Double-Stranded RNA Uptake through Topical Application, Mediates Silencing of Five CYP4 Genes and Suppresses Insecticide Resistance in Diaphorina citri
Identifieur interne : 000B27 ( Main/Exploration ); précédent : 000B26; suivant : 000B28Double-Stranded RNA Uptake through Topical Application, Mediates Silencing of Five CYP4 Genes and Suppresses Insecticide Resistance in Diaphorina citri
Auteurs : Nabil Killiny [États-Unis] ; Subhas Hajeri [États-Unis] ; Siddharth Tiwari [États-Unis] ; Siddarame Gowda [États-Unis] ; Lukasz L. Stelinski [États-Unis]Source :
- PLoS ONE [ 1932-6203 ] ; 2014.
Abstract
Silencing of genes through RNA interference (RNAi) in insects has gained momentum during the past few years. RNAi has been used to cause insect mortality, inhibit insect growth, increase insecticide susceptibility, and prevent the development of insecticide resistance. We investigated the efficacy of topically applied dsRNA to induce RNAi for five Cytochrome P450 genes family 4 (
Url:
DOI: 10.1371/journal.pone.0110536
PubMed: 25330026
PubMed Central: 4203802
Affiliations:
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Double-Stranded RNA Uptake through Topical Application, Mediates Silencing of Five CYP4 Genes and Suppresses Insecticide Resistance in <italic>Diaphorina citri</italic></title><author><name sortKey="Killiny, Nabil" sort="Killiny, Nabil" uniqKey="Killiny N" first="Nabil" last="Killiny">Nabil Killiny</name><affiliation wicri:level="2"><nlm:aff id="aff1"><addr-line>Department of Entomology and Nematology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Entomology and Nematology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida</wicri:regionArea><placeName><region type="state">Floride</region></placeName></affiliation></author><author><name sortKey="Hajeri, Subhas" sort="Hajeri, Subhas" uniqKey="Hajeri S" first="Subhas" last="Hajeri">Subhas Hajeri</name><affiliation wicri:level="2"><nlm:aff id="aff2"><addr-line>Department of Plant Pathology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Plant Pathology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida</wicri:regionArea><placeName><region type="state">Floride</region></placeName></affiliation></author><author><name sortKey="Tiwari, Siddharth" sort="Tiwari, Siddharth" uniqKey="Tiwari S" first="Siddharth" last="Tiwari">Siddharth Tiwari</name><affiliation wicri:level="2"><nlm:aff id="aff1"><addr-line>Department of Entomology and Nematology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Entomology and Nematology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida</wicri:regionArea><placeName><region type="state">Floride</region></placeName></affiliation></author><author><name sortKey="Gowda, Siddarame" sort="Gowda, Siddarame" uniqKey="Gowda S" first="Siddarame" last="Gowda">Siddarame Gowda</name><affiliation wicri:level="2"><nlm:aff id="aff2"><addr-line>Department of Plant Pathology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Plant Pathology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida</wicri:regionArea><placeName><region type="state">Floride</region></placeName></affiliation></author><author><name sortKey="Stelinski, Lukasz L" sort="Stelinski, Lukasz L" uniqKey="Stelinski L" first="Lukasz L." last="Stelinski">Lukasz L. Stelinski</name><affiliation wicri:level="2"><nlm:aff id="aff1"><addr-line>Department of Entomology and Nematology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Entomology and Nematology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida</wicri:regionArea><placeName><region type="state">Floride</region></placeName></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">25330026</idno><idno type="pmc">4203802</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4203802</idno><idno type="RBID">PMC:4203802</idno><idno type="doi">10.1371/journal.pone.0110536</idno><date when="2014">2014</date><idno type="wicri:Area/Pmc/Corpus">000213</idno><idno type="wicri:Area/Pmc/Curation">000213</idno><idno type="wicri:Area/Pmc/Checkpoint">000857</idno><idno type="wicri:Area/Ncbi/Merge">001867</idno><idno type="wicri:Area/Ncbi/Curation">001867</idno><idno type="wicri:Area/Ncbi/Checkpoint">001867</idno><idno type="wicri:Area/Main/Merge">000B29</idno><idno type="wicri:Area/Main/Curation">000B27</idno><idno type="wicri:Area/Main/Exploration">000B27</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Double-Stranded RNA Uptake through Topical Application, Mediates Silencing of Five CYP4 Genes and Suppresses Insecticide Resistance in <italic>Diaphorina citri</italic></title><author><name sortKey="Killiny, Nabil" sort="Killiny, Nabil" uniqKey="Killiny N" first="Nabil" last="Killiny">Nabil Killiny</name><affiliation wicri:level="2"><nlm:aff id="aff1"><addr-line>Department of Entomology and Nematology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Entomology and Nematology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida</wicri:regionArea><placeName><region type="state">Floride</region></placeName></affiliation></author><author><name sortKey="Hajeri, Subhas" sort="Hajeri, Subhas" uniqKey="Hajeri S" first="Subhas" last="Hajeri">Subhas Hajeri</name><affiliation wicri:level="2"><nlm:aff id="aff2"><addr-line>Department of Plant Pathology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Plant Pathology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida</wicri:regionArea><placeName><region type="state">Floride</region></placeName></affiliation></author><author><name sortKey="Tiwari, Siddharth" sort="Tiwari, Siddharth" uniqKey="Tiwari S" first="Siddharth" last="Tiwari">Siddharth Tiwari</name><affiliation wicri:level="2"><nlm:aff id="aff1"><addr-line>Department of Entomology and Nematology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Entomology and Nematology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida</wicri:regionArea><placeName><region type="state">Floride</region></placeName></affiliation></author><author><name sortKey="Gowda, Siddarame" sort="Gowda, Siddarame" uniqKey="Gowda S" first="Siddarame" last="Gowda">Siddarame Gowda</name><affiliation wicri:level="2"><nlm:aff id="aff2"><addr-line>Department of Plant Pathology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Plant Pathology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida</wicri:regionArea><placeName><region type="state">Floride</region></placeName></affiliation></author><author><name sortKey="Stelinski, Lukasz L" sort="Stelinski, Lukasz L" uniqKey="Stelinski L" first="Lukasz L." last="Stelinski">Lukasz L. Stelinski</name><affiliation wicri:level="2"><nlm:aff id="aff1"><addr-line>Department of Entomology and Nematology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Entomology and Nematology, Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, Florida</wicri:regionArea><placeName><region type="state">Floride</region></placeName></affiliation></author></analytic><series><title level="j">PLoS ONE</title><idno type="eISSN">1932-6203</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Silencing of genes through RNA interference (RNAi) in insects has gained momentum during the past few years. RNAi has been used to cause insect mortality, inhibit insect growth, increase insecticide susceptibility, and prevent the development of insecticide resistance. We investigated the efficacy of topically applied dsRNA to induce RNAi for five Cytochrome P<sub>450</sub> genes family 4 (<italic>CYP4</italic>) in <italic>Diaphorina citri</italic>. We previously reported that these <italic>CYP4</italic> genes are associated with the development of insecticide resistance in <italic>D. citri</italic>. We targeted five <italic>CYP4</italic> genes that share a consensus sequence with one dsRNA construct. Quantitative PCR confirmed suppressed expression of the five <italic>CYP4</italic> genes as a result of dsRNA topically applied to the thoracic region of <italic>D. citri</italic> when compared to the expression levels in a control group. Western blot analysis indicated a reduced signal of cytochrome P<sub>450</sub> proteins (45 kDa) in adult <italic>D. citri</italic> treated with the dsRNA. In addition, oxidase activity and insecticide resistance were reduced for <italic>D. citri</italic> treated with dsRNA that targeted specific <italic>CYP4</italic> genes. Mortality was significantly higher in adults treated with dsRNA than in adults treated with water. Our results indicate that topically applied dsRNA can penetrate the cuticle of <italic>D. citri</italic> and induce RNAi. These results broaden the scope of RNAi as a mechanism to manage pests by targeting a broad range of genes. The results also support the application of RNAi as a viable tool to overcome insecticide resistance development in <italic>D. citri</italic> populations. However, further research is needed to develop grower-friendly delivery systems for the application of dsRNA under field conditions. Considering the high specificity of dsRNA, this tool can also be used for management of <italic>D. citri</italic> by targeting physiologically critical genes involved in growth and development.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Huvenne, H" uniqKey="Huvenne H">H Huvenne</name></author><author><name sortKey="Smagghe, G" uniqKey="Smagghe G">G Smagghe</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Izant, Jg" uniqKey="Izant J">JG Izant</name></author><author><name sortKey="Weintraub, H" uniqKey="Weintraub H">H Weintraub</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Fire, A" uniqKey="Fire A">A Fire</name></author><author><name sortKey="Xu, S" uniqKey="Xu S">S Xu</name></author><author><name sortKey="Montgomery, Mk" uniqKey="Montgomery M">MK Montgomery</name></author><author><name sortKey="Kostas, Sa" uniqKey="Kostas S">SA 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Tao</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Baum, Ja" uniqKey="Baum J">JA Baum</name></author><author><name sortKey="Bogaert, T" uniqKey="Bogaert T">T Bogaert</name></author><author><name sortKey="Clinton, W" uniqKey="Clinton W">W Clinton</name></author><author><name sortKey="Heck, Gr" uniqKey="Heck G">GR Heck</name></author><author><name sortKey="Feldmann, P" uniqKey="Feldmann P">P Feldmann</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Clemens, Mj" uniqKey="Clemens M">MJ Clemens</name></author><author><name sortKey="Elia, A" uniqKey="Elia A">A Elia</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Floride</li></region></list><tree><country name="États-Unis"><region name="Floride"><name sortKey="Killiny, Nabil" sort="Killiny, Nabil" uniqKey="Killiny N" first="Nabil" last="Killiny">Nabil Killiny</name></region><name sortKey="Gowda, Siddarame" sort="Gowda, Siddarame" uniqKey="Gowda S" first="Siddarame" last="Gowda">Siddarame Gowda</name><name sortKey="Hajeri, Subhas" sort="Hajeri, Subhas" uniqKey="Hajeri S" first="Subhas" last="Hajeri">Subhas Hajeri</name><name sortKey="Stelinski, Lukasz L" sort="Stelinski, Lukasz L" uniqKey="Stelinski L" first="Lukasz L." last="Stelinski">Lukasz L. Stelinski</name><name sortKey="Tiwari, Siddharth" sort="Tiwari, Siddharth" uniqKey="Tiwari S" first="Siddharth" last="Tiwari">Siddharth Tiwari</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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