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. 1997 Mar;63(3):1095–1101. doi: 10.1128/aem.63.3.1095-1101.1997

Influence of Exposure to Single versus Multiple Toxins of Bacillus thuringiensis subsp. israelensis on Development of Resistance in the Mosquito Culex quinquefasciatus (Diptera: Culicidae)

G P Georghiou, M C Wirth
PMCID: PMC1389136  PMID: 16535542

Abstract

The impending widespread use of transgenic crop plants encoding a single insecticidal toxin protein of Bacillus thuringiensis has focused attention on the perceived risk of rapid selection of resistance in target insects. We have used Bacillus thuringiensis subsp. israelensis toxins as a model system and determined the speed and magnitude of evolution of resistance in colonies of the mosquito Culex quinquefasciatus during selection for 28 consecutive generations with single or multiple toxins. The parental strain was synthesized by combining approximately 500 larvae from each of 19 field collections obtained from the states of California, Oregon, Louisiana, and Tennessee. At least 10,000 larvae were selected in each generation of each line at an average mortality level of 84%. The susceptibilities of the parental and selected lines were compared in parallel tests in every third generation by using fresh suspensions of toxin powders. The normal toxin complement of B. thuringiensis subsp. israelensis consists of four toxins, CryIVA, CryIVB, CryIVD, and CytA. Resistance became evident first in the line that was selected with a single toxin (CryIVD), attaining the highest level (resistance ratio [RR], >913 at 95% lethal concentration) by generation F(inf28) when the study was completed. Resistance evolved more slowly and to a lower level (RR, >122 by F(inf25)) in the line selected with two toxins (CryIVA+CryIVB) and lower still (RR, 91 by F(inf28)) in the line selected with three toxins (CryIVA+CryIVB+ CryIVD). Resistance was remarkably low (RR, 3.2) in the line selected with all four toxins. The results reveal the importance of the full complement of toxins found in natural populations of B. thuringiensis subsp. israelensis as an effective approach to resistance management.

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Selected References

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  1. Alstad D. N., Andow D. A. Managing the evolution of insect resistance to transgenic plants. Science. 1995 Jun 30;268(5219):1894–1896. doi: 10.1126/science.268.5219.1894. [DOI] [PubMed] [Google Scholar]
  2. Becker N., Ludwig M. Investigations on possible resistance in Aedes vexans field populations after a 10-year application of Bacillus thuringiensis israelensis. J Am Mosq Control Assoc. 1993 Jun;9(2):221–224. [PubMed] [Google Scholar]
  3. Chang C., Dai S. M., Frutos R., Federici B. A., Gill S. S. Properties of a 72-kilodalton mosquitocidal protein from Bacillus thuringiensis subsp. morrisoni PG-14 expressed in B. thuringiensis subsp. kurstaki by using the shuttle vector pHT3101. Appl Environ Microbiol. 1992 Feb;58(2):507–512. doi: 10.1128/aem.58.2.507-512.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chilcott C. N., Ellar D. J. Comparative toxicity of Bacillus thuringiensis var. israelensis crystal proteins in vivo and in vitro. J Gen Microbiol. 1988 Sep;134(9):2551–2558. doi: 10.1099/00221287-134-9-2551. [DOI] [PubMed] [Google Scholar]
  5. Delécluse A., Charles J. F., Klier A., Rapoport G. Deletion by in vivo recombination shows that the 28-kilodalton cytolytic polypeptide from Bacillus thuringiensis subsp. israelensis is not essential for mosquitocidal activity. J Bacteriol. 1991 Jun;173(11):3374–3381. doi: 10.1128/jb.173.11.3374-3381.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Delécluse A., Poncet S., Klier A., Rapoport G. Expression of cryIVA and cryIVB Genes, Independently or in Combination, in a Crystal-Negative Strain of Bacillus thuringiensis subsp. israelensis. Appl Environ Microbiol. 1993 Nov;59(11):3922–3927. doi: 10.1128/aem.59.11.3922-3927.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Denolf P., Jansens S., Peferoen M., Degheele D., Van Rie J. Two Different Bacillus thuringiensis Delta-Endotoxin Receptors in the Midgut Brush Border Membrane of the European Corn Borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Pyralidae). Appl Environ Microbiol. 1993 Jun;59(6):1828–1837. doi: 10.1128/aem.59.6.1828-1837.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gasser C. S., Fraley R. T. Genetically engineering plants for crop improvement. Science. 1989 Jun 16;244(4910):1293–1299. doi: 10.1126/science.244.4910.1293. [DOI] [PubMed] [Google Scholar]
  9. Goldman I. F., Arnold J., Carlton B. C. Selection for resistance to Bacillus thuringiensis subspecies israelensis in field and laboratory populations of the mosquito Aedes aegypti. J Invertebr Pathol. 1986 May;47(3):317–324. doi: 10.1016/0022-2011(86)90102-3. [DOI] [PubMed] [Google Scholar]
  10. Gould F., Martinez-Ramirez A., Anderson A., Ferre J., Silva F. J., Moar W. J. Broad-spectrum resistance to Bacillus thuringiensis toxins in Heliothis virescens. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):7986–7990. doi: 10.1073/pnas.89.17.7986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hama H., Sakurai T., Kasuya Y., Fujiki M., Masaki T., Goto K. Action of endothelin-1 on rat astrocytes through the ETB receptor. Biochem Biophys Res Commun. 1992 Jul 15;186(1):355–362. doi: 10.1016/s0006-291x(05)80815-0. [DOI] [PubMed] [Google Scholar]
  12. Höfte H., Whiteley H. R. Insecticidal crystal proteins of Bacillus thuringiensis. Microbiol Rev. 1989 Jun;53(2):242–255. doi: 10.1128/mr.53.2.242-255.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ibarra J. E., Federici B. A. Isolation of a relatively nontoxic 65-kilodalton protein inclusion from the parasporal body of Bacillus thuringiensis subsp. israelensis. J Bacteriol. 1986 Feb;165(2):527–533. doi: 10.1128/jb.165.2.527-533.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lecadet M. M., Blondel M. O., Ribier J. Generalized transduction in Bacillus thuringiensis var. berliner 1715 using bacteriophage CP-54Ber. J Gen Microbiol. 1980 Nov;121(1):203–212. doi: 10.1099/00221287-121-1-203. [DOI] [PubMed] [Google Scholar]
  15. Mani G. S. Evolution of resistance in the presence of two insecticides. Genetics. 1985 Apr;109(4):761–783. doi: 10.1093/genetics/109.4.761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. McGaughey W. H. Insect Resistance to the Biological Insecticide Bacillus thuringiensis. Science. 1985 Jul 12;229(4709):193–195. doi: 10.1126/science.229.4709.193. [DOI] [PubMed] [Google Scholar]
  17. Moar W. J., Pusztai-Carey M., Van Faassen H., Bosch D., Frutos R., Rang C., Luo K., Adang M. J. Development of Bacillus thuringiensis CryIC Resistance by Spodoptera exigua (Hubner) (Lepidoptera: Noctuidae). Appl Environ Microbiol. 1995 Jun;61(6):2086–2092. doi: 10.1128/aem.61.6.2086-2092.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Murphy R. C., Stevens S. E., Jr Cloning and expression of the cryIVD gene of Bacillus thuringiensis subsp. israelensis in the cyanobacterium Agmenellum quadruplicatum PR-6 and its resulting larvicidal activity. Appl Environ Microbiol. 1992 May;58(5):1650–1655. doi: 10.1128/aem.58.5.1650-1655.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rao D. R., Mani T. R., Rajendran R., Joseph A. S., Gajanana A., Reuben R. Development of a high level of resistance to Bacillus sphaericus in a field population of Culex quinquefasciatus from Kochi, India. J Am Mosq Control Assoc. 1995 Mar;11(1):1–5. [PubMed] [Google Scholar]
  20. Tabashnik B. E. Managing resistance with multiple pesticide tactics: theory, evidence, and recommendations. J Econ Entomol. 1989 Oct;82(5):1263–1269. doi: 10.1093/jee/82.5.1263. [DOI] [PubMed] [Google Scholar]
  21. Wu D., Johnson J. J., Federici B. A. Synergism of mosquitocidal toxicity between CytA and CryIVD proteins using inclusions produced from cloned genes of Bacillus thuringiensis. Mol Microbiol. 1994 Sep;13(6):965–972. doi: 10.1111/j.1365-2958.1994.tb00488.x. [DOI] [PubMed] [Google Scholar]

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