Cry1Ac and Vip3Aa proteins from Bacillus thuringiensis targeting Cry toxin resistance in Diatraea flavipennella and Elasmopalpus lignosellus from sugarcane

The biological potential of Vip and Cry proteins from Bacillus is well known and widely established. Thus, it is important to look for new genes showing different modes of action, selecting those with differentiated entomotoxic activity against Diatraea flavipennella and Elasmopalpus lignosellus, wh...

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Main Authors: Ana Rita Nunes Lemes, Camila Soares Figueiredo, Isis Sebastião, Liliane Marques da Silva, Rebeka da Costa Alves, Herbert Álvaro Abreu de Siqueira, Manoel Victor Franco Lemos, Odair Aparecido Fernandes, Janete Apparecida Desidério
Format: Article
Language:English
Published: PeerJ Inc. 2017-01-01
Series:PeerJ
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Online Access:https://peerj.com/articles/2866.pdf
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author Ana Rita Nunes Lemes
Camila Soares Figueiredo
Isis Sebastião
Liliane Marques da Silva
Rebeka da Costa Alves
Herbert Álvaro Abreu de Siqueira
Manoel Victor Franco Lemos
Odair Aparecido Fernandes
Janete Apparecida Desidério
author_facet Ana Rita Nunes Lemes
Camila Soares Figueiredo
Isis Sebastião
Liliane Marques da Silva
Rebeka da Costa Alves
Herbert Álvaro Abreu de Siqueira
Manoel Victor Franco Lemos
Odair Aparecido Fernandes
Janete Apparecida Desidério
author_sort Ana Rita Nunes Lemes
collection DOAJ
description The biological potential of Vip and Cry proteins from Bacillus is well known and widely established. Thus, it is important to look for new genes showing different modes of action, selecting those with differentiated entomotoxic activity against Diatraea flavipennella and Elasmopalpus lignosellus, which are secondary pests of sugarcane. Therefore, Cry1 and Vip3 proteins were expressed in Escherichia coli, and their toxicities were evaluated based on bioassays using neonate larvae. Of those, the most toxic were Cry1Ac and Vip3Aa considering the LC50 values. Toxins from E. coli were purified, solubilized, trypsinized, and biotinylated. Brush Border Membrane Vesicles (BBMVs) were prepared from intestines of the two species to perform homologous and heterologous competition assays. The binding assays demonstrated interactions between Cry1Aa, Cry1Ac, and Vip3Aa toxins and proteins from the BBMV of D. flavipennella and E. lignosellus. Homologous competition assays demonstrated that binding to one of the BBMV proteins was specific for each toxin. Heterologous competition assays indicated that Vip3Aa was unable to compete for Cry1Ac toxin binding. Our results suggest that Cry1Ac and Vip3Aa may have potential in future production of transgenic sugarcane for control of D. flavipennella and E. lignosellus, but more research is needed on the potential antagonism or synergism of the toxins in these pests.
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spelling doaj.art-68adc584af09423f97cad30bb29825102023-12-03T10:56:28ZengPeerJ Inc.PeerJ2167-83592017-01-015e286610.7717/peerj.2866Cry1Ac and Vip3Aa proteins from Bacillus thuringiensis targeting Cry toxin resistance in Diatraea flavipennella and Elasmopalpus lignosellus from sugarcaneAna Rita Nunes Lemes0Camila Soares Figueiredo1Isis Sebastião2Liliane Marques da Silva3Rebeka da Costa Alves4Herbert Álvaro Abreu de Siqueira5Manoel Victor Franco Lemos6Odair Aparecido Fernandes7Janete Apparecida Desidério8Department of Biologia Aplicada á Agropecuária, Universidade Estadual Paulista, Jaboticabal, São Paulo, BrazilDepartment of Biologia Aplicada á Agropecuária, Universidade Estadual Paulista, Jaboticabal, São Paulo, BrazilDepartment of Biologia Aplicada á Agropecuária, Universidade Estadual Paulista, Jaboticabal, São Paulo, BrazilDepartment of Agronomia, Universidade Federal Rural de Pernambuco, Recife, Pernambuco, BrazilDepartment of Agronomia, Universidade Federal Rural de Pernambuco, Recife, Pernambuco, BrazilDepartment of Agronomia, Universidade Federal Rural de Pernambuco, Recife, Pernambuco, BrazilDepartment of Biologia Aplicada á Agropecuária, Universidade Estadual Paulista, Jaboticabal, São Paulo, BrazilDepartamento de Fitossanidade/FCAV/UNESP, Universidade Estadual Paulista, Jaboticabal, São Paulo, BrazilDepartment of Biologia Aplicada á Agropecuária, Universidade Estadual Paulista, Jaboticabal, São Paulo, BrazilThe biological potential of Vip and Cry proteins from Bacillus is well known and widely established. Thus, it is important to look for new genes showing different modes of action, selecting those with differentiated entomotoxic activity against Diatraea flavipennella and Elasmopalpus lignosellus, which are secondary pests of sugarcane. Therefore, Cry1 and Vip3 proteins were expressed in Escherichia coli, and their toxicities were evaluated based on bioassays using neonate larvae. Of those, the most toxic were Cry1Ac and Vip3Aa considering the LC50 values. Toxins from E. coli were purified, solubilized, trypsinized, and biotinylated. Brush Border Membrane Vesicles (BBMVs) were prepared from intestines of the two species to perform homologous and heterologous competition assays. The binding assays demonstrated interactions between Cry1Aa, Cry1Ac, and Vip3Aa toxins and proteins from the BBMV of D. flavipennella and E. lignosellus. Homologous competition assays demonstrated that binding to one of the BBMV proteins was specific for each toxin. Heterologous competition assays indicated that Vip3Aa was unable to compete for Cry1Ac toxin binding. Our results suggest that Cry1Ac and Vip3Aa may have potential in future production of transgenic sugarcane for control of D. flavipennella and E. lignosellus, but more research is needed on the potential antagonism or synergism of the toxins in these pests.https://peerj.com/articles/2866.pdfBiological controlInsecticidal proteinsCompetition assay
spellingShingle Ana Rita Nunes Lemes
Camila Soares Figueiredo
Isis Sebastião
Liliane Marques da Silva
Rebeka da Costa Alves
Herbert Álvaro Abreu de Siqueira
Manoel Victor Franco Lemos
Odair Aparecido Fernandes
Janete Apparecida Desidério
Cry1Ac and Vip3Aa proteins from Bacillus thuringiensis targeting Cry toxin resistance in Diatraea flavipennella and Elasmopalpus lignosellus from sugarcane
PeerJ
Biological control
Insecticidal proteins
Competition assay
title Cry1Ac and Vip3Aa proteins from Bacillus thuringiensis targeting Cry toxin resistance in Diatraea flavipennella and Elasmopalpus lignosellus from sugarcane
title_full Cry1Ac and Vip3Aa proteins from Bacillus thuringiensis targeting Cry toxin resistance in Diatraea flavipennella and Elasmopalpus lignosellus from sugarcane
title_fullStr Cry1Ac and Vip3Aa proteins from Bacillus thuringiensis targeting Cry toxin resistance in Diatraea flavipennella and Elasmopalpus lignosellus from sugarcane
title_full_unstemmed Cry1Ac and Vip3Aa proteins from Bacillus thuringiensis targeting Cry toxin resistance in Diatraea flavipennella and Elasmopalpus lignosellus from sugarcane
title_short Cry1Ac and Vip3Aa proteins from Bacillus thuringiensis targeting Cry toxin resistance in Diatraea flavipennella and Elasmopalpus lignosellus from sugarcane
title_sort cry1ac and vip3aa proteins from bacillus thuringiensis targeting cry toxin resistance in diatraea flavipennella and elasmopalpus lignosellus from sugarcane
topic Biological control
Insecticidal proteins
Competition assay
url https://peerj.com/articles/2866.pdf
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