Cry1Ac and Vip3Aa proteins from Bacillus thuringiensis targeting Cry toxin resistance in Diatraea flavipennella and Elasmopalpus lignosellus from sugarcane
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
Affiliations
Ana Rita Nunes Lemes
Department of Biologia Aplicada á Agropecuária, Universidade Estadual Paulista, Jaboticabal, São Paulo, Brazil
Camila Soares Figueiredo
Department of Biologia Aplicada á Agropecuária, Universidade Estadual Paulista, Jaboticabal, São Paulo, Brazil
Isis Sebastião
Department of Biologia Aplicada á Agropecuária, Universidade Estadual Paulista, Jaboticabal, São Paulo, Brazil
Liliane Marques da Silva
Department of Agronomia, Universidade Federal Rural de Pernambuco, Recife, Pernambuco, Brazil
Rebeka da Costa Alves
Department of Agronomia, Universidade Federal Rural de Pernambuco, Recife, Pernambuco, Brazil
Herbert Álvaro Abreu de Siqueira
Department of Agronomia, Universidade Federal Rural de Pernambuco, Recife, Pernambuco, Brazil
Manoel Victor Franco Lemos
Department of Biologia Aplicada á Agropecuária, Universidade Estadual Paulista, Jaboticabal, São Paulo, Brazil
Odair Aparecido Fernandes
Departamento de Fitossanidade/FCAV/UNESP, Universidade Estadual Paulista, Jaboticabal, São Paulo, Brazil
Janete Apparecida Desidério
Department of Biologia Aplicada á Agropecuária, Universidade Estadual Paulista, Jaboticabal, São Paulo, Brazil
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.
