Management of virus diseases in vegetables

• To undertake an audit of management systems used for tomato spotted wilt virus (TSWV) in greenhouse and field production with the aim of improving disease management determining knowledge gaps in virus-vector relationships. • To investigate the basis for the development of resistance breaking strains of TSWV in capsicums and apply this to virus management in capsicums. • To further develop effective virus management systems in vegetable cucurbit crops. Aspects to be investigated include value of barrier crops, non-insecticide products and cultivar tolerance to virus. • To further develop and assess the adoption and impact of integrated viral disease management systems in field grown and protected cropping systems as part of the vegetable industry development plan.

Reducing the impact of pulse diseases in the northern region

Diseases remain a significant impediment to the achievement of maximum yield potential of pulses (chickpea, peanut and mungbean) and sunflowers in the GRDC northern region. This project worked closely with public and private breeding programs to identify sources of resistance to the major diseases of pulses and sunflower that dominate in the region. Through varied surveillance activities, a watching brief on pulse and sunflower diseases was maintained and a timely and appropriate response was made to several significant disease outbreaks. Information on the biology and management of diseases was extended to clients in a wide variety of ways.

Management of mirids and stinkbugs in Bollgard II

The 'insurance' sprays by the growers run the risk of flaring secondary pests such as whitefly, mites and aphids. Although recent research on mirids has addressed some aspects of this issue, further effort is needed to refine mirid management.

Inheritance and relative dominance, expressed as toxicity response and delayed development, of phosphine resistance in immature stages of Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae)

Fumigation of stored grain with phosphine (PH 3) is used widely to control the lesser grain borer Rhyzopertha dominica. However, development of high level resistance to phosphine in this species threatens control. Effective resistance management relies on knowledge of the expression of resistance in relation to dosage at all life stages. Therefore, we determined the mode of inheritance of phosphine resistance and strength of the resistance phenotype at each developmental stage. We achieved this by comparing mortality and developmental delay between a strongly resistant strain (R-strain), a susceptible strain (S-strain) and their F 1 progenies. Resistance was a maternally inherited, semi-dominant trait in the egg stage but was inherited as an autosomal, incompletely recessive trait in larvae and pupae. The rank order of developmental tolerance in both the sensitive and resistant strains was eggs > pupae > larvae. Comparison of published values for the response of adult R. dominica relative to our results from immature stages reveals that the adult stage of the S-strain is more sensitive to phosphine than are larvae. This situation is reversed in the R-strain as the adult stage is much more resistant to phosphine than even the most tolerant immature stage. Phosphine resistance factors at LC 50 were eggs 400×, larvae 87× and pupae 181× with respect to reference susceptible strain (S-strain) adults indicating that tolerance conferred by a particular immature stage neither strongly nor reliably interacts with the genetic resistance element. Developmental delay relative to unfumigated control insects was observed in 93% of resistant pupae, 86% of resistant larvae and 41% of resistant eggs. Increased delay in development and the toxicity response to phosphine exposure were both incompletely recessive. We show that resistance to phosphine has pleiotropic effects and that the expression of these effects varies with genotype and throughout the life history of the insect. © 2012.

Individual-based modelling of the efficacy of fumigation tactics to control lesser grain borer (Rhyzopertha dominica) in stored grain

Increasing resistance to phosphine (PH 3) in insect pests, including lesser grain borer (Rhyzopertha dominica) has become a critical issue, and development of effective and sustainable strategies to manage resistance is crucial. In practice, the same grain store may be fumigated multiple times, but usually for the same exposure period and concentration. Simulating a single fumigation allows us to look more closely at the effects of this standard treatment.We used an individual-based, two-locus model to investigate three key questions about the use of phosphine fumigant in relation to the development of PH 3 resistance. First, which is more effective for insect control; long exposure time with a low concentration or short exposure period with a high concentration? Our results showed that extending exposure duration is a much more efficient control tactic than increasing the phosphine concentration. Second, how long should the fumigation period be extended to deal with higher frequencies of resistant insects in the grain? Our results indicated that if the original frequency of resistant insects is increased n times, then the fumigation needs to be extended, at most, n days to achieve the same level of insect control. The third question is how does the presence of varying numbers of insects inside grain storages impact the effectiveness of phosphine fumigation? We found that, for a given fumigation, as the initial population number was increased, the final survival of resistant insects increased proportionally. To control initial populations of insects that were n times larger, it was necessary to increase the fumigation time by about n days. Our results indicate that, in a 2-gene mediated resistance where dilution of resistance gene frequencies through immigration of susceptibles has greater effect, extending fumigation times to reduce survival of homozygous resistant insects will have a significant impact on delaying the development of resistance. © 2012 Elsevier Ltd.

Spatial and temporal foraging patterns of Queensland fruit fly, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), for protein and implications for management

Fruit flies require protein for reproductive development and actively feed upon protein sources in the field. Liquid protein baits mixed with insecticide are used routinely to manage pest fruit flies, such as Bactrocera tryoni (Froggatt). However, there are still some gaps in the underpinning science required to improve the efficacy of bait spray technology. The spatial and temporal foraging behaviour of B. tryoni in response to protein was investigated in the field. A series of linked trials using either wild flies in the open field or laboratory-reared flies in field cages and a netted orchard were undertaken using nectarines and guavas. Key questions investigated were the fly's response to protein relative to: height of protein within the canopy, fruiting status of the tree, time of day, season and size of the experimental arena. Canopy height had a significant response on B. tryoni foraging, with more flies foraging on protein in the mid to upper canopy. Fruiting status also had a significant effect on foraging, with most flies responding to protein when applied to fruiting hosts. B. tryoni demonstrated a repeatable diurnal response pattern to protein, with the peak response being between 12:0016:00 h. Season showed significant but unpredictable effects on fruit fly response to protein in the subtropical environment where the work was undertaken. Relative humidity, but not temperature or rainfall, was positively correlated with protein response. The number of B. tryoni responding to protein decreased dramatically as the spatial scale increased from field cage through to the open field. Based on these results, it is recommend that, to be most effective, protein bait sprays should be applied to the mid to upper canopies of fruiting hosts. Overall, the results show that the protein used, an industry standard, has very low attractancy to B. tryoni and that further work is urgently needed to develop more volatile protein baits.

Pheromone of the banana-spotting bug, Amblypelta lutescens lutescens Distant (Heteroptera: Coreidae): Identification, synthesis, and field bioassay

The banana-spotting bug, Amblypelta lutescens lutescens Distant (Heteroptera: Coreidae), is one of the principal pests of tree fruits and nuts across northern and eastern Australia. Apart from visual damage assessment, there are currently no reliable methods for monitoring bug activity to aid management decisions. An attractant pheromone for this species that could be used as a trap lure could potentially fill this void. Earlier, two male-specific compounds were identified in airborne extracts from A. lutescens lutescens, (E,E)-α-farnesene and (R,E)-nerolidol; an unknown compound with a molecular weight 220 was also detected. We now report the identification of this hitherto unknown compound as (R,E,E)-α-farnesene-10,11-oxide. Synthesis of this epoxide was conducted using a regioselective asymmetric dihydroxylation of a sulfolene. A blend mimicking the natural proportions of (E,E)-α-farnesene, (R,E)-nerolidol, and (R,E,E)-α-farnesene-10,11- oxide attracted male and female A. lutescens lutescens as well as nymphs in the field, verifying that the aggregation pheromone comprises or is contained within this group of compounds. Copyright © 2012 Ashot Khrimian et al.

Mitochondrial DNA diversity of Cleruchoides noackae (Hymenoptera: Mymaridae): a potential biological control agent for Thaumastocoris peregrinus (Hemiptera: Thaumastocoridae)

Carpintero and Dellap, (Hemiptera: Thaumastocoridae) is a native Australian sap-feeding insect that has become invasive and seriously damaging to commercially grown in the Southern Hemisphere. Lin and Huber (Hymenoptera: Mymaridae) was recently discovered as an egg parasitoid of the Thaumastocoridae in Australia. Mitochondrial DNA (mtDNA; cytochrome oxidase subunit I, COI) sequence diversity amongst 104 individuals from these native populations revealed 24 sequence haplotypes. The COI haplotypes of individuals collected from the Sydney and Southeast Queensland clustered in distinct groups, indicating limited spread of the insect between the regions. Individuals collected from Perth in Western Australia were represented by four COI haplotypes. Although this population is geographically more isolated from other populations, two COI haplotypes were identical to haplotypes found in the Sydney region. The results suggest that has recently been introduced into Perth, possibly from the Sydney area. The high mtDNA diversity and limited spread that is suggested for is in contrast to the lack of geographic associated mtDNA diversity and extensive spread of . If implemented as a biological control agent, this factor will need to be considered in collecting and releasing .

Managing glyphosate resistance in Australian cotton farming: modelling shows how to delay evolution and maintain long-term population control

Glyphosate resistance is a rapidly developing threat to profitability in Australian cotton farming. Resistance causes an immediate reduction in the effectiveness of in-crop weed control in glyphosate-resistant transgenic cotton and summer fallows. Although strategies for delaying glyphosate resistance and those for managing resistant populations are qualitatively similar, the longer resistance can be delayed, the longer cotton growers will have choice over which tactics to apply and when to apply them. Effective strategies to avoid, delay, and manage resistance are thus of substantial value. We used a model of glyphosate resistance dynamics to perform simulations of resistance evolution in Sonchus oleraceus (common sowthistle) and Echinochloa colona (awnless barnyard grass) under a range of resistance prevention, delaying, and management strategies. From these simulations, we identified several elements that could contribute to effective glyphosate resistance prevention and management strategies. (i) Controlling glyphosate survivors is the most robust approach to delaying or preventing resistance. High-efficacy, high-frequency survivor control almost doubled the useful lifespan of glyphosate from 13 to 25 years even with glyphosate alone used in summer fallows. (ii) Two non-glyphosate tactics in-crop plus two in-summer fallows is the minimum intervention required for long-term delays in resistance evolution. (iii) Pre-emergence herbicides are important, but should be backed up with non-glyphosate knockdowns and strategic tillage; replacing a late-season, pre-emergence herbicide with inter-row tillage was predicted to delay glyphosate resistance by 4 years in awnless barnyard grass. (iv) Weed species' ecological characteristics, particularly seed bank dynamics, have an impact on the effectiveness of resistance strategies; S. oleraceus, because of its propensity to emerge year-round, was less exposed to selection with glyphosate than E. colona, resulting in an extra 5 years of glyphosate usefulness (18 v. 13 years) even in the most rapid cases of resistance evolution. Delaying tactics are thus available that can provide some or many years of continued glyphosate efficacy. If glyphosate-resistant cotton cropping is to remain profitable in Australian farming systems in the long-term, however, growers must adapt to the probability that they will have to deal with summer weeds that are no longer susceptible to glyphosate. Robust resistance management systems will need to include a diversity of weed control options, used appropriately.

Wheat biomass and yield increased when populations of the root-lesion nematode (Pratylenchus thornei) were reduced through sequential rotation of partially resistant winter and summer crops

The root-lesion nematode, Pratylenchus thornei, can reduce wheat yields by >50%. Although this nematode has a broad host range, crop rotation can be an effective tool for its management if the host status of crops and cultivars is known. The summer crops grown in the northern grain region of Australia are poorly characterised for their resistance to P. thornei and their role in crop sequencing to improve wheat yields. In a 4-year field experiment, we prepared plots with high or low populations of P. thornei by growing susceptible wheat or partially resistant canaryseed (Phalaris canariensis); after an 11-month, weed-free fallow, several cultivars of eight summer crops were grown. Following another 15-month, weed-free fallow, P. thornei-intolerant wheat cv. Strzelecki was grown. Populations of P. thornei were determined to 150 cm soil depth throughout the experiment. When two partially resistant crops were grown in succession, e.g. canaryseed followed by panicum (Setaria italica), P. thornei populations were <739/kg soil and subsequent wheat yields were 3245 kg/ha. In contrast, after two susceptible crops, e.g. wheat followed by soybean, P. thornei populations were 10 850/kg soil and subsequent wheat yields were just 1383 kg/ha. Regression analysis showed a linear, negative response of wheat biomass and grain yield with increasing P. thornei populations and a predicted loss of 77% for biomass and 62% for grain yield. The best predictor of wheat yield loss was P. thornei populations at 0-90 cm soil depth. Crop rotation can be used to reduce P. thornei populations and increase wheat yield, with greatest gains being made following two partially resistant crops grown sequentially.

Panel of real-time PCRs for the multiplexed detection of two tomato-infecting begomoviruses and their cognate whitefly vector species

A new approach for the simultaneous identification of the viruses and vectors responsible for tomato yellow leaf curl disease (TYLCD) epidemics is presented. A panel of quantitative multiplexed real-time PCR assays was developed for the sensitive and reliable detection of Tomato yellow leaf curl virus-Israel (TYLCV-IL), Tomato leaf curl virus (ToLCV), Bemisia tabaci Middle East Asia Minor 1 species (MEAM1, B biotype) and B.tabaci Mediterranean species (MED, Q biotype) from either plant or whitefly samples. For quality-assurance purposes, two internal control assays were included in the assay panel for the co-amplification of solanaceous plant DNA or B.tabaci DNA. All assays were shown to be specific and reproducible. The multiplexed assays were able to reliably detect as few as 10 plasmid copies of TYLCV-IL, 100 plasmid copies of ToLCV, 500fg B.tabaci MEAM1 and 300fg B.tabaci MED DNA. Evaluated methods for routine testing of field-collected whiteflies are presented, including protocols for processing B.tabaci captured on yellow sticky traps and for bulking of multiple B.tabaci individuals prior to DNA extraction. This work assembles all of the essential features of a validated and quality-assured diagnostic method for the identification and discrimination of tomato-infecting begomovirus and B.tabaci vector species in Australia. This flexible panel of assays will facilitate improved quarantine, biosecurity and disease-management programmes both in Australia and worldwide.

Optimisation of resin extraction from an Australian arid grass ‘Triodia pungens’ and its preliminary evaluation as an anti-termite timber coating

Spinifex grasses are the dominant vegetative component in Australian grassland habitats, covering approximately 26% of the Australian landmass. Our ongoing work explores the utility of both the cellulosic and resinous components of this abundant biomass for modern applications and a potential economy for our Aboriginal collaborators. This study is focused on the optimisation of a resin extraction process using solvent, and the subsequent evaluation, via a field trial, of the potential use and efficacy of the resin as an anti-termite coating material. Termiticidal performance was evaluated by re-dissolving the extracted resin in acetone and coating on pine timber blocks. The resin-coated and control blocks were then exposed to a colony of Mastotermes darwiniensis’ (Froggatt) termites, which are the most primitive alive and destructive species in subterranean area, at a trial site in northeast Australia, for six months. The results clearly showed that spinifex resin effectively protected the timber from termite attack, while the uncoated control samples were extensively damaged. By demonstrating an enhanced termite resistance, we here report that plant resins that are produced by arid/semi-arid grasses could be potentially used as treatments to prevent termite attack.

Biodiversity, biosecurity and integrated pest management

Breaches of biosecurity, leading to incursions by invasive species, have the potential to cause substantial economic, social and environmental losses, including drastic reduction in biodiversity. It is argued that improving biosecurity reduces risk to biodiversity, while maintaining stable ecosystems through biodiversity can be a safeguard against biosecurity breaches. The global costs of invasive alien species (IAS) have been estimated at around US$350 billion, while alien invertebrate and vertebrate pests and weeds are estimated to cost Australia at least $7 billion a year. A striking, current, example is the incursion by Myrtle Rust (Puccinia psidii) an organism which can infect all members of the Myrtaceae, the most important family in the Australian flora. Myrtle rust was first detected on a property on the central coast of New South Wales in late April 2010. Two years later the disease has been detected in numerous locations in Queensland and New South Wales ranging from commercial plant nurseries and public amenities to large areas of bushland. This particular breach of biosecurity will, inevitably, diminish biodiversity of flora and fauna over large areas of the continent. Integrated pest management (IPM), an enrichment of diversity in managing invasive and other pest species, offers the best opportunity to address problems such as these. Australia's response to increasing biosecurity risk is comprehensive and includes national networking of scientists engaged in a complex program of biosecurity research and development, including studies of IPM. This network is being enhanced by the development of international linkages.