Timothy J. Dennehy

Development time and resistance to Bt crops

Crop plants genetically engineered to produce insecticidal toxins derived from the bacterium Bacillus thuringiensis (Bt) are being grown on millions of hectares, but their success will be short-lived if pests adapt to them quickly,. The primary strategy for delaying insect resistance to transgenic Bt plants is to provide refuges of host plants that do not produce Bt toxins. This potentially delays the development of insect resistance to Bt crops by providing susceptible insects for mating with resistant insects. But our laboratory results with a worldwide pest of cotton, pink bollworm moths (Pectinophora gossypiella), contradict an important assumption of the refuge strategy. We find that a resistant strain of larvae on Bt cotton takes longer to develop than susceptible larvae on non-Bt cotton. This developmental asynchrony favours non-random mating that could reduce the expected benefits of the refuge strategy.

Three cadherin alleles associated with resistance to Bacillus thuringiensis in pink bollworm

Evolution of resistance by pests is the main threat to long-term insect control by transgenic crops that produce Bacillus thuringiensis (Bt) toxins. Because inheritance of resistance to the Bt toxins in transgenic crops is typically recessive, DNA-based screening for resistance alleles in heterozygotes is potentially much more efficient than detection of resistant homozygotes with bioassays. Such screening, however, requires knowledge of the resistance alleles in field populations of pests that are associated with survival on Bt crops. Here we report that field populations of pink bollworm (Pectinophora gossypiella), a major cotton pest, harbored three mutant alleles of a cadherin-encoding gene linked with resistance to Bt toxin Cry1Ac and survival on transgenic Bt cotton. Each of the three resistance alleles has a deletion expected to eliminate at least eight amino acids upstream of the putative toxin-binding region of the cadherin protein. Larvae with two resistance alleles in any combination were resistant, whereas those with one or none were susceptible to Cry1Ac. Together with previous evidence, the results reported here identify the cadherin gene as a leading target for DNA-based screening of resistance to Bt crops in lepidopteran pests.

Rapid Spread of a Bacterial Symbiont in an Invasive Whitefly Is Driven by Fitness Benefits and Female Bias

A Rickettsia bacterium promotes its own geographical spread by manipulating its insect host’s sex ratio and fecundity. Maternally inherited bacterial symbionts of arthropods are common, yet symbiont invasions of host populations have rarely been observed. Here, we show that Rickettsia sp. nr. bellii swept into a population of an invasive agricultural pest, the sweet potato whitefly, Bemisia tabaci, in just 6 years. Compared with uninfected whiteflies, Rickettsia-infected whiteflies produced more offspring, had higher survival to adulthood, developed faster, and produced a higher proportion of daughters. The symbiont thus functions as both mutualist and reproductive manipulator. The observed increased performance and sex-ratio bias of infected whiteflies are sufficient to explain the spread of Rickettsia across the southwestern United States. Symbiont invasions such as this represent a sudden evolutionary shift for the host, with potentially large impacts on its ecology and invasiveness.

Long-term regional suppression of pink bollworm by Bacillus thuringiensis cotton

Despite the potentially profound impact of genetically modified crops on agriculture and the environment, we know little about their long-term effects. Transgenic crops that produce toxins from Bacillus thuringiensis (Bt) to control insects are grown widely, but rapid evolution of resistance by pests could nullify their benefits. Here, we present theoretical analyses showing that long-term suppression of pest populations is governed by interactions among reproductive rate, dispersal propensity, and regional abundance of a Bt crop. Supporting this theory, a 10-year study in 15 regions across Arizona shows that Bt cotton suppressed a major pest, pink bollworm (Pectinophora gossypiella), independent of demographic effects of weather and variation among regions. Pink bollworm population density declined only in regions where Bt cotton was abundant. Such long-term suppression has not been observed with insecticide sprays, showing that transgenic crops open new avenues for pest control. The debate about putative benefits of Bt crops has focused primarily on short-term decreases in insecticide use. The present findings suggest that long-term regional pest suppression after deployment of Bt crops may also contribute to reducing the need for insecticide sprays.

Delayed resistance to transgenic cotton in pink bollworm

Transgenic crops producing Bacillus thuringiensis (Bt) toxins kill some key insect pests and thus can reduce reliance on insecticides. Widespread planting of such Bt crops increased concerns that their usefulness would be cut short by rapid evolution of resistance to Bt toxins by pests. Pink bollworm (Pectinophora gossypiella) is a major pest that has experienced intense selection for resistance to Bt cotton in Arizona since 1997. We monitored pink bollworm resistance to Bt toxin for 8 years with laboratory bioassays of strains derived annually from 10-17 cotton fields statewide. Bioassay results show no net increase from 1997 to 2004 in the mean frequency of pink bollworm resistance to Bt toxin. A synthesis of experimental and modeling results suggests that this delay in resistance can be explained by refuges of cotton without Bt toxin, recessive inheritance of resistance, incomplete resistance, and fitness costs associated with resistance.

Mutations in the Bemisia tabaci para sodium channel gene associated with resistance to a pyrethroid plus organophosphate mixture.

The voltage-gated sodium channel is the primary target site of pyrethroid insecticides. In some insects, super knockdown resistance (super-kdr) to pyrethroids is caused by point mutations in the linker fragment between transmembrane segments 4 and 5 of the para-type sodium channel protein domain II (IIS4-5). Here, we identify two mutations in the IIS4-5 linker of the para-type sodium channel of the whitefly, BEMISIA TABACI: methionine to valine at position 918 (M918V) and leucine to isoleucine at position 925 (L925I). Although each mutation was isolated independently from strains >100-fold resistant to a pyrethroid (fenpropathrin) plus organophosphate (acephate) mixture, only L925I was associated with resistance in strains derived from the field in 2000 and 2001. The L925I mutation occurred in all individuals from nine different field collections that survived exposure to a discriminating concentration of fenpropathrin plus acephate. Linkage analysis of hemizygous male progeny of unmated heterozygous F1 females (L925Ixwild-type) shows that the observed resistance is tightly linked to the voltage-gated sodium channel locus. The results provide a molecular tool for better understanding, monitoring and managing pyrethroid resistance in B. tabaci.

Arthropod Abundance and Diversity in Bt and Non-Bt Cotton Fields

Abstract The widespread planting of crops genetically modified to produce Bacillus thuringeinsis Berliner (Bt) toxins for pest control may affect nontarget arthropods. To address this issue, we compared the abundance and diversity of arthropods on plants in field plots of Bt cotton, non-Bt cotton, and a row mixture of 75% Bt cotton and 25% non-Bt cotton at two sites in Arizona. Over three sampling dates during 2 yr, we recorded all of the arthropods found on 120 cotton plants per treatment. This yielded 3,309 individual arthropods from 69 families. Excluding pink bollworm, Pectinophora gossypiella Saunders, the pest targeted by Bt cotton, we compared the abundance and diversity of all arthropods, chewing herbivores, sucking herbivores, rasping-sucking arthropods, and natural enemies. Arthropod abundance was significantly affected by site, plant height, and cotton type. More arthropods were collected from row mixture plots than Bt plots, but arthropod abundance did not differ significantly between Bt plots and non-Bt plots. The number of families collected was 57 for row mixture plots, 55 for non-Bt plots, and 47 for Bt plots. The number of families increased as arthropod abundance increased, suggesting that the differences in diversity among treatments were caused by differences in abundance. Within row mixture plots, arthropod abundance and diversity did not differ significantly between Bt plants and non-Bt plants. We conclude that the differences between Bt and non-Bt cultivars had relatively minor effects on the arthropod community on cotton plants.

Beet armyworm (Spodoptera exigua) resistance to spinosad.

Susceptibility to spinosad (Success®/Tracer®) of beet armyworm (Spodoptera exigua) from the southern USA and Southeast Asia was determined through exposure of second- and third-instar larvae to dipped cotton leaves. LC50 estimates of susceptibility of second- and third-instar larvae of field populations ranged from 0.279 to 6.14 and 0.589 to 14.0 mg spinosad litre−1, respectively. A Thailand population was 22- and 24-fold less susceptible than the six other US field populations evaluated, and 85- and 58-fold less susceptible than a reference laboratory population, respectively. From these results, we initiated experiments to test the hypothesis that the Thailand population was resistant to spinosad. F1 crosses between the resistant Thailand population and a susceptible reference strain yielded individuals that were 22-fold less sensitive to spinosad than the susceptible parent. This same resistant strain exhibited significantly greater survivorship on plants treated with spinosad in the field. Lastly, selection of an Arizona population resulted in a significant reduction in susceptibility to spinosad, further substantiating the hypothesis of a genetic basis for resistance to spinosad. These findings indicate a vulnerability of this new insecticide to resistance development in beet armyworm and should serve as a warning against excessive use of it. © 2000 Society of Chemical Industry

Large-Scale Management of Insect Resistance to Transgenic Cotton in Arizona: Can Transgenic Insecticidal Crops be Sustained?

Abstract A major challenge for agriculture is management of insect resistance to toxins from Bacillus thuringiensis (Bt) produced by transgenic crops. Here we describe how a large-scale program is being developed in Arizona for management of resistance to Bt cotton in the pink bollworm, Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae), and other insect pests of cotton. Financial support from growers makes this program possible. Collaboration between the Arizona Cotton Research and Protection Council, the University of Arizona, and government agencies has led to development of resistance management guidelines, a remedial action plan, and tools for monitoring compliance with the proposed guidelines. Direct participation in development of resistance management policies is a strong incentive for growers to invest in resistance management research. However, more research, regularly updated regulations, and increased collaboration between stakeholders are urgently needed to maintain efficacy of Bt toxins in transgenic crops.

Effects of Bt Cotton and Cry1Ac Toxin on Survival and Development of Pink Bollworm (Lepidoptera: Gelechiidae)

Abstract We evaluated the effects of Bacillus thuringiensis (Bt) toxin Cry1Ac on survival and development of a susceptible strain and laboratory-selected resistant strains of pink bollworm, Pectinophora gossypiella (Saunders). For susceptible and resistant strains tested on artificial diet, increases in Cry1Ac concentration reduced developmental rate and pupal weight. In greenhouse tests, survival of resistant larvae on transgenic cotton that produces Cry1Ac (Bt cotton) was 46% relative to their survival on non-Bt cotton. In contrast, Bt cotton killed all susceptible larvae tested. F1 hybrid progeny of resistant and susceptible adults did not survive on Bt cotton, which indicates recessive inheritance of resistance. Compared with resistant or susceptible larvae reared on non-Bt cotton, resistant larvae reared on Bt cotton had lower survival and slower development, and achieved lower pupal weight and fecundity. Recessive resistance to Bt cotton is consistent with one of the basic assumptions of the refuge strategy for delaying resistance to Bt cotton. Whereas slower development of resistant insects on Bt cotton could increase the probability of mating between resistant adults and accelerate resistance, negative effects of Bt cotton on the survival and development of resistant larvae could delay evolution of resistance.