Binding of Bacillus thuringiensis Cry1A toxins to brush border membrane vesicles of midgut from Cry1Ac susceptible and resistant Plutella xylostella

Masahiro Higuchi, Kohsuke Haginoya, Takanori Yamazaki, Kazuhisa Miyamoto, Takahiro Katagiri, Kazuya Tomimoto, Yasuyuki Shitomi, Tohru Hayakawa, Ryoichi Sato, Hidetaka Hori

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Plutella xylostella strain resistant (PXR) to Bacillus thuringiensis Cry1Ac toxin was not killed at even more than 1000 μg Cry1Ac/g diet but killed by Cry1Ab at 0.5 μg/g diet. In contrast, susceptible strain (PXS) was killed by Cry1Ac at 1 μg/g diet. Cy3-labeld Cry1A(s) binding to brush border membrane vesicles (BBMV) prepared from both strains were analyzed with direct binding assay. The Kd value of Cry1Aa to both BBMV was almost identical: 213.2 and 205.8 nM, and 263.5 and 265.0 nM for Cry1Ac. The highest Kd values were in Cry1Ab which showed most effective insecticidal activity in PXS and PXR, 2126 and 2463 nM, respectively. These results clearly showed that the BBMV from PXR and PXS could equally bind to Cry1Ac. The binding between BBMV and Cy3-labeled Cry1Ac was inhibited only by anti-175 kDa cadherin-like protein (CadLP) and -252 kDa protein antisera, but not by anti-120 kDa aminopeptidase. This supports that resistance in PXR resulted from the abortion of pore formation after the binding of Cry1Ac to the BBMV. And furthermore, the importance of 175K CadLP and P252 proteins in those bindings was suggested. We briefly discuss possible mechanisms of the resistance.

Original languageEnglish
Pages (from-to)716-724
Number of pages9
JournalComparative Biochemistry and Physiology - B Biochemistry and Molecular Biology
Volume147
Issue number4
DOIs
Publication statusPublished - Aug 2007
Externally publishedYes

Keywords

  • BBMV
  • Bacillus thuringiensis
  • Cry1Ac toxin
  • Mechanism of Cry toxin resistance
  • Midgut brush border membrane vesicle
  • Plutella xylostella

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Molecular Biology

Fingerprint Dive into the research topics of 'Binding of Bacillus thuringiensis Cry1A toxins to brush border membrane vesicles of midgut from Cry1Ac susceptible and resistant Plutella xylostella'. Together they form a unique fingerprint.

  • Cite this