Understanding successful resistance management: the European corn borer and Bt corn in the United States

The European corn borer, Ostrinia nubilalis Hübner (Lepidoptera: Crambidae) has been a major pest of corn and other crops in North America since its accidental introduction nearly a hundred years ago. Wide adoption of transgenic corn hybrids that express toxins from Bacillus thuringiensis, referred to as Bt corn, has suppressed corn borer populations and reduced the pest status of this insect in parts of the Corn Belt. Continued suppression of this pest, however, will depend on managing potential resistance to Bt corn, currently through the high-dose refuge (HDR) strategy. In this review, we describe what has been learned with regard to O. nubilalis resistance to Bt toxins either through laboratory selection experiments or isolation of resistance from field populations. We also describe the essential components of the HDR strategy as they relate to O. nubilalis biology and ecology. Additionally, recent developments in insect resistance management (IRM) specific to O. nubilalis that may affect the continued sustainability of this technology are considered.     

Regulation of Bt crops in Canada

The Canadian Food Inspection Agency (CFIA) regulates environmental releases of plants with novel traits, which include transgenic plants such as Bt crops. Bt crops are regulated in Canada because they express insect resistance novel to their species. Commercialization of crops with novel traits such as the production of insecticidal Bt proteins requires an approval for environmental release, as well as approvals for use as feed and food. Environmental factors such as potential impacts on non-target species are considered. Insect resistance management (IRM) may be imposed as a condition for environmental release of Bt crops to delay the development of resistance in the target insect. Bt potato and European corn borer-resistant Bt corn have been released with mandatory IRM. The CFIA imposes an IRM plan consisting of appropriate refugia, education of farmers and seed dealers, and monitoring and mitigation. Industry, regulators, government extension staff and public researchers provide expert advice on IRM.     

The design and implementation of insect resistance management programs for Bt crops

Cotton and corn plants with insect resistance traits introduced through biotechnological methods and derived from the bacterium Bacillus thuringiensis (Bt) have been widely adopted since they were first introduced in 1996. Because of concerns about resistance evolving to these Bt crops, they have been released with associated IRM programs that employ multiple components and reflect the input of academic, industrial and regulatory experts. This paper summarizes the current status of Bt crop technologies in cotton and corn, the principles of IRM for Bt crops and what they mean for the design of IRM programs. It describes how these IRM programs have been implemented and some of the key factors affecting successful implementation. Finally, it suggests how they may evolve to properly steward these traits in different geographies around the world. The limited number of reported cases of resistance after more than 15 years of intensive global use of Bt crops suggest that this exercise has been broadly successful. Where resistance issues have been observed, they have been associated with first generation technologies and incomplete or compromised IRM programs (i.e., inadequate structured refuge). Next generation technologies with multiple pyramided modes of action, together with the implementation of IRM strategies that are more dependent upon manufacturing and less dependent upon grower behavior, such as seed mixes, should further enhance IRM programs for Bt crops.     

A global approach to resistance monitoring

Transgenic crops producing insecticidal toxins from the bacterium Bacillus thuringiensis (Bt) have been grown in many parts of the world since 1996. In the United States, the Environmental Protection Agency (EPA) has required that industry submit insect resistance management (IRM) plans for each Bt corn and cotton product commercialized. A coalition of stakeholders including the EPA, USDA, academic scientists, industry, and grower organizations have cooperated in developing specific IRM strategies. Resistance monitoring (requiring submission of annual reports to the EPA), and a remedial action plan addressing any contingency if resistance should occur, are important elements of these strategies. At a global level, Monsanto conducts baseline susceptibility studies (prior to commercialization), followed by monitoring studies on target pest populations, for all of its commercialized Bt crop products. To date, Monsanto has conducted baseline/monitoring studies in Argentina, Australia, Brazil, Canada, China, Colombia, India, Mexico, the Philippines, South Africa, Spain, and the United States. Examples of pests on which resistance monitoring has been conducted include cotton bollworm, Helicoverpa zea, European corn borer, Ostrinia nubilalis, pink bollworm, Pectinophora gossypiella, Southwestern corn borer, Diatraea grandiosella, tobacco budworm, Heliothis virescens, and western corn rootworm, Diabrotica virgifera virgifera, in the United States, cotton bollworm, Helicoverpa armigera, in China, India and Australia, and H. virescens and H. zea in Mexico. No field-selected resistance to Bt crops has been documented.     

Recent developments and future prospects in insect pest control in transgenic crops

The adoption of insect-resistant transgenic crops has been increasing annually at double-digit rates since the commercial release of first-generation maize and cotton expressing a single modified Bacillus thuringiensis toxin (Bt) nine years ago. Studies have shown that these Bt crops can be successfully deployed in agriculture, which has led to a decrease in pesticide usage, and that they are environmentally benign. However, the sustainability and durability of pest resistance continues to be discussed. In this review, we focus on the science that underpins second- and third-generation insect-resistant transgenic plants and examine the appropriateness and relevance of models that are currently being used to determine deployment strategies to maximize sustainability and durability. We also review strategies that are being developed for novel approaches to transgenic insect pest control.     

Planting transgenic insecticidal corn based on economic thresholds: consequences for integrated pest management and insect resistance management

A simulation model of the western corn rootworm, Diabrotica virgifera virgifera LeConte, was used to investigate whether sampling and economic thresholds can improve integrated pest management (IPM) and insect resistance management (IRM) when transgenic insecticidal crops are used for insect pest management. When transgenic corn killed at least 80% of susceptible larvae, the calculated economic threshold increased linearly as the proportion of susceptible beetles surviving the toxin increased. The use of economic thresholds slightly slowed the evolution of resistance to transgenic insecticidal crops. In areas with or without rotation-resistant western corn rootworm phenotypes, the use of sampling and economic thresholds generated similar returns compared with strategies of planting transgenic corn, Zea mays L., every season. Because transgenic crops are extremely effective, farmers may be inclined to plant transgenic crops every season rather than implementing costly and time-consuming sampling protocols.     

Fees or refuges: which is better for the sustainable management of insect resistance to transgenic Bt corn?

The evolution of resistance in insect pests will imperil the efficiency of transgenic insect-resistant crops. The currently advised strategy to delay resistance evolution is to plant non-toxic crops (refuges) in close proximity to plants engineered to express the toxic protein of the bacterium Bacillus thuringiensis (Bt). We seek answers to the question of how to induce growers to plant non-toxic crops. A first strategy, applied in the United States, is to require Bt growers to plant non-Bt refuges and control their compliance with requirements. We suggest that an alternative strategy is to make Bt seed more expensive by instituting a user fee, and we compare both strategies by integrating economic processes into a spatially explicit, population genetics model. Our results indicate that although both strategies may allow the sustainable management of the common pool of Bt-susceptibility alleles in pest populations, for the European corn borer (Ostrinia nubilalis) one of the most serious pests in the US corn belt, the fee strategy is less efficient than refuge requirements.     

Insect resistance management in GM crops: past, present and future

Transgenic plants expressing insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) were first commercialized in 1996 amid concern from some scientists, regulators and environmentalists that the widespread use of Bt crops would inevitably lead to resistance and the loss of a 'public good,' specifically, the susceptibility of insect pests to Bt proteins. Eight years later, Bt corn and cotton have been grown on a cumulative area >80 million ha worldwide. Despite dire predictions to the contrary, resistance to a Bt crop has yet to be documented, suggesting that resistance management strategies have been effective thus far. However, current strategies to delay resistance remain far from ideal. Eight years without resistance provides a timely opportunity for researchers, regulators and industry to reassess the risk of resistance and the most effective strategies to preserve Bt and other novel insect-resistant crops in development.     

Transgenic Plants for Insect Pest Control: A Forward Looking Scientific Perspective

One of the first successes of plant biotechnology has been the creation and commercialisation of transgenic crops exhibiting resistance to major insect pests. First generation products encompassed plants with single insecticidal Bt genes with resistance against major pests of corn and cotton. Modelling studies predicted that usefulness of these resistant plants would be short-lived, as a result of the ability of insects to develop resistance against single insecticidal gene products. However, despite such dire predictions no such collapse has taken place and the acreage of transgenic insect resistance crops has been increasing at a steady rate over the 9 years since the deployment of the first transgenic insect resistant plant. However, in order to assure durability and sustainability of resistance, novel strategies have been contemplated and are being developed. This perspective addresses a number of potentially useful strategies to assure the longevity of second and third generation insect resistant plants.     

Detection and monitoring of insect resistance to transgenic Bt crops

Transgenic crops expressing Bacillus thuringiensis （Bt） endotoxins havebecome one of the most important tools for managing corn and cotton insect pests in the US and other countries. The widespread adoption of transgenic Bt crops could place a high degree of selection pressure on the target insect populations and accelerate development of resistance, raising concerns about the long-term durability of Bt plants as an effective pest management tool. Conservation of Bt susceptibility in insects has become one of the most active research areas in modern agriculture. One of the key factors for a successful Bt resistance management plan is to have a cost-effective monitoring system that can provide information on. （i） the initial Bt resistance allele frequencies at low levels in field insect populations; and （ii） early shifts in Bt resistance allele frequencies so that proactive measures for managing resistance can be deployed well before field control failures. Developing such a monitoring program has been difficult because： （i） resistance traits that occur at very low frequencies are hard to detect; （ii） many factors affect the sensitivity and accuracy of a Bt resistance monitoring program; and （iii） monitoring resistance is costly. Several novel methods for detecting Bt resistance alleles developed during the last decade have made a cost-effective monitoring system possible. Future studies should focus on how to improve and standardize the methodologies for insect sampling and Bt resistance detection.     

Large-scale management of insect resistance to transgenic cotton in Arizona: can transgenic insecticidal crops be sustained?

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.     

In silico designing of insecticidal small interfering RNA (siRNA) for Helicoverpa armigera control

Helicoverpa armigera, a polyphagous lepidopteron insect pest causes severe yield loss in cotton, legumes, tomato, okra and other crops. Application of chemical pesticides although effective, has human health and environmental safety concerns. Moreover, development of resistance against most of the available pesticides is compelling to look for alternative strategies. Adoption of Bt transgenic crops have resulted in reduction in pesticide consumption and increasing crop productivity. However, sustainability of Bt transgenic crops is threatened by the emergence of insect resistance. In the present study potential insecticidal siRNA were identified in six H. armigera horrhonal pathway genes. Out of over 2000 computationally identified siRNA, 16 most promising siRNA were selected that address the biosafety concerns and have high potential of targeted gene silencing. These siRNA will be useful for chemical synthesis, in insect feeding assays and knockdown the target H. armigera hormone biosynthesis, consequently obstructing the completion of insect life cycle. The siRNA have a great potential of deployment to control H. annrmigera alone as well as with Bt for insect resistance management.     

转Bt基因抗虫水稻的研究进展与生态安全评价

表达杀虫蛋白的转Bt基因植物正在改革着现代农业．综述了国内外转Bt基因水稻及其抗虫性的研究进展及水稻害虫对Bt水稻的抗性风险及抗性管理策略,提出了对转基因Bt水稻进行生态安全风险评价的具体内容．     

Assessment of fitness costs in Cry3Bb1‐resistant and susceptible western corn rootworm (Coleoptera: Chrysomelidae) laboratory colonies

Abstract Maize production in the United States is dominated by plants genetically modified with transgenes from Bacillus thuringiensis (Bt). Cry3Bb delta endotoxins expressed by Bt maize specifically target corn rootworms (genus Diabrotica) and have proven highly efficacious. However, development of resistance to Bt maize, especially among western corn rootworm (Diabrotica virgifera virgifera) populations, poses a significant threat to the future viability of this pest control biotechnology. The structured refuge insect resistance management (IRM) strategy implemented in the United States for Bt maize adopts a conservative approach to managing resistance by assuming no fitness costs of Bt resistance, even though these trade‐offs strongly influence the dynamics of Bt resistance within numerous agricultural pest species. To investigate the effects of Bt resistance on fitness components of western corn rootworm, we compared survivorship, fecundity and viability of five Bt‐resistant laboratory lines reared on MON863 (YieldGard Rootworm), a Bt maize product that expresses Cry3Bb1 delta endotoxin, and on its non‐transgenic isoline. Analysis of performance on the isoline maize demonstrated no fitness costs associated with Bt resistance. In fact, resistant lines emerged approximately 2–3 days earlier than control lines when reared on both MON863 and the isoline, indicating that selection for Bt resistance resulted in a general increase in the rate of larval development. In addition, resistant lines reared on Bt maize displayed higher fecundity than those reared on the isoline, which may have significant management implications. These data will be valuable for formulating improved IRM strategies for a principal agricultural pest of maize.     

Insect resistance to Bt crops: evidence versus theory

Evolution of insect resistance threatens the continued success of transgenic crops producing Bacillus thuringiensis (Bt) toxins that kill pests. The approach used most widely to delay insect resistance to Bt crops is the refuge strategy, which requires refuges of host plants without Bt toxins near Bt crops to promote survival of susceptible pests. However, large-scale tests of the refuge strategy have been problematic. Analysis of more than a decade of global monitoring data reveals that the frequency of resistance alleles has increased substantially in some field populations of Helicoverpa zea, but not in five other major pests in Australia, China, Spain and the United States. The resistance of H. zea to Bt toxin Cry1Ac in transgenic cotton has not caused widespread crop failures, in part because other tactics augment control of this pest. The field outcomes documented with monitoring data are consistent with the theory underlying the refuge strategy, suggesting that refuges have helped to delay resistance.     

Field tests on managing resistance to Bt-engineered plants

Several important crops have been engineered to express toxins of Bacillus thuringiensis (Bt) for insect control. In 1999, US farmers planted nearly 8 million hectares (nearly 20 million acres) of transgenic Bt crops approved by the EPA. Bt-transgenic plants can greatly reduce the use of broader spectrum insecticides, but insect resistance may hinder this technology. Present resistance management strategies rely on a "refuge" composed of non-Bt plants to conserve susceptible alleles. We have used Bt-transgenic broccoli plants and the diamondback moth as a model system to examine resistance management strategies. The higher number of larvae on refuge plants in our field tests indicate that a "separate refuge" will be more effective at conserving susceptible larvae than a "mixed refuge" and would thereby reduce the number of homozygous resistant (RR) offspring. Our field tests also examined the strategy of spraying the refuge to prevent economic loss to the crop while maintaining susceptible alleles in the population. Results indicate that great care must be taken to ensure that refuges, particularly those sprayed with efficacious insecticides, produce adequate numbers of susceptible alleles. Each insect/Bt crop system may have unique management requirements because of the biology of the insect, but our studies validate the need for a refuge. As we learn more about how to refine our present resistance management strategies, it is important to also develop the next generation of technology and implementation strategies.     

Resistance of the fall armyworm,Spodoptera frugiperda,to transgenic Bacillus thuringiensis Cry1F corn in the Americas: lessons and implications for Bt corn IRM in China

The fall armyworm, Spodoptera frugiperda (J.E. Smith), is a major target pest of Bt crops (e.g., corn, cotton, and soybean) in North and South America. This pest has recently invaded Africa and Asia including China and the invasion has placed a great threat to the food security in many countries of these two continents. Due to the extensive use of Bt crops, practical resistance of S. frugiperda to Cry1F corn (TC 1507) with field control problems has widely occurred in Puerto Rico, Brazil, Argentina, and the mainland United States. Analyzing data generated from decade-long studies showed that several factors might have contributed to the wide development of the resistance. These factors include (1) limited modes of action of Bt proteins used in Bt crops; (2) cross-resistance among Cry1 proteins; (3) use of nonhigh dose Bt crop traits; (4) that the resistance is complete on Bt corn plants; (5) abundant in initial Cry1F resistance alleles; and (6) lack of fitness costs/recessive fitness costs of the resistance. The long-term use of Bt crop technology in the Americas suggests that Bt corn can be an effective tool for controlling S. frugiperda in China. IRM programs for Bt corn in China should be as simple as possible to be easily adopted by small-scale growers. The following aspects may be considered in its Bt corn IRM programs: (1) use of only “high dose” traits for both S. frugiperda and stalk borers; (2) developing and implementing a combined resistance monitoring program; (3) use “gene pyramiding” as a primary IRM strategy; and (4) if possible, Bt corn may not be planted in the areas where S. frugiperda overwinters. Lessons and experience gained from the global long-term use of Bt crops should have values in improving IRM programs in the Americas, as well as for a sustainable use of Bt corn technology in China.     

基于微生物源的抗虫基因发掘

植物转基因抗虫技术在害虫控制方面取得了巨大成功。商业化运用的抗虫基因目前全部来源于苏云金杆菌（Bacillus thuringiensis,Bt）的杀虫晶体蛋白基因,存在抗虫谱较窄及害虫逐渐产生抗性等问题,表明新型抗虫基因的筛选尤为重要。已有的文献研究表明,除了继续发掘Bt来源的新型杀虫蛋白基因以外,非Bt杀虫细菌及杀虫真菌也具有重要的发掘价值。     

Delaying corn rootworm resistance to Bt corn

Transgenic crops producing Bacillus thuringiensis (Bt) toxins for insect control have been successful, but their efficacy is reduced when pests evolve resistance. To delay pest resistance to Bt crops, the U.S. Environmental Protection Agency (EPA) has required refuges of host plants that do not produce Bt toxins to promote survival of susceptible pests. Such refuges are expected to be most effective if the Bt plants deliver a dose of toxin high enough to kill nearly all hybrid progeny produced by matings between resistant and susceptible pests. In 2003, the EPA first registered corn, Zea mays L., producing a Bt toxin (Cry3Bb1) that kills western corn rootworm, Diabrotica virgifera virgifera LeConte, one of the most economically important crop pests in the United States. The EPA requires minimum refuges of 20% for Cry3Bb1 corn and 5% for corn producing two Bt toxins active against corn rootworms. We conclude that the current refuge requirements are not adequate, because Bt corn hybrids active against corn rootworms do not meet the high-dose standard, and western corn rootworm has rapidly evolved resistance to Cry3Bb1 corn in the laboratory, greenhouse, and field. Accordingly, we recommend increasing the minimum refuge for Bt corn targeting corn rootworms to 50% for plants producing one toxin active against these pests and to 20% for plants producing two toxins active against these pests. Increasing the minimum refuge percentage can help to delay pest resistance, encourage integrated pest management, and promote more sustainable crop protection.     

Beyond the spore--past and future developments of Bacillus thuringiensis as a biopesticide

Formulated and sporulated cultures of Bacillus thuringiensis (Bt) are widely used as foliar sprays as part of integrated pest management strategies against insect pests of agricultural crops. Although in several cases the presence of the spore has been shown to improve the activity of the product, other Bt-based insecticides have been developed in which the spore is absent. The most notable of these are transgenic plants expressing just the insect toxin gene from the bacterium. This paper will discuss these developments, and the advantages and disadvantages of having the spore present.