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RNAi in Insect Pest Control

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A special issue of Insects (ISSN 2075-4450). This special issue belongs to the section "Insect Molecular Biology and Genomics".

Deadline for manuscript submissions: closed (31 December 2019) | Viewed by 37703

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Special Issue Editors

Dr. Olivier Christiaens

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Guest Editor

Department of Plants and Crops, Faculty of Bioscience Engineering, Ghent University, Coupure Links, 653 9000 Ghent, Belgium
Interests: genes; gene-expression; biosafety; ecdysone recepto; insect pests; insects
Special Issues, Collections and Topics in MDPI journals

Prof. Dr. Jinzhi Niu

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Guest Editor

College of Plant Protection, Southwest University, Chongqing, China
Interests: RNAi-based pest control; insect symbiont virus; insect-plant interaction; spider mites; invasive species

Dr. Clauvis Nji Tizi Taning

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Guest Editor

Department of Plants and Crops, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, 9000 Ghent, Belgium
Interests: molecular genetics; gene expression; RNAi; pest control; gene editing; biosafety; gene drives
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear colleagues,

Due to the increasing occurrence of resistance against the currently used insect pest control products for crop protection and, also, environmental concerns, the search for more environmentally friendly pest management products with novel modes of action is ongoing.

RNA interference (RNAi) is a natural gene-silencing mechanism triggered by double-stranded RNA (dsRNA) homologous to the target gene and can be exploited for the management of insect pests. While the first products, targeting RNAi-sensitive beetle species, are coming to the market soon, physiological and cellular barriers in many other insects have proven to be a barrier towards achieving strong enough RNAi gene silencing through oral uptake of dsRNA. This Special Issue will report on the recent advances in insect RNAi and RNAi-based insecticides, novel delivery methods for dsRNA, target gene selection studies, as well as research investigating the environmental safety aspects of this novel pest control strategy.

Dr. Olivier Christiaens
Prof. Dr. Jinzhi Niu
Dr. Clauvis Nji Tizi Taning
Guest Editors

Manuscript Submission Information

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Keywords

RNA interference
insects
crop protection
dsRNA delivery
RNAi biosafety

Published Papers (10 papers)

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Editorial

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7 pages, 220 KiB

Open AccessEditorial

RNAi in Insects: A Revolution in Fundamental Research and Pest Control Applications

by Olivier Christiaens, Jinzhi Niu and Clauvis Nji Tizi Taning

Insects 2020, 11(7), 415; https://doi.org/10.3390/insects11070415 - 03 Jul 2020

Cited by 38 | Viewed by 5209

Abstract

In this editorial for the Special Issue on ‘RNAi in insect pest control’, three important applications of RNA interference (RNAi) in insects are briefly discussed and linked to the different studies published in this Special Issue. The discovery of the RNAi mechanism revolutionized entomological research, as it presented researchers with a tool to knock down genes, which is easily applicable in a wide range of insect species. Furthermore, RNAi also provides crop protection with a novel and promising pest control mode-of-action. The sequence-dependent nature allows RNAi-based control strategies to be highly species selective and the active molecule, a natural biological molecule known as double-stranded RNA (dsRNA), has a short environmental persistence. However, more research is needed to investigate different cellular and physiological barriers, such as cellular uptake and dsRNA degradation in the digestive system in insects, in order to provide efficient control methods against a wide range of insect pest species. Finally, the RNAi pathway is an important part of the innate antiviral immune defence of insects, and could even lead to applications targeting viruses in beneficial insects such as honeybees in the future. Full article

(This article belongs to the Special Issue RNAi in Insect Pest Control)

Research

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16 pages, 3545 KiB

Open AccessArticle

Silencing of Double-Stranded Ribonuclease Improves Oral RNAi Efficacy in Southern Green Stinkbug Nezara viridula

by Rohit Sharma, Clauvis Nji Tizi Taning, Guy Smagghe and Olivier Christiaens

Insects 2021, 12(2), 115; https://doi.org/10.3390/insects12020115 - 28 Jan 2021

Cited by 16 | Viewed by 3076

Abstract

Variability in RNA-interference (RNAi) efficacy among different insect orders poses a big hurdle in the development of RNAi-based pest control strategies. The activity of double-stranded ribonucleases (dsRNases) in the digestive canal of insects can be one of the critical factors affecting oral RNAi efficacy. Here, the involvement of these dsRNases in the southern green stinkbug Nezara viridula was investigated. First, the full sequence of the only dsRNase (NvdsRNase) in the transcriptome of N. viridula was obtained, followed by an oral feeding bioassay to evaluate the effect of NvdsRNase-silencing on oral RNAi efficacy. The NvdsRNase was first silenced in nymphs by NvdsRNase-dsRNA injections, followed by exposure to an artificial diet containing a lethal αCop-specific dsRNA. A significantly higher mortality was observed in the NvdsRNase-silenced nymphs when placed on the dsαCop-containing diet (65%) than in the dsGFP injected and dsαCop fed control (46.67%). Additionally, an ex vivo dsRNA degradation assay showed a higher stability of dsRNA in the saliva and midgut juice of NvdsRNase-silenced adults. These results provide evidence for the involvement of NvdsRNase in the reduction of oral RNAi efficacy in N. viridula. This information will be useful in further improving potential RNAi-based strategies to control this pest. Full article

(This article belongs to the Special Issue RNAi in Insect Pest Control)

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13 pages, 1833 KiB

Open AccessCommunication

RNAi and CRISPR/Cas9 as Functional Genomics Tools in the Neotropical Stink Bug, Euschistus heros

by Deise Cagliari, Guy Smagghe, Moises Zotti and Clauvis Nji Tizi Taning

Insects 2020, 11(12), 838; https://doi.org/10.3390/insects11120838 - 27 Nov 2020

Cited by 12 | Viewed by 3172

Abstract

The Neotropical brown stink bug, Euschistus heros, is one of the most important stink bug pests in leguminous plants in South America. RNAi and CRISPR/Cas9 are important and useful tools in functional genomics, as well as in the future development of new integrated pest management strategies. Here, we explore the use of these technologies as complementing functional genomic tools in E. heros. Three genes, abnormal wing disc (awd), tyrosine hydroxylase (th) and yellow (yel), known to be involved in wing development (awd) and the melanin pathway (th and yel) in other insects, were chosen to be evaluated using RNAi and CRISPR/Cas9 as tools. First, the genes were functionally characterized using RNAi knockdown technology. The expected phenotype of either deformed wing or lighter cuticle pigmentation/defects in cuticle sclerotization was observed for awd and th, respectively. However, for yel, no obvious phenotype was observed. Based on this, yel was selected as a target for the development of a CRISPR/Cas9 workflow to study gene knockout in E. heros. A total of 719 eggs were injected with the Cas9 nuclease (300 ng/µL) together with the sgRNA (300 ng/µL) targeting yel. A total of six insects successfully hatched from the injected eggs and one of the insects showed mutation in the target region, however, the phenotype was still not obvious. Overall, this study for the first time provides a useful CRISPR/Cas9 gene editing methodology to complement RNAi for functional genomic studies in one of the most important and economically relevant stink bug species. Full article

(This article belongs to the Special Issue RNAi in Insect Pest Control)

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12 pages, 1764 KiB

Open AccessArticle

RNA Interference Is Enhanced by Knockdown of Double-Stranded RNases in the Yellow Fever Mosquito Aedes aegypti

by David Giesbrecht, Daniel Heschuk, Ian Wiens, David Boguski, Parker LaChance and Steve Whyard

Insects 2020, 11(6), 327; https://doi.org/10.3390/insects11060327 - 27 May 2020

Cited by 22 | Viewed by 4083

Abstract

RNA interference (RNAi) techniques are being developed for a range of pest insect control technologies, including the sterile insect technique (SIT) and double-stranded RNA (dsRNA)-based insecticides. In SIT applications, where >99% of the released males should be sterile to meet industry standards, the efficiency of RNAi will need to be improved for many insect species if this technology is to be adopted. Endogenous dsRNases can impede dsRNA delivery in some insects, and, here, we investigated whether dsRNases in the midgut could limit RNAi efficacy in the mosquito Aedes aegypti. Ten putative dsRNases were identified in the Ae. aegypti genome, with two highly expressed in the midguts of larvae. Using an ex vivo assay, we observed that dsRNA was rapidly degraded within the mosquito larva’s gut. Double-stranded RNA targeting these two dsRNases, when fed to the larvae, effectively reduced gut dsRNase activity. When these dsRNase-specific dsRNAs were co-delivered with dsRNA targeting a cyan fluorescent protein (CFP) reporter gene, greater knockdown of CFP fluorescence was observed. These results suggest that inhibiting dsRNase activity could enable the implementation of RNAi-based mosquito control methods. Full article

(This article belongs to the Special Issue RNAi in Insect Pest Control)

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17 pages, 4050 KiB

Open AccessArticle

Using RNA Interference to Reveal the Function of Chromatin Remodeling Factor ISWI in Temperature Tolerance in Bemisia tabaci Middle East–Asia Minor 1 Cryptic Species

by Shun-Xia Ji, Xiao-Di Wang, Xiao-Na Shen, Lin Liang, Wan-Xue Liu, Fang-Hao Wan and Zhi-Chuang Lü

Insects 2020, 11(2), 113; https://doi.org/10.3390/insects11020113 - 10 Feb 2020

Cited by 13 | Viewed by 2874

Abstract

Invasive species often encounter rapid environmental changes during invasions and only the individuals that successfully overcome environmental stresses can colonize and spread. Chromatin remodeling may be essential in environmental adaptation. To assess the functions of imitation switch (ISWI) in invasive Bemisia tabaci Middle East–Asia Minor 1 (MEAM1) cryptic species, we cloned and characterized the MEAM1 BtISWI gene and determined its functions in response to thermal stress. The full-length cDNA of BtISWI was 3712 bp, with a 3068 bp open reading frame (ORF) encoding a 118.86 kDa protein. BtISWI mRNA expression was significantly up-regulated after exposure to heat shock or cold shock conditions, indicating that BtISWI expression can be induced by thermal stress. After feeding double-stranded RNA (dsRNA), specifically for BtISWI, resistance to both heat and cold decreased significantly, suggesting that BtISWI may function directly in the thermal tolerance of MEAM1. Moreover, the preferred temperature of MEAM1 adults fed dsRNA was 1.9–3.5 °C higher than the control groups. Taken together, our findings highlight the importance of epigenetic gene regulation in the thermal response or thermal adaptation of invasive Bemisia tabaci (B. tabaci), and provide a new potential target for establishing sustainable control strategies for B. tabaci. Full article

(This article belongs to the Special Issue RNAi in Insect Pest Control)

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14 pages, 2667 KiB

Open AccessArticle

Expression Dynamics of Core RNAi Machinery Genes in Pea Aphids Upon Exposure to Artificially Synthesized dsRNA and miRNAs

by Li Yang, Yuan Tian, Yuan-Yuan Peng, Jinzhi Niu and Jin-Jun Wang

Insects 2020, 11(2), 70; https://doi.org/10.3390/insects11020070 - 21 Jan 2020

Cited by 10 | Viewed by 3350

Abstract

The pea aphid is an important pest of vegetables and causes serious losses worldwide. RNA interference (RNAi) is an effective pest control tool, and three sub-pathways have been described: The miRNA pathway, siRNA pathway, and piRNA pathway. A large number of genes in miRNA pathway and piRNA pathway are found to be expanded. To study the roles of these genes, the expression of 25 core RNAi genes was screened in spatiotemporal samples, artificially synthesized dsRNA and miRNA treated samples. The 25 genes were all expressed during different development stages and in different tissues. In dsRNA-treated samples and miRNA-treated samples, the expressions of genes in these three pathways were induced, especially the expanded genes. This suggests a complex network of RNAi core genes in the three sub-pathways. Treatment of miRNA seems to induce gene expression in a dosage-dependent manner. These results increase our knowledge of the siRNA pathway and related factors from RNAi pathway in aphids and promote the use of RNAi for the control of aphid pests. Full article

(This article belongs to the Special Issue RNAi in Insect Pest Control)

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12 pages, 2765 KiB

Open AccessArticle

Feeding Delivery of dsHvSnf7 Is a Promising Method for Management of the Pest Henosepilachna vigintioctopunctata (Coleoptera: Coccinellidae)

by Jing Lü, Zhuoqi Liu, Wei Guo, Mujuan Guo, Shimin Chen, Huali Li, Chunxiao Yang, Youjun Zhang and Huipeng Pan

Insects 2020, 11(1), 34; https://doi.org/10.3390/insects11010034 - 31 Dec 2019

Cited by 24 | Viewed by 2727

Abstract

RNA interference (RNAi) techniques have emerged as powerful tools in the development of novel management strategies for the control of insect pests, such as Henosepilachna vigintioctopunctata, which is a major solanaceous pest in Asia. Our results showed that levels of HvSnf7 expression were greater in larval midguts than in other tissues. Silencing of HvSnf7 led to greater H. vigintioctopunctata mortality rates and appeared to be time- and partially dose-dependent. Bacterially expressed dsHvSnf7 that was applied to detached plant leaves caused 98, 88, and 60% mortality in 1st and 3rd instars, and adults after 10, 12, and 14 d, respectively; when applied to living plants, bacterially expressed dsHvSnf7 led to mortality in 1st and 3rd instars, with no effect on adults. Bacterially expressed dsHvSnf7 led to improved plant protection against H. vigintioctopunctata. Ultrastructural changes caused by HvSnf7-RNAi in larval midguts showed extensive loss of cellular contents that indicate loss of membrane integrity. This study indicate that HvSnf7 potentially can be used as RNAi target gene for controlling of H. vigintioctopunctata. Full article

(This article belongs to the Special Issue RNAi in Insect Pest Control)

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11 pages, 1770 KiB

Open AccessArticle

RNAi-Mediated Knockdown of Chitin Synthase 1 (CHS1) Gene Causes Mortality and Decreased Longevity and Fecundity in Aphis gossypii

by Farman Ullah, Hina Gul, Xiu Wang, Qian Ding, Fazal Said, Xiwu Gao, Nicolas Desneux and Dunlun Song

Insects 2020, 11(1), 22; https://doi.org/10.3390/insects11010022 - 26 Dec 2019

Cited by 29 | Viewed by 4189

Abstract

Chitin is a vital part of the insect exoskeleton and peritrophic membrane, synthesized by chitin synthase (CHS) enzymes. Chitin synthase 1 (CHS1) is a crucial enzyme in the final step of chitin biosynthetic pathway and consequently plays essential role towards insect growth and molting. RNA interference (RNAi) is an agent that could be used as an extremely target-specific and ecologically innocuous tactic to control different insect pests associated with economically important crops. The sole purpose of the current study is to use CHS1 as the key target gene against the cotton-melon aphid, Aphis gossypii, via oral feeding on artificial diets mixed with dsRNA-CHS1. Results revealed that the expression level of CHS1 gene significantly decreased after the oral delivery of dsRNA-CHS1. The knockdown of CHS1 gene caused up to 43%, 47%, and 59% mortality in third-instar nymph after feeding of dsCHS1 for 24, 48, and 72 h, respectively, as compared to the control. Consistent with this, significantly lower longevity (approximately 38%) and fecundity (approximately 48%) were also found in adult stage of cotton-melon aphids that were fed with dsCHS1 for 72 h at nymphal stage. The qRT-PCR analysis of gene expression demonstrated that the increased mortality rates and lowered longevity and fecundity of A. gossypii were attributed to the downregulation of CHS1 gene via oral-delivery-mediated RNAi. The results of current study confirm that CHS1 could be an appropriate candidate target gene for the RNAi-based control of cotton-melon aphids. Full article

(This article belongs to the Special Issue RNAi in Insect Pest Control)

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9 pages, 1412 KiB

Open AccessArticle

Bacteria-Mediated RNA Interference for Management of Plagiodera versicolora (Coleoptera: Chrysomelidae)

by Yiqiu Zhang, Letian Xu, Shengchun Li and Jiang Zhang

Insects 2019, 10(12), 415; https://doi.org/10.3390/insects10120415 - 21 Nov 2019

Cited by 29 | Viewed by 4180

Abstract

RNA interference (RNAi) has emerged as a novel and feasible strategy for pest management. Methods for cost-effective production and stable delivery of double-stranded RNA (dsRNA) to the target insects are crucial for the wide application of RNAi for pest control. In this study, we tested the expression of dsRNA in RNaseIII-deficient Escherichia coli HT115 which was then fed to Plagiodera versicolora larvae, an insect pest of Salicaceae plants worldwide. By targeting six potential genes, including actin (ACT), signal recognition particle protein 54k (SRP54), heat shock protein 70 (HSC70), shibire (SHI), cactus (CACT), and soluble N-ethylmaleimide-sensitive fusion attachment proteins (SNAP), we found that feeding bacteria-expressed dsRNA successfully triggered the silencing of the five target genes tested and the suppression of ACT and SRP54 genes caused significant mortality. Our results suggest that the oral delivery of bacteria-expressed dsRNA is a potential alternative for the control of P. versicolora, and that ACT and SRP54 genes are the potent targets. Full article

(This article belongs to the Special Issue RNAi in Insect Pest Control)

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13 pages, 1678 KiB

Open AccessArticle

Knockdown of β-N-acetylglucosaminidase 2 Impairs Molting and Wing Development in Lasioderma serricorne (Fabricius)

by Wen-Jia Yang, Kang-Kang Xu, Xin Yan and Can Li

Insects 2019, 10(11), 396; https://doi.org/10.3390/insects10110396 - 08 Nov 2019

Cited by 13 | Viewed by 2823

Abstract

β-N-acetylglucosaminidases (NAGs) are carbohydrate enzymes that degrade chitin oligosaccharides into N-acetylglucosamine monomers. This process is important for chitin degradation during insect development and metamorphosis. We identified and evaluated a β-N-acetylglucosaminidase 2 gene (LsNAG2) from the cigarette beetle, Lasioderma serricorne (Fabricius). The full-length open reading frame of LsNAG2 was 1776 bp and encoded a 591 amino acid protein. The glycoside hydrolase family 20 (GH20) catalytic domain and an additional GH20b domain of the LsNAG2 protein were highly conserved. Phylogenetic analysis revealed that LsNAG2 clustered with the group II NAGs. Quantitative real-time PCR analyses showed that LsNAG2 was expressed in all developmental stages and was most highly expressed in the late larval and late pupal stages. In the larval stage, LsNAG2 was predominantly expressed in the integument. Knockdown of LsNAG2 in fifth instar larvae disrupted larval–pupal molting and reduced the expression of four chitin synthesis genes (trehalase 1 (LsTRE1), UDP-N-acetylglucosamine pyrophosphorylase 1 and 2 (LsUAP1 and LsUAP2), and chitin synthase 1 (LsCHS1)). In late pupae, LsNAG2 depletion resulted in abnormal adult eclosion and wing deformities. The expression of five wing development-related genes (teashirt (LsTSH), vestigial (LsVG), wingless (LsWG), ventral veins lacking (LsVVL), and distal-less (LsDLL)) significantly declined in the LsNAG2-depleted beetles. These findings suggest that LsNAG2 is important for successful molting and wing development of L. serricorne. Full article

(This article belongs to the Special Issue RNAi in Insect Pest Control)

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