Knock down of Whitefly Gut Gene Expression and Mortality by Orally Delivered Gut Gene-Specific dsRNAs
Control of the whitefly Bemisia tabaci (Genn.) agricultural pest and plant virus vector relies on the use of chemical insecticides. RNA-interference (RNAi) is a homology-dependent innate immune response in eukaryotes, including insects, which results in degradation of the corresponding transcript fo...
Main Authors: | , , , , , , |
---|---|
Other Authors: | |
Language: | en |
Published: |
PUBLIC LIBRARY SCIENCE
2017
|
Online Access: | http://hdl.handle.net/10150/622632 http://arizona.openrepository.com/arizona/handle/10150/622632 |
id |
ndltd-arizona.edu-oai-arizona.openrepository.com-10150-622632 |
---|---|
record_format |
oai_dc |
spelling |
ndltd-arizona.edu-oai-arizona.openrepository.com-10150-6226322017-02-23T03:00:38Z Knock down of Whitefly Gut Gene Expression and Mortality by Orally Delivered Gut Gene-Specific dsRNAs Vyas, Meenal Raza, Amir Ali, Muhammad Yousaf Ashraf, Muhammad Aleem Mansoor, Shahid Shahid, Ahmad Ali Brown, Judith K. Univ Arizona, Sch Plant Sci Control of the whitefly Bemisia tabaci (Genn.) agricultural pest and plant virus vector relies on the use of chemical insecticides. RNA-interference (RNAi) is a homology-dependent innate immune response in eukaryotes, including insects, which results in degradation of the corresponding transcript following its recognition by a double-stranded RNA (dsRNA) that shares 100% sequence homology. In this study, six whitefly `gut' genes were selected from an in silico-annotated transcriptome library constructed from the whitefly alimentary canal or 'gut' of the B biotype of B. tabaci, and tested for knock down efficacy, post-ingestion of dsRNAs that share 100% sequence homology to each respective gene target. Candidate genes were: Acetylcholine receptor subunit a, Alpha glucosidase 1, Aquaporin 1, Heat shock protein 70, Trehalasel, and Trehalose transported. The efficacy of RNAi knock down was further tested in a gene-specific functional bioassay, and mortality was recorded in 24 hr intervals, six days, post-treatment. Based on qPCR analysis, all six genes tested showed significantly reduced gene expression. Moderate-to-high whitefly mortality was associated with the down-regulation of osmoregulation, sugar metabolism and sugar transport -associated genes, demonstrating that whitefly survivability was linked with RNAi results. Silenced Acetylcholine receptor subunit a and Heat shock protein 70 genes showed an initial low whitefly mortality, however, following insecticide or high temperature treatments, respectively, significantly increased knockdown efficacy and death was observed, indicating enhanced post-knockdown sensitivity perhaps related to systemic silencing. The oral delivery of gut-specific dsRNAs, when combined with qPCR analysis of gene expression and a corresponding gene-specific bioassay that relates knockdown and mortality, offers a viable approach for functional genomics analysis and the discovery of prospective dsRNA biopesticide targets. The approach can be applied to functional genomics analyses to facilitate, species-specific dsRNA-mediated control of other non-model hemipterans. 2017-01-03 Article Knock down of Whitefly Gut Gene Expression and Mortality by Orally Delivered Gut Gene-Specific dsRNAs 2017, 12 (1):e0168921 PLOS ONE 1932-6203 10.1371/journal.pone.0168921 http://hdl.handle.net/10150/622632 http://arizona.openrepository.com/arizona/handle/10150/622632 PLOS ONE en http://dx.plos.org/10.1371/journal.pone.0168921 © 2017 Vyas et al. This is an open access article distributed under the terms of the Creative Commons Attribution License PUBLIC LIBRARY SCIENCE |
collection |
NDLTD |
language |
en |
sources |
NDLTD |
description |
Control of the whitefly Bemisia tabaci (Genn.) agricultural pest and plant virus vector relies on the use of chemical insecticides. RNA-interference (RNAi) is a homology-dependent innate immune response in eukaryotes, including insects, which results in degradation of the corresponding transcript following its recognition by a double-stranded RNA (dsRNA) that shares 100% sequence homology. In this study, six whitefly `gut' genes were selected from an in silico-annotated transcriptome library constructed from the whitefly alimentary canal or 'gut' of the B biotype of B. tabaci, and tested for knock down efficacy, post-ingestion of dsRNAs that share 100% sequence homology to each respective gene target. Candidate genes were: Acetylcholine receptor subunit a, Alpha glucosidase 1, Aquaporin 1, Heat shock protein 70, Trehalasel, and Trehalose transported. The efficacy of RNAi knock down was further tested in a gene-specific functional bioassay, and mortality was recorded in 24 hr intervals, six days, post-treatment. Based on qPCR analysis, all six genes tested showed significantly reduced gene expression. Moderate-to-high whitefly mortality was associated with the down-regulation of osmoregulation, sugar metabolism and sugar transport -associated genes, demonstrating that whitefly survivability was linked with RNAi results. Silenced Acetylcholine receptor subunit a and Heat shock protein 70 genes showed an initial low whitefly mortality, however, following insecticide or high temperature treatments, respectively, significantly increased knockdown efficacy and death was observed, indicating enhanced post-knockdown sensitivity perhaps related to systemic silencing. The oral delivery of gut-specific dsRNAs, when combined with qPCR analysis of gene expression and a corresponding gene-specific bioassay that relates knockdown and mortality, offers a viable approach for functional genomics analysis and the discovery of prospective dsRNA biopesticide targets. The approach can be applied to functional genomics analyses to facilitate, species-specific dsRNA-mediated control of other non-model hemipterans. |
author2 |
Univ Arizona, Sch Plant Sci |
author_facet |
Univ Arizona, Sch Plant Sci Vyas, Meenal Raza, Amir Ali, Muhammad Yousaf Ashraf, Muhammad Aleem Mansoor, Shahid Shahid, Ahmad Ali Brown, Judith K. |
author |
Vyas, Meenal Raza, Amir Ali, Muhammad Yousaf Ashraf, Muhammad Aleem Mansoor, Shahid Shahid, Ahmad Ali Brown, Judith K. |
spellingShingle |
Vyas, Meenal Raza, Amir Ali, Muhammad Yousaf Ashraf, Muhammad Aleem Mansoor, Shahid Shahid, Ahmad Ali Brown, Judith K. Knock down of Whitefly Gut Gene Expression and Mortality by Orally Delivered Gut Gene-Specific dsRNAs |
author_sort |
Vyas, Meenal |
title |
Knock down of Whitefly Gut Gene Expression and Mortality by Orally Delivered Gut Gene-Specific dsRNAs |
title_short |
Knock down of Whitefly Gut Gene Expression and Mortality by Orally Delivered Gut Gene-Specific dsRNAs |
title_full |
Knock down of Whitefly Gut Gene Expression and Mortality by Orally Delivered Gut Gene-Specific dsRNAs |
title_fullStr |
Knock down of Whitefly Gut Gene Expression and Mortality by Orally Delivered Gut Gene-Specific dsRNAs |
title_full_unstemmed |
Knock down of Whitefly Gut Gene Expression and Mortality by Orally Delivered Gut Gene-Specific dsRNAs |
title_sort |
knock down of whitefly gut gene expression and mortality by orally delivered gut gene-specific dsrnas |
publisher |
PUBLIC LIBRARY SCIENCE |
publishDate |
2017 |
url |
http://hdl.handle.net/10150/622632 http://arizona.openrepository.com/arizona/handle/10150/622632 |
work_keys_str_mv |
AT vyasmeenal knockdownofwhiteflygutgeneexpressionandmortalitybyorallydeliveredgutgenespecificdsrnas AT razaamir knockdownofwhiteflygutgeneexpressionandmortalitybyorallydeliveredgutgenespecificdsrnas AT alimuhammadyousaf knockdownofwhiteflygutgeneexpressionandmortalitybyorallydeliveredgutgenespecificdsrnas AT ashrafmuhammadaleem knockdownofwhiteflygutgeneexpressionandmortalitybyorallydeliveredgutgenespecificdsrnas AT mansoorshahid knockdownofwhiteflygutgeneexpressionandmortalitybyorallydeliveredgutgenespecificdsrnas AT shahidahmadali knockdownofwhiteflygutgeneexpressionandmortalitybyorallydeliveredgutgenespecificdsrnas AT brownjudithk knockdownofwhiteflygutgeneexpressionandmortalitybyorallydeliveredgutgenespecificdsrnas |
_version_ |
1718416512878903296 |