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Open AccessArticle

Reduced Membrane-Bound Alkaline Phosphatase Does Not Affect Binding of Vip3Aa in a Heliothis virescens Resistant Colony

1
Department of Genetics, Instituto de Biotecnología y Biomedicina (BIOTECMED), Universitat de València, 46100 Burjassot, Spain
2
Department of Entomology and Plant Pathology, University of Tennessee, Knoxville, TN 37996, USA
3
Department of Life Sciences, Imperial College London, Silwood Park Campus, Ascot, Berks SL5 7PY, UK
*
Author to whom correspondence should be addressed.
Current address: Centre de Biotechnologie de Sfax, Sfax, Tunisia.
Toxins 2020, 12(6), 409; https://doi.org/10.3390/toxins12060409
Received: 7 May 2020 / Revised: 29 May 2020 / Accepted: 17 June 2020 / Published: 19 June 2020
The Vip3Aa insecticidal protein from Bacillus thuringiensis (Bt) is produced by specific transgenic corn and cotton varieties for efficient control of target lepidopteran pests. The main threat to this technology is the evolution of resistance in targeted insect pests and understanding the mechanistic basis of resistance is crucial to deploy the most appropriate strategies for resistance management. In this work, we tested whether alteration of membrane receptors in the insect midgut might explain the >2000-fold Vip3Aa resistance phenotype in a laboratory-selected colony of Heliothis virescens (Vip-Sel). Binding of 125I-labeled Vip3Aa to brush border membrane vesicles (BBMV) from 3rd instar larvae from Vip-Sel was not significantly different from binding in the reference susceptible colony. Interestingly, BBMV from Vip-Sel larvae showed dramatically reduced levels of membrane-bound alkaline phosphatase (mALP) activity, which was further confirmed by a strong downregulation of the membrane-bound alkaline phosphatase 1 (HvmALP1) gene. However, the involvement of HvmALP1 as a receptor for the Vip3Aa protein was not supported by results from ligand blotting and viability assays with insect cells expressing HvmALP1. View Full-Text
Keywords: Bacillus thuringiensis; insecticidal proteins; insect resistance; tobacco budworm Bacillus thuringiensis; insecticidal proteins; insect resistance; tobacco budworm
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Pinos, D.; Chakroun, M.; Millán-Leiva, A.; Jurat-Fuentes, J.L.; Wright, D.J.; Hernández-Martínez, P.; Ferré, J. Reduced Membrane-Bound Alkaline Phosphatase Does Not Affect Binding of Vip3Aa in a Heliothis virescens Resistant Colony. Toxins 2020, 12, 409.

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