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The insecticidal activity of recombinant garlic lectins towards aphids

Research output: Contribution to journalArticle

Published copy (DOI)

Author(s)

  • Elaine Fitches
  • Duncan Wiles
  • Angela E. Douglas
  • Gareth Hinchliffe
  • Neil Audsley
  • John A. Gatehouse

Department/unit(s)

Publication details

JournalInsect Biochemistry and Molecular Biology
DatePublished - Oct 2008
Issue number10
Volume38
Number of pages11
Pages (from-to)905-915
Original languageEnglish

Abstract

The heterodimeric and homodimeric garlic lectins ASAI and ASAII were produced as recombinant proteins in the yeast Pichia pastoris. The proteins were purified as functional dimeric lectins, but underwent post-translational proteolysis. Recombinant ASAII was a single homogenous polypeptide which had undergone C-terminal processing similar to that occurring in planta. The recombinant ASAI was glycosylated and subject to variable and heterogenous proteolysis. Both lectins showed insecticidal effects when fed to pea aphids (Acyrthosiphon pisum) in artificial diet, ASAII being more toxic than ASAI at the same concentration. Acute toxicity (mortality at <= 48 h exposure; similar timescale to starvation) was only apparent at the highest lectin concentrations tested (2.0 mg ml(-1)), but dose-dependent chronic toxicity (mortality at >3 d exposure) was observed over the concentration range 0.125-2.0 mg ml(-1). The recombinant lectins caused mortality in both symbiotic and antibiotic-treated aphids, showing that toxicity is not dependent on the presence of the bacterial symbiont (Buchnera aphidicola), or on interaction with symbiont proteins, such as the previously identified lectin "receptor" symbionin. A pulldown assay coupled with peptide mass fingerprinting identified two abundant membrane-associated aphid gut proteins, alanyl aminopeptidase N and sucrase, as "receptors" for lectin binding. (C) 2008 Elsevier Ltd. All rights reserved.

    Research areas

  • Pichia pastoris, Lectin receptors, Symbiosis, Proteolytic processing, Insect midgut, GALANTHUS-NIVALIS AGGLUTININ, MANNOSE-BINDING LECTINS, RICE BROWN PLANTHOPPER, SATIVUM LEAF AGGLUTININ, LARGE-SCALE PRODUCTION, YEAST PICHIA-PASTORIS, SNOWDROP LECTIN, ACYRTHOSIPHON-PISUM, ALLIUM-SATIVUM, PEA APHID

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