Lisa J. Bird

Relative fitness of Cry1a-resistant and -susceptible Helicoverpa armigera (Lepidoptera: Noctuidae) on conventional and transgenic cotton

Abstract Glasshouse and laboratory experiments were conducted to evaluate the relative fitness of Cry1A-susceptible and laboratory-selected resistant strains of Helicoverpa armigera (Hübner). Life history parameters of H. armigera larvae feeding on young cotton plants showed a significant developmental delay of up to 7 d for the resistant strain compared with the susceptible strain on non-Bacillus thuringiensis (Bt) cotton. This fitness cost was not evident on artificial diet. There was no developmental delay in the F1 hybrid progeny from the reciprocal backcross of the resistant and susceptible strains, indicating that the fitness cost is recessive. In two cohorts tested, survival to pupation of resistant larvae on Bt cotton expressing Cry1Ac was 54 and 51% lower than on non-Bt cotton, whereas all susceptible and F1 larvae tested on Cry1Ac cotton were killed. Mortality of susceptible larvae occurred in the first or second instar, whereas the F1 larvae were able to develop to later instars before dying, demonstrating that resistance is incompletely recessive. The intrinsic rate of increase was reduced by >50% in the resistant strain on Cry1Ac cotton compared with the susceptible strain on non-Bt cotton. There was a significant reduction in the survival of postdiapausal adults from the resistant strain and the F1 strains, indicating that there is a nonrecessive overwintering cost associated with Cry1A resistance in H. armigera.

Fitness of Cry1A-Resistant and -Susceptible Helicoverpa armigera (Lepidoptera: Noctuidae) on Transgenic Cotton with Reduced Levels of Cry1Ac

Abstract The performance of Helicoverpa armigera (Hübner) on 15-wk-old cotton plants was compared for a susceptible strain, a near-isogenic laboratory-selected strain, and F1 progeny of the two strains. Glasshouse experiments were conducted to test the three insect types on conventional plants and transgenic plants that produced the Bacillus thuringiensis (Bt) toxin Cry1Ac. At the time of testing (15 wk), the Cry1Ac concentration in cotton leaves was 75% lower than at 4 wk. On these plants, <10% of susceptible larvae reached the fifth instar, and none survived to pupation. In contrast, survival to adulthood on Cry1Ac cotton was 62% for resistant larvae and 39% for F1 larvae. These results show that inheritance of resistance to 15-wk-old Cry1Ac cotton is partially dominant, in contrast to results previously obtained on 4-wk-old Cry1Ac cotton. Growth and survival of resistant insects were similar on Cry1Ac cotton and on non-Bt cotton, but F1 insects developed more slowly on Cry1Ac cotton than on non-Bt cotton. Survival was lower and development was slower for resistant larvae than for susceptible and F1 larvae on non-Bt cotton. These results show recessive fitness costs are associated with resistance to Cry1Ac.

Baseline Susceptibility of Helicoverpa armigera (Lepidoptera: Noctuidae) to Indoxacarb, Emamectin Benzoate, and Chlorantraniliprole in Australia

ABSTRACT Baseline susceptibility of Helicoverpa armigera (Hübner) to emamectin benzoate, chlorantraniliprole, and indoxacarb was determined in feeding assays on insecticide-incorporated artificial diet in the laboratory. The intraspecific variation of H. armigera was established from field populations collected between September 2012 and March 2013, primarily from commercial farms across eastern Australia. Emamectin benzoate had the highest toxicity with a median lethal concentration (LC50) of 0.01 µg/ml diet (n = 20 strains). The LC50 for chlorantraniliprole was 0.03 µg/ml diet (n = 21 strains), while indoxacarb had the lowest relative toxicity with an average LC50 of 0.3 µg/ml diet (n = 22 strains). Variation in susceptibility amongst field strains was 2.3-fold for emamectin benzoate and 2.9-fold for chlorantraniliprole and indoxacarb. Discriminating concentrations of 0.2, 1, and 12 µg of insecticide per milliliter of diet for emamectin benzoate, chlorantraniliprole, and indoxacarb, respectively, were calculated from toxicological data from field H. armigera strains as a first step in resistance management of these classes of insecticide in Australia. The low intraspecific tolerance, high slope values, and goodnessof-fit to a probit binomial model obtained in this study suggest that a feeding assay using diet incorporated insecticide is an effective laboratory method for measuring the dose—responses of these classes of insecticides in H. armigera.

Toxicity and Cross-Resistance of Insecticides to Cry2Ab-Resistant and Cry2Ab-Susceptible Helicoverpa armigera and Helicoverpa punctigera (Lepidoptera: Noctuidae)

ABSTRACT Since 2004–2005 cotton expressing Cry1Ac and Cry2Ab insecticidal proteins from the bacterium Bacillus thuringiensis has been commercially available in Australia to manage the target pests Helicoverpa armigera (Hübner) and Helicoverpa punctigera (Wallengren). In both target species, the frequency of alleles conferring resistance to Cry2Ab is unexpectedly high in field populations. A significant challenge for managing these pests would occur if resistance to Cry2Ab toxins inadvertently selected for resistance to other insecticides used to control them. Dose-response bioassays were performed to measure the toxicity of currently registered insecticide sprays on isogenic strains of Cry2Ab-resistant and Cry2Ab-susceptible H. armigera and H. punctigera. Within-species comparisons of Cry2Ab-resistant and Cry2Ab-susceptible strains of H. armigera and H. punctigera indicate no cross-resistance with pyrethroid insecticides. Additionally, Cry2Ab-resistant strains were not cross-resistant to the following selective insecticides: indoxacarb, chlorantraniliprole, and avermectins. In both H. armigera and H. punctigera, Cry2Ab-resistant colonies exhibited a small, but significant, degree of enhanced susceptibility in response to chlorpyrifos and methomyl. We report higher tolerance to conventional insecticides in H. armigera compared with H. punctigera. Our results indicate that there is no significant interplay between Cry2Ab resistance frequencies in H. armigera and H. punctigera and frequencies of resistance to a range of insecticide sprays currently registered for cotton. Therefore, we conclude that any increases in frequencies of the common Cry2Ab resistance phenotypes identified in Australian populations of Helicoverpa spp. are unlikely to increase resistance risk for the indoxacarb, chlorantraniliprole, or avermectin classes of insecticide.

Rapid antibody-based field test to distinguish between Helicoverpa armigera (Lepidoptera: Noctuidae) and Helicoverpa punctigera (Lepidoptera: Noctuidae).

Abstract Helicoverpa armigera (Hübner) and Helicoverpa punctigera (Wallengren) are the two most important insect pests of cotton production in Australia and require application of insecticides to control them. H. armigera has developed resistance to several insecticides but H. punctigera has not. Cost-effective management of insecticide resistance requires that growers be able to determine the proportion of H. armigera eggs or young larvae present on their crop before applying insecticides. This is impossible visually. We generated two monoclonal antibodies that reacted with the insect protein “lipophorin” and were capable of discriminating individuals of the two species at all life-stages. The antibodies were incorporated into a rapid test kit that was tested under field conditions over two growing seasons. Results obtained with the kit agreed closely with those obtained by rearing larvae through to second instar.