Towards an understanding of the molecular basis of effective RNAi against a global insect pest, the whitefly Bemisia tabaci
In planta RNAi against essential insect genes offers a promising route to control insect crop pests, but is constrained for many insect groups, notably phloem sap-feeding hemipterans, by poor RNAi efficacy. This study conducted on the phloem-feeding whitefly Bemisia tabaci reared on tomato plants in...
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| Veröffentlicht in: | Insect biochemistry and molecular biology Jg. 88; S. 21 - 29 |
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| Format: | Journal Article |
| Sprache: | Englisch |
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England
Elsevier Ltd
01.09.2017
Elsevier Science |
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| ISSN: | 0965-1748, 1879-0240, 1879-0240 |
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| Abstract | In planta RNAi against essential insect genes offers a promising route to control insect crop pests, but is constrained for many insect groups, notably phloem sap-feeding hemipterans, by poor RNAi efficacy. This study conducted on the phloem-feeding whitefly Bemisia tabaci reared on tomato plants investigated the causes of low RNAi efficacy and routes to ameliorate the problem. Experiments using tomato transgenic lines containing ds-GFP (green fluorescent protein) revealed that full-length dsRNA is phloem-mobile, ingested by the insects, and degraded in the insect. We identified B. tabaci homologs of nuclease genes (dsRNases) in other insects that degrade dsRNA, and demonstrated that degradation of ds-GFP in B. tabaci is suppressed by administration of dsRNA against these genes. dsRNA against the nuclease genes was co-administered with dsRNA against two insect genes, an aquaporin AQP1 and sucrase SUC1, that are predicted to protect B. tabaci against osmotic collapse. When dsRNA constructs for AQP1, SUC1, dsRNase1 and dsRNase2 were stacked, insect mortality was significantly elevated to 50% over 6 days on artificial diets. This effect was accompanied by significant reduction in gene expression of the target genes in surviving diet-fed insects. This study offers proof-of-principle that the efficacy of RNAi against insect pests can be enhanced by using dsRNA to suppress the activity of RNAi-suppressing nuclease genes, especially where multiple genes with related physiological function but different molecular function are targeted.
[Display omitted]
•Tomato plants transformed with dsRNA deliver full-length dsRNA to the phloem-feeding whitefly Bemisia tabaci.•Orally-acquired dsRNA is degraded in the body of the whitefly B. tabaci.•The B. tabaci genome encodes three non-specific nucleases, two of which are expressed in the gut.•RNAi-mediated suppression of nuclease expression reduces degradation of other dsRNA molecules ingested by whiteflies.•Whitefly mortality on diets is enhanced by stacking oral RNAi against insect osmoregulation genes and nucleases. |
|---|---|
| AbstractList | In planta RNAi against essential insect genes offers a promising route to control insect crop pests, but is constrained for many insect groups, notably phloem sap-feeding hemipterans, by poor RNAi efficacy. This study conducted on the phloem-feeding whitefly Bemisia tabaci reared on tomato plants investigated the causes of low RNAi efficacy and routes to ameliorate the problem. Experiments using tomato transgenic lines containing ds-GFP (green fluorescent protein) revealed that full-length dsRNA is phloem-mobile, ingested by the insects, and degraded in the insect. We identified B. tabaci homologs of nuclease genes (dsRNases) in other insects that degrade dsRNA, and demonstrated that degradation of ds-GFP in B. tabaci is suppressed by administration of dsRNA against these genes. dsRNA against the nuclease genes was co-administered with dsRNA against two insect genes, an aquaporin AQP1 and sucrase SUC1, that are predicted to protect B. tabaci against osmotic collapse. When dsRNA constructs for AQP1, SUC1, dsRNase1 and dsRNase2 were stacked, insect mortality was significantly elevated to 50% over 6 days on artificial diets. This effect was accompanied by significant reduction in gene expression of the target genes in surviving diet-fed insects. This study offers proof-of-principle that the efficacy of RNAi against insect pests can be enhanced by using dsRNA to suppress the activity of RNAi-suppressing nuclease genes, especially where multiple genes with related physiological function but different molecular function are targeted.
Image 1
•
Tomato plants transformed with dsRNA deliver full-length dsRNA to the phloem-feeding whitefly Bemisia tabaci.
•
Orally-acquired dsRNA is degraded in the body of the whitefly B. tabaci.
•
The B. tabaci genome encodes three non-specific nucleases, two of which are expressed in the gut.
•
RNAi-mediated suppression of nuclease expression reduces degradation of other dsRNA molecules ingested by whiteflies.
•
Whitefly mortality on diets is enhanced by stacking oral RNAi against insect osmoregulation genes and nucleases. In planta RNAi against essential insect genes offers a promising route to control insect crop pests, but is constrained for many insect groups, notably phloem sap-feeding hemipterans, by poor RNAi efficacy. This study conducted on the phloem-feeding whitefly Bemisia tabaci reared on tomato plants investigated the causes of low RNAi efficacy and routes to ameliorate the problem. Experiments using tomato transgenic lines containing ds-GFP (green fluorescent protein) revealed that full-length dsRNA is phloem-mobile, ingested by the insects, and degraded in the insect. We identified B. tabaci homologs of nuclease genes (dsRNases) in other insects that degrade dsRNA, and demonstrated that degradation of ds-GFP in B. tabaci is suppressed by administration of dsRNA against these genes. dsRNA against the nuclease genes was co-administered with dsRNA against two insect genes, an aquaporin AQP1 and sucrase SUC1, that are predicted to protect B. tabaci against osmotic collapse. When dsRNA constructs for AQP1, SUC1, dsRNase1 and dsRNase2 were stacked, insect mortality was significantly elevated to 50% over 6 days on artificial diets. This effect was accompanied by significant reduction in gene expression of the target genes in surviving diet-fed insects. This study offers proof-of-principle that the efficacy of RNAi against insect pests can be enhanced by using dsRNA to suppress the activity of RNAi-suppressing nuclease genes, especially where multiple genes with related physiological function but different molecular function are targeted. In planta RNAi against essential insect genes offers a promising route to control insect crop pests, but is constrained for many insect groups, notably phloem sap-feeding hemipterans, by poor RNAi efficacy. This study conducted on the phloem-feeding whitefly Bemisia tabaci reared on tomato plants investigated the causes of low RNAi efficacy and routes to ameliorate the problem. Experiments using tomato transgenic lines containing ds-GFP (green fluorescent protein) revealed that full-length dsRNA is phloem-mobile, ingested by the insects, and degraded in the insect. We identified B. tabaci homologs of nuclease genes (dsRNases) in other insects that degrade dsRNA, and demonstrated that degradation of ds-GFP in B. tabaci is suppressed by administration of dsRNA against these genes. dsRNA against the nuclease genes was co-administered with dsRNA against two insect genes, an aquaporin AQP1 and sucrase SUC1, that are predicted to protect B. tabaci against osmotic collapse. When dsRNA constructs for AQP1, SUC1, dsRNase1 and dsRNase2 were stacked, insect mortality was significantly elevated to 50% over 6 days on artificial diets. This effect was accompanied by significant reduction in gene expression of the target genes in surviving diet-fed insects. This study offers proof-of-principle that the efficacy of RNAi against insect pests can be enhanced by using dsRNA to suppress the activity of RNAi-suppressing nuclease genes, especially where multiple genes with related physiological function but different molecular function are targeted. In planta RNAi against essential insect genes offers a promising route to control insect crop pests, but is constrained for many insect groups, notably phloem sap-feeding hemipterans, by poor RNAi efficacy. This study conducted on the phloem-feeding whitefly Bemisia tabaci reared on tomato plants investigated the causes of low RNAi efficacy and routes to ameliorate the problem. Experiments using tomato transgenic lines containing ds-GFP (green fluorescent protein) revealed that full-length dsRNA is phloem-mobile, ingested by the insects, and degraded in the insect. We identified B. tabaci homologs of nuclease genes (dsRNases) in other insects that degrade dsRNA, and demonstrated that degradation of ds-GFP in B. tabaci is suppressed by administration of dsRNA against these genes. dsRNA against the nuclease genes was co-administered with dsRNA against two insect genes, an aquaporin AQP1 and sucrase SUC1, that are predicted to protect B. tabaci against osmotic collapse. When dsRNA constructs for AQP1, SUC1, dsRNase1 and dsRNase2 were stacked, insect mortality was significantly elevated to 50% over 6 days on artificial diets. This effect was accompanied by significant reduction in gene expression of the target genes in surviving diet-fed insects. This study offers proof-of-principle that the efficacy of RNAi against insect pests can be enhanced by using dsRNA to suppress the activity of RNAi-suppressing nuclease genes, especially where multiple genes with related physiological function but different molecular function are targeted. [Display omitted] •Tomato plants transformed with dsRNA deliver full-length dsRNA to the phloem-feeding whitefly Bemisia tabaci.•Orally-acquired dsRNA is degraded in the body of the whitefly B. tabaci.•The B. tabaci genome encodes three non-specific nucleases, two of which are expressed in the gut.•RNAi-mediated suppression of nuclease expression reduces degradation of other dsRNA molecules ingested by whiteflies.•Whitefly mortality on diets is enhanced by stacking oral RNAi against insect osmoregulation genes and nucleases. In planta RNAi against essential insect genes offers a promising route to control insect crop pests, but is constrained for many insect groups, notably phloem sap-feeding hemipterans, by poor RNAi efficacy. This study conducted on the phloem-feeding whitefly Bemisia tabaci reared on tomato plants investigated the causes of low RNAi efficacy and routes to ameliorate the problem. Experiments using tomato transgenic lines containing ds-GFP (green fluorescent protein) revealed that full-length dsRNA is phloem-mobile, ingested by the insects, and degraded in the insect. We identified B. tabaci homologs of nuclease genes (dsRNases) in other insects that degrade dsRNA, and demonstrated that degradation of ds-GFP in B. tabaci is suppressed by administration of dsRNA against these genes. dsRNA against the nuclease genes was co-administered with dsRNA against two insect genes, an aquaporin AQP1 and sucrase SUC1, that are predicted to protect B. tabaci against osmotic collapse. When dsRNA constructs for AQP1, SUC1, dsRNase1 and dsRNase2 were stacked, insect mortality was significantly elevated to 50% over 6 days on artificial diets. This effect was accompanied by significant reduction in gene expression of the target genes in surviving diet-fed insects. This study offers proof-of-principle that the efficacy of RNAi against insect pests can be enhanced by using dsRNA to suppress the activity of RNAi-suppressing nuclease genes, especially where multiple genes with related physiological function but different molecular function are targeted.In planta RNAi against essential insect genes offers a promising route to control insect crop pests, but is constrained for many insect groups, notably phloem sap-feeding hemipterans, by poor RNAi efficacy. This study conducted on the phloem-feeding whitefly Bemisia tabaci reared on tomato plants investigated the causes of low RNAi efficacy and routes to ameliorate the problem. Experiments using tomato transgenic lines containing ds-GFP (green fluorescent protein) revealed that full-length dsRNA is phloem-mobile, ingested by the insects, and degraded in the insect. We identified B. tabaci homologs of nuclease genes (dsRNases) in other insects that degrade dsRNA, and demonstrated that degradation of ds-GFP in B. tabaci is suppressed by administration of dsRNA against these genes. dsRNA against the nuclease genes was co-administered with dsRNA against two insect genes, an aquaporin AQP1 and sucrase SUC1, that are predicted to protect B. tabaci against osmotic collapse. When dsRNA constructs for AQP1, SUC1, dsRNase1 and dsRNase2 were stacked, insect mortality was significantly elevated to 50% over 6 days on artificial diets. This effect was accompanied by significant reduction in gene expression of the target genes in surviving diet-fed insects. This study offers proof-of-principle that the efficacy of RNAi against insect pests can be enhanced by using dsRNA to suppress the activity of RNAi-suppressing nuclease genes, especially where multiple genes with related physiological function but different molecular function are targeted. |
| Author | Turgeon, R. Douglas, Angela E. Chung, Seung Ho Luo, Yuan Chen, Qingguo Van Eck, Joyce Luan, Junbo |
| AuthorAffiliation | b Plant Biology Section, School of Integrative Plant Science, Cornell University, Ithaca, NY 14853, USA d Department of Molecular Biology & Genetics, Cornell University, Ithaca, NY 14853, USA a Department of Entomology, Cornell University, Ithaca, NY 14853, USA c Boyce Thompson Institute, Ithaca, NY 14853, USA |
| AuthorAffiliation_xml | – name: d Department of Molecular Biology & Genetics, Cornell University, Ithaca, NY 14853, USA – name: b Plant Biology Section, School of Integrative Plant Science, Cornell University, Ithaca, NY 14853, USA – name: a Department of Entomology, Cornell University, Ithaca, NY 14853, USA – name: c Boyce Thompson Institute, Ithaca, NY 14853, USA |
| Author_xml | – sequence: 1 givenname: Yuan surname: Luo fullname: Luo, Yuan organization: Department of Entomology, Cornell University, Ithaca, NY 14853, USA – sequence: 2 givenname: Qingguo surname: Chen fullname: Chen, Qingguo organization: Plant Biology Section, School of Integrative Plant Science, Cornell University, Ithaca, NY 14853, USA – sequence: 3 givenname: Junbo surname: Luan fullname: Luan, Junbo organization: Department of Entomology, Cornell University, Ithaca, NY 14853, USA – sequence: 4 givenname: Seung Ho surname: Chung fullname: Chung, Seung Ho organization: Department of Entomology, Cornell University, Ithaca, NY 14853, USA – sequence: 5 givenname: Joyce orcidid: 0000-0002-8005-365X surname: Van Eck fullname: Van Eck, Joyce organization: Boyce Thompson Institute, Ithaca, NY 14853, USA – sequence: 6 givenname: R. surname: Turgeon fullname: Turgeon, R. organization: Plant Biology Section, School of Integrative Plant Science, Cornell University, Ithaca, NY 14853, USA – sequence: 7 givenname: Angela E. surname: Douglas fullname: Douglas, Angela E. email: aes326@cornell.edu organization: Department of Entomology, Cornell University, Ithaca, NY 14853, USA |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/28736300$$D View this record in MEDLINE/PubMed |
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| Keywords | RNAi efficacy RNA interference Bemisia tabaci dsRNA degradation |
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| SubjectTerms | Animals aquaporins artificial diets Bemisia tabaci double-stranded RNA dsRNA degradation Female genes genetically modified organisms Genome, Insect green fluorescent protein Hemiptera - genetics Hemiptera - metabolism Insect Control - methods insect pests Male mortality Osmoregulation - genetics phloem plant pests rearing RNA Interference RNA, Double-Stranded - pharmacokinetics RNAi efficacy sucrose alpha-glucosidase tomatoes |
| Title | Towards an understanding of the molecular basis of effective RNAi against a global insect pest, the whitefly Bemisia tabaci |
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