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Draft genome sequences of two bacillus thuringiensis strains and characterization of a putative 41.9-kDa insecticidal toxin

dc.contributor.authorPalma Dovis, Leopoldo
dc.contributor.authorMuñoz Labiano, Delia
dc.contributor.authorBerry, Colin
dc.contributor.authorMurillo Martínez, Jesús
dc.contributor.authorCaballero Murillo, Primitivo
dc.contributor.departmentNekazaritza Ekoizpenaeu
dc.contributor.departmentProducción Agrariaes_ES
dc.contributor.departmentIdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutuaes_ES
dc.contributor.funderUniversidad Pública de Navarra / Nafarroako Unibertsitate Publikoaes
dc.date.accessioned2014-06-30T12:15:39Z
dc.date.available2014-06-30T12:15:39Z
dc.date.issued2014
dc.description.abstractIn this work, we report the genome sequencing of two Bacillus thuringiensis strains using Illumina next-generation sequencing technology (NGS). Strain Hu4-2, toxic to many lepidopteran pest species and to some mosquitoes, encoded genes for two insecticidal crystal (Cry) proteins, cry1Ia and cry9Ea, and a vegetative insecticidal protein (Vip) gene, vip3Ca2. Strain Leapi01 contained genes coding for seven Cry proteins (cry1Aa, cry1Ca, cry1Da, cry2Ab, cry9Ea and two cry1Ia gene variants) and a vip3 gene (vip3Aa10). A putative novel insecticidal protein gene 1143 bp long was found in both strains, whose sequences exhibited 100% nucleotide identity. The predicted protein showed 57 and 100% pairwise identity to protein sequence 72 from a patented Bt strain (US8318900) and to a putative 41.9-kDa insecticidal toxin from Bacillus cereus, respectively. The 41.9-kDa protein, containing a C-terminal 6× HisTag fusion, was expressed in Escherichia coli and tested for the first time against four lepidopteran species (Mamestra brassicae, Ostrinia nubilalis, Spodoptera frugiperda and S. littoralis) and the green-peach aphid Myzus persicae at doses as high as 4.8 μg/cm2 and 1.5 mg/mL, respectively. At these protein concentrations, the recombinant 41.9-kDa protein caused no mortality or symptoms of impaired growth against any of the insects tested, suggesting that these species are outside the protein’s target range or that the protein may not, in fact, be toxic. While the use of the polymerase chain reaction has allowed a significant increase in the number of Bt insecticidal genes characterized to date, novel NGS technologies promise a much faster, cheaper and efficient screening of Bt pesticidal proteins.en
dc.description.sponsorshipThis research was supported by the Spanish Ministry of Science and Innovation (grant ref. AGL2009-13340-C02) and by the Universidad Pública de Navarra (PhD contract awarded to L.P.).en
dc.format.mimetypeapplication/pdfen
dc.identifier.doi10.3390/toxins6051490
dc.identifier.issn2072-6651
dc.identifier.urihttps://academica-e.unavarra.es/handle/2454/11044
dc.language.isoengen
dc.publisherMDPIen
dc.relation.ispartofToxins, 6 (5). Págs. 1490-1504en
dc.relation.projectIDinfo:eu-repo/grantAgreement/MICINN//AGL2009-13340-C02-02/ES/en
dc.relation.publisherversionhttps://doi.org/10.3390/toxins6051490
dc.rights© 2014 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license.en
dc.rights.accessRightsinfo:eu-repo/semantics/openAccessen
dc.rights.accessRightsAcceso abierto / Sarbide irekiaes
dc.rights.urihttp://creativecommons.org/licenses/by/3.0/
dc.subjectBacillus thuringiensisen
dc.subjectInsecticidal toxinsen
dc.subjectNext-generacion sequencingen
dc.subjectGenome annotationen
dc.subjectMicrobial controlen
dc.subjectInsecticidal activityen
dc.titleDraft genome sequences of two bacillus thuringiensis strains and characterization of a putative 41.9-kDa insecticidal toxinen
dc.typeArtículo / Artikuluaes
dc.typeinfo:eu-repo/semantics/articleen
dc.type.versionVersión publicada / Argitaratu den bertsioaes
dc.type.versioninfo:eu-repo/semantics/publishedVersionen
dspace.entity.typePublication
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relation.isAuthorOfPublicationcc878517-e4e6-491d-a424-b6e9ada5a61e
relation.isAuthorOfPublication3f350c70-2e68-4b9b-95d6-26e9cc1540a0
relation.isAuthorOfPublicationecde8e03-14c2-46a3-9e89-ae3b2c668297
relation.isAuthorOfPublication.latestForDiscoveryb2cb88ba-4e7e-4129-8d02-5f6fcf26d797

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