Agricultural Insect Research Articles

Flexible metabolism in Metarhizium anisopliae and Beauveria bassiana: role of the glyoxylate cycle during insect pathogenesis

Insect pathogenic fungi such as Metarhizium anisopliae and Beauveria bassiana have an increasing role in the control of agricultural insect pests and vectors of human diseases. Many of the virulence factors are well studied but less is known of the metabolism of these fungi during the course of insect infection or saprobic growth. Here, we assessed enzyme activity and gene expression in the central carbon metabolic pathway, including isocitrate dehydrogenase, aconitase, citrate synthase, malate synthase (MLS) and isocitrate lyase (ICL), with particular attention to the glyoxylate cycle when M. anisopliae and B. bassiana were grown under various conditions. We observed that ICL and MLS, glyoxylate cycle intermediates, were upregulated during growth on 2-carbon compounds (acetate and ethanol) as well as in insect haemolymph. We fused the promoter of the M. anisopliae ICL gene (Ma-icl) to a marker gene (mCherry) and showed that Ma-icl was upregulated when M. anisopliae was grown in the presence of acetate. Furthermore, Ma-icl was upregulated when fungi were engulfed by insect haemocytes as well as during appressorium formation. Addition of the ICL inhibitor 3-nitroproprionate delayed conidial germination and inhibited appressorium formation. These results show that these insect pathogenic fungi have a flexible metabolism that includes the glyoxylate cycle as an integral part of germination, pathogenesis and saprobic growth.

Novaluron nanoparticles: Formation and potential use in controlling agricultural insect pests

Nanoparticles of novaluron, a water-insoluble insecticide, were prepared by a novel method, based on a direct conversion of O/W microemulsions containing pesticide and volatile solvents, into powders. The conversion of nanoparticles into powder was achieved by rapid evaporation of all the liquids in the microemulsion by spray drying. The microemulsions were evaluated by SAXS, self diffusion NMR, conductivity, and viscosity. The droplet size was approximately 6 nm, and the novaluron particle size, after redispersion and evaluation by DLS, was 200 ± 50 nm. These particles consisted of aggregates of nanoparticles (30–100 nm), as viewed by Cryo-TEM. Electron diffraction and XRD showed that the nanoparticles were amorphous indicating a possible improved bioactivity. The stability of the dispersed nanoparticles was evaluated by following particle size by DLS for a period of time, revealing a slight increase in particle size despite the high value of zeta potential. In vivo experiments carried out with Egyptian cotton leafworm Spodoptera littoralis larvae indicated that the toxicity of nanoparticles of novaluron resembled that of the commercial formulation.

Entomotoxic action of Sambucus nigra agglutinin i in Acyrthosiphon pisum aphids and Spodoptera exigua caterpillars through caspase‐3‐like‐dependent apoptosis

In this project, the toxicity and mechanism of action of the ricin-B-related lectin SNA-I from elderberry (Sambucus nigra) in the pea aphid (Acyrthosiphon pisum) and the beet armyworm (Spodoptera exigua), two important pest insects in agriculture, were studied. SNA-I is a chimeric lectin belonging to the class of ribosome-inactivating proteins and consists of an A-chain with N-glycosidase activity and a carbohydrate-binding B-chain. Incorporation of 2 mg/ml of SNA-I in the diet of neonates and adults of A. pisum caused 40-46% mortality within 2 days, while in third instars of S. exigua, the larval biomass was significantly reduced by 12% after feeding for 3 days on a diet containing 5 mg/g of SNA-I. Interestingly, extracts of the (mid)gut of treated A. pisum and S. exigua demonstrated DNA fragmentation and this was accompanied with an increase in caspase-3-like activity. The involvement of cell death or apoptosis in the entomotoxicity of SNA-I through induction of caspase-3-like activity was also confirmed by addition of the permeable caspase-3 inhibitor III in the diet, leading to a rescue of the treated aphid neonates. Finally, similar to the chimeric lectin SNA-I, the hololectin SNA-II, consisting of two carbohydrate-binding B-chains caused high mortality to neonate A. pisum aphids with an LC₅₀ of 1.59 mg/ml, suggesting that the entomotoxic action of the lectins under study mainly relies on their carbohydrate-binding activity.

From microbial sprays to insect-resistant transgenic plants: history of the biospesticideBacillus thuringiensis. A review

Bacillus thuringiensis, known as Bt, is a spore-forming bacterium that occurs naturally in soil and that produces highly specific insecticidal proteins called Cry proteins. These proteins are stomach poisons that specifically affect insects. Today, Bt preparations are considered as the most effective, specific and environmentally-friendly bioinsecticides; they have been used as biological pesticides in agriculture, forestry and in human health for the elimination of vectors of diseases for more than 60 years and their implementation far exceeds other microbial agents such as fungi, protozoa or viruses. This review on the use of this entomopathogenic bacterium in crop protection is not intended to be a compilation of the results of all the investigations made in this field. Instead, it is an attempt to provide an overview of the major trends and developments of Bt for the control of agricultural insect pests and to describe the main approaches that have been used to improve this natural bioinsecticide. Bt-based insecticides are considered safe for mammals and birds, and are safer for non-target insects than conventional insecticides; they have become the most widely used microbial insecticides. However, Bt products have several limitations, such as a narrow activity spectrum, instability in rain and sunlight, and inefficiency against pest feeding on internal tissues of the plants. The first step towards improving Bt has involved the isolation of new strains with higher and broader insecticidal activity against targeted insect pests and the cloning of cry genes encoding new insecticidal crystal proteins. A second strategy was to increase the persistence of its toxins in the field by encapsulation in recombinant asporogenic Bt strains or other heterologous recombinant microbial hosts; this protected the toxins against UV degradation and had the advantage that the transgenic microorganisms released into the environment were non-viable. Bt has also become a key source of genes for transgenic expression to provide pest resistance in plants and in so-called genetically modified plants. The engineering of plants to express Bt cry genes has been especially helpful against pests that attack parts of the plant that are usually not well protected by conventional insecticide application. The potential effects on human health and the environment of the large-scale use of these Bt crops are also in the scope of this review.

Effective liquid–liquid extraction method for analysis of pyrethroid and phenylpyrazole pesticides in emulsion-prone surface water samples

The distribution of pyrethroid and phenylpyrazole pesticides in the water environment has raised public concerns because of their potential risks to ecosystem and human health. However, co-extraction of emulsifier type compounds (by liquid–liquid extraction, LLE) present in environmental samples can present a challenge for quantifying typically low concentrations of pesticides. Several methods were evaluated for breaking emulsions in problematic environmental surface water samples extracted by LLE using methylene chloride. Target pesticides included 11 typical pyrethroid and phenylpyrazole pesticides commonly used in agricultural and landscape insect pest control. The most effective method was selected for validation in fortification studies with GC-ECD analysis. The average recoveries of spiked pyrethroid and phenylpyrazole pesticides were 88.2–123.4% for water samples with moderate emulsions and 93.0–117.4% for water samples with severe emulsions. Recoveries of the pesticides ranged 81.0–126.4% (water samples with moderate emulsions) and 95.9–110.6% (water samples with severe emulsions) for lowest fortification level (5–20 ng L −1), 88.2–123.4% (water samples with moderate emulsions) and 93.0–117.4% (water samples with severe emulsions) for middle fortification level (10–40 ng L −1), and 90.2–119.9% (water samples with moderate emulsions) and 91.2–105.9% (water samples with severe emulsions) for highest fortification level (50–200 ng L −1). Relative standard deviations of pesticide recoveries were usually <10%. Results indicate that this method is a robust and reproducible option for LLE of pyrethroid and phenylpyrazole pesticides from emulsion-prone surface water samples.

전자빔 조사가 담배가루이, 복숭아혹진딧물, 배추좀나방과 점박이응애의 발육과 생식에 미치는 영향

Four major agricultural insect pests, Bemisia tabaci, Myzus persicae, Plutella xylostella and Tetranychus urticae, were irradiated with 30, 50, 70, or 100 Gy electron-beam. Longevity, egg hatching, emergence, and fecundity of the test insects were measured. Hatchability of B. tabaci, P. xylostella and T. urticae declined with increasing irradiation doses, and all B. tabaci and T. urticae eggs were dead at 100 Gy. When eggs of B. tabaci, P. xylostella and T. urticae were irradiated, hatch was inhibited. B. tabaci adults grown from 70 Gy irradiated eggs did not lay eggs. Fecundity of P. xylostella from the 100 Gy irradiated eggs decreased. When B. tabaci, P. xylostella, M. persicae and T. urticae nymphs/larvae were irradiated, the results were similar as those of the hatched eggs. When P. xylostella pupae were irradiated with 100 Gy, fecundity of emerged adults decreased and no eggs hatched. When B. tabaci, P. xylostella, M. persicae and T. urticae adults were irradiated with 70 and 100 Gy, fecundity decreased and egg hatch of B. tabaci was inhibited. However, the longevity of adults did not change and electron-beam irradiation of all developmental stages had no effect on the longevity of adults.

Genomic structures of Cyp9m10 in pyrethroid resistant and susceptible strains of Culex quinquefasciatus

Development of insecticide resistance reduces the efficacy of controlling measures against the medical and agricultural insect pests. Cytochrome P450s are one of the major detoxification enzymes involved in insecticide metabolisms. Previously, we have reported that the P450 gene Cyp9m10 is about 260-fold overexpressed in a pyrethroid-resistant strain of Culex quinquefasciatus compared to a susceptible strain. In this study, we obtained direct evidence that the Cyp9m10 overexpression is caused by a cis-acting mutation. Additionally, a region of approximately 100 kb in length including the Cyp9m10 locus was specifically duplicated in the resistant strain. The two duplicated Cyp9m10 copies shared a completely identical sequence within the transcribed region and the flanking region up to the breakpoint located 1.1 kb upstream of the transcriptional start site. A Miniature Inverted-repeat Transposable Element (MITE)-like element was specifically inserted 0.2 kb upstream of both Cyp9m10 copies in the resistant strain. In backcross experiment, a haplotype containing the two duplicated Cyp9m10 copies was strongly associated with the pyrethroid resistance.

Degree-days units and expected generation numbers of peach fruit fly Bactrocera zonata (Saunders) (Diptera: Tephritidae) under climate change in

Climate change could profoundly affect the status of agricultural insect pests. This study predicts expected peach fruit fly (PFF) annual generations under current and future climate by using the accumulated thermal heat units expressed as degree-days unit (DDU). We evaluated how temperature expected to influence the annual generation numbers in three governorates of Egypt using the climate change data output from the HadCM3 model for A1 scenarios proposed by the Intergovernmental Panel on Climate Change. Results indicated that PFF in Asyout have the highest number of possible generations as compared to other locations (North Sinai and EL Beheira) under current climate to be 8 compared with 6 and 7 for other locations, respectively. Generation numbers of PFF under climate change conditions would be increased especially in Asyout governorate to 9 and 10 by 2050 and 2100, respectively. However, the expected generation numbers of PFF at 2050 and 2100 will be 8-9 and 9-10 generations per year for other two locations, respectively.

Resource concentration dilutes a key pest in indigenous potato agriculture

Modern restructuring of agricultural landscapes, due to the expansion of monocultures and the resulting elimination of non-crop habitat, is routinely blamed for rising populations of agricultural insect pests. However, landscape studies demonstrating a positive correlation between pest densities and the spatial extent of crop monocultures are rare. We test this hypothesis with a data set from 140 subsistence farms in the Andes and find the inverse correlation. Infestations by the Andean potato weevil (Premnotrypes spp.), the most important pest in Andean potato agriculture, decrease with increasing amounts of potato in the landscape. A statistical model predicts that aggregating potato fields may outperform the management of Andean potato weevils by IPM and chemical control. We speculate that the strong pest suppression generated by aggregating potato fields may partly explain why indigenous potato farmers cluster their potato fields under a traditional rotation system common in Andean agriculture (i.e., “sectoral fallow”). Our results suggest that some agricultural pests may also respond negatively to the expansion of monocultures, and that manipulating the spatial arrangement of host crops may offer an important tool for some IPM programs.

Sterile-insect methods for control of mosquito-borne diseases: an analysis.

Effective vector control, and more specifically mosquito control, is a complex and difficult problem, as illustrated by the continuing prevalence (and spread) of mosquito-transmitted diseases. The sterile insect technique and similar methods control certain agricultural insect pest populations in a species-specific, environmentally sound, and effective manner; there is increased interest in applying this approach to vector control. Such an approach, like all others in use and development, is not a one-size-fits-all solution, and will be more appropriate in some situations than others. In addition, the proposed release of pest insects, and more so genetically modified pest insects, is bound to raise questions in the general public and the scientific community as to such a method's efficacy, safety, and sustainability. This article attempts to address these concerns and indicate where sterile-insect methods are likely to be useful for vector control.

NMR Structure of Navel Orangeworm Moth Pheromone-Binding Protein (AtraPBP1): Implications for pH-Sensitive Pheromone Detection,

The navel orangeworm, Amyelois transitella (Walker), is an agricultural insect pest that can be controlled by disrupting male-female communication with sex pheromones, a technique known as mating disruption. Insect pheromone-binding proteins (PBPs) provide fast transport of hydrophobic pheromones through the aqueous sensillar lymph and promote sensitive delivery of pheromones to receptors. Here we present the three-dimensional structure of a PBP from A. transitella (AtraPBP1) in solution at pH 4.5 determined by nuclear magnetic resonance (NMR) spectroscopy. Pulsed-field gradient NMR diffusion experiments, multiangle light scattering, and (15)N NMR relaxation analysis indicate that AtraPBP1 forms a stable monomer in solution at pH 4.5 in contrast to forming mostly dimers at pH 7. The NMR structure of AtraPBP1 at pH 4.5 contains seven alpha-helices (alpha1, L8-L23; alpha2, D27-F36; alpha3, R46-V62; alpha4, A73-M78; alpha5, D84-S100; alpha6, R107-L125; alpha7, M131-E141) that adopt an overall main-chain fold similar to that of PBPs found in Antheraea polyphemus and Bombyx mori. The AtraPBP1 structure is stabilized by three disulfide bonds formed by C19/C54, C50/C108, and C97/C117 and salt bridges formed by H69/E60, H70/E57, H80/E132, H95/E141, and H123/D40. All five His residues are cationic at pH 4.5, whereas H80 and H95 become neutral at pH 7.0. The C-terminal helix (alpha7) contains hydrophobic residues (M131, V133, V134, V135, V138, L139, and A140) that contact conserved residues (W37, L59, A73, F76, A77, I94, V111, and V115) suggested to interact with bound pheromone. Our NMR studies reveal that acid-induced formation of the C-terminal helix at pH 4.5 is triggered by a histidine protonation switch that promotes rapid release of bound pheromone under acidic conditions.

Sex Pheromones and Their Impact on Pest Management

The idea of using species-specific behavior-modifying chemicals for the management of noxious insects in agriculture, horticulture, forestry, stored products, and for insect vectors of diseases has been a driving ambition through five decades of pheromone research. Hundreds of pheromones and other semiochemicals have been discovered that are used to monitor the presence and abundance of insects and to protect plants and animals against insects. The estimated annual production of lures for monitoring and mass trapping is on the order of tens of millions, covering at least 10 million hectares. Insect populations are controlled by air permeation and attract-and-kill techniques on at least 1 million hectares. Here, we review the most important and widespread practical applications. Pheromones are increasingly efficient at low population densities, they do not adversely affect natural enemies, and they can, therefore, bring about a long-term reduction in insect populations that cannot be accomplished with conventional insecticides. A changing climate with higher growing season temperatures and altered rainfall patterns makes control of native and invasive insects an increasingly urgent challenge. Intensified insecticide use will not provide a solution, but pheromones and other semiochemicals instead can be implemented for sustainable area-wide management and will thus improve food security for a growing population. Given the scale of the challenges we face to mitigate the impacts of climate change, the time is right to intensify goal-oriented interdisciplinary research on semiochemicals, involving chemists, entomologists, and plant protection experts, in order to provide the urgently needed, and cost-effective technical solutions for sustainable insect management worldwide.

SIT for African malaria vectors: Epilogue

As a result of increased support and the diligent application of new and conventional anti-malaria tools, significant reductions in malaria transmission are being accomplished. Historical and current evolutionary responses of vectors and parasites to malaria interventions demonstrate that it is unwise to assume that a limited suite of tools will remain effective indefinitely, thus efforts to develop new interventions should continue. This collection of manuscripts surveys the prospects and technical challenges for applying a novel tool, the sterile insect technique (SIT), against mosquitoes that transmit malaria. The method has been very successful against many agricultural pest insects in area-wide programs, but demonstrations against malaria vectors have not been sufficient to determine its potential relative to current alternatives, much of which will hinge ultimately upon cost. These manuscripts provide an overview of current efforts to develop SIT and identify key research issues that remain.

Comparing the refuge strategy for managing the evolution of insect resistance under different reproductive strategies

Genetically modified (GM) crops are used extensively worldwide to control diploid agricultural insect pests that reproduce sexually. However, future GM crops will likely soon target haplodiploid and parthenogenetic insects. As rapid pest adaptation could compromise these novel crops, strategies to manage resistance in haplodiploid and parthenogenetic pests are urgently needed. Here, we developed models to characterize factors that could delay or prevent the evolution of resistance to GM crops in diploid, haplodiploid, and parthenogenetic insect pests. The standard strategy for managing resistance in diploid pests relies on refuges of non-GM host plants and GM crops that produce high toxin concentrations. Although the tenets of the standard refuge strategy apply to all pests, this strategy does not greatly delay the evolution of resistance in haplodiploid or parthenogenetic pests. Two additional factors are needed to effectively delay or prevent the evolution of resistance in such pests, large recessive or smaller non-recessive fitness costs must reduce the fitness of resistance individuals in refuges (and ideally also on GM crops), and resistant individuals must have lower fitness on GM compared to non-GM crops (incomplete resistance). Recent research indicates that the magnitude and dominance of fitness costs could be increased by using specific host–plants, natural enemies, or pathogens. Furthermore, incomplete resistance could be enhanced by engineering desirable traits into novel GM crops. Thus, the sustainability of GM crops that target haplodiploid or parthenogenetic pests will require careful consideration of the effects of reproductive mode, fitness costs, and incomplete resistance.

Predicting insect pest status under climate change scenarios: combining experimental data and population dynamics modelling

AbstractClimate change could profoundly affect the status of agricultural insect pests. Several approaches have been used to predict how the temperature and precipitation changes could modify the abundances, distributions or status of insect pests. In this article it is demonstrated how the use of simple models, such as Ricker’s classic equation, including a mechanistic representation of the influence of exogenous forces may improve our predictive capacity of the dynamic behaviour of insect populations. Using data from classical experiments in population ecology, we evaluate how temperature and humidity influence the density of two stored grain insect pest, Tribolium confusum and Callosobruchus chinensis, and then, using the A2 and B2 scenarios proposed by the Intergovernmental Panel on Climate Change and the previous modelling, we develop predictions over the future pest status of T. confusum along South America austral region, and specifically for eight cities in the continental Chilean territory. Tribolium confusum and C. chinensis show qualitatively different responses to the exogenous forcing of temperature and humidity, respectively. Our simulations predict a change in the equilibrium density of T. confusum from 10 to 14% under the moderate B2 scenario and 12 to 22% under the extreme A2 scenario to the period, 2071–2100. Both results imply a severe change in the pest status of this species in the southern region. This study illustrates how the use of theoretically based models may improve our predictive capacity. This approach provides an opportunity to examine the link between invasive species and climate change and how new suitable habitat may become available for species whose niche space is limited in some degree by climatic conditions. The use of different scenarios allows us to examine the sensitivity of the predictions, and to improve the communication with the general public and decision‐makers; a key aspect in integrated pest management.

Enantioselectivity in chronic toxicology and accumulation of the synthetic pyrethroid insecticide bifenthrin in Daphnia magna

Introduced in the early 1980s, synthetic pyrethroids (SPs) are widely used in controlling a multitude of agricultural and domestic insect species, but they are highly toxic to fish and aquatic invertebrates. To date, however, studies have not adequately associated enantioselectivity in biological behavior with toxicity, especially regarding uptake and chronic toxicity. This study aimed to elucidate the relationship between chronic enantioselective toxicity and enantioselective accumulation of cis-bifenthrin (cis-BF) in Daphnia magna by 14C-labeled assay. The results demonstrated a clear enantioselectivity in chronic toxicity, for which 1R-cis-BF was evidently more toxic to D. magna than 1S-cis-BF. The lowest-observed-effective concentration in terms of survival and fecundity for 1R-cis-BF were almost 40- and 80-fold higher than that of 1S-cis-BF on day 7 and day 14, respectively. Further studies indicated that the accumulation of 1R-cis-BF was approximately 14- to 40-fold more than that of 1S-cis-BF in D. magna. This enantioselective accumulation also was consistent with the enantioselectivity in chronic toxicity of cis-BF. Thus, our results support the notion that enantioselectivity in chronic toxicity may be caused primarily by an enantiomer-specific biological process, such as uptake in aquatic organisms. This suggests that to properly assess the risk associated with use of SPs, the potential for enantioselectivity in their biotransformation and toxicity should be evaluated concurrently.

Bacillus thuringiensisBel Protein Enhances the Toxicity of Cry1Ac Protein toHelicoverpa armigeraLarvae by Degrading Insect Intestinal Mucin

Bacillus thuringiensis has been used as a bioinsecticide to control agricultural insects. Bacillus cereus group genomes were found to have a Bacillus enhancin-like (bel) gene, encoding a peptide with 20 to 30% identity to viral enhancin protein, which can enhance viral infection by degradation of the peritrophic matrix (PM) of the insect midgut. In this study, the bel gene was found to have an activity similar to that of the viral enhancin gene. A bel knockout mutant was constructed by using a plasmid-free B. thuringiensis derivative, BMB171. The 50% lethal concentrations of this mutant plus the cry1Ac insecticidal protein gene were about 5.8-fold higher than those of the BMB171 strain. When purified Bel was mixed with the Cry1Ac protein and fed to Helicoverpa armigera larvae, 3 mug/ml Cry1Ac alone induced 34.2% mortality. Meanwhile, the mortality rate rose to 74.4% when the same amount of Cry1Ac was mixed with 0.8 mug/ml of Bel. Microscopic observation showed a significant disruption detected on the midgut PM of H. armigera larvae after they were fed Bel. In vitro degradation assays showed that Bel digested the intestinal mucin (IIM) of Trichoplusia ni and H. armigera larvae to various degrading products, similar to findings for viral enhancin. These results imply Bel toxicity enhancement depends on the destruction of midgut PM and IIM, similar to the case with viral enhancin. This discovery showed that Bel has the potential to enhance insecticidal activity of B. thuringiensis-based biopesticides and transgenic crops.

Manduca sexta (Lepidoptera: Sphingidae) cadherin fragments function as synergists for Cry1A and Cry1C Bacillus thuringiensis toxins against noctuid moths Helicoverpa zea, Agrotis ipsilon and Spodoptera exigua

Specific Bacillus thuringiensis Berliner (Bt) toxins are effective against a narrow spectrum of species. While specificity is an advantage for limiting adverse effects on non-target organisms, it is also the primary drawback of Bt's application for controlling multiple pest species in agriculture, forestry and other areas. Recently, it was reported that a small toxin-binding fragment of Manduca sexta (Joh.) cadherin acts as a synergist of Bt toxins to M. sexta, Heliothis virescens F. and Helicoverpa zea (Boddie). These insects are quite susceptible to the Cry1A toxins. The first aim of the present study was to determine if longer-sized fragments of M. sexta cadherin differed in the level of toxin enhancement. The second aim was to examine enhancement of Bt toxins against relatively Bt-tolerant insects Agrotis ipsilon (Hufn.) and Spodoptera exigua (Hübner). Cadherin fragments longer than previously reported had improved synergistic properties. Significant enhancement of Bt Cry1A toxins against A. ipsilon and S. exigua was found. A cadherin fragment also increased Cry1C toxicity to S. exigua. The commercial development of this synergist has the potential to widen the spectrum of Bt toxicity to other important agricultural lepidopteran insect pests and thus increase its usefulness in agriculture.

A review of chemical control options for invasive social insects in island ecosystems

AbstractSocial insects present unique challenges to chemically based management strategies, especially because fast‐acting compounds commonly applied for many pest insects may not be the most effective for colony elimination. The reproductive caste of a colony is the most protected from direct damage by insecticides, and compounds that cause rapid mortality among foragers frequently do not impact the reproductive members or even markedly reduce overall colony size. With recent bans on persistent insecticides that previously have been used to control social insects, especially termites, new compounds must be used. Island and coastal ecosystems are particularly sensitive to the effects of widespread pesticide use and concerns about unintentional water pollution and runoff are common, and international attention is being paid to developing sustainable pesticide options for agricultural and urban pest insects in particularly sensitive environments. Given the precarious status of many native insects and arthropods care must be taken to minimize exposure to potentially harmful insecticides and the non‐target impacts of these chemicals. However, recent developments in the synthesis and discovery of highly selective insecticides with low mammalian and non‐target toxicity provide viable alternatives to the broad‐spectrum persistent organochlorine insecticides that have been largely deregistered. Novel technologies, particularly synthetic analogues of biologically active compounds, yield new chemical control options and management strategies for island and other sensitive ecosystems; case studies from Australia, the Galapagos Islands and New Zealand highlight current challenges and successes.

Efficient screening and breeding of Bacillus thuringiensis subsp. kurstaki for high toxicity against Spodoptera exigua and Heliothis armigera

Spodoptera exigua is one of the most renowned agricultural pest insects and relatively insensitive to Bacillus thuringiensis subsp. kurstaki strains which are widely used commercial products to control lepidopterans such as Heliothis armigera. In the current study, we have developed a new and efficient approach to screen and breed a B. thuringiensis subsp. kurstaki strain exhibiting high toxicity against S. exigua while retaining its high toxicity against H. armigera. UV and diethyl sulfate methods were used for mutagenesis, followed by an agar plug plate diffusion assay for preliminary screening of Zwittermicin A over-producing mutants, from which we obtained a mutant strain, designated here as B. thuringiensis subsp. kurstaki D1-23, with high toxicity against S. exigua. The toxicity of D1-23 against S. exigua and H. armigera was improved by 115.4 and 25.9%, respectively, compared to its parental commercial strain BMB005.