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Development of RNAi Methods for Peregrinus maidis, the Corn Planthopper
The corn planthopper, Peregrinus maidis, is a major pest of agronomically-important crops. Peregrinus maidis has a large geographical distribution and transmits Maize mosaic rhabdovirus (MMV) and Maize stripe tenuivirus (MSpV). The objective of this study was to develop effective RNAi methods for P....
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3737204/ https://www.ncbi.nlm.nih.gov/pubmed/23950915 http://dx.doi.org/10.1371/journal.pone.0070243 |
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author | Yao, Jianxiu Rotenberg, Dorith Afsharifar, Alireza Barandoc-Alviar, Karen Whitfield, Anna E. |
author_facet | Yao, Jianxiu Rotenberg, Dorith Afsharifar, Alireza Barandoc-Alviar, Karen Whitfield, Anna E. |
author_sort | Yao, Jianxiu |
collection | PubMed |
description | The corn planthopper, Peregrinus maidis, is a major pest of agronomically-important crops. Peregrinus maidis has a large geographical distribution and transmits Maize mosaic rhabdovirus (MMV) and Maize stripe tenuivirus (MSpV). The objective of this study was to develop effective RNAi methods for P. maidis. Vacuolar-ATPase (V-ATPase) is an essential enzyme for hydrolysis of ATP and for transport of protons out of cells thereby maintaining membrane ion balance, and it has been demonstrated to be an efficacious target for RNAi in other insects. In this study, two genes encoding subunits of P. maidis V-ATPase (V-ATPase B and V-ATPase D) were chosen as RNAi target genes. The open reading frames of V-ATPase B and D were generated and used for constructing dsRNA fragments. Experiments were conducted using oral delivery and microinjection of V-ATPase B and V-ATPase D dsRNA to investigate the effectiveness of RNAi in P. maidis. Real-time quantitative reverse transcriptase-PCR (qRT-PCR) analysis indicated that microinjection of V-ATPase dsRNA led to a minimum reduction of 27-fold in the normalized abundance of V-ATPase transcripts two days post injection, while ingestion of dsRNA resulted in a two-fold reduction after six days of feeding. While both methods of dsRNA delivery resulted in knockdown of target transcripts, the injection method was more rapid and effective. The reduction in V-ATPase transcript abundance resulted in observable phenotypes. Specifically, the development of nymphs injected with 200 ng of either V-ATPase B or D dsRNA was impaired, resulting in higher mortality and lower fecundity than control insects injected with GFP dsRNA. Microscopic examination of these insects revealed that female reproductive organs did not develop normally. The successful development of RNAi in P. maidis to target specific genes will enable the development of new insect control strategies and functional analysis of vital genes and genes associated with interactions between P. maidis and MMV. |
format | Online Article Text |
id | pubmed-3737204 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-37372042013-08-15 Development of RNAi Methods for Peregrinus maidis, the Corn Planthopper Yao, Jianxiu Rotenberg, Dorith Afsharifar, Alireza Barandoc-Alviar, Karen Whitfield, Anna E. PLoS One Research Article The corn planthopper, Peregrinus maidis, is a major pest of agronomically-important crops. Peregrinus maidis has a large geographical distribution and transmits Maize mosaic rhabdovirus (MMV) and Maize stripe tenuivirus (MSpV). The objective of this study was to develop effective RNAi methods for P. maidis. Vacuolar-ATPase (V-ATPase) is an essential enzyme for hydrolysis of ATP and for transport of protons out of cells thereby maintaining membrane ion balance, and it has been demonstrated to be an efficacious target for RNAi in other insects. In this study, two genes encoding subunits of P. maidis V-ATPase (V-ATPase B and V-ATPase D) were chosen as RNAi target genes. The open reading frames of V-ATPase B and D were generated and used for constructing dsRNA fragments. Experiments were conducted using oral delivery and microinjection of V-ATPase B and V-ATPase D dsRNA to investigate the effectiveness of RNAi in P. maidis. Real-time quantitative reverse transcriptase-PCR (qRT-PCR) analysis indicated that microinjection of V-ATPase dsRNA led to a minimum reduction of 27-fold in the normalized abundance of V-ATPase transcripts two days post injection, while ingestion of dsRNA resulted in a two-fold reduction after six days of feeding. While both methods of dsRNA delivery resulted in knockdown of target transcripts, the injection method was more rapid and effective. The reduction in V-ATPase transcript abundance resulted in observable phenotypes. Specifically, the development of nymphs injected with 200 ng of either V-ATPase B or D dsRNA was impaired, resulting in higher mortality and lower fecundity than control insects injected with GFP dsRNA. Microscopic examination of these insects revealed that female reproductive organs did not develop normally. The successful development of RNAi in P. maidis to target specific genes will enable the development of new insect control strategies and functional analysis of vital genes and genes associated with interactions between P. maidis and MMV. Public Library of Science 2013-08-07 /pmc/articles/PMC3737204/ /pubmed/23950915 http://dx.doi.org/10.1371/journal.pone.0070243 Text en © 2013 Yao et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Yao, Jianxiu Rotenberg, Dorith Afsharifar, Alireza Barandoc-Alviar, Karen Whitfield, Anna E. Development of RNAi Methods for Peregrinus maidis, the Corn Planthopper |
title | Development of RNAi Methods for Peregrinus maidis, the Corn Planthopper |
title_full | Development of RNAi Methods for Peregrinus maidis, the Corn Planthopper |
title_fullStr | Development of RNAi Methods for Peregrinus maidis, the Corn Planthopper |
title_full_unstemmed | Development of RNAi Methods for Peregrinus maidis, the Corn Planthopper |
title_short | Development of RNAi Methods for Peregrinus maidis, the Corn Planthopper |
title_sort | development of rnai methods for peregrinus maidis, the corn planthopper |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3737204/ https://www.ncbi.nlm.nih.gov/pubmed/23950915 http://dx.doi.org/10.1371/journal.pone.0070243 |
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