Mode of action of Bacillus thuringiensis PS86Q3 strain in hymenopteran forest pests.
The mode of action of Cry toxins has been described principally in lepidopteran insects as a multistep process. In this work we describe the mode of action of a Cry toxin active in the common pine sawfly Diprion pini (Hymenoptera, Diprionidae), considered a major forest pest in Europe. Strain PS86Q3 contains a long bipyramidal crystal composed of five major proteins. The N-terminal sequence shows that the 155 kDa protein corresponds to Cry5B toxin and the other proteins belong to the Cry5A subgroup. PCR analysis indicates the presence of cry5Ac and cry5Ba genes, suggesting that Cry5A protein should be Cry5Ac. Activation of protoxins with trypsin or with midgut content from D. pini and Cephacia abietis (Hymenoptera, Pamphiliidae) (spruce webspinning sawfly), another important hymenopteran forest pest, produced a single 75 kDa toxin that corresponded to Cry5A by N-terminal sequence and is responsible for the insecticidal activity. Homologous competition experiments with D. pini and C. abietis brush border membrane vesicles (BBMV) showed that the binding interaction of Cry5A is specific. Membrane potential measurements using a fluorescent dye indicate that Cry5A toxin at nM concentration caused immediate permeability changes in the BBMV isolated from both hymenopteran larvae. The initial response and the sustained permeability change are cationic as previously shown for Cry1 toxins. These results indicate that the hymenopteran specific Cry5A toxin exerts toxicity by a similar mechanism as Cry1 toxins.
Garcia-Robles
I
I
Departamento de Genética, Universidad de Valencia, Dr. Moliner, 50, 46100, Valencia, Burjassot, Spain.
Sánchez
J
J
Gruppe
A
A
Martínez-Ramírez
A C
AC
Rausell
C
C
Real
M D
MD
Bravo
A
A
D000818:Animals; D001413:Bacillus thuringiensis / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000378:metabolism*; D001427:Bacterial Toxins; D001710:Biotin; D010450:Endopeptidases / Q000378:metabolism; D004731:Endotoxins / Q000378:metabolism*; D004789:Enzyme Activation; D006460:Hemolysin Proteins; D006927:Hymenoptera / Q000382:microbiology*; D007814:Larva
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D001710:Biotin; D010450:Endopeptidases