The ‘natural refuge strategy” for delaying insect resistance to transgenic cotton that produces insecticidal proteins from Bacillus thuringiensis (Bt) relies on refuges of host plants other than cotton that do not make Bt toxins. We tested this widely adopted strategy by comparing predictions from modeling with data from a four-year field study of cotton bollworm (Helicoverpa armigera) resistance to transgenic cotton producing Bt toxin Cry1Ac in six provinces of northern China. Bioassay data revealed that the percentage of resistant insects increased from 0.93% in 2010 to 5.5% in 2013. Modeling predicted that the percentage of resistant insects would exceed 98% in 2013 without natural refuges, but would increase to only 1.1% if natural refuges were as effective as non-Bt cotton refuges. Therefore, the results imply that natural refuges delayed resistance, but were not as effective as an equivalent area of non-Bt cotton refuges. The percentage of resistant insects with nonrecessive inheritance of resistance increased from 37% in 2010 to 84% in 2013. Switching to Bt cotton producing two or more toxins and integrating other control tactics could slow further increases in resistance.
This is a preview of subscription content
Subscribe to Journal
Get full journal access for 1 year
only $8.25 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Get time limited or full article access on ReadCube.
All prices are NET prices.
James, C. Global status of commercialized biotech/GM Crops: 2013. ISAAA Briefs 46 (ISAAA, Ithaca, NY, 2013).
Bagla, P. Hardy cotton-munching pests are latest blow to GM crops. Science 327, 1439 (2010).
Alyokhin, A. Scant evidence supports EPA's pyramided Bt corn refuge size of 5%. Nat. Biotechnol. 29, 577–578 (2011).
Gilbert, N. A hard look at GM crops. Nature 497, 24–26 (2013).
Sanahuja, G., Banakar, R., Twyman, R., Capell, T. & Christou, P. Bacillus thuringiensis: a century of research, development and commercial applications. Plant Biotechnol. J. 9, 283–300 (2011).
Pardo-López, L., Soberón, M. & Bravo, A. Bacillus thuringiensis insecticidal three-domain Cry toxins: mode of action, insect resistance and consequences for crop protection. FEMS Microbiol. Rev. 37, 3–22 (2013).
Mendelsohn, M., Kough, J., Vaituzis, Z. & Matthews, K. Are Bt crops safe? Nat. Biotechnol. 21, 1003–1009 (2003).
Comas, C., Lumbierres, B., Pons, X. & Albajes, R. No effects of Bacillus thuringiensis maize on nontarget organisms in the field in southern Europe: a meta-analysis of 26 arthropod taxa. Transgenic Res. 23, 135–143 (2014).
Carpenter, J.E. Peer-reviewed surveys indicate positive impact of commercialized GM crops. Nat. Biotechnol. 28, 319–321 (2010).
Hutchison, W.D. et al. Areawide suppression of European corn borer with Bt maize reaps savings to non-Bt maize growers. Science 330, 222–225 (2010).
Tabashnik, B.E. et al. Suppressing resistance to Bt cotton with sterile insect releases. Nat. Biotechnol. 28, 1304–1307 (2010).
Edgerton, M.D. et al. Transgenic insect resistance traits increase corn yield and yield stability. Nat. Biotechnol. 30, 493–496 (2012).
Kathage, J. & Qaim, M. Economic impacts and impact dynamics of Bt (Bacillus thuringiensis) cotton in India. Proc. Natl. Acad. Sci. USA 109, 11652–11656 (2012).
Lu, Y., Wu, K., Jiang, Y., Guo, Y. & Desneux, N. Widespread adoption of Bt cotton and insecticide decrease promotes biocontrol services. Nature 487, 362–365 (2012).
Storer, N.P., Kubiszak, M.E., Ed King, J., Thompson, G.D. & Santos, A.C. Status of resistance to Bt maize in Spodoptera frugiperda: Lessons from Puerto Rico. J. Invertebr. Pathol. 110, 294–300 (2012).
Tabashnik, B.E., Brévault, T. & Carrière, Y. Insect resistance to Bt crops: lessons from the first billion acres. Nat. Biotechnol. 31, 510–521 (2013).
Van den Berg, J., Hilbeck, A. & Bøhn, T. Pest resistance to Cry1Ab Bt maize: Field resistance, contributing factors and lessons from South Africa. Crop Prot. 54, 154–160 (2013).
Gassmann, A.J. et al. Field-evolved resistance by western corn rootworm to multiple Bacillus thuringiensis toxins in transgenic maize. Proc. Natl. Acad. Sci. USA 111, 5141–5146 (2014).
Gould, F. Sustainability of transgenic insecticidal cultivars: integrating pest genetics and ecology. Annu. Rev. Entomol. 43, 701–726 (1998).
Tabashnik, B.E. Delaying insect resistance to transgenic crops. Proc. Natl. Acad. Sci. USA 105, 19029–19030 (2008).
Tabashnik, B.E., Gassmann, A.J., Crowder, D.W. & Carrière, Y. Insect resistance to Bt crops: evidence versus theory. Nat. Biotechnol. 26, 199–202 (2008).
Tabashnik, B.E., Van Rensburg, J.B.J. & Carrière, Y. Field-evolved insect resistance to Bt crops: definition, theory, and data. J. Econ. Entomol. 102, 2011–2025 (2009).
Downes, S. et al. Adaptive management of pest resistance by Helicoverpa species (Noctuidae) in Australia to the Cry2Ab Bt toxin in Bollgard II® cotton. Evol. Appl. 3, 574–584 (2010).
Wu, K., Lu, Y., Feng, H., Jiang, Y. & Zhao, J.Z. Suppression of cotton bollworm in multiple crops in China in areas with Bt toxin-containing cotton. Science 321, 1676–1678 (2008).
Wu, K. & Guo, Y. The evolution of cotton pest management practices in China. Annu. Rev. Entomol. 50, 31–52 (2005).
Wu, K., Guo, Y. & Gao, S. Evaluation of the natural refuge function for Helicoverpa armigera (Lepidoptera: Noctuidae) within Bacillus thuringiensis transgenic cotton growing areas in north China. J. Econ. Entomol. 95, 832–837 (2002).
Huang, J.K., Rozelle, S., Pray, C. & Wang, Q.F. Plant biotechnology in China. Science 295, 674–676 (2002).
Li, G.-P. et al. Increasing tolerance to Cry1Ac cotton from cotton bollworm, Helicoverpa armigera, was confirmed in Bt cotton farming area of China. Ecol. Entomol. 32, 366–375 (2007).
Yang, Y., Chen, H., Wu, Y., Yang, Y. & Wu, S. Mutated cadherin alleles from a field population of Helicoverpa armigera confer resistance to Bacillus thuringiensis toxin Cry1Ac. Appl. Environ. Microbiol. 73, 6939–6944 (2007).
An, J. et al. Vip3Aa tolerance response of Helicoverpa armigera populations from a Cry1Ac cotton planting region. J. Econ. Entomol. 103, 2169–2173 (2010).
Li, G., Feng, H., Gao, Y., Wyckhuys, K.A. & Wu, K. Frequency of Bt resistance alleles in Helicoverpa armigera in the Xinjiang cotton-planting region of China. Environ. Entomol. 39, 1698–1704 (2010).
Liu, F. et al. Evidence of field-evolved resistance to Cry1Ac-expressing Bt cotton in Helicoverpa armigera (Lepidoptera: Noctuidae) in northern China. Pest Manag. Sci. 66, 155–161 (2010).
Zhang, H. et al. Early warning of cotton bollworm resistance associated with intensive planting of Bt cotton in China. PLoS ONE 6, e22874 (2011).
Zhang, H. et al. Diverse genetic basis of field-evolved resistance to Bt cotton in cotton bollworm from China. Proc. Natl. Acad. Sci. USA 109, 10275–10280 (2012).
Jin, L. et al. Dominant resistance to Bt cotton and minor cross-resistance to Bt toxin Cry2Ab in cotton bollworm from China. Evol. Appl. 6, 1222–1235 (2013).
Zhang, H., Tang, M., Yang, F., Yang, Y. & Wu, Y. DNA-based screening for an intracellular cadherin mutation conferring non-recessive Cry1Ac resistance in field populations of Helicoverpa armigera . Pestic. Biochem. Physiol. 107, 148–152 (2013).
Behere, G.T. et al. Mitochondrial DNA analysis of field populations of Helicoverpa armigera (Lepidoptera: Noctuidae) and of its relationship to H. zea . BMC Evol. Biol. 7, 117 (2007).
Zhang, H., Wu, S., Yang, Y., Tabashnik, B.E. & Wu, Y. Non-recessive Bt toxin resistance conferred by an intracellular cadherin mutation in field-selected populations of cotton bollworm. PLoS ONE 7, e53418 (2012).
Ravi, K.C. et al. Relative abundance of Helicoverpa armigera (Lepidoptera: Noctuidae) on different host crops in India and the role of these crops as natural refuge for Bacillus thuringiensis cotton. Environ. Entomol. 34, 59–69 (2005).
Gould, F. et al. Bacillus thuringiensis-toxin resistance management: stable isotope assessment of alternate host use by Helicoverpa zea . Proc. Natl. Acad. Sci. USA 99, 16581–16586 (2002).
Brévault, T., Nibouche, S., Achaleke, J. & Carrière, Y. Assessing the role of non-cotton refuges in delaying Helicoverpa armigera resistance to Bt cotton in West Africa. Evol. Appl. 5, 53–65 (2012).
US Environmental Protection Agency. Pesticides news story: EPA approves natural refuges for insect resistance management in Bollgard II cotton. http://www.epa.gov/oppfead1/cb/csb_page/updates/2007/bollgard-cotton.htm (EPA, 2007).
Wan, P. et al. Increased frequency of pink bollworm resistance to Bt toxin Cry1Ac in China. PLoS ONE 7, e29975 (2012).
Liu, Y. & Tabashnik, B.E. Inheritance of resistance to the Bacillus thuringiensis toxin Cry1C in the diamondback moth. Appl. Environ. Microbiol. 63, 2218–2223 (1997).
Gustafson, D.I., Head, G.P. & Caprio, M.A. Modeling the impact of alternative hosts on Helicoverpa zea adaptation to Bollgard cotton. J. Econ. Entomol. 99, 2116–2124 (2006).
Carrière, Y. & Tabashnik, B.E. Reversing insect adaptation to transgenic insecticidal plants. Proc. R. Sci. 268, 1475–1480 (2001).
Tabashnik, B.E., Dennehy, T.J. & Carrière, Y. Delayed resistance to transgenic cotton in pink bollworm. Proc. Natl. Acad. Sci. USA 102, 15389–15393 (2005).
Tabashnik, B.E. & Croft, B.A. Managing pesticide resistance in crop-arthropod complexes: interactions between biological and operational factors. Environ. Entomol. 11, 1137–1144 (1982).
Tay, W.T. et al. A brave new world for an Old World pest: Helicoverpa armigera (Lepidoptera: Noctuidae) in Brazil. PLoS ONE 8, e80134 (2013).
Mahon, R.J., Downes, S.J. & James, B. Vip3A resistance alleles exist at high levels in Australian targets before release of cotton expressing this toxin. PLoS ONE 7, e39192 (2012).
Tabashnik, B.E., Crowder, D.W. & Carrière, Y. Modeling evolution of insect resistance to genetically modified crops. Nat. Protoc. Network doi:10.1038/nprot.2008.125, http://www.nature.com/protocolexchange/protocols/462 (19 June 2008).
Ge, F., Chen, F., Parajulee, M.N. & Yardim, E.N. Quantification of diapausing fourth generation and suicidal fifth generation cotton bollworm, Helicoverpa armigera, in cotton and corn in northern China. Entomol. Exp. Appl. 116, 1–7 (2005).
Feng, H., Gould, F., Huang, Y., Jiang, Y. & Wu, K. Modeling the population dynamics of cotton bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) over a wide area in northern China. Ecol. Modell. 221, 1819–1830 (2010).
Feng, H., Wu, X., Wu, B. & Wu, K. Seasonal migration of Helicoverpa armigera over the Bohai Sea. J. Econ. Entomol. 102, 95–104 (2009).
Hartl, D.L. & Clark, A.G. Principles of Population Genetics edn. 2 (Sinauer, Sunderland, MA, 1989).
Liu, Y.B. et al. Effects of Bt cotton and CrylAc toxin on survival and development of pink bollworm (Lepidoptera: Gelechiidae). J. Econ. Entomol. 94, 1237–1242 (2001).
Bird, L.J. & Akhurst, R.J. Effects of host plant species on fitness costs of Bt resistance in Helicoverpa armigera (Lepidoptera: Noctuidae). Biol. Control 40, 196–203 (2007).
Liang, G.M. et al. Changes of inheritance mode and fitness in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) along with its resistance evolution to Cry1Ac toxin. J. Invertebr. Pathol. 97, 142–149 (2008).
Cao, G. et al. Quantitative analysis of fitness costs associated with the development of resistance to the Bt toxin Cry1Ac in Helicoverpa armigera . Sci. Rep. 4, 5629 (2014).
Wu, K., Feng, H. & Guo, Y. Evaluation of maize as a refuge for management of resistance to Bt cotton by Helicoverpa armigera (Hübner) in the Yellow River cotton-farming region of China. Crop Prot. 23, 523–530 (2004).
Yang, Y.-H. et al. Introgression of a disrupted cadherin gene enables susceptible Helicoverpa armigera to obtain resistance to Bacillus thuringiensis toxin Cry1Ac. Bull. Entomol. Res. 99, 175–181 (2009).
Wan, P., Zhang, Y.J., Wu, K.M. & Huang, M.S. Seasonal expression profiles of insecticidal protein and control efficacy against Helicoverpa armigera for Bt cotton in the Yangtze River valley of China. J. Econ. Entomol. 98, 195–201 (2005).
Newcombe, R.G. Two-sided confidence intervals for the single proportion: Comparison of seven methods. Stat. Med. 17, 857–872 (1998).
We thank M. Sisterson and three anonymous reviewers for their comments that improved the manuscript. This work was funded by grants from the Ministry of Agriculture of China (2014ZX08012-004), the National Natural Science Foundation of China (31071983 and 31321004), the 111 program of China (B07030) and the US Department of Agriculture Biotechnology Risk Assessment Grants program (2011-33522-30729). We thank M.P. Carey (Case Western Reserve University, USA) for providing activated Cry1Ac toxin.
B.E.T. is coauthor of a patent on modified Bt toxins, “Suppression of Resistance in Insects to Bacillus thuringiensis Cry Toxins, Using Toxins that do not Require the Cadherin Receptor” (patent numbers: CA2690188A1, CN101730712A, EP2184293A2, EP2184293A4, EP2184293B1, WO2008150150A2, WO2008150150A3). Pioneer, Dow AgroSciences, Monsanto and Bayer CropScience did not provide funding to support this work, but may be affected financially by publication of this paper and have funded other work by B.E.T.
About this article
Cite this article
Jin, L., Zhang, H., Lu, Y. et al. Large-scale test of the natural refuge strategy for delaying insect resistance to transgenic Bt crops. Nat Biotechnol 33, 169–174 (2015). https://doi.org/10.1038/nbt.3100
Need for growing non-Bt cotton refugia to overcome Bt resistance problem in targeted larvae of the cotton bollworms, Helicoverpa armigera and Pectinophora gossypiella
Egyptian Journal of Biological Pest Control (2021)
Insect resistance management in Bacillus thuringiensis cotton by MGPS (multiple genes pyramiding and silencing)
Journal of Cotton Research (2020)
On application of the precautionary principle to ban GMVs: an evolutionary model of new seed technology integration
Journal of Evolutionary Economics (2020)
Transgenic Research (2019)