Molecular Characterizations of Double-Stranded RNA Degrading Nuclease Genes from <i>Ostrinia nubilalis</i>
Variable RNA interference (RNAi) efficiencies limit RNAi-based pest management strategies for many pests. Previous efforts to understand mechanisms contributing to low RNAi efficiency indicate that double-stranded RNA (dsRNA) is degraded in the European corn borer (ECB), <i>Ostrinia nubilalis,...
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| Format: | Article |
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MDPI AG
2020-09-01
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| Series: | Insects |
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| Online Access: | https://www.mdpi.com/2075-4450/11/10/652 |
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| author | Anastasia M. W. Cooper Huifang Song Xuekai Shi Zhitao Yu Marcé Lorenzen Kristopher Silver Jianzhen Zhang Kun Yan Zhu |
| author_facet | Anastasia M. W. Cooper Huifang Song Xuekai Shi Zhitao Yu Marcé Lorenzen Kristopher Silver Jianzhen Zhang Kun Yan Zhu |
| author_sort | Anastasia M. W. Cooper |
| collection | DOAJ |
| description | Variable RNA interference (RNAi) efficiencies limit RNAi-based pest management strategies for many pests. Previous efforts to understand mechanisms contributing to low RNAi efficiency indicate that double-stranded RNA (dsRNA) is degraded in the European corn borer (ECB), <i>Ostrinia nubilalis,</i> due to nuclease activity. To investigate the contribution of dsRNA-degrading endonucleases (dsRNases) and lepidopteran-specific RNAi efficiency-related nucleases (REases) to dsRNA instability and low RNAi efficiency in ECB, five complementary DNAs putatively encoding four dsRNases (<i>OndsRNase1, 2, 3,</i> and <i>4</i>) and one REase (<i>OnREase</i>) were sequenced. Characterization of these transcripts revealed that substrate specificity might vary among the four dsRNases due to different amino acid combinations in the substrate-binding sites. Gene expression analysis indicated that <i>OndsRNase2</i> and <i>OnREase</i> were highly expressed in the larval gut, and <i>OndsRNase1</i> showed the highest expression in hemolymph, especially in older developmental stages. Transcript level analysis after dsRNA exposure revealed that expression of <i>OnREase</i> rapidly increased upon dsRNA ingestion or injection, whereas <i>OndsRNase4</i> expression only increased after long-term ingestion of dsRNA. While the biological function of these nucleases remains to be verified, our results suggest that OnREase and OndsRNase2, and OndsRNase1 and OndsRNase4 may be responsible for degradation of dsRNAs in the ECB gut and hemolymph, respectively, thereby contributing to low RNAi efficiency. |
| first_indexed | 2024-03-10T16:06:18Z |
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| institution | Directory Open Access Journal |
| issn | 2075-4450 |
| language | English |
| last_indexed | 2024-03-10T16:06:18Z |
| publishDate | 2020-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Insects |
| spelling | doaj.art-0e143fa540c4482494a788e6caecdce52023-11-20T14:48:19ZengMDPI AGInsects2075-44502020-09-01111065210.3390/insects11100652Molecular Characterizations of Double-Stranded RNA Degrading Nuclease Genes from <i>Ostrinia nubilalis</i>Anastasia M. W. Cooper0Huifang Song1Xuekai Shi2Zhitao Yu3Marcé Lorenzen4Kristopher Silver5Jianzhen Zhang6Kun Yan Zhu7Department of Entomology, 123 Waters Hall, Kansas State University, Manhattan, KS 66506, USADepartment of Entomology, 123 Waters Hall, Kansas State University, Manhattan, KS 66506, USADepartment of Entomology, 123 Waters Hall, Kansas State University, Manhattan, KS 66506, USADepartment of Entomology, 123 Waters Hall, Kansas State University, Manhattan, KS 66506, USADepartment of Entomology and Plant Pathology, Campus Box 7613, North Carolina State University, Raleigh, NC 27695, USADepartment of Entomology, 123 Waters Hall, Kansas State University, Manhattan, KS 66506, USADepartment of Entomology, 123 Waters Hall, Kansas State University, Manhattan, KS 66506, USADepartment of Entomology, 123 Waters Hall, Kansas State University, Manhattan, KS 66506, USAVariable RNA interference (RNAi) efficiencies limit RNAi-based pest management strategies for many pests. Previous efforts to understand mechanisms contributing to low RNAi efficiency indicate that double-stranded RNA (dsRNA) is degraded in the European corn borer (ECB), <i>Ostrinia nubilalis,</i> due to nuclease activity. To investigate the contribution of dsRNA-degrading endonucleases (dsRNases) and lepidopteran-specific RNAi efficiency-related nucleases (REases) to dsRNA instability and low RNAi efficiency in ECB, five complementary DNAs putatively encoding four dsRNases (<i>OndsRNase1, 2, 3,</i> and <i>4</i>) and one REase (<i>OnREase</i>) were sequenced. Characterization of these transcripts revealed that substrate specificity might vary among the four dsRNases due to different amino acid combinations in the substrate-binding sites. Gene expression analysis indicated that <i>OndsRNase2</i> and <i>OnREase</i> were highly expressed in the larval gut, and <i>OndsRNase1</i> showed the highest expression in hemolymph, especially in older developmental stages. Transcript level analysis after dsRNA exposure revealed that expression of <i>OnREase</i> rapidly increased upon dsRNA ingestion or injection, whereas <i>OndsRNase4</i> expression only increased after long-term ingestion of dsRNA. While the biological function of these nucleases remains to be verified, our results suggest that OnREase and OndsRNase2, and OndsRNase1 and OndsRNase4 may be responsible for degradation of dsRNAs in the ECB gut and hemolymph, respectively, thereby contributing to low RNAi efficiency.https://www.mdpi.com/2075-4450/11/10/652dsRNaseEuropean corn borergene expressionREaseRNAi efficiencysubstrate specificity |
| spellingShingle | Anastasia M. W. Cooper Huifang Song Xuekai Shi Zhitao Yu Marcé Lorenzen Kristopher Silver Jianzhen Zhang Kun Yan Zhu Molecular Characterizations of Double-Stranded RNA Degrading Nuclease Genes from <i>Ostrinia nubilalis</i> Insects dsRNase European corn borer gene expression REase RNAi efficiency substrate specificity |
| title | Molecular Characterizations of Double-Stranded RNA Degrading Nuclease Genes from <i>Ostrinia nubilalis</i> |
| title_full | Molecular Characterizations of Double-Stranded RNA Degrading Nuclease Genes from <i>Ostrinia nubilalis</i> |
| title_fullStr | Molecular Characterizations of Double-Stranded RNA Degrading Nuclease Genes from <i>Ostrinia nubilalis</i> |
| title_full_unstemmed | Molecular Characterizations of Double-Stranded RNA Degrading Nuclease Genes from <i>Ostrinia nubilalis</i> |
| title_short | Molecular Characterizations of Double-Stranded RNA Degrading Nuclease Genes from <i>Ostrinia nubilalis</i> |
| title_sort | molecular characterizations of double stranded rna degrading nuclease genes from i ostrinia nubilalis i |
| topic | dsRNase European corn borer gene expression REase RNAi efficiency substrate specificity |
| url | https://www.mdpi.com/2075-4450/11/10/652 |
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