A holistic approach for determining the entomopathogenic potential of Bacillus thuringiensis strains

Masson Luke, Erlandson M., Puzstai Carey M., Brousseau Roland, Juarez-Perez Victor-Manuel, Frutos Roger. 1998. A holistic approach for determining the entomopathogenic potential of Bacillus thuringiensis strains. Applied and Environmental Microbiology, 64 (12) : 4782-4788.

Autre titre : Approche holistique afin de déterminer le potentiel entomopathogène de souches de Bacillus thuringiensis

Résumé : The #cry# gene content of #Bacillus thuringiensis# subsp. #aizawai# HD-133 was analyzed by a combination of highpressure liquid chromatography (HPLC) and exclusive PCR. A total of six cry genes were detected in genomic DNA purified from HD-133, four from the #cry#1 family (#cry#1Aa, #cry#1Ab, #cry#1C, and #cry#1D) as well as a gene each from the #cry#2 (#cry#2B) and the #cry#1l families. To directly determine which genes were expressed and crystallized in the purified parasporal inclusions, solubilized and trypsinized HD-133 crystals were subjected to chromatographic separation by HPLC. Only three proteins, #Cry#1Ab, #Cry#1C, and #Cry#1D, were found, in a 60/37/3 ratio. Dot blot analysis of total mRNA purified from HD-133 showed that both the #cry#2B and #cry#1I genes, but not the #cry#1Aa gene, were transcribed. Cloning and sequencing of the #cry#1Aa gene revealed an inserted DNA sequence within the cry coding sequence, resulting in a disrupted reading frame. Taken together, our results show that combining crystal protein analysis with a genetic approach is a highly complementary and powerful way to assess the potential of #B. thuringiensis# isolates for new insecticidal genes and specificities. Furthermore, based on the number of cryptic genes found in HD-133, the total #cry# gene content of #B. thuringiensis# strains may be higher than previously thought.

Mots-clés Agrovoc : Bacillus thuringiensis, pouvoir pathogène, bactérie entomopathogène, biotechnologie, génie génétique, résistance aux organismes nuisibles

Classification Agris : H10 - Ravageurs des plantes

Auteurs et affiliations

• Masson Luke
• Erlandson M.
• Puzstai Carey M.
• Brousseau Roland
• Juarez-Perez Victor-Manuel
• Frutos Roger, CIRAD-AMIS-PROTECTION DES CULTURES (FRA)

Source : Cirad - Agritrop (https://agritrop.cirad.fr/401138/)

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