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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/168486
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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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1844613115450228736 |
score |
13.070432 |