Abstract
In order to isolate novel delta-endotoxins fromBacillus thuringiensis Berliner, a total of 426 native isolates (in varying numbers for each pest) were screened against four major maize pests: corn earworm,Helicoverpa zea; fall armyworm,Spodoptera frugiperda; southwestern corn borer,Diatraea graridiosella, and sugarcane borer,Diatraea saccharalis. Spore-crystal complexes from the isolates were integrated into semi-artificial diets of each pest and mortality was assessed 7 days after treatment. A total of 25 isolates were selected on the basis of highest larval mortality against at least one insect species. There was no correspondence of the most toxic isolates when tested against the four different insect species. Most of the 25 selected isolates caused higher toxicities against all four pests than the standard strain HD-1, regardless of not achieving 100% mortality in any bioassay.H. zea demonstrated the highest level of mortality (96%) and was susceptible to the largest number of isolates (98). None of the other insect species were found susceptible at levels greater than 60%. All the selected active strains were isolated from stored grain dusts (except for LBIT-167), and had bipyramidal crystals with Cry I-like proteins. Most isolates also formed an associated square (cubic) inclusion, with Cry Il-like proteins according to SDS-PAGE analysis of their parasporal bodies. The most active isolates will be subjected to further studies, in order to identify putative novel genes to be expressed in transgenic maize
Résumé
Les 426 souches deBacillus thuringiensis Berliner isolées ont été testées sur quatre ravageurs importants du maïs,Helicoverpa zea, Spodoptera frugiperda, Diatraea grandiosella, Diatraea saccharalis dans le but d’identifier de nouvelles delta endotoxines. Le complexe spore-cristal obtenu à partir de chacun des isolats a été incorporé dans un milieu nutritif artificiel pour insecte et la mortalité a été relevée après sept jours de traitements. Vingt-cinq isolats ont été sélectionnés sur la base d’une toxicité élevée sur les larves d’au moins une espèce. Aucun des isolats actifs ne s’est révélé actif à la fois contre les quatre espèces d’insectes étudiées. La plupart des 25 isolats sélectionnés ont montré une toxicité supérieure à celle de la souche de reférence HD- 1, que la mortalité ait atteint ou non 100 %.H. zea s’est révélée être l’espèce la plus sensible (96 % de mortalité) au plus grand nombre d’isolats (98). Aucune des trois autre espèces n’a montré plus de 60 % mortalité. Toutes les souches actives ont été isolées à partir de poussières de graines stockées (excepté l’isolat LBIT-167) et produisaient des cristaux bipyramidaux. L’analyse du contenu protéique des cristaux en gel de polyacrylamide-SDS a montré que les cristaux bipyramidaux contenaient des protéines de type Cryl et que les cristaux cubiques associés produits par la plupart des isolats contenaient des protéines de type Cryll. Les souches les plus actives seront analysées de façon plus approfondie afin de détecter la présence potentielle de nouveaux gènes pouvant être exprimés chez les maïs transgéniques
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Bohorova, N., Maciel, A.M., Brito, R.M. et al. Selection and characterization of mexican strains ofBacillus thuringiensis active against four major lepidopteran maize pests. Entomophaga 41, 153–165 (1996). https://doi.org/10.1007/BF02764243
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DOI: https://doi.org/10.1007/BF02764243