Presence and significance of Bacillus thuringiensis Cry proteins associated with the Andean weevil Premnotrypes vorax (Coleoptera: Curculionidae)

Presence and significance of Bacillus thuringiensis Cry proteins associated with the Andean weevil Premnotrypes vorax (Coleoptera: Curculionidae)

The Andean weevil Premnotrypes vorax represents an important cause of damage to Colombian potato crops. Due to the impact of this plague on the economy of the country, we searched for new alternatives for its biological control, based on the entomopathogenic bacteria Bacillus thuringiensis. A total...

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Autores principales: López-Pazos, Silvio Alejandro, Wilson Martínez, John, Ximena Castillo, Adriana, Cerón Salamanca, Jairo Alonso
Formato: Online
Idioma:eng
Publicado: Universidad de Costa Rica 2009
Acceso en línea:https://revistas.ucr.ac.cr/index.php/rbt/article/view/5460
id RBT5460
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spelling RBT54602022-06-06T18:20:04Z Presence and significance of Bacillus thuringiensis Cry proteins associated with the Andean weevil Premnotrypes vorax (Coleoptera: Curculionidae) Presence and significance of Bacillus thuringiensis Cry proteins associated with the Andean weevil Premnotrypes vorax (Coleoptera: Curculionidae) López-Pazos, Silvio Alejandro Wilson Martínez, John Ximena Castillo, Adriana Cerón Salamanca, Jairo Alonso cepa nativa gorgojo andino proteína Cry3Aa cultivo de papa mutagénesis sitio-dirigida native strain Andean weevil Cry3Aa protein potato crop site-directed mutagenesis The Andean weevil Premnotrypes vorax represents an important cause of damage to Colombian potato crops. Due to the impact of this plague on the economy of the country, we searched for new alternatives for its biological control, based on the entomopathogenic bacteria Bacillus thuringiensis. A total of 300 B. thuringiensis strains obtained from potato plantations infested with P. vorax were analyzed through crystal morphology, SDS-PAGE, PCR and bioassays. We used site-directed mutagenesis to modify the Cry3Aa protein. Most of the B. thuringiensis isolates had a bipyramidal crystal morphology. SDS-PAGE analyses had seven strains groups with ?-endotoxins from 35 to 135 kDa. The genes cry 2 and cry 1 were significantly more frequent in the P. vorax habitat (PCR analyses). Three mutant toxins, 1 (D354E), 2 (R345A, ?Y350, ?Y351), and 3 (Q482A, S484A, R485A), were analyzed to assess their activity against P. vorax larvae. Toxicity was low, or absent, against P. vorax for isolates, wild type cry 3Aa and cry 3Aa mutants. The genetic characterization of the collection provides opportunities for the selection of strains to be tested in bioassays against other insect pests of agricultural importance, and for designing Cry proteins with improved insecticidal toxicity. The Andean weevil Premnotrypes vorax represents an important cause of damage to Colombian potato crops. Due to the impact of this plague on the economy of the country, we searched for new alternatives for its biological control, based on the entomopathogenic bacteria Bacillus thuringiensis. A total of 300 B. thuringiensis strains obtained from potato plantations infested with P. vorax were analyzed through crystal morphology, SDS-PAGE, PCR and bioassays. We used site-directed mutagenesis to modify the Cry3Aa protein. Most of the B. thuringiensis isolates had a bipyramidal crystal morphology. SDS-PAGE analyses had seven strains groups with ?-endotoxins from 35 to 135 kDa. The genes cry 2 and cry 1 were significantly more frequent in the P. vorax habitat (PCR analyses). Three mutant toxins, 1 (D354E), 2 (R345A, ?Y350, ?Y351), and 3 (Q482A, S484A, R485A), were analyzed to assess their activity against P. vorax larvae. Toxicity was low, or absent, against P. vorax for isolates, wild type cry 3Aa and cry 3Aa mutants. The genetic characterization of the collection provides opportunities for the selection of strains to be tested in bioassays against other insect pests of agricultural importance, and for designing Cry proteins with improved insecticidal toxicity. Universidad de Costa Rica 2009-12-01 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Article Text application/pdf https://revistas.ucr.ac.cr/index.php/rbt/article/view/5460 10.15517/rbt.v57i4.5460 Revista de Biología Tropical; Vol. 57 No. 4 (2009): Volume 57 – Regular number 4 – December 2009; 1235–1243 Revista de Biología Tropical; Vol. 57 Núm. 4 (2009): Volumen 57 – Número regular 4 – Diciembre 2009; 1235–1243 Revista Biología Tropical; Vol. 57 N.º 4 (2009): Volume 57 – Regular number 4 – December 2009; 1235–1243 2215-2075 0034-7744 10.15517/rbt.v57i4 eng https://revistas.ucr.ac.cr/index.php/rbt/article/view/5460/5206 Copyright (c) 2009 Revista de Biología Tropical http://creativecommons.org/licenses/by/4.0
institution Universidad de Costa Rica
collection Revista de Biología Tropical
language eng
format Online
author López-Pazos, Silvio Alejandro
Wilson Martínez, John
Ximena Castillo, Adriana
Cerón Salamanca, Jairo Alonso
spellingShingle López-Pazos, Silvio Alejandro
Wilson Martínez, John
Ximena Castillo, Adriana
Cerón Salamanca, Jairo Alonso
Presence and significance of Bacillus thuringiensis Cry proteins associated with the Andean weevil Premnotrypes vorax (Coleoptera: Curculionidae)
author_facet López-Pazos, Silvio Alejandro
Wilson Martínez, John
Ximena Castillo, Adriana
Cerón Salamanca, Jairo Alonso
author_sort López-Pazos, Silvio Alejandro
description The Andean weevil Premnotrypes vorax represents an important cause of damage to Colombian potato crops. Due to the impact of this plague on the economy of the country, we searched for new alternatives for its biological control, based on the entomopathogenic bacteria Bacillus thuringiensis. A total of 300 B. thuringiensis strains obtained from potato plantations infested with P. vorax were analyzed through crystal morphology, SDS-PAGE, PCR and bioassays. We used site-directed mutagenesis to modify the Cry3Aa protein. Most of the B. thuringiensis isolates had a bipyramidal crystal morphology. SDS-PAGE analyses had seven strains groups with ?-endotoxins from 35 to 135 kDa. The genes cry 2 and cry 1 were significantly more frequent in the P. vorax habitat (PCR analyses). Three mutant toxins, 1 (D354E), 2 (R345A, ?Y350, ?Y351), and 3 (Q482A, S484A, R485A), were analyzed to assess their activity against P. vorax larvae. Toxicity was low, or absent, against P. vorax for isolates, wild type cry 3Aa and cry 3Aa mutants. The genetic characterization of the collection provides opportunities for the selection of strains to be tested in bioassays against other insect pests of agricultural importance, and for designing Cry proteins with improved insecticidal toxicity.
title Presence and significance of Bacillus thuringiensis Cry proteins associated with the Andean weevil Premnotrypes vorax (Coleoptera: Curculionidae)
title_short Presence and significance of Bacillus thuringiensis Cry proteins associated with the Andean weevil Premnotrypes vorax (Coleoptera: Curculionidae)
title_full Presence and significance of Bacillus thuringiensis Cry proteins associated with the Andean weevil Premnotrypes vorax (Coleoptera: Curculionidae)
title_fullStr Presence and significance of Bacillus thuringiensis Cry proteins associated with the Andean weevil Premnotrypes vorax (Coleoptera: Curculionidae)
title_full_unstemmed Presence and significance of Bacillus thuringiensis Cry proteins associated with the Andean weevil Premnotrypes vorax (Coleoptera: Curculionidae)
title_sort presence and significance of bacillus thuringiensis cry proteins associated with the andean weevil premnotrypes vorax (coleoptera: curculionidae)
title_alt Presence and significance of Bacillus thuringiensis Cry proteins associated with the Andean weevil Premnotrypes vorax (Coleoptera: Curculionidae)
publisher Universidad de Costa Rica
publishDate 2009
url https://revistas.ucr.ac.cr/index.php/rbt/article/view/5460
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