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Analyses of Cry1Ab binding in resistant and susceptible strains of the European corn borer, Ostrinia nubilalis (Hubner) (Lepidoptera: Crambidae).
Appl Environ Microbiol. 2006 Aug; 72(8):5318-24.AE

Abstract

Cry1Ab toxin binding analysis was performed to determine whether resistance in laboratory-selected Ostrinia nubilalis strains is associated with target site alteration. Brush border membrane vesicles were prepared using dissected midguts from late instars of susceptible and resistant strains (Europe-R and RSTT) of O. nubilalis. Immunoblot analysis indicated that three different proteins bound to Cry1Ab toxin and were recognized by an anticadherin serum. In a comparison of resistant and susceptible strains, reduced Cry1Ab binding was apparent for all three bands corresponding to cadherin-like proteins in the Europe-R strain, while reduced binding was apparent in only one band for the RSTT strain. Real-time analysis of Cry1Ab binding to gut receptors using surface plasmon resonance suggested slight differences in affinity in both resistant strains. Additional binding analysis was conducted using 125I-labeled Cry1Ab, Cry1Ac, and Cry1Aa. Slight differences were again observed between the resistant and susceptible strains for Cry1Ab binding. However, when binding of 125I-labeled Cry1Aa was tested, a 10-fold reduction in the concentration of binding sites was observed in the Europe-R strain. Expression of the O. nubilalis cadherin gene was similar in both the resistant and susceptible strains and did not account for differences in binding. In combination, the results of the present work suggest that differences in susceptibility to Cry1A toxins in the Europe-R strain of O. nubilalis are associated with altered receptor binding, although the precise nature of this mechanism is still uncertain.

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Authors+Show Affiliations

Siqueira HA
Department of Entomology, University of Nebraska, Lincoln, NE 68583-0816, USA.
González-Cabrera J
No affiliation info available
Ferré J
No affiliation info available
Flannagan R
No affiliation info available
Siegfried BD
No affiliation info available

MeSH

AnimalsBacillus thuringiensisBacillus thuringiensis ToxinsBacterial ProteinsBacterial ToxinsDrug ResistanceEndotoxinsHemolysin ProteinsInsecticidesMothsPest Control, BiologicalProtein BindingZea mays

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

16885282

Citation

Siqueira, Herbert A A., et al. "Analyses of Cry1Ab Binding in Resistant and Susceptible Strains of the European Corn Borer, Ostrinia Nubilalis (Hubner) (Lepidoptera: Crambidae)." Applied and Environmental Microbiology, vol. 72, no. 8, 2006, pp. 5318-24.
Siqueira HA, González-Cabrera J, Ferré J, et al. Analyses of Cry1Ab binding in resistant and susceptible strains of the European corn borer, Ostrinia nubilalis (Hubner) (Lepidoptera: Crambidae). Appl Environ Microbiol. 2006;72(8):5318-24.
Siqueira, H. A., González-Cabrera, J., Ferré, J., Flannagan, R., & Siegfried, B. D. (2006). Analyses of Cry1Ab binding in resistant and susceptible strains of the European corn borer, Ostrinia nubilalis (Hubner) (Lepidoptera: Crambidae). Applied and Environmental Microbiology, 72(8), 5318-24.
Siqueira HA, et al. Analyses of Cry1Ab Binding in Resistant and Susceptible Strains of the European Corn Borer, Ostrinia Nubilalis (Hubner) (Lepidoptera: Crambidae). Appl Environ Microbiol. 2006;72(8):5318-24. PubMed PMID: 16885282.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Analyses of Cry1Ab binding in resistant and susceptible strains of the European corn borer, Ostrinia nubilalis (Hubner) (Lepidoptera: Crambidae). AU - Siqueira,Herbert A A, AU - González-Cabrera,Joel, AU - Ferré,Juan, AU - Flannagan,Ronald, AU - Siegfried,Blair D, PY - 2006/8/4/pubmed PY - 2006/10/6/medline PY - 2006/8/4/entrez SP - 5318 EP - 24 JF - Applied and environmental microbiology JO - Appl Environ Microbiol VL - 72 IS - 8 N2 - Cry1Ab toxin binding analysis was performed to determine whether resistance in laboratory-selected Ostrinia nubilalis strains is associated with target site alteration. Brush border membrane vesicles were prepared using dissected midguts from late instars of susceptible and resistant strains (Europe-R and RSTT) of O. nubilalis. Immunoblot analysis indicated that three different proteins bound to Cry1Ab toxin and were recognized by an anticadherin serum. In a comparison of resistant and susceptible strains, reduced Cry1Ab binding was apparent for all three bands corresponding to cadherin-like proteins in the Europe-R strain, while reduced binding was apparent in only one band for the RSTT strain. Real-time analysis of Cry1Ab binding to gut receptors using surface plasmon resonance suggested slight differences in affinity in both resistant strains. Additional binding analysis was conducted using 125I-labeled Cry1Ab, Cry1Ac, and Cry1Aa. Slight differences were again observed between the resistant and susceptible strains for Cry1Ab binding. However, when binding of 125I-labeled Cry1Aa was tested, a 10-fold reduction in the concentration of binding sites was observed in the Europe-R strain. Expression of the O. nubilalis cadherin gene was similar in both the resistant and susceptible strains and did not account for differences in binding. In combination, the results of the present work suggest that differences in susceptibility to Cry1A toxins in the Europe-R strain of O. nubilalis are associated with altered receptor binding, although the precise nature of this mechanism is still uncertain. SN - 0099-2240 UR - https://www.unboundmedicine.com/medline/citation/16885282/Analyses_of_Cry1Ab_binding_in_resistant_and_susceptible_strains_of_the_European_corn_borer_Ostrinia_nubilalis__Hubner___Lepidoptera:_Crambidae__ DB - PRIME DP - Unbound Medicine ER -
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