Improving RNA interference in the southern green stink bug, Nezara viridula

Dhandapani Gurusamy; Jeffrey L. Howell; Shankar C.R.R. Chereddy; Kanakachari Mogilicherla; Subba Reddy Palli

doi:10.1007/s10340-021-01358-3

Improving RNA interference in the southern green stink bug, Nezara viridula

Dhandapani Gurusamy, Jeffrey L. Howell, Shankar C.R.R. Chereddy, Kanakachari Mogilicherla, Subba Reddy Palli

Research output: Contribution to journal › Article › peer-review

13 Scopus citations

Abstract

The southern green stink bug, Nezara viridula, is a worldwide pest of many important crops. RNA from N. viridula nymphs and adults was sequenced and assembled into a transcriptome. Orthologs of genes coding for proteins involved in RNA interference (RNAi) pathways and potential target genes for RNAi-mediated control of this pest were identified by searching the transcriptome. The watery saliva, lumen and hemolymph collected from N. viridula showed lower dsRNase activity than that in the lumen and hemolymph collected from the fall armyworm, Spodoptera frugiperda, which is refractory to RNAi. Seven housekeeping genes, 60S RP L12, 40S RP S17, 18S rRNA, EF1-α, Actin, Ubiquitin and β-Tubulin, were evaluated across developmental stages and dsRNA treatments and identified 60S RP L12 and 40S RP S17 as the best reference genes for use in reverse transcriptase quantitative PCR (RT-qPCR). One microgram of dsRNA targeting the orthologs of seven known RNAi target genes (SNF7, IAP1, IAP2, IAP5, PP1, GPCR and ATPase1) was tested in N. viridula. Three out of seven dsRNAs tested induced 67–100% mortality. Injection of dsRNA targeting SNF7 into N. viridula adults caused 100% mortality, while feeding dsSNF7 induced 60% mortality in both adults and nymphs. Delivery of dsRNA by either feeding or injection efficiently knocked down target gene expression. These data demonstrate that RNAi pathway components are present in N. viridula and delivering dsRNA by either injection or feeding induces silencing of target gene expression and mortality, suggesting that RNAi-based methods could be developed to manage this pest.

Original language	English
Pages (from-to)	1461-1472
Number of pages	12
Journal	Journal of Pest Science
Volume	94
Issue number	4
DOIs	https://doi.org/10.1007/s10340-021-01358-3
State	Published - Sep 2021

Bibliographical note

Publisher Copyright:
© 2021, The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.

Funding

The authors thank Dr. Amit Sethi from Corteva Agriscience, Dr. Guo-Qing Tang from Syngenta and Dr. Sven Geibel from Bayer Crop Science for discussions and advice. This material is based upon work supported by the National Science Foundation I/UCRC, the Center for Arthropod Management Technologies under Grant IIP-1821936 and by industry partners and the USDA/NIFA (under Hatch Project 2351177000).

Funders	Funder number
Corteva Agriscience
UCRC
U.S. Department of Agriculture
U.S. Department of Energy Chinese Academy of Sciences Guangzhou Municipal Science and Technology Project Oak Ridge National Laboratory Extreme Science and Engineering Discovery Environment National Science Foundation National Energy Research Scientific Computing Center National Natural Science Foundation of China	1821936
Center for Arthropod Management Technologies	IIP-1821936
US Department of Agriculture National Institute of Food and Agriculture, Agriculture and Food Research Initiative	2351177000

Keywords

Double-stranded RNA
DsRNase
RT-qPCR
SNF7
Transcriptome

ASJC Scopus subject areas

Agronomy and Crop Science

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10.1007/s10340-021-01358-3

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abstract = "The southern green stink bug, Nezara viridula, is a worldwide pest of many important crops. RNA from N. viridula nymphs and adults was sequenced and assembled into a transcriptome. Orthologs of genes coding for proteins involved in RNA interference (RNAi) pathways and potential target genes for RNAi-mediated control of this pest were identified by searching the transcriptome. The watery saliva, lumen and hemolymph collected from N. viridula showed lower dsRNase activity than that in the lumen and hemolymph collected from the fall armyworm, Spodoptera frugiperda, which is refractory to RNAi. Seven housekeeping genes, 60S RP L12, 40S RP S17, 18S rRNA, EF1-α, Actin, Ubiquitin and β-Tubulin, were evaluated across developmental stages and dsRNA treatments and identified 60S RP L12 and 40S RP S17 as the best reference genes for use in reverse transcriptase quantitative PCR (RT-qPCR). One microgram of dsRNA targeting the orthologs of seven known RNAi target genes (SNF7, IAP1, IAP2, IAP5, PP1, GPCR and ATPase1) was tested in N. viridula. Three out of seven dsRNAs tested induced 67–100\% mortality. Injection of dsRNA targeting SNF7 into N. viridula adults caused 100\% mortality, while feeding dsSNF7 induced 60\% mortality in both adults and nymphs. Delivery of dsRNA by either feeding or injection efficiently knocked down target gene expression. These data demonstrate that RNAi pathway components are present in N. viridula and delivering dsRNA by either injection or feeding induces silencing of target gene expression and mortality, suggesting that RNAi-based methods could be developed to manage this pest.",

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Improving RNA interference in the southern green stink bug, Nezara viridula

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

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