Cloning, ligand-binding, and temporal expression of ecdysteroid receptors in the diamondback moth, Plutella xylostella

Baozhen Tang, Wei Dong, Pei Liang, Xuguo Zhou, Xiwu Gao

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

Background: The diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), is a devastating pest of cruciferous crops worldwide, and has developed resistance to a wide range of insecticides, including diacylhydrazine-based ecdysone agonists, a highly selective group of molt-accelerating biopesticides targeting the ecdysone receptors.Result: In this study, we cloned and characterized the ecdysone receptors from P. xylostella, including the two isoforms of EcR and a USP. Sequence comparison and phylogenetic analysis showed striking conservations among insect ecdysone receptors, especially between P. xylostella and other lepidopterans. The binding affinity of ecdysteroids to in vitro-translated receptor proteins indicated that PxEcRB isoform bound specifically to ponasterone A, and the binding affinity was enhanced by co-incubation with PxUSP (Kd =3.0±1.7 nM). In contrast, PxEcRA did not bind to ponasterone A, even in the presence of PxUSP. The expression of PxEcRB were consistently higher than that of PxEcRA across each and every developmental stage, while the pattern of PxUSP expression is more or less ubiquitous.Conclusions: Target site insensitivity, in which the altered binding of insecticides (ecdysone agonists) to their targets (ecdysone receptors) leads to an adaptive response (resistance), is one of the underlying mechanisms of diacylhydrazine resistance. Given the distinct differences at expression level and the ligand-binding capacity, we hypothesis that PxEcRB is the ecdysone receptor that controls the remodeling events during metamorphosis. More importantly, PxEcRB is the potential target site which is modified in the ecdysone agonist-resistant P. xylostella.

Original languageEnglish
Article number32
JournalBMC Molecular Biology
Volume13
DOIs
StatePublished - Oct 19 2012

Bibliographical note

Funding Information:
This research is supported by grants from the National Natural Science Foundation of China (Nos. 30971941 and 31171873), the Special Fund for Agroscientific Research in the Public Interest (No. 201103021), the National 863 plans project (No. 2012AA101502). Special thanks go to John J. Obrycki (University of Kentucky) for his comments on an earlier draft.

Keywords

  • Binding affinity, Expression profiling, Ecdysone agonist
  • Plutella xylostella, Ecdysone receptor (EcR)

ASJC Scopus subject areas

  • Molecular Biology

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