Inoculation of Nicotiana tabacum with recombinant potato virus X induces RNA interference in the solenopsis mealybug, Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae)

Khan, Arif Muhammad; Ashfaq, Muhammad; Khan, Azhar Abbas; Rasool, Akhtar; Iqbal, Javed; Mansoor, Shahid

doi:10.1007/s10529-015-1880-7

Cited by 15 publications

(16 citation statements)

References 26 publications

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“…Therefore, given the level of sequence divergence between the corresponding M. persicae genes and the Be. tabaci ecdysone receptor and P. solenopsis chitin synthase constructs (Table S1), the RNAi effects on the expression levels of these genes are likely to be more minor than those observed with M. persicae cathepsin L. Moreover, we did not observe the morphological phenotypes that were previously reported when silencing ecdysone receptor or chitin synthase expression (Zhu et al , ; Khan et al , ; Shang et al , ; Yan et al , ; Vellichirammal et al , ; Zhao et al , ), and the identified protein accumulation deficit is more similar to what one would expect from silencing the expression of a gut protease.…”

Section: Discussionmentioning

confidence: 50%

“…Moreover, a C ‐> U mismatch at such a position is predicted to have a severe effect on RNAi efficiency (Ahmed and Raghava, ). Previous experiments involving expression silencing of chitin synthase in aphids and mealybugs resulted in physical deformities and/or the inability to moult (Khan et al , ; Shang et al , ; Zhao et al , ). We did not observe such morphological effects in our M. persicae experiments.…”

Section: Resultsmentioning

confidence: 99%

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Silencing cathepsin L expression reduces Myzus persicae protein content and the nutritional value as prey for Coccinella septempunctata

Rauf

Asif

Amin

et al. 2019

Insect Molecular Biology

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Gut‐expressed aphid genes, which may be more easily inhibited by RNA interference (RNAi) constructs, are attractive targets for pest control efforts involving transgenic plants. Here we show that expression of cathepsin L, which encodes a cysteine protease that functions in aphid guts, can be reduced by expression of an RNAi construct in transgenic tobacco. The effectiveness of this approach is demonstrated by up to 80% adult mortality, reduced fecundity, and delayed nymph production of Myzus persicae (green peach aphids) when cathepsin L expression was reduced by plant‐mediated RNAi. Consistent with the function of cathepsin L as a gut protease, M. persicae fed on the RNAi plants had a lower protein content in their bodies and excreted more protein and/or free amino acids in their honeydew. Larvae of Coccinella septempunctata (seven‐spotted ladybugs) grew more slowly on aphids having reduced cathepsin L expression, suggesting that prey insect nutritive value, and not just direct negative effects of the RNAi construct, needs to be considered when producing transgenic plants for RNAi‐mediated pest control.

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Section: Discussionmentioning

confidence: 50%

Section: Resultsmentioning

confidence: 99%

Silencing cathepsin L expression reduces Myzus persicae protein content and the nutritional value as prey for Coccinella septempunctata

Rauf

Asif

Amin

et al. 2019

Insect Molecular Biology

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“…Phenotypic effects and mortality in feeding mealy bugs indicated that RNAi triggers were present in the inoculated plants in sufficient amount to trigger RNAi when expressed through recombinant PVX. This observation is consistent with the previous studies (Wuriyanghan & Falk, ; Khan et al ., ) where insect siRNA from the phloem sap was recovered when expressed through recombinant plant virus (Pitino et al ., ).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of potential RNA‐interference‐target genes to control cotton mealybug, Phenacoccus solenopsis (Hemiptera: Pseudococcuidae)

et al. 2017

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RNA interference (RNAi) of vital insect genes is a potential tool for targeted pest control. However, selection of the right target genes is a challenge because the RNAi efficacy is known to vary among insect species. Cotton mealybug, Phenacoccus solenopsis, is a phloem-feeding economically important crop pest. We evaluated the RNAi of 2 vital genes, Bursicon (PsBur) and V-ATPase (PsV-ATPase) as potential targets in P. solenopsis for its control. PCR fragments of PsBur and PsV-ATPase were amplified using cDNA synthesized from the total RNA. The PCR amplicons were cloned into Potato virus X (PVX) to develop recombinant PVX for the inoculation of Nicotiana tabacum plants for bioassays with healthy P. solenopsis. Reverse-transcription-polymerase chain reaction (RT-PCR) was used to validate the expression of transgenes in the recombinant-PVX-inoculated plants (treated), and suppression of the target genes in the mealybugs exposed to them. The RT-PCR confirmed the expression of transgenes in the treated plants. Mealybug individuals on treated plants either died or showed physical deformities. Further, the population of mealybug was significantly reduced by feeding on N. tabacum expressing RNAi triggers against PsBur and PsV-ATPase. The results conclude that RNAi is activated in P. solenopsis by feeding on N. tabacum expressing RNAi triggering elements of PsBur and PsV-ATPase genes through recombinant PVX vector. Further, V-ATPase and Bursicon genes are potential targets for RNAi-mediated control of P. solenopsis.

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“…Interestingly, Eph silencing was reproducibly obtained in whole aphids and in the guts after feeding the insects on N. benthamiana infected with TRV-Eph. To our knowledge, this technique has never been tested before to silence genes in aphids, but was successfully developed to silence genes in some phloem feeding insects as the potato psyllid Bactericera cockerelli , and the mealybugs Planococcus citri and Phenacoccus solenopsis [67,68,69]. In these previous experiments, a Tobacco mosaic virus- or a Potato virus X -derived virus was used to silence the insect genes.…”

Section: Discussionmentioning

confidence: 99%

Comparative Analysis of RNAi-Based Methods to Down-Regulate Expression of Two Genes Expressed at Different Levels in Myzus persicae

Mulot

Boissinot

Monsion

et al. 2016

Viruses

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With the increasing availability of aphid genomic data, it is necessary to develop robust functional validation methods to evaluate the role of specific aphid genes. This work represents the first study in which five different techniques, all based on RNA interference and on oral acquisition of double-stranded RNA (dsRNA), were developed to silence two genes, ALY and Eph, potentially involved in polerovirus transmission by aphids. Efficient silencing of only Eph transcripts, which are less abundant than those of ALY, could be achieved by feeding aphids on transgenic Arabidopsis thaliana expressing an RNA hairpin targeting Eph, on Nicotiana benthamiana infected with a Tobacco rattle virus (TRV)-Eph recombinant virus, or on in vitro-synthesized Eph-targeting dsRNA. These experiments showed that the silencing efficiency may differ greatly between genes and that aphid gut cells seem to be preferentially affected by the silencing mechanism after oral acquisition of dsRNA. In addition, the use of plants infected with recombinant TRV proved to be a promising technique to silence aphid genes as it does not require plant transformation. This work highlights the need to pursue development of innovative strategies to reproducibly achieve reduction of expression of aphid genes.

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Inoculation of Nicotiana tabacum with recombinant potato virus X induces RNA interference in the solenopsis mealybug, Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae)

Abstract: chitin synthase 1 is a potential RNAi target in P. solenopsis and the recombinant PVX can be used as a tool to evaluate candidate RNAi triggering elements in plants.

Cited by 15 publications

References 26 publications

Silencing cathepsin L expression reduces Myzus persicae protein content and the nutritional value as prey for Coccinella septempunctata

Silencing cathepsin L expression reduces Myzus persicae protein content and the nutritional value as prey for Coccinella septempunctata

Evaluation of potential RNA‐interference‐target genes to control cotton mealybug, Phenacoccus solenopsis (Hemiptera: Pseudococcuidae)

Comparative Analysis of RNAi-Based Methods to Down-Regulate Expression of Two Genes Expressed at Different Levels in Myzus persicae

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