Enhanced virulence of Beauveria bassiana against Diatraea saccharalis using a soluble recombinant enzyme with endo- and exochitinase activity

Autores
Lovera, Andrea; Belaich, Mariano Nicolas; Villamizar, Laura; Patarroyo, Manuel A.; Barrera, Gloria
Año de publicación
2020
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Beauveria bassiana chitinases are involved in degrading the chitin in insects’ exoskeletons and internal structures, and thus are important virulence factors as they participate in initial to final steps of infection. In this work, the B. bassiana (Bv062 isolate) open reading frame (ORF) encoding a chitinase identified as Chit37 (orthologous Bvchit1) was molecularly cloned and expressed in Escherichia coli, and the potential of the recombinant protein (rChit37) to enhance the insecticidal activity of Bv062 conidia against second instar Diatraea saccharalis larvae was studied. rChit37 was produced in both soluble and insoluble fractions of an E. coli culture. Both fractions expressing endo- (90 mU/µL) and exochitinase (170 mU/µL) enzymatic activity, with optimum conditions for enzyme activity of 45 °C and pH 5.0. His-tag affinity chromatography was used to purify the rChit37 from the soluble fraction. Purified rChit37 was then diluted to 200 and 300 µg/mL for use as Bv062 conidia additive (1x106con/mL) in a laboratory bioassay against D. saccharalis larvae. No significant differences were observed between the efficacy of Bv062 conidia applied alone or mixed with 200 µg/mL purified rChit37. However, 300 µg/mL rChit37 increased BV062 conidia insecticidal activity, achieving 96.7% efficacy (14 days post-infection) and 6.2 days median lethal time (LT50), compared to 60% efficacy and 8.9 days for conidia alone. rChit37 addition did not affect conidial viability in terms of germination (96.6% after 24 h) or vigour estimated as germ-tube elongation rate. This work provides proof of concept about soluble recombinant chitinase as an additive to enhance B. bassiana virulence against D. saccharalis.
Fil: Lovera, Andrea. Universidad Nacional de Colombia; Colombia. Corporación Colombiana de Investigación Agropecuaria; Colombia
Fil: Belaich, Mariano Nicolas. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular. Área Virus de Insectos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Villamizar, Laura. No especifíca;
Fil: Patarroyo, Manuel A.. Fundación Instituto de Inmunología; Colombia. Universidad Colegio Mayor de Nuestra Señora del Rosario; Colombia
Fil: Barrera, Gloria. Corporación Colombiana de Investigación Agropecuaria; Colombia
Materia
BEAUVERIA BASSIANA
CHITINASE
DIATRAEA SACCHARALIS
RECOMBINANT PROTEIN
VIRULENCE
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/168486

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network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Enhanced virulence of Beauveria bassiana against Diatraea saccharalis using a soluble recombinant enzyme with endo- and exochitinase activityLovera, AndreaBelaich, Mariano NicolasVillamizar, LauraPatarroyo, Manuel A.Barrera, GloriaBEAUVERIA BASSIANACHITINASEDIATRAEA SACCHARALISRECOMBINANT PROTEINVIRULENCEhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Beauveria bassiana chitinases are involved in degrading the chitin in insects’ exoskeletons and internal structures, and thus are important virulence factors as they participate in initial to final steps of infection. In this work, the B. bassiana (Bv062 isolate) open reading frame (ORF) encoding a chitinase identified as Chit37 (orthologous Bvchit1) was molecularly cloned and expressed in Escherichia coli, and the potential of the recombinant protein (rChit37) to enhance the insecticidal activity of Bv062 conidia against second instar Diatraea saccharalis larvae was studied. rChit37 was produced in both soluble and insoluble fractions of an E. coli culture. Both fractions expressing endo- (90 mU/µL) and exochitinase (170 mU/µL) enzymatic activity, with optimum conditions for enzyme activity of 45 °C and pH 5.0. His-tag affinity chromatography was used to purify the rChit37 from the soluble fraction. Purified rChit37 was then diluted to 200 and 300 µg/mL for use as Bv062 conidia additive (1x106con/mL) in a laboratory bioassay against D. saccharalis larvae. No significant differences were observed between the efficacy of Bv062 conidia applied alone or mixed with 200 µg/mL purified rChit37. However, 300 µg/mL rChit37 increased BV062 conidia insecticidal activity, achieving 96.7% efficacy (14 days post-infection) and 6.2 days median lethal time (LT50), compared to 60% efficacy and 8.9 days for conidia alone. rChit37 addition did not affect conidial viability in terms of germination (96.6% after 24 h) or vigour estimated as germ-tube elongation rate. This work provides proof of concept about soluble recombinant chitinase as an additive to enhance B. bassiana virulence against D. saccharalis.Fil: Lovera, Andrea. Universidad Nacional de Colombia; Colombia. Corporación Colombiana de Investigación Agropecuaria; ColombiaFil: Belaich, Mariano Nicolas. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular. Área Virus de Insectos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Villamizar, Laura. No especifíca;Fil: Patarroyo, Manuel A.. Fundación Instituto de Inmunología; Colombia. Universidad Colegio Mayor de Nuestra Señora del Rosario; ColombiaFil: Barrera, Gloria. Corporación Colombiana de Investigación Agropecuaria; ColombiaAcademic Press Inc Elsevier Science2020-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/168486Lovera, Andrea; Belaich, Mariano Nicolas; Villamizar, Laura; Patarroyo, Manuel A.; Barrera, Gloria; Enhanced virulence of Beauveria bassiana against Diatraea saccharalis using a soluble recombinant enzyme with endo- and exochitinase activity; Academic Press Inc Elsevier Science; Biological Control; 144; 5-2020; 1-111049-9644CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S1049964419308163info:eu-repo/semantics/altIdentifier/doi/10.1016/j.biocontrol.2020.104211info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:35:43Zoai:ri.conicet.gov.ar:11336/168486instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:35:43.855CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Enhanced virulence of Beauveria bassiana against Diatraea saccharalis using a soluble recombinant enzyme with endo- and exochitinase activity
title Enhanced virulence of Beauveria bassiana against Diatraea saccharalis using a soluble recombinant enzyme with endo- and exochitinase activity
spellingShingle Enhanced virulence of Beauveria bassiana against Diatraea saccharalis using a soluble recombinant enzyme with endo- and exochitinase activity
Lovera, Andrea
BEAUVERIA BASSIANA
CHITINASE
DIATRAEA SACCHARALIS
RECOMBINANT PROTEIN
VIRULENCE
title_short Enhanced virulence of Beauveria bassiana against Diatraea saccharalis using a soluble recombinant enzyme with endo- and exochitinase activity
title_full Enhanced virulence of Beauveria bassiana against Diatraea saccharalis using a soluble recombinant enzyme with endo- and exochitinase activity
title_fullStr Enhanced virulence of Beauveria bassiana against Diatraea saccharalis using a soluble recombinant enzyme with endo- and exochitinase activity
title_full_unstemmed Enhanced virulence of Beauveria bassiana against Diatraea saccharalis using a soluble recombinant enzyme with endo- and exochitinase activity
title_sort Enhanced virulence of Beauveria bassiana against Diatraea saccharalis using a soluble recombinant enzyme with endo- and exochitinase activity
dc.creator.none.fl_str_mv Lovera, Andrea
Belaich, Mariano Nicolas
Villamizar, Laura
Patarroyo, Manuel A.
Barrera, Gloria
author Lovera, Andrea
author_facet Lovera, Andrea
Belaich, Mariano Nicolas
Villamizar, Laura
Patarroyo, Manuel A.
Barrera, Gloria
author_role author
author2 Belaich, Mariano Nicolas
Villamizar, Laura
Patarroyo, Manuel A.
Barrera, Gloria
author2_role author
author
author
author
dc.subject.none.fl_str_mv BEAUVERIA BASSIANA
CHITINASE
DIATRAEA SACCHARALIS
RECOMBINANT PROTEIN
VIRULENCE
topic BEAUVERIA BASSIANA
CHITINASE
DIATRAEA SACCHARALIS
RECOMBINANT PROTEIN
VIRULENCE
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Beauveria bassiana chitinases are involved in degrading the chitin in insects’ exoskeletons and internal structures, and thus are important virulence factors as they participate in initial to final steps of infection. In this work, the B. bassiana (Bv062 isolate) open reading frame (ORF) encoding a chitinase identified as Chit37 (orthologous Bvchit1) was molecularly cloned and expressed in Escherichia coli, and the potential of the recombinant protein (rChit37) to enhance the insecticidal activity of Bv062 conidia against second instar Diatraea saccharalis larvae was studied. rChit37 was produced in both soluble and insoluble fractions of an E. coli culture. Both fractions expressing endo- (90 mU/µL) and exochitinase (170 mU/µL) enzymatic activity, with optimum conditions for enzyme activity of 45 °C and pH 5.0. His-tag affinity chromatography was used to purify the rChit37 from the soluble fraction. Purified rChit37 was then diluted to 200 and 300 µg/mL for use as Bv062 conidia additive (1x106con/mL) in a laboratory bioassay against D. saccharalis larvae. No significant differences were observed between the efficacy of Bv062 conidia applied alone or mixed with 200 µg/mL purified rChit37. However, 300 µg/mL rChit37 increased BV062 conidia insecticidal activity, achieving 96.7% efficacy (14 days post-infection) and 6.2 days median lethal time (LT50), compared to 60% efficacy and 8.9 days for conidia alone. rChit37 addition did not affect conidial viability in terms of germination (96.6% after 24 h) or vigour estimated as germ-tube elongation rate. This work provides proof of concept about soluble recombinant chitinase as an additive to enhance B. bassiana virulence against D. saccharalis.
Fil: Lovera, Andrea. Universidad Nacional de Colombia; Colombia. Corporación Colombiana de Investigación Agropecuaria; Colombia
Fil: Belaich, Mariano Nicolas. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular. Área Virus de Insectos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Villamizar, Laura. No especifíca;
Fil: Patarroyo, Manuel A.. Fundación Instituto de Inmunología; Colombia. Universidad Colegio Mayor de Nuestra Señora del Rosario; Colombia
Fil: Barrera, Gloria. Corporación Colombiana de Investigación Agropecuaria; Colombia
description Beauveria bassiana chitinases are involved in degrading the chitin in insects’ exoskeletons and internal structures, and thus are important virulence factors as they participate in initial to final steps of infection. In this work, the B. bassiana (Bv062 isolate) open reading frame (ORF) encoding a chitinase identified as Chit37 (orthologous Bvchit1) was molecularly cloned and expressed in Escherichia coli, and the potential of the recombinant protein (rChit37) to enhance the insecticidal activity of Bv062 conidia against second instar Diatraea saccharalis larvae was studied. rChit37 was produced in both soluble and insoluble fractions of an E. coli culture. Both fractions expressing endo- (90 mU/µL) and exochitinase (170 mU/µL) enzymatic activity, with optimum conditions for enzyme activity of 45 °C and pH 5.0. His-tag affinity chromatography was used to purify the rChit37 from the soluble fraction. Purified rChit37 was then diluted to 200 and 300 µg/mL for use as Bv062 conidia additive (1x106con/mL) in a laboratory bioassay against D. saccharalis larvae. No significant differences were observed between the efficacy of Bv062 conidia applied alone or mixed with 200 µg/mL purified rChit37. However, 300 µg/mL rChit37 increased BV062 conidia insecticidal activity, achieving 96.7% efficacy (14 days post-infection) and 6.2 days median lethal time (LT50), compared to 60% efficacy and 8.9 days for conidia alone. rChit37 addition did not affect conidial viability in terms of germination (96.6% after 24 h) or vigour estimated as germ-tube elongation rate. This work provides proof of concept about soluble recombinant chitinase as an additive to enhance B. bassiana virulence against D. saccharalis.
publishDate 2020
dc.date.none.fl_str_mv 2020-05
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/168486
Lovera, Andrea; Belaich, Mariano Nicolas; Villamizar, Laura; Patarroyo, Manuel A.; Barrera, Gloria; Enhanced virulence of Beauveria bassiana against Diatraea saccharalis using a soluble recombinant enzyme with endo- and exochitinase activity; Academic Press Inc Elsevier Science; Biological Control; 144; 5-2020; 1-11
1049-9644
CONICET Digital
CONICET
url http://hdl.handle.net/11336/168486
identifier_str_mv Lovera, Andrea; Belaich, Mariano Nicolas; Villamizar, Laura; Patarroyo, Manuel A.; Barrera, Gloria; Enhanced virulence of Beauveria bassiana against Diatraea saccharalis using a soluble recombinant enzyme with endo- and exochitinase activity; Academic Press Inc Elsevier Science; Biological Control; 144; 5-2020; 1-11
1049-9644
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S1049964419308163
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.biocontrol.2020.104211
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Academic Press Inc Elsevier Science
publisher.none.fl_str_mv Academic Press Inc Elsevier Science
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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