetic analysis indicates at least three separate introductions of ZYMV into Australia, with new introductions rarely occurring. ZYMV isolates clustered into three groups according to collection location i) Kununurra, ii) Northern Territory and iii) Carnarvon, Qld and Vic. A multiplex Real-Time PCR was developed which distinguished between the three groups of Australian isolates. Integrated disease management (IDM) strategies for virus diseases of vegetable cucurbit crops grown in the field were improved incorporating the new information gathered. These strategies are aimed at causing using minimal extra expense, labour demands and disruption to normal practices.

Strategies for collecting screen-less oscillation data

Screen-less oscillation photography is the method of choice for recording three-dimensional X-ray diffraction data for crystals of biological macromolecules. The geometry of an oscillation camera is extremely simple. However, the manner in which the reciprocal lattice is recorded in any experiment is fairly complex. This depends on the Laue symmetry of the reciprocal lattice, the lattice type, the orientation of the crystal on the camera and to a lesser extent on the unit-cell dimensions. Exploring the relative efficiency of collecting X-ray diffraction data for different crystal orientations prior to data collection might reduce the number of films required to record most of the unique data and the consequent amount of time required for processing these films. Here algorithms are presented suitable for this purpose and results are reported for the 11 Laue groups, different lattice types and crystal orientations often employed in data collection.

Participatory adaptation and mitigation strategies for climate change on mixed farms

The project uses participatory methods to engage primary producers and advisers in central Queensland, southern Queensland, and north east New South Wales on-farm trials and demonstrations to adapt mixed farming systems to changed climate conditions. The focus is adaptation to climate change but will support abatement of greenhouse gas emissions by building soil carbon, better managing soil nitrogen and soil organic carbon. Data will be collected and integrated with data from Round 1 of the Climate Change Research Program to extend industry understanding beyond a general awareness of ‘climate change’. Nitrous oxide and soil carbon data will help farmers/advisers understand the implications of climate change and develop adaptation strategies for a more sustainable, climate sensitive future.

Risk assessment and preventive strategies for herbicide resistance in NR (phase III)

The threat and management of glyphosate# resistant weeds are major issues facing northern region growers. At present five weeds are confirmed glyphosate-resistant: barnyard grass, liverseed grass, windmill grass, annual ryegrass and flaxleaf fleabane. This project used 25 experiments to investigate the ecology of the grass weeds, plus new or improved chemical and non-chemical control tactics for them. The refined glyphosate resistance model developed in this project used the experiments' findings to predict the long-term impacts on evolution of resistance and on seed bank numbers of resistant weeds. These data led to revised management and resistance avoidance strategies, which were published in the Reporter newsletter, and via an on-line risk assessment tool. - See more at: http://finalreports.grdc.com.au/UQ00054#sthash.oTkCN4Sk.dpuf

Insights into the molecular genetics of celiac disease : Applying family- and population-based strategies

Celiac disease, or gluten intolerance, is triggered by dietary glutens in genetically susceptible individuals and it affects approximately 1% of the Caucasian population. The best known genetic risk factors for celiac disease are HLA DQ2 and DQ8 heterodimers, which are necessary for the development of the disease. However, they alone are not sufficient for disease induction, other risk factors are required. This thesis investigated genetic factors for celiac disease, concentrating on susceptibility loci on chromosomes 5q31-q33, 19p13 and 2q12 previously reported in genome-wide linkage and association studies. In addition, a novel genotyping method for the detection of HLA DQ2 and DQ8 coding haplotypes was validated. This study was conducted using Finnish and Hungarian family materials, and Finnish, Hungarian and Italian case-control materials. Genetic linkage and association were analysed in these materials using candidate gene and fine-mapping approaches. The results confirmed linkage to celiac disease on the chromosomal regions 5q31-q33 and 19p13. Fine-mapping on chromosome 5q31-q33 revealed several modest associations in the region, and highlighted the need for further investigations to locate the causal risk variants. The MYO9B gene on chromosome 19p13 showed evidence for linkage and association particularly with dermatitis herpetiformis, the skin manifestation of celiac disease. This implies a potential difference in the genetic background of the intestinal and skin forms of the disease, although studies on larger samplesets are required. The IL18RAP locus on chromosome 2q12, shown to be associated with celiac disease in a previous genome-wide association study and a subsequent follow-up, showed association in the Hungarian population in this study. The expression of IL18RAP was further investigated in small intestinal tissue and in peripheral blood mononuclear cells. The results showed that IL18RAP is expressed in the relevant tissues. Two putative isoforms of IL18RAP were detected by Western blot analysis, and the results suggested that the ratios and total levels of these isoforms may contribute to the aetiology of celiac disease. A novel genotyping method for celiac disease-associated HLA haplotypes was also validated in this thesis. The method utilises single-nucleotide polymorphisms tagging these HLA haplotypes with high sensitivity and specificity. Our results suggest that this method is transferable between populations, and it is suitable for large-scale analysis. In conclusion, this doctorate study provides an insight into the roles of the 5q31-q33, MYO9B, IL18RAP and HLA loci in the susceptibility to celiac disease in the Finnish, Hungarian and Italian populations, highlighting the need for further studies at these genetic loci and examination of the function of the candidate genes.

2.2.2.2 Improved Management Summer Weeds: Best weed management strategies dryland cropping systems

To undertake a scoping study to identify the major issues in weed management in dryland cropping systems with cotton.

Weed management strategies for farming systems with herbicide tolerant cotton.

The introduction of glyphosate tolerant cotton has significantly improved the flexibility and management of a number of problem weeds in cotton systems. However, reliance on glyphosate poses risks to the industry in term of glyphosate resistance and species shift. The aims of this project were to identify these risks, and determine strategies to prevent and mitigate the potential for resistance evolution. Field surveys identified fleabane as the most common weed now in both irrigated and dryland system. Sowthistle has also increased in prevalence, and bladder ketmia and peachvine remained common. The continued reliance on glyphosate has favoured small seeded, and glyphosate tolerant species. Fleabane is both of these, with populations confirmed resistant in grains systems in Queensland and NSW. When species were assessed for their resistance risk, fleabane, liverseed grass, feathertop Rhodes grass, sowthistle and barnyard grass were determined to have high risk ratings. Management practices were also determined to rely heavily on glyphosate and therefore be high risk in summer fallows, and dryland glyphosate tolerant and conventional cotton. Situations were these high risk species are present in high risk cropping phases need particular attention. The confirmation of a glyphosate resistance barnyard grass population in a dryland glyphosate tolerant cotton system means resistance is now a reality for the cotton industry. However, experiments have shown that resistant populations can be managed with other herbicide options currently available. However, the options for fleabane management in cotton are still limited. Although some selective residual herbicides are showing promise, the majority of fleabane control tactics can only be used in other phases of the cotton rotation. An online glyphosate resistance tool has been developed. This tool allows growers to assess their individual glyphosate resistance risks, and how they can adjust their practices to reduce their risks. It also provides researchers with current information on weed species present and practices used across the industry. This tool will be extremely useful in tailoring future research and extension efforts. Simulations from the expanded glyphosate resistance model have shown that glyphosate resistance can be prevented and managed in glyphosate-tolerant cotton farming systems. However, for strategies to be successful, some effort is required. Simulations have shown the importance of controlling survivors of glyphosate applications, using effective glyphosate alternatives in fallows, and combining several effective glyphosate alternatives in crop, and these are the key to the prevention and management of glyphosate resistance.

Efficient organogenesis of an Australian passionfruit hybrid (Passiflora edulis x Passiflora edulis var. flavicarpa) suitable for gene delivery

An efficient regeneration protocol based on organogenesis from cotyledon explants and suitable for gene delivery has been developed for an Australian passionfruit hybrid. Multiple shoots were regenerated from 30-day-old cotyledon explants on Murashige and Skoog (MS) medium containing 6-benzylvaminopurine (BAP) and coconut water. Media pulsing experiments were conducted to investigate the effect on organogenesis of exposure time of the explants to MS containing 10 mu M BAP and 10% (v/v) coconut water, i.e. passionfruit regeneration medium (PRM). Continuous exposure of these explants to PRM maximised the number of shoots produced to 12.1 per explant. However, periods on hormone-free medium improved the appearance of the shoots and increased the number of explants with shoots from 75 to 84.6%. Further, shoots exposed for 7 days to half-strength MS supplemented with 10 mu M NAA (1-napthalene acetic acid) produced twice as many plantlets than those on half-strength MS alone. Transient GUS histochemical assays indicated delivery of the uidA gene via Agrobacterium tumefaciens.

New molecular strategies for prevention of fibroproliferative vascular disease : An experimental approach to restenosis

Vascular intimal hyperplasia is a major complication following angioplasty. The hallmark feature of this disorder is accumulation of dedifferentiated smooth muscle cells (SMCs) to the luminal side of the injured artery, cellular proliferation, migration, and synthesis of extracellular matrix. This finally results in intimal hyperplasia, which is currently considered an untreatable condition. According to current knowledge, a major part of neointimal cells derive from circulating precursor cells. This has outdated the traditional in vitro cell culture methods of studying neointimal cell migration and proliferation using cultured medial SMCs. Somatostatin and some of its analogs with different selectivity for the five somatostatin receptors (sst1 through sst5) have been shown to have vasculoprotective properties in animal studies. However, clinical trials using analogs selective for sst2/sst3/sst5 to prevent restenosis after percutaneous transluminal coronary angioplasty (PTCA) have failed to show any major benefits. Sirolimus is a cell cycle inhibitor that has been suggested to act synergistically with the protein-tyrosine kinase inhibitor imatinib to inhibit intimal hyperplasia in rat already at well-tolerated submaximal oral doses. The mechanisms behind this synergy and its long-term efficacy are not known. The aim of this study was to set up an ex vivo vascular explant culture model to measure neointimal cell activity without excluding the participation of circulating progenitor cells. Furthermore, two novel potential vasculoprotective treatment strategies were evaluated in detail in rat models of intimal hyperplasia and in the ex vivo explant model: sst1/sst4-selective somatostatin receptor analogs and combination treatment with sirolimus and imatinib. This study shows how whole vessel explants can be used to study the kinetics of neointimal cells and their progenitors, and to evaluate the anti-migratory and anti-proliferative properties of potential vasculoprotective compounds. It also shows how the influx of neointimal progenitor cells occurs already during the first days after vascular injury, how the contribution of cell migration is more important in the injury response than cell proliferation, and how the adventitia actively contribute in vascular repair. The vasculoprotective effect of somatostatin is mediated preferentially through sst4, and through inhibition of cell migration rather than of proliferation, which may explain why sst2/sst3/sst5-selective analogs have failed in clinical trials. Furthermore, a brief early oral treatment with the combination of sirolimus and imatinib at submaximal doses results in long-term synergistic suppression of intimal hyperplasia. The synergy is a result of inhibition of post-operative thrombocytosis and leukocytosis, inhibition of neointimal cell migration to the injury-site, and maintenance of cell integrity by inhibition of apoptosis and SMC dedifferentiation. In conclusion, the influx of progenitor cells already during the first days after injury and the high neointimal cell migratory activity underlines the importance of early therapeutic intervention with anti-migratory compounds to prevent neointimal hyperplasia. Sst4-selective analogs and the combination therapy with sirolimus and imatinib represent potential targets for the development of such vasculoprotective therapies.

A hybrid clustering procedure for concentric and chain-like clusters

K-means algorithm is a well known nonhierarchical method for clustering data. The most important limitations of this algorithm are that: (1) it gives final clusters on the basis of the cluster centroids or the seed points chosen initially, and (2) it is appropriate for data sets having fairly isotropic clusters. But this algorithm has the advantage of low computation and storage requirements. On the other hand, hierarchical agglomerative clustering algorithm, which can cluster nonisotropic (chain-like and concentric) clusters, requires high storage and computation requirements. This paper suggests a new method for selecting the initial seed points, so that theK-means algorithm gives the same results for any input data order. This paper also describes a hybrid clustering algorithm, based on the concepts of multilevel theory, which is nonhierarchical at the first level and hierarchical from second level onwards, to cluster data sets having (i) chain-like clusters and (ii) concentric clusters. It is observed that this hybrid clustering algorithm gives the same results as the hierarchical clustering algorithm, with less computation and storage requirements.

Reproductive strategies of two invasive tilapia species Oreochromis mossambicus and Tilapia mariae in northern Australia

The reproductive biology of two invasive tilapia species, Oreochromis mossambicus and Tilapia mariae, resident in freshwater habitats in north-eastern Australia was investigated. Oreochromis mossambicus exhibited plasticity in some of its life-history characteristics that enhanced its ability to occupy a range of habitats. These included a shallow, weed-choked, freshwater coastal drain that was subject to temperature and dissolved oxygen extremes and water-level fluctuations to cooler, relatively high-altitude impoundments. Adaptations to harsher conditions included a decreased total length (LT) and age ( A) at 50% maturity (m50), short somatic growth intervals, early maturation and higher relative fecundities. Potential fecundity in both species was relatively low, but parental care ensured high survival rates of both eggs and larvae. No significant difference in the relative fecundity of T. mariae populations in a large impoundment and a coastal river was found, but there were significant differences in relative fecundities between several of the O. mossambicus populations sampled. Total length ( LT) and age at 50% maturity of O. mossambicus populations varied considerably depending on habitat. The LTm50 and Am50 values for male and female O. mossambicus in a large impoundment were considerably greater than for those resident in a small coastal drain. Monthly gonad developmental stages and gonado-somatic indices suggested that in coastal areas, spawning of O. mossambicus and T. mariae occurred throughout most of the year while in cooler, high-altitude impoundments, spawning peaked in the warmer, summer months. The contribution these reproductive characteristics make to the success of both species as colonizers is discussed in the context of future control and management options for tilapia incursions in Australia.

Corymbia leaf oils, latitude, hybrids and herbivory: a test using common-garden field trials

Foliar oils, particularly monoterpenes, can influence the susceptibility of plants to herbivory. In plants, including eucalypts, monoterpenes are often associated with plant defence. A recent analysis revealed an increase in foliar oil content with increasing latitudinal endemism, and we tested this pattern using three eucalypt taxa comprising a latitudinal replacement cline. We also examined the relative concentrations of two monoterpenes (alpha-pinene and 1,8-cineole), for which meta-analyses also showed latitudinal variation, using hybrids of these three taxa with Corymbia torelliana. These, and pure C. torelliana, were then assessed in common-garden field plots for the abundance and distribution of herbivory by four distinct herbivore taxa. Differing feeding strategies among these herbivores allowed us to test hypotheses regarding heritability of susceptibility and relationships to alpha-pinene and 1,8-cineole. We found no support for an increase in foliar oil content with increasing latitude, nor did our analysis support predictions for consistent variation in alpha-pinene and 1,8-cineole contents with latitude. However, herbivore species showed differential responses to different taxa and monoterpene contents. For example, eriophyid mites, the most monophagous of our censused herbivores, avoided the pure species, but fed on hybrid taxa, supporting hypotheses on hybrid susceptibility. The most polyphagous herbivore (leaf blister sawfly Phylacteophaga froggatti) showed no evidence of response to plant secondary metabolites, while the distribution and abundance patterns of Paropsis atomaria showed some relationship to monoterpene yields.