Host-delivered RNAi: An effective strategy to silence genes in plant parasitic nematodes

Root-knot nematodes (Meloidogyne spp.) are obligate, sedentary endoparasites that infect many plant species causing large economic losses worldwide. Available nematicides are being banned due to their toxicity or ozone-depleting properties and alternative control strategies are urgently required. We have produced transgenic tobacco (Nicotiana tabacum) plants expressing different dsRNA hairpin structures targeting a root-knot nematode (Meloidogyne javanica) putative transcription factor, MjTis11. We provide evidence that MjTis11 was consistently silenced in nematodes feeding on the roots of transgenic plants. The observed silencing was specific for MjTis11, with other sequence-unrelated genes being unaffected in the nematodes. Those transgenic plants able to induce silencing of MjTis11, also showed the presence of small interfering RNAs. Even though down-regulation of MjTis11 did not result in a lethal phenotype, this study demonstrates the feasibility of silencing root-knot nematode genes by expressing dsRNA in the host plant. Host-delivered RNA interference-triggered (HD-RNAi) silencing of parasite genes provides a novel disease resistance strategy with wide biotechnological applications. The potential of HD-RNAi is not restricted to parasitic nematodes but could be adapted to control other plant-feeding pests.

Simulating crop-parasitic weed interactions using APSIM: Model evaluation and application

The parasitic weed Orobanche crenata inflicts major damage on faba bean, lentil, pea and other crops in Mediterranean environments. The development of methods to control O. crenata is to a large extent hampered by the complexity of host-parasite systems. Using a model of host-parasite interactions can help to explain and understand this intricacy. This paper reports on the evaluation and application of a model simulating host-parasite competition as affected by environment and management that was implemented in the framework of the Agricultural Production Systems Simulator (APSIM). Model-predicted faba bean and O. crenata growth and development were evaluated against independent data. The APSIM-Fababean and -Parasite modules displayed a good capability to reproduce effects of pedoclimatic conditions, faba bean sowing date and O. crenata infestation on host-parasite competition. The r(2) values throughout exceeded 0.84 (RMSD: 5.36 days) for phenological, 0.85 (RMSD: 223.00 g m(-2)) for host growth and 0.78 (RMSD: 99.82 g m(-2)) for parasite growth parameters. Inaccuracies of simulated faba bean root growth that caused some bias of predicted parasite number and host yield loss may be dealt with by more flexibly simulating vertical root distribution. The model was applied in simulation experiments to determine optimum sowing windows for infected and non-infected faba bean in Mediterranean environments. Simulation results proved realistic and testified to the capability of APSIM to contribute to the development of tactical approaches in parasitic weed control.

Beyond chemical dependency for managing plant-parasitic nematodes: Examples from the banana, pineapple and vegetable industries of tropical and subtropical Australia

Plant-parasitic nematodes are important pests of horticultural crops grown in tropical and subtropical regions of Australia. Burrowing nematode (Radopholus similis) is a major impediment to banana production and root-knot nematodes (predominantly Meloidogyne javanica and M. incognita) cause problems on pineapple and a range of annual vegetables, including tomato, capsicum, zucchini, watermelon, rockmelon, potato and sweet potato. In the early 1990s, nematode control in these industries was largely achieved with chemicals, with methyl bromide widely used on some subtropical vegetable crops, ethylene dibromide applied routinely to pineapples and non-volatile nematicides such as fenamiphos applied up to four times a year in banana plantations. This paper discusses the research and extension work done over the last 15 years to introduce an integrated pest management approach to nematode control in tropical and subtropical horticulture. It then discusses various components of current integrated pest management programs, including crop rotation, nematode monitoring, clean planting material, organic amendments, farming systems to enhance biological suppression of nematodes and judicious use of nematicides. Finally, options for improving current management practices are considered.

Do parasitic flies attack mites? Evidence in Baltic amber

We provide the first evidence of a small-headed fly planidium (first instar larva; Diptera: Acroceridae) associated with a whirligig mite (Acari: Acariformes: Prostigmata: Anystina: Anystidae) in Baltic amber. This fossil is surprising as parasitic nematodes are the only metazoans known to successfully attack acariform mites, and Acroceridae are believed to be host-restricted parasitoids of spiders. The fossil corroborates a previously published, but widely dismissed, paper that first reported parasitism of parasitengone mites by acrocerid planidia. The possible natural history implications of this find are discussed.

Critically endangered Pristis microdon (Elasmobranchii), as a host for the Indian parasitic copepod, Caligus furcisetifer Redkar, Rangnekar et Murti, 1949 (Siphonostomatoida): New records from northern Australia.

This paper presents the first records of the parasitic copepod Caligus furcisetifer Redkar, Rangnekar et Murti, 1949 beyond Indian waters, specifically, on the body surface and head of the critically endangered largetooth sawfish (commonly referred to as the freshwater sawfish in Australia), Pristis microdon Latham, 1794 (Elasmobranchii, Pristidae), in brackish tidal waters of the Fitzroy River in the Kimberley region of Western Australia and the Leichhardt River in the Gulf of Carpentaria in northern Queensland. This represents a geographic range extension of similar to 8000 km for this parasite. Further, it is only the second member of the genus Caligus to be found on an elasmobranch host in Western Australia and it is the first time this species has been reported from the Southern Hemisphere. Male biased dispersal of P microdon may be the vector in which the parasite has dispersed from India across to northern Australia, or vice versa. A decline in populations of the critically endangered P microdon (and possibly other pristid species) in these regions may lead to a concomitant decline in their parasite fauna.

Extent of dense native woodland and exotic weed infestation in the extensive grazing lands of the Upper Herbert and Upper Burdekin River Catchments of far north Queensland: results of a producer survey

The Mt Garnet Landcare Group commissioned a survey of landholders within the Upper Herbert and Upper Burdekin River Catchments to assess the density of native woodlands and to gauge the extent of exotic weed infestation. Twenty-four of 49 landholders responded, representing an area of nearly 500 000 ha or 47% of the total area. Dense native woodland covers 24% (>117 000 ha) of the area surveyed, while a further 30% (140 000 ha) supports moderately dense stands. The dense stands are largely confined to the highly fertile alluvial soils (26% dense woodland) and the lower fertility sandy-surfaced soils (33% or >96 000 ha). Moderate and dense infestations of exotic weeds, principally Lantana camara, occur on 54% (20 000 ha) of alluvial soils and on 13% of sandy-surfaced soils (39 000 ha), where praxelis (Praxelis clematidia) is the major weed. Basaltic soils have low levels of both dense woodland and exotic weed infestation. Some implications of the results are discussed.

Selective herbicide strategies for use in Australian desmanthus seed crops

Grass and broad-leaved weeds can reduce both yields and product marketability of desmanthus (Desmanthus virgatus) seed crops, even when cultural control strategies are used. Selective herbicides might economically control these weeds, but, prior to this study, the few herbicides tolerated by desmanthus did not control key weed contaminants of desmanthus seed crops. In this study, the tolerance of desmanthus cv. Marc to 55 herbicides used for selective weed control in other leguminous crops was assessed in 1 pot trial and 3 Queensland field trials. One field trial assessed the tolerance of desmanthus seedlings to combinations of the most promising pre-emergent and post emergent herbicides. The pre-emergent herbicides, imazaquin, imazethapyr, pendimethalin, oryzalin and trifluralin, gave useful weed control with very little crop damage. The post-emergent herbicides, haloxyfop, clethodim, propyzamide, carbetamide and dalapon, were safe for controlling grass weeds in desmanthus. Selective post-emergence control of broad-leaved weeds was achieved using bentazone, bromoxynil and imazethapyr. One trial investigated salvaging second-year desmanthus crops from mature perennial weeds, and atrazine, terbacil and hexazinone showed some potential in this role. Overall, our results show that desmanthus tolerates herbicides which collectively control a wide range of weeds encountered in Queensland. These, in combination with cultural weed control strategies, should control most weeds in desmanthus seed crops.

Effect of microwave radiation on seed mortality of rubber vine (Cryptostegia grandiflora R.Br.), parthenium (Parthenium hysterophorous L.) and bellyache bush (Jatropha gossypiifolia L.)

A trial was undertaken to evaluate the effect of microwaves on seed mortality of three weed species. Seeds of rubber vine (Cryptostegia grandiflora R.Br.), parthenium (Parthenium hysterophorous L.) and bellyache bush (Jatropha gossypiifolia L.) were buried at six depths (0, 2.5, 5, 10, 20 and 40 cm) in coarse sand maintained at one of two moisture levels, oven dry or wet (field capacity), and then subjected to one of five microwave radiation durations of (0, 2, 4, 8 and 16 min). Significant interactions between soil moisture level, microwave radiation duration, seed burial depth and species were detected for mortality of seeds of all three species. Maximum seed mortality of rubber vine (88%), parthenium (67%) and bellyache bush (94%) occurred in wet soil irradiated for 16 min. Maximum seed mortality of rubber vine and bellyache bush seeds occurred in seeds buried at 2.5 cm depth whereas that of parthenium occurred in seeds buried at 10 cm depth. Maximum soil temperatures of 114.1 and 87.5°C in dry and wet soil respectively occurred at 2.5 cm depth following 16 min irradiation. Irrespective of the greater soil temperatures recorded in dry soil, irradiating seeds in wet soil generally increased seed mortality 2.9-fold compared with dry soil. Moisture content of wet soil averaged 5.7% compared with 0.1% for dry soil. Results suggest that microwave radiation has the potential to kill seeds located in the soil seed bank. However, many factors, including weed species susceptibility, determine the effectiveness of microwave radiation on buried seeds. Microwave radiation may be an alternative to conventional methods at rapidly depleting soil seed banks in the field, particularly in relatively wet soils that contain long lived weed seeds.

Damage potential of an introduced biological control agent Agonosoma trilineatum (F.) on bellyache bush (Jatropha gossypiifolia L.)

The seed-feeding jewel bug, Agonosoma trilineatum (F.), is an introduced biological control agent for bellyache bush, Jatropha gossypiifolia L. To quantify the damage potential of this agent, shadehouse experiments were conducted with individual bellyache bush plants exposed to a range of jewel bug densities (0, 6 or 24 jewel bugs/plant). The level of abortion of both immature and mature seed capsules and impacts on seed weight and seed viability were recorded in an initial short-term study. The ability of the jewel bug to survive and cause sustained damage was then investigated by measuring seed production, the survival of adults and nymph density across three 6-month cycles. The level of seed capsule abortion caused by the jewel bug was significantly affected by the maturity status of capsules and the density of insects present. Immature capsules were most susceptible and capsule abortion increased with jewel bug density. Similarly, on average, the insects reduced the viability of bellyache bush seeds by 79% and 89% at low and high densities, respectively. However, sustaining jewel bug populations for prolonged periods proved difficult. Adult survival at the end of three 6-month cycles averaged 11% and associated reductions in viable seed production ranged between 55% and 77%. These results suggest that the jewel bug has the potential to reduce the number of viable seeds entering the soil seed bank provided populations can be established and maintained at sufficiently high densities.

Evaluation of the Australian branched broomrape (Orobanche ramosa) eradication program

Because weed eradication programs commonly take 10 or more years to complete, there is a need to evaluate progress toward the eradication objective. We present a simple model, based on information that is readily obtainable, that assesses conformity to the delimitation and extirpation criteria for eradication. It is applied to the program currently targeting the annual parasitic weed, branched broomrape, in South Australia. The model consists of delimitation and extirpation (E) measures plotted against each other to form an 'eradograph.' Deviations from the 'ideal' eradograph plot can inform tactical responses, e.g., increases in survey and/or control effort. Infestations progress from the active phase to the monitoring phase when no plants have been detected for at least 12 mo. They revert to the active phase upon further detection of plants. We summarize this process for the invasion as a whole in a state-and-transition model. Using this model we demonstrate that the invasion is unlikely to be delimited unless the amount of newly detected infested area decreases, on average, by at least 50% per annum. As a result of control activities implemented, on average approximately 70% (range, 44 to 86%) of active infestations progressed to the monitoring phase in the year following their detection. Simulations suggest that increasing this rate of transition will not increase E to a significant extent. The rate of reversion of infestations from the monitoring phase to the active phase decreased logarithmically with time since last detection, but it is likely that lower rates of reversion would accelerate the trend toward extirpation. Program performance with respect to the delimitation criterion has been variable; performance with respect to the extirpation criterion would be improved considerably by the development and application of cost-effective methods for eliminating branched broomrape soil seed populations.

Development of key soil health indicators for the Australian banana industry

To improve the sustainability and environmental accountability of the banana industry there is a need to develop a set of soil health indicators that integrate physical, chemical and biological soil properties. These indicators would allow banana growers, extension and research workers to improve soil health management practices. To determine changes in soil properties due to the cultivation of bananas, a paired site survey was conducted comparing soil properties under conventional banana systems to less intensively managed vegetation systems, such as pastures and forest. Measurements were made on physical, chemical and biological soil properties at seven locations in tropical and sub-tropical banana producing areas. Soil nematode community composition was used as a bioindicator of the biological properties of the soil. Soils under conventional banana production tended to have a greater soil bulk density, with less soil organic carbon (C) (both total C and labile C), greater exchangeable cations, higher extractable P, greater numbers of plant-parasitic nematodes and less nematode diversity, relative to less intensively managed plant systems. The organic banana production systems at two locations had greater labile C, relative to conventional banana systems, but there was no significant change in nematode community composition. There were significant interactions between physical, chemical and nematode community measurements in the soil, particularly with soil C measurements, confirming the need for a holistic set of indicators to aid soil management. There was no single indicator of soil health for the Australian banana industry, but a set of soil health indicators, which would allow the measurement of soil improvements should include: bulk density, soil C, pH, EC, total N, extractable P, ECEC and soil nematode community structure.

Resistance of Brassicaceae plants to root-knot nematode (Meloidogyne spp.) in northern Australia

Brassicaceae plants have the potential as part of an integrated approach to replace fumigant nematicides, providing the biofumigation response following their incorporation is not offset by reproduction of plant-parasitic nematodes on their roots. Forty-three Brassicaceae cultivars were screened in a pot trial for their ability to reduce reproduction of three root-knot nematode isolates from north Queensland, Australia: M. arenaria (NQ1), M. javanica (NQ2) and M. arenaria race 2 (NQ5/7). No cultivar was found to consistently reduce nematode reproduction relative to forage sorghum, the current industry standard, although a commercial fodder radish (Raphanus sativus) and a white mustard (Sinapis alba) line were consistently as resistant to the formation of galls as forage sorghum. A second pot trial screened five commercially available Brassicaceae cultivars, selected for their biofumigation potential, for resistance to two nematode species, M. javanica (NQ2) and M. arenaria (NQ5/7). The fodder radish cv. Weedcheck, was found to be as resistant as forage sorghum to nematode reproduction. A multivariate cluster analysis using the resistance measurements, gall index, nematode number per g of root and multiplication for two nematode species (NQ2 and NQ5/7) confirmed the similarity in resistance between the radish cultivar and forage sorghum. A field trial confirmed the resistance of the fodder radish cv. Weedcheck, with a similar reduction in the number of Meloidogyne spp. juveniles recovered from the roots 8 weeks after planting. The use of fodder radish cultivars as biofumigation crops to manage root-knot nematodes in tropical vegetable production systems deserves further investigation.

A blowfly strike vaccine requires an understanding of host-pathogen interactions

The phase-out of Mulesing by 2010 means the Australian wool industry requires immediate and viable alternatives for the control and prevention of blowfly strike, an economically important parasitic disease of sheep. In this review we have analysed previous research aimed toward the development of a vaccine against blowfly strike and the reasons why the approaches taken were unsuccessful at the time. Close scrutiny has provided new insight into this host-parasite interaction and identified new opportunities for the development of a vaccine. Here we propose that addressing immunosuppression together with the induction of cellular immunity is likely to result in an anti-blowfly strike vaccine, as opposed to the use of "standard" approaches aimed at inducing humoral immunity.

Managing the risk of glyphosate resistance in Australian glyphosate-resistant cotton production systems

BACKGROUND: Glyphosate-resistant cotton varieties are an important tool for weed control in Australian cotton production systems. To increase the sustainability of this technology and to minimise the likelihood of resistance evolving through its use, weed scientists, together with herbicide regulators, industry representatives and the technology owners, have developed a framework that guides the use of the technology. Central to this framework is a crop management plan (CMP) and grower accreditation course. A simulation model that takes into account the characteristics of the weed species, initial gene frequencies and any associated fitness penalties was developed to ensure that the CMP was sufficiently robust to minimise resistance risks. RESULTS: The simulations showed that, when a combination of weed control options was employed in addition to glyphosate, resistance did not evolve over the 30 year period of the simulation. CONCLUSION: These simulations underline the importance of maintaining an integrated system for weed management to prevent the evolution of glyphosate resistance, prolonging the use of glyphosate-resistant cotton.