Construction of an improved mycoinsecticide overexpressing a toxic protease
Mycoinsecticides are being used for the control of many insect pests as an environmentally acceptable alternative to chemical insecticides. A key aim of much recent work has been to increase the speed of kill and so improve commercial efficacy of these biocontrol agents. This might be achieved by ad...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1996-06, Vol.93 (13), p.6349-6354 |
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| container_title | Proceedings of the National Academy of Sciences - PNAS |
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| creator | St Leger, R.J. (Cornell University, Ithaca, NY.) Joshi, L Bidochka, M.J Roberts, D.W |
| description | Mycoinsecticides are being used for the control of many insect pests as an environmentally acceptable alternative to chemical insecticides. A key aim of much recent work has been to increase the speed of kill and so improve commercial efficacy of these biocontrol agents. This might be achieved by adding insecticidal genes to the fungus, an approach considered to have enormous potential for the improvement of biological pesticides. We report here the development of a genetically improved entomopathogenic fungus. Additional copies of the gene encoding a regulated cuticle-degrading protease (Pr1) from Metarhizium anisopliae were inserted into the genome of M. anisopliae such that Pr1 was constitutively overproduced in the hemolymph of Manduca sexta, activating the prophenoloxidase system. The combined toxic effects of Pr1 and the reaction products of phenoloxidase caused larvae challenged with the engineered fungus to exhibit a 25% reduction in time of death and reduced food consumption by 40% compared to infections by the wild-type fungus. In addition, infected insects were rapidly melanized, and the resulting cadavers were poor substrates for fungal sporulation. Thus, environmental persistence of the genetically engineered fungus is reduced, thereby providing biological containment |
| doi_str_mv | 10.1073/pnas.93.13.6349 |
| format | Article |
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(Cornell University, Ithaca, NY.) ; Joshi, L ; Bidochka, M.J ; Roberts, D.W</creator><creatorcontrib>St Leger, R.J. (Cornell University, Ithaca, NY.) ; Joshi, L ; Bidochka, M.J ; Roberts, D.W</creatorcontrib><description>Mycoinsecticides are being used for the control of many insect pests as an environmentally acceptable alternative to chemical insecticides. A key aim of much recent work has been to increase the speed of kill and so improve commercial efficacy of these biocontrol agents. This might be achieved by adding insecticidal genes to the fungus, an approach considered to have enormous potential for the improvement of biological pesticides. We report here the development of a genetically improved entomopathogenic fungus. Additional copies of the gene encoding a regulated cuticle-degrading protease (Pr1) from Metarhizium anisopliae were inserted into the genome of M. anisopliae such that Pr1 was constitutively overproduced in the hemolymph of Manduca sexta, activating the prophenoloxidase system. The combined toxic effects of Pr1 and the reaction products of phenoloxidase caused larvae challenged with the engineered fungus to exhibit a 25% reduction in time of death and reduced food consumption by 40% compared to infections by the wild-type fungus. In addition, infected insects were rapidly melanized, and the resulting cadavers were poor substrates for fungal sporulation. Thus, environmental persistence of the genetically engineered fungus is reduced, thereby providing biological containment</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.93.13.6349</identifier><identifier>PMID: 8692818</identifier><language>eng</language><publisher>United States: National Academy of Sciences of the United States of America</publisher><subject>ADN RECOMBINADO ; ADN RECOMBINE ; Amino Acid Sequence ; Animals ; Base Sequence ; Biochemistry ; DNA, Complementary ; DOSAGE BIOLOGIQUE ; ENSAYO BIOLOGICO ; Enzyme Activation ; Enzyme Precursors - metabolism ; EXPRESION GENICA ; EXPRESSION DES GENES ; Fungal Proteins - genetics ; Fungal Proteins - toxicity ; Fungi ; Fungi - genetics ; GENE ; GENES ; HEMOLINFA ; Hemolymph ; HEMOLYMPHE ; Infections ; Insect genetics ; Insect larvae ; Insect pathology ; Insect pests ; Insecticides ; Instars ; Larval development ; LARVAS ; LARVE ; Lepidoptera - enzymology ; MANDUCA SEXTA ; METARHIZIUM ANISOPLIAE ; Molecular Sequence Data ; Monophenol Monooxygenase - metabolism ; MORTALIDAD ; MORTALITE ; OXIDORREDUCTASAS ; OXYDOREDUCTASE ; PATOGENICIDAD ; Pest Control, Biological ; Pesticides ; PIGMENT ; PIGMENTOS ; POUVOIR PATHOGENE ; PROTEASAS ; PROTEASE ; Serine Endopeptidases - genetics ; Serine Endopeptidases - toxicity ; SUPERVIVENCIA ; SURVIE ; Transcription, Genetic ; TRANSFERENCIA DE GENES ; TRANSFERT DE GENE ; TRANSFORMACION GENETICA ; TRANSFORMATION GENETIQUE ; Transgenic insects</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 1996-06, Vol.93 (13), p.6349-6354</ispartof><rights>Copyright 1996 National Academy of Sciences</rights><rights>Copyright National Academy of Sciences Jun 25, 1996</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c573t-2bf536f0656b6175486909a4f46d971ee72ab8d4270f0ab63382249ed7b78c503</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/93/13.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/39308$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/39308$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,723,776,780,799,881,27903,27904,53770,53772,57996,58229</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8692818$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>St Leger, R.J. (Cornell University, Ithaca, NY.)</creatorcontrib><creatorcontrib>Joshi, L</creatorcontrib><creatorcontrib>Bidochka, M.J</creatorcontrib><creatorcontrib>Roberts, D.W</creatorcontrib><title>Construction of an improved mycoinsecticide overexpressing a toxic protease</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>Mycoinsecticides are being used for the control of many insect pests as an environmentally acceptable alternative to chemical insecticides. A key aim of much recent work has been to increase the speed of kill and so improve commercial efficacy of these biocontrol agents. This might be achieved by adding insecticidal genes to the fungus, an approach considered to have enormous potential for the improvement of biological pesticides. We report here the development of a genetically improved entomopathogenic fungus. Additional copies of the gene encoding a regulated cuticle-degrading protease (Pr1) from Metarhizium anisopliae were inserted into the genome of M. anisopliae such that Pr1 was constitutively overproduced in the hemolymph of Manduca sexta, activating the prophenoloxidase system. The combined toxic effects of Pr1 and the reaction products of phenoloxidase caused larvae challenged with the engineered fungus to exhibit a 25% reduction in time of death and reduced food consumption by 40% compared to infections by the wild-type fungus. In addition, infected insects were rapidly melanized, and the resulting cadavers were poor substrates for fungal sporulation. Thus, environmental persistence of the genetically engineered fungus is reduced, thereby providing biological containment</description><subject>ADN RECOMBINADO</subject><subject>ADN RECOMBINE</subject><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Base Sequence</subject><subject>Biochemistry</subject><subject>DNA, Complementary</subject><subject>DOSAGE BIOLOGIQUE</subject><subject>ENSAYO BIOLOGICO</subject><subject>Enzyme Activation</subject><subject>Enzyme Precursors - metabolism</subject><subject>EXPRESION GENICA</subject><subject>EXPRESSION DES GENES</subject><subject>Fungal Proteins - genetics</subject><subject>Fungal Proteins - toxicity</subject><subject>Fungi</subject><subject>Fungi - genetics</subject><subject>GENE</subject><subject>GENES</subject><subject>HEMOLINFA</subject><subject>Hemolymph</subject><subject>HEMOLYMPHE</subject><subject>Infections</subject><subject>Insect genetics</subject><subject>Insect larvae</subject><subject>Insect pathology</subject><subject>Insect pests</subject><subject>Insecticides</subject><subject>Instars</subject><subject>Larval development</subject><subject>LARVAS</subject><subject>LARVE</subject><subject>Lepidoptera - enzymology</subject><subject>MANDUCA SEXTA</subject><subject>METARHIZIUM ANISOPLIAE</subject><subject>Molecular Sequence Data</subject><subject>Monophenol Monooxygenase - metabolism</subject><subject>MORTALIDAD</subject><subject>MORTALITE</subject><subject>OXIDORREDUCTASAS</subject><subject>OXYDOREDUCTASE</subject><subject>PATOGENICIDAD</subject><subject>Pest Control, Biological</subject><subject>Pesticides</subject><subject>PIGMENT</subject><subject>PIGMENTOS</subject><subject>POUVOIR PATHOGENE</subject><subject>PROTEASAS</subject><subject>PROTEASE</subject><subject>Serine Endopeptidases - genetics</subject><subject>Serine Endopeptidases - toxicity</subject><subject>SUPERVIVENCIA</subject><subject>SURVIE</subject><subject>Transcription, Genetic</subject><subject>TRANSFERENCIA DE GENES</subject><subject>TRANSFERT DE GENE</subject><subject>TRANSFORMACION GENETICA</subject><subject>TRANSFORMATION GENETIQUE</subject><subject>Transgenic insects</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9ks1rFDEYh4Moda2eBUEZPOhptvn-gF5kaVUseNCeQ2Yms2aZScYkU7b_vRl2Xa0HT4E8z-_lDb8A8BLBNYKCXEzepLUia0TWnFD1CKwQVKjmVMHHYAUhFrWkmD4Fz1LaQQgVk_AMnEmusERyBb5sgk85zm12wVehr4yv3DjFcGe7arxvg_PJFti6zlblMtr9FG1Kzm8rU-Wwd21V7GxNss_Bk94Myb44nufg9vrq--ZTffP14-fNh5u6ZYLkGjc9I7yHnPGGI8FoWQYqQ3vKOyWQtQKbRnYUC9hD03BCJMZU2U40QrYMknNweZg7zc1ou9b6HM2gp-hGE-91ME4_JN790Ntwp4mCmJX4u2M8hp-zTVmPLrV2GIy3YU4aMc4pZaqIb_8Rd2GOvjxNY4gIZkrQIl0cpDaGlKLtT3sgqJeK9FKRVkQjopeKSuL13-uf_GMnhb858iX4mz4Y8P6_gu7nYch2n4v56mDuUg7xpBJFoPwDexO02UaX9O03xfHyd8gvO8a3RQ</recordid><startdate>19960625</startdate><enddate>19960625</enddate><creator>St Leger, R.J. (Cornell University, Ithaca, NY.)</creator><creator>Joshi, L</creator><creator>Bidochka, M.J</creator><creator>Roberts, D.W</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</general><general>National Academy of Sciences</general><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QG</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7QO</scope><scope>7T7</scope><scope>5PM</scope></search><sort><creationdate>19960625</creationdate><title>Construction of an improved mycoinsecticide overexpressing a toxic protease</title><author>St Leger, R.J. (Cornell University, Ithaca, NY.) ; Joshi, L ; Bidochka, M.J ; Roberts, D.W</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c573t-2bf536f0656b6175486909a4f46d971ee72ab8d4270f0ab63382249ed7b78c503</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>ADN RECOMBINADO</topic><topic>ADN RECOMBINE</topic><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Base Sequence</topic><topic>Biochemistry</topic><topic>DNA, Complementary</topic><topic>DOSAGE BIOLOGIQUE</topic><topic>ENSAYO BIOLOGICO</topic><topic>Enzyme Activation</topic><topic>Enzyme Precursors - metabolism</topic><topic>EXPRESION GENICA</topic><topic>EXPRESSION DES GENES</topic><topic>Fungal Proteins - genetics</topic><topic>Fungal Proteins - toxicity</topic><topic>Fungi</topic><topic>Fungi - genetics</topic><topic>GENE</topic><topic>GENES</topic><topic>HEMOLINFA</topic><topic>Hemolymph</topic><topic>HEMOLYMPHE</topic><topic>Infections</topic><topic>Insect genetics</topic><topic>Insect larvae</topic><topic>Insect pathology</topic><topic>Insect pests</topic><topic>Insecticides</topic><topic>Instars</topic><topic>Larval development</topic><topic>LARVAS</topic><topic>LARVE</topic><topic>Lepidoptera - enzymology</topic><topic>MANDUCA SEXTA</topic><topic>METARHIZIUM ANISOPLIAE</topic><topic>Molecular Sequence Data</topic><topic>Monophenol Monooxygenase - metabolism</topic><topic>MORTALIDAD</topic><topic>MORTALITE</topic><topic>OXIDORREDUCTASAS</topic><topic>OXYDOREDUCTASE</topic><topic>PATOGENICIDAD</topic><topic>Pest Control, Biological</topic><topic>Pesticides</topic><topic>PIGMENT</topic><topic>PIGMENTOS</topic><topic>POUVOIR PATHOGENE</topic><topic>PROTEASAS</topic><topic>PROTEASE</topic><topic>Serine Endopeptidases - genetics</topic><topic>Serine Endopeptidases - toxicity</topic><topic>SUPERVIVENCIA</topic><topic>SURVIE</topic><topic>Transcription, Genetic</topic><topic>TRANSFERENCIA DE GENES</topic><topic>TRANSFERT DE GENE</topic><topic>TRANSFORMACION GENETICA</topic><topic>TRANSFORMATION GENETIQUE</topic><topic>Transgenic insects</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>St Leger, R.J. 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(Cornell University, Ithaca, NY.)</au><au>Joshi, L</au><au>Bidochka, M.J</au><au>Roberts, D.W</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Construction of an improved mycoinsecticide overexpressing a toxic protease</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1996-06-25</date><risdate>1996</risdate><volume>93</volume><issue>13</issue><spage>6349</spage><epage>6354</epage><pages>6349-6354</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>Mycoinsecticides are being used for the control of many insect pests as an environmentally acceptable alternative to chemical insecticides. A key aim of much recent work has been to increase the speed of kill and so improve commercial efficacy of these biocontrol agents. This might be achieved by adding insecticidal genes to the fungus, an approach considered to have enormous potential for the improvement of biological pesticides. We report here the development of a genetically improved entomopathogenic fungus. Additional copies of the gene encoding a regulated cuticle-degrading protease (Pr1) from Metarhizium anisopliae were inserted into the genome of M. anisopliae such that Pr1 was constitutively overproduced in the hemolymph of Manduca sexta, activating the prophenoloxidase system. The combined toxic effects of Pr1 and the reaction products of phenoloxidase caused larvae challenged with the engineered fungus to exhibit a 25% reduction in time of death and reduced food consumption by 40% compared to infections by the wild-type fungus. In addition, infected insects were rapidly melanized, and the resulting cadavers were poor substrates for fungal sporulation. Thus, environmental persistence of the genetically engineered fungus is reduced, thereby providing biological containment</abstract><cop>United States</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>8692818</pmid><doi>10.1073/pnas.93.13.6349</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Proceedings of the National Academy of Sciences - PNAS, 1996-06, Vol.93 (13), p.6349-6354 |
| issn | 0027-8424 1091-6490 |
| language | eng |
| recordid | cdi_proquest_journals_201325974 |
| source | MEDLINE; Jstor Complete Legacy; PubMed Central; Alma/SFX Local Collection; Free Full-Text Journals in Chemistry |
| subjects | ADN RECOMBINADO ADN RECOMBINE Amino Acid Sequence Animals Base Sequence Biochemistry DNA, Complementary DOSAGE BIOLOGIQUE ENSAYO BIOLOGICO Enzyme Activation Enzyme Precursors - metabolism EXPRESION GENICA EXPRESSION DES GENES Fungal Proteins - genetics Fungal Proteins - toxicity Fungi Fungi - genetics GENE GENES HEMOLINFA Hemolymph HEMOLYMPHE Infections Insect genetics Insect larvae Insect pathology Insect pests Insecticides Instars Larval development LARVAS LARVE Lepidoptera - enzymology MANDUCA SEXTA METARHIZIUM ANISOPLIAE Molecular Sequence Data Monophenol Monooxygenase - metabolism MORTALIDAD MORTALITE OXIDORREDUCTASAS OXYDOREDUCTASE PATOGENICIDAD Pest Control, Biological Pesticides PIGMENT PIGMENTOS POUVOIR PATHOGENE PROTEASAS PROTEASE Serine Endopeptidases - genetics Serine Endopeptidases - toxicity SUPERVIVENCIA SURVIE Transcription, Genetic TRANSFERENCIA DE GENES TRANSFERT DE GENE TRANSFORMACION GENETICA TRANSFORMATION GENETIQUE Transgenic insects |
| title | Construction of an improved mycoinsecticide overexpressing a toxic protease |
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