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Identification of two Bacillus thuringiensis Cry3Aa toxin-binding aminopeptidase N from Rhynchophorus ferrugineus (Coleoptera: Curculionidae)

Published online by Cambridge University Press:  19 July 2023

Shaozhen Wang ,
Yajie Guo ,
Yunzhu Sun ,
Mingqing Weng ,
Qiliao Liao ,
Ru Qiu ,
Shuangquan Zou  and
Songqing Wu Open the ORCID record for Songqing Wu [Opens in a new window]
Shaozhen Wang
Affiliation:
College of Forestry, Fujian Agriculture and Forestry University, Fuzhou 350000, China CAS Key Laboratory of Insect Developmental and Evolutionary Biology, CAS Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai 200032, China
Yajie Guo
Affiliation:
College of Forestry, Fujian Agriculture and Forestry University, Fuzhou 350000, China Key Laboratory of Integrated Pest Management in Ecological Forests, Fujian Province University, Fujian Agriculture and Forestry University, Fuzhou 350000, China Asian Research Center for Bioresource and Environmental Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 188-0002, Japan
Yunzhu Sun
Affiliation:
College of Forestry, Fujian Agriculture and Forestry University, Fuzhou 350000, China Key Laboratory of Integrated Pest Management in Ecological Forests, Fujian Province University, Fujian Agriculture and Forestry University, Fuzhou 350000, China
Mingqing Weng
Affiliation:
College of Forestry, Fujian Agriculture and Forestry University, Fuzhou 350000, China Key Laboratory of Integrated Pest Management in Ecological Forests, Fujian Province University, Fujian Agriculture and Forestry University, Fuzhou 350000, China
Qiliao Liao
Affiliation:
College of Forestry, Fujian Agriculture and Forestry University, Fuzhou 350000, China
Ru Qiu
Affiliation:
College of Forestry, Fujian Agriculture and Forestry University, Fuzhou 350000, China
Shuangquan Zou
Affiliation:
College of Forestry, Fujian Agriculture and Forestry University, Fuzhou 350000, China
Songqing Wu*
Affiliation:
College of Forestry, Fujian Agriculture and Forestry University, Fuzhou 350000, China Key Laboratory of Integrated Pest Management in Ecological Forests, Fujian Province University, Fujian Agriculture and Forestry University, Fuzhou 350000, China State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, College of Forestry, Fujian Agriculture and Forestry University, Fuzhou, China, 350002
*
Corresponding author: Songqing Wu; Email: dabinyang@126.com
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Abstract

Rhynchophorus ferrugineus is a quarantine pest that mainly damages plants in tropical regions, which are essential economic resources. Cry3Aa has been used to control coleopteran pests and is known to be toxic to R. ferrugineus. The binding of the Cry toxin to specific receptors on the target insect plays a crucial role in the toxicological mechanism of Cry toxins. However, in the case of R. ferrugineus, the nature and identity of the receptor proteins involved remain unknown. In the present study, pull-down assays and mass spectrometry were used to identify two proteins of aminopeptidase N proteins (RfAPN2a and RfAPN2b) in the larval midguts of R. ferrugineus. Cry3Aa was able to bind to RfAPN2a (Kd = 108.5 nM) and RfAPN2b (Kd = 68.2 nM), as well as midgut brush border membrane vesicles (Kd = 482.5 nM). In silico analysis of both RfAPN proteins included the signal peptide and anchored sites for glycosyl phosphatidyl inositol. In addition, RfAPN2a and RfAPN2b were expressed in the human embryonic kidney 293T cell line, and cytotoxicity assays showed that the transgenic cells were not susceptible to activated Cry3Aa. Our results show that RfAPN2a and RfAPN2b are Cry3Aa-binding proteins involved in the Cry3Aa toxicity of R. ferrugineus. This study deepens our understanding of the action mechanism of Cry3Aa in R. ferrugineus larvae.

Keywords

Aminopeptidase NCry3Aa toxininteraction mechanismRhynchophorus ferrugineus
Type
Research Paper
Information
Bulletin of Entomological Research , Volume 113 , Issue 5 , October 2023 , pp. 615 - 625
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Copyright
Copyright © The Author(s), 2023. Published by Cambridge University Press

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Footnotes

*

These authors contributed equally.

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