Cat's claw creeper leaf-mining jewel beetle Hylaeogena jureceki Obenberger (Coleoptera: Buprestidae), a host-specific biological control agent for Dolichandra unguis-cati (Bignoniaceae) in Australia

Cat's claw creeper, Dolichandra unguis-cati (L.) L.G. Lohman (syn: Macfadyena unguis-cati (L.) A.H. Gentry) (Bignoniaceae), a major environmental weed in Queensland and New South Wales, is a Weed of National Significance and an approved target for biological control. A leaf-mining jewel beetle, Hylaeogena jureceki Obenberger (Coleoptera: Buprestidae), first collected in 2002 from D. unguis-cati in Brazil and Argentina, was imported from South Africa into a quarantine facility in Brisbane in 2009 for host-specificity testing. H. jureceki adults chew holes in leaves and lay eggs on leaf margins and the emerging larvae mine within the leaves of D. unguis-cati. The generation time (egg to adult) of H. jureceki under quarantine conditions was 55.4 ± 0.2 days. Host-specificity trials conducted in Australia on 38 plant species from 11 families supplement and support South African studies which indicated that H. jureceki is highly host-specific and does not pose a risk to any non-target plant species in Australia. In no-choice tests, adults survived significantly longer (>32 weeks) on D. unguis-cati than on non-target test plant species (<3 weeks). Oviposition occurred on D. unguis-cati and one non-target test plant species, Citharexylum spinosum (Verbenaceae), but no larval development occurred on the latter species. In choice tests involving D. unguis-cati, C. spinosum and Avicennia marina (Avicenniaceae), feeding and oviposition were evident only on D. unguis-cati. The insect was approved for field release in Australia in May 2012.

Predicting risk and benefit a priori in biological control of invasive plant species: a systems modelling approach

While the method using specialist herbivores in managing invasive plants (classical biological control) is regarded as relatively safe and cost-effective in comparison to other methods of management, the rarity of strict monophagy among insect herbivores illustrates that, like any management option, biological control is not risk-free. The challenge for classical biological control is therefore to predict risks and benefits a priori. In this study we develop a simulation model that may aid in this process. We use this model to predict the risks and benefits of introducing the chrysomelid beetle Charidotis auroguttata to manage the invasive liana Macfadyena unguis-cati in Australia. Preliminary host-specificity testing of this herbivore indicated that there was limited feeding on a non-target plant, although the non-target was only able to sustain some transitions of the life cycle of the herbivore. The model includes herbivore, target and non-target life history and incorporates spillover dynamics of populations of this herbivore from the target to the non-target under a variety of scenarios. Data from studies of this herbivore in the native range and under quarantine were used to parameterize the model and predict the relative risks and benefits of this herbivore when the target and non-target plants co-occur. Key model outputs include population dynamics on target (apparent benefit) and non-target (apparent risk) and fitness consequences to the target (actual benefit) and non-target plant (actual risk) of herbivore damage. The model predicted that risk to the non-target became unacceptable (i.e. significant negative effects on fitness) when the ratio of target to non-target in a given patch ranged from 1:1 to 3:2. By comparing the current known distribution of the non-target and the predicted distribution of the target we were able to identify regions in Australia where the agent may be pose an unacceptable risk. By considering risk and benefit simultaneously, we highlight how such a simulation modelling approach can assist scientists and regulators in making more objective decisions a priori, on the value of releasing specialist herbivores as biological control agents.

How widespread is Parthenium hysterophorus and its biological control agent Zygogramma bicolorata in South Asia?

Parthenium hysterophorus is a weed of global significance causing severe economic, environmental, human and animal health problems in Asia, Africa, Australia and the Pacific. In South Asia, P. hysterophorus occurs in India, Pakistan, Sri Lanka, Bangladesh and Nepal. A host-specific leaf-feeding beetle Zygogramma bicolorata from Mexico was introduced into India in 1984, as a biological control agent for P. hysterophorus. In this study, a GIS-based distribution map of P. hysterophorus and its biological control agent Z. bicolorata in South Asia based on meta-analysis is presented. The map highlights the limited published information on P. hysterophorus incidence in many of the states and territories in India, as well as in neighbouring Bangladesh, Bhutan, Nepal and Pakistan. Incidence of Z. bicolorata was recorded as three distinct clusters, covering many states in India. In Pakistan, Z. bicolorata was recorded in the Punjab region bordering India. A CLIMEX model based on the current distribution of Z. bicolorata in India suggests that the geographic range of this agent in India and Pakistan can extend to other P. hysterophorus-infested areas in the region. The CLIMEX model also suggests that all of Bangladesh and Sri Lanka, and parts of Nepal are climatically suitable for Z. bicolorata.

Competition of sorghum cultivars and densities with Japanese millet (Echinochloa esculenta).

Field studies were conducted at two locations in southern Queensland, Australia during the 2003-2004 and 2004-2005 growing seasons to determine the differential competitiveness of sorghum (Sorghum bicolor L. Moench) cultivars and crop densities against weeds and the sorghum yield loss due to weeds. Weed competition was investigated by growing sorghum in the presence or absence of a model grass weed, Japanese millet (Echinochloa esculenta). The correlation analyses showed that the early growth traits (height, shoot biomass, and daily growth rate of the shoot biomass) of sorghum adversely affected the height, biomass, and seed production of millet, as measured at maturity. "MR Goldrush" and "Bonus MR" were the most competitive cultivars, resulting in reduced weed biomass, weed density, and weed seed production. The density of sorghum also had a significant effect on the crop's ability to compete with millet. When compared to the density of 4.5 plants per m2, sorghum that was planted at 7.5 plants per m2 suppressed the density, biomass, and seed production of millet by 22%, 27% and 38%, respectively. Millet caused a significant yield loss in comparison with the weed-free plots. The combined weed-suppressive effects of the competitive cultivars, such as MR Goldrush, and high crop densities minimized the yield losses from the weeds. These results indicate that sorghum competition against grass weeds can be improved by choosing competitive cultivars and by using a high crop density of > 7.5 plants per m2. These non-chemical options should be included in an integrated weed management program for better weed management, particularly where the control options are limited by the evolution of herbicide resistance.

Feasiblity Study into fungal biocontrol of small hive beetle

Investigation of potential for fungal control of small hive beetles.

Flat grain beetle fumigation protocol

Flat grain beetle (FGB) is a major emergency plant pest (EPP) of stored grain in Australia. Populations of FGB have recently developed high level resistance to phosphine (the only viable fumigant available for non-quarantine use) resulting in control failures with current dosage regimes. As there is no practical alternative to phosphine, failure to control FGB with phosphine places at risk market access for Australian grain worth up to $7 billion in annual trade. Therefore there is an urgent need to develop appropriate phosphine fumigation protocols to eradicate outbreaks of strongly resistant FGB. Research outcomes: - Characterisation of high resistance to phosphine in flat grain beetles (FGB) for the first time internationally. - Establishment of fumigation protocols and an eradication strategy that will enable industry to eradicate infestations of phosphine-resistant flat grain beetle and prevent or delay further selection for resistance to phosphine. - Development of a rapid test to detect highly resistant FGB. -Facilitate continued market access of Australian grain.

Life history and host range of Alagoasa extrema (Coleoptera: Chrysomelidae: Alticinae), a potential biological control agent of Lantana spp. (Verbenaceae) in Australia

The life history and host range of the lantana beetle, Alagoasa extrema, a potential biocontrol agent for Lantana spp. were investigated in a quarantine unit at the Alan Fletcher Research Station, Brisbane, Australia. Adults feed on leaves and females lay batches of about 17 eggs on the soil surface around the stems of plants. The eggs take 16 days to hatch and newly emerged larvae move up the stem to feed on young leaves. Larvae feed for about 23 days and there are three instars. There is a prepupal non-feeding stage that lasts about 12 days and the pupal stage, which occurs in a cocoon in the soil, lasts 16 days. Teneral adults remain in the cocoon for 3 days to harden prior to emergence. Males live for about 151 days while females live for about 127 days. The pre-oviposition period is 19 days. In no-choice larval feeding trials, nine plant species, representing three families, supported development to adult. Three species, Aloysia triphylla, Citharexylum spinosum and Pandorea pandorana were able to support at least two successive generations. These results confirm those reported in South Africa and suggest that A. extrema is not sufficiently specific for release in Australia. Furthermore, it is not recommended for release in any other country which is considering biological control of lantana.

Strong resistance to phosphine in the rusty grain beetle, Cryptolestes ferrugineus (Stephens) (Coleoptera: Laemophloeidae): its characterisation, a rapid assay for diagnosis and its distribution in Australia

BACKGROUND: The recent development of very high resistance to phosphine in rusty grain beetle, Cryptolestes ferrugineus (Stephens), seriously threatens stored-grain biosecurity. The aim was to characterise this resistance, to develop a rapid bioassay for its diagnosis to support pest management and to document the distribution of resistance in Australia in 20072011. RESULTS: Bioassays of purified laboratory reference strains and field-collected samples revealed three phenotypes: susceptible, weakly resistant and strongly resistant. With resistance factors of > 1000 x , resistance to phosphine expressed by the strong resistance phenotype was higher than reported for any stored-product insect species. The new time-to-knockdown assay rapidly and accurately diagnosed each resistance phenotype within 6 h. Although less frequent in western Australia, weak resistance was detected throughout all grain production regions. Strong resistance occurred predominantly in central storages in eastern Australia. CONCLUSION: Resistance to phosphine in the rusty grain beetle is expressed through two identifiable phenotypes: weak and strong. Strong resistance requires urgent changes to current fumigation dosages. The development of a rapid assay for diagnosis of resistance enables the provision of same-day advice to expedite resistance management decisions. (c) 2012 Commonwealth of Australia. Published by John Wiley & Sons, Ltd.

Rapid genome wide mapping of phosphine resistance loci by a simple regional averaging analysis in the red flour beetle, Tribolium castaneum

Background Next-generation sequencing technology is an important tool for the rapid, genome-wide identification of genetic variations. However, it is difficult to resolve the ‘signal’ of variations of interest and the ‘noise’ of stochastic sequencing and bioinformatic errors in the large datasets that are generated. We report a simple approach to identify regional linkage to a trait that requires only two pools of DNA to be sequenced from progeny of a defined genetic cross (i.e. bulk segregant analysis) at low coverage (<10×) and without parentage assignment of individual SNPs. The analysis relies on regional averaging of pooled SNP frequencies to rapidly scan polymorphisms across the genome for differential regional homozygosity, which is then displayed graphically. Results Progeny from defined genetic crosses of Tribolium castaneum (F4 and F19) segregating for the phosphine resistance trait were exposed to phosphine to select for the resistance trait while the remainders were left unexposed. Next generation sequencing was then carried out on the genomic DNA from each pool of selected and unselected insects from each generation. The reads were mapped against the annotated T. castaneum genome from NCBI (v3.0) and analysed for SNP variations. Since it is difficult to accurately call individual SNP frequencies when the depth of sequence coverage is low, variant frequencies were averaged across larger regions. Results from regional SNP frequency averaging identified two loci, tc_rph1 on chromosome 8 and tc_rph2 on chromosome 9, which together are responsible for high level resistance. Identification of the two loci was possible with only 5-7× average coverage of the genome per dataset. These loci were subsequently confirmed by direct SNP marker analysis and fine-scale mapping. Individually, homozygosity of tc_rph1 or tc_rph2 results in only weak resistance to phosphine (estimated at up to 1.5-2.5× and 3-5× respectively), whereas in combination they interact synergistically to provide a high-level resistance >200×. The tc_rph2 resistance allele resulted in a significant fitness cost relative to the wild type allele in unselected beetles over eighteen generations. Conclusion We have validated the technique of linkage mapping by low-coverage sequencing of progeny from a simple genetic cross. The approach relied on regional averaging of SNP frequencies and was used to successfully identify candidate gene loci for phosphine resistance in T. castaneum. This is a relatively simple and rapid approach to identifying genomic regions associated with traits in defined genetic crosses that does not require any specialised statistical analysis.

Host Tree Influences on Longicorn Beetle (Coleoptera: Cerambycidae) Attack in Subtropical Corymbia (Myrtales: Myrtaceae)

Phoracantha longicorn beetles are endemic to Australia, and some species have become significant pests of eucalypts worldwide, yet little is known about their host plant interactions and factors influencing tree susceptibility in Australia. Here, we investigate the host relationships of Phoracantha solida (Blackburn, 1894) on four eucalypt taxa (one pure species and three hybrid families), examining feeding site physical characteristics including phloem thickness, density, and moisture content, and host tree factors such as diameter, height, growth, taper, and survival. We also determine the cardinal and vertical (within-tree) and horizontal (between-tree) spatial distribution of borers. Fewer than 10% of P. solida attacks were recorded from the pure species (Corymbia citriodora subsp. variegate (Hook)), and this taxon also showed the highest survival, phloem thickness, relative growth rate, and bark:wood area. For the two most susceptible taxa, borer severity was negatively correlated with moisture content, and positively related to phloem density. Borers were nonrandomly and nonuniformly distributed within trees, and were statistically aggregated in 32% of plots. More attacks were situated on the northern side of the tree than the other aspects, and most larvae fed within the lower 50 cm of the bole, with attack height positively correlated with severity. Trees with borers had more dead neighbors, and more bored neighbors, than trees without borers, while within plots, borer incidence and severity were positively correlated. Because the more susceptible taxa overlapped with less susceptible taxa for several physical tree factors, the role of primary and secondary chemistries in determining host suitability needs to be investigated. Nevertheless, taxon, moisture content, phloem density, tree size, and mortality of neighboring trees appeared the most important physical characteristics influencing host suitability for P. solida at this site.

Aiming for the management of the small hive beetle, Aethina tumida, using relative humidity and diatomaceous earth

Small hive beetles (SHBs) are a global pest of European honeybee colonies. In the laboratory, the survival of adult SHBs was evaluated in relation to relative humidity (RH = 56, 64, 73, 82 and 96 %) and treatment with diatomaceous earth (DE) across 4 days. Low RH reduced survival. The application of DE reduced survival in addition to RH. Adults treated with corn flour (control) showed no difference in survival from untreated beetles. Scanning electron microscopy images showed no scarification of adult beetle cuticle after exposure to DE; therefore, water loss is likely facilitated through non-abrasive means such as the adsorption of cuticular lipids. The data agree with the hypothesis that DE causes mortality through water loss from treated insects. Egress, ingress, mortality and the egg-laying behaviours of beetles were observed in relation to a popular in-hive trench trap with and without the addition of DE. Traps filled with DE resulted in 100 % mortality of beetles compared with 8.6 % mortality when no DE was present. A simple method for visually determining beetle sex was used and documented.

The Jewel Beetle (Hylaeogena jureceki); a new biocontrol for cat’s claw creeper (Dolichandra unguis-cati) in Queensland

Cat’s claw creeper (Dolichandra unguis-cati (Bignoniaceae) is a serious environmental weed in Queensland and New South Wales. It presents a threat to riparian and rainforest ecosystems and is often found in inaccessible locations that are not suitable for chemical or physical control methods. This makes biological control an important tool for managing this weed. The jewel beetle Hylaeo¬gena jureceki was approved for release in Australia in May 2012. Since approval, approximately 35,000 insects have been released at 53 sites. Multiple and single releases have been made at sites with the number of insects released ranging from 20 to 1590. Post-release monitoring before and after winter found the beetle persisting at 73% of release sites in southeast Queensland. Within the release sites, the beetle appears to disperse widely, up to 100 m over a 15 month period. Based on these early field results, it appears that the beetle will establish and spread in Queensland and New South Wales. In addition to direct field releases, the beetle has been supplied to various community and Landcare groups for breeding and field release. This will hasten the spread of the insect to a wider area. It is expected that the jewel beetle will complement the leaf-sucking tingid (Carvalhotingis visenda) and leaf-tying moth (Hypocosmia pyrochroma) that were released in 2007.

Biological control of prickly acacia: Current research and future prospects

Prickly acacia, Vachellia nilotica subsp. indica (syn. Acacia nilotica subsp. indica) (Fabaceae), a major weed in the natural grasslands of western Queensland, has been a target of biological control since the 1980s with limited success to date. Surveys in India, based on genetic and climate matching, identified five insects and two rust pathogens as potential agents. Host-specificity tests were conducted for the insects in India and under quarantine conditions in Australia, and for the rust pathogens under quarantine conditions at CABI in the UK. In no-choice tests, the brown leaf-webber, Phycita sp. A, (Lepidoptera: Pyralidae) completed development on 17 non-target plant species. Though the moth showed a clear preference for prickly acacia in oviposition choice trials screening of additional test-plant species was terminated in view of the potential non-target risk. The scale insect Anomalococcus indicus (Hemiptera: Lecanodiaspididae) developed into mature gravid females on 13 out of 58 non-target plant species tested. In the majority of cases very few female scales matured but development was comparable to that on prickly acacia on four of the non-target species. In multiple choice tests, the scale insect showed a significant preference for the target weed over non-target species tested. In a paired-choice trial under field conditions in India, crawler establishment occurred only on prickly acacia and not on the non-target species tested. Further choice trials are to be conducted under natural field conditions in India. A colony of the green leaf-webber Phycita sp. B has been established in quarantine facilities in Australia and host-specificity testing has commenced. The gall-rust Ravenelia acaciae-arabicae and the leaf-rust Ravenelia evansii (Puccineales: Raveneliaceae) both infected and produced viable urediniospores on Vachellia sutherlandii (Fabaceae), a non-target Australian native plant species. Hence, no further testing with the two rust species was pursued. Inoculation trials using the gall mite Aceria liopeltus (Acari: Eriophyidae) from V. nilotica subsp. kraussiana in South Africa resulted in no gall induction on V. nilotica subsp. indica. Future research will focus on the leaf-weevil Dereodus denticollis (Coleoptera: Curculionidae) and the leaf-beetle Pachnephorus sp. (Coleoptera: Chrysomelidae) under quarantine conditions in Australia. Native range surveys for additional potential biological control agents will also be pursued in northern and western Africa.

Genetic characterization of field-evolved resistance to phosphine in the rusty grain beetle, Cryptolestes ferrugineus (Laemophloeidae: Coleoptera)

Inheritance of resistance to phosphine fumigant was investigated in three field-collected strains of rusty grain beetle, Cryptolestes ferrugineus, Susceptible (S-strain), Weakly Resistant (Weak-R) and Strongly Resistant (Strong-R). The strains were purified for susceptibility, weak resistance and strong resistance to phosphine, respectively, to ensure homozygosity of resistance genotype. Crosses were established between S-strain × Weak-R, S-strain × Strong-R and Weak-R × Strong-R, and the dose mortality responses to phosphine of these strains and their F1, F2 and F1-backcross progeny were obtained. The fumigations were undertaken at 25 °C and 55% RH for 72 h. Weak-R and Strong-R showed resistance factors of 6.3 × and 505 × compared with S-strain at the LC50. Both weak and strong resistances were expressed as incompletely recessive with degrees of dominance of − 0.48 and − 0.43 at the LC50, respectively. Responses of F2 and F1-backcross progeny indicated the existence of one major gene in Weak-R, and at least two major genes in Strong-R, one of which was allelic with the major factor in Weak-R. Phenotypic variance analyses also estimated that the number of independently segregating genes conferring weak resistance was 1 (nE = 0.89) whereas there were two genes controlling strong resistance (nE = 1.2). The second gene, unique to Strong-R, interacted synergistically with the first gene to confer a very high level of resistance (~ 80 ×). Neither of the two major resistance genes was sex linked. Despite the similarity of the genetics of resistance to that previously observed in other pest species, a significant proportion (~ 15 to 30%) of F1 individuals survived at phosphine concentrations higher than predicted. Thus it is likely that additional dominant heritable factors, present in some individuals in the population, also influenced the resistance phenotype. Our results will help in understanding the process of selection for phosphine resistance in the field which will inform resistance management strategies. In addition, this information will provide a basis for the identification of the resistance genes.