Evaluation of a cuticle protein gene as a potential RNAi target in aphids

Shang, Feng; Ding, Bi‐Yue; Ye, Chao; Yang, Li; Chang, T.-H.; Xie, Jiaqin; Tang, Liangde; Niu, Jinzhi; Wang, Jinjun

doi:10.1002/ps.5599

Cited by 50 publications

(39 citation statements)

References 49 publications

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“…As considered in the Introduction, the number and length of exactly matching siRNA sequences derived from a heterologous dsRNA that are required for successful expression knockdown can vary with the target gene and insect species. Although the cross-reactivity of dsRNAs in plant sapfeeding insects has not been investigated extensively, evidence that heterologous dsRNA can confer effective RNAi is provided by a study of CP19, an essential gene coding aphid cuticle protein 19 which protects the insects against cuticular water loss and desiccation [60]. Across the three aphid species studied, CP19 had 94% sequence identity (the number of matching 21-mer sequences was not reported) and dsCP19 matching the CP19 sequence of Aphis citricidus reduced CP19 expression in both A. pisum and M. persicae, with attendant mortality effects.…”

Section: Discussionmentioning

confidence: 99%

Non-Target Effects of dsRNA Molecules in Hemipteran Insects

Arora

Chung

Douglas

2021

Genes

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Insect pest control by RNA interference (RNAi)-mediated gene expression knockdown can be undermined by many factors, including small sequence differences between double-stranded RNA (dsRNA) and the target gene. It can also be compromised by effects that are independent of the dsRNA sequence on non-target organisms (known as sequence-non-specific effects). This study investigated the species-specificity of RNAi in plant sap-feeding hemipteran pests. We first demonstrated sequence-non-specific suppression of aphid feeding by dsRNA at dietary concentrations ≥0.5 µg µL−1. Then we quantified the expression of NUC (nuclease) genes in insects administered homologous dsRNA (with perfect sequence identity to the target species) or heterologous dsRNA (generated against a related gene of non-identical sequence in a different insect species). For the aphids Acyrthosiphon pisum and Myzus persicae, significantly reduced NUC expression was obtained with the homologous but not heterologous dsRNA at 0.2 µg µL−1, despite high dsNUC sequence identity. Follow-up experiments demonstrated significantly reduced expression of NUC genes in the whitefly Bemisia tabaci and mealybug Planococcus maritimus administered homologous dsNUCs, but not heterologous aphid dsNUCs. Our demonstration of inefficient expression knockdown by heterologous dsRNA in these insects suggests that maximal dsRNA sequence identity is required for RNAi targeting of related pest species, and that heterologous dsRNAs at appropriate concentrations may not be a major risk to non-target sap-feeding hemipterans.

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Section: Discussionmentioning

confidence: 99%

Non-Target Effects of dsRNA Molecules in Hemipteran Insects

Arora

Chung

Douglas

2021

Genes

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“…Cuticle protein genes are involved in the cuticle formation, and they are necessary for cuticle development, flexibility, and metamorphosis [45]. Moreover, expression of the cuticle protein genes is also related with the survival of insects [45,46]. Transcriptional patterns of genes encoding cuticle proteins in the water flea Daphnia pulex have responses to the interaction between biotic (predator presence) and abiotic (low calcium concentration) environmental stresses [47].…”

Section: Discussionmentioning

confidence: 99%

Comparative transcriptome analysis of the rice leaf folder (Cnaphalocrocis medinalis) to heat acclimation

Quan

Wang

et al. 2020

BMC Genomics

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Background: The rice leaf folder Cnaphalocrocis medinalis Güenée is a serious insect pest of rice in Asia. This pest occurs in summer, and it is sensitive to high temperature. However, the larvae exhibit heat acclimation/adaptation. To understand the underlying mechanisms, we established a heat-acclimated strain via multigenerational selection at 39°C. After heat shock at 41°C for 1 h, the transcriptomes of the heat-acclimated (S-39) and unacclimated (S-27) larvae were sequenced, using the unacclimated larvae without exposure to 41°C as the control. Results: Five generations of selection at 39°C led larvae to acclimate to this heat stress. Exposure to 41°C induced 1160 differentially expressed genes (DEGs) between the heat-acclimated and unacclimated larvae. Both the heatacclimated and unacclimated larvae responded to heat stress via upregulating genes related to sensory organ development and structural constituent of eye lens, whereas the unacclimated larvae also upregulated genes related to structural constituent of cuticle. Compared to unacclimated larvae, heat-acclimated larvae downregulated oxidoreductase activity-related genes when encountering heat shock. Both the acclimated and unacclimated larvae adjusted the longevity regulating, protein processing in endoplasmic reticulum, antigen processing and presentation, MAPK and estrogen signaling pathway to responsed to heat stress. Additionally, the unacclimated larvae also adjusted the spliceosome pathway, whereas the heat-acclimated larvae adjusted the biosynthesis of unsaturated fatty acids pathway when encountering heat stress. Although the heat-acclimated and unacclimated larvae upregulated expression of heat shock protein genes under heat stress including HSP70, HSP27 and CRYAB, their biosynthesis, metabolism and detoxification-related genes expressed differentially. Conclusions: The rice leaf folder larvae could acclimate to a high temperature via multigenerational heat selection. The heat-acclimated larvae induced more DEGs to response to heat shock than the unacclimated larvae. The changes in transcript level of genes were related to heat acclimation of larvae, especially these genes in sensory organ development, structural constituent of eye lens, and oxidoreductase activity. The DEGs between heat-acclimated and unacclimated larvae after heat shock were enriched in the biosynthesis and metabolism pathways. These results are helpful to understand the molecular mechanism underlying heat acclimation of insects.

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“…An alternative, yet complementary approach for the assessment off-targets in NTOs is to conduct laboratory bioassays with representative NTOs that are exposed to the dsRNA (Whyard et al, 2009;Bachman et al, 2013Bachman et al, , 2016Pan et al, 2017;Haller et al, 2019;Shang et al, 2019). Representative NTOs can include surrogate species that are selected based on their sensitivity to the dsRNA, reliability and relevance (Romeis et al, 2013).…”

Section: Environmental Risk Assessmentmentioning

confidence: 99%

Risk Assessment Considerations for Genetically Modified RNAi Plants: EFSA’s Activities and Perspective

et al. 2020

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Genetically modified plants (GMPs) intended for market release can be designed to induce "gene silencing" through RNA interference (RNAi). The European Food Safety Authority (EFSA) and other international risk assessment bodies/regulatory agencies have taken several actions to determine whether the existing risk assessment approaches for GMPs are appropriate for the risk assessment of RNAi-based GMPs or require complementary or alternative approaches. To our knowledge, at the international level, no dedicated guidelines have been developed for the risk assessment and regulation of RNAi-based GMPs, confirming that existing science-based risk assessment approaches for GMPs are generally considered suitable for RNAi-based GMPs. However, some specificities have been identified for the risk assessment of RNAi-based GMPs. Here, we report on some of these specificities as identified and addressed by the EFSA GMO Panel for the molecular characterisation, food/feed safety assessment and environmental risk assessment of RNAi-based GMPs, using the DvSnf7 dsRNA-expressing maize MON87411 as a case study.

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Evaluation of a cuticle protein gene as a potential RNAi target in aphids

Cited by 50 publications

References 49 publications

Non-Target Effects of dsRNA Molecules in Hemipteran Insects

Non-Target Effects of dsRNA Molecules in Hemipteran Insects

Comparative transcriptome analysis of the rice leaf folder (Cnaphalocrocis medinalis) to heat acclimation

Risk Assessment Considerations for Genetically Modified RNAi Plants: EFSA’s Activities and Perspective

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