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The brown marmorated stink bug (BMSB) is native to Asia and recently invaded the USA. RNA interference (RNAi) is a gene silencing mechanism in which the introduction of double-stranded RNA (dsRNA) inhibits gene function by degrading target mRNA. In dsRNA stability assays, the dsRNases present in the hemolymph and salivary gland secretions of BMSB showed lower activity than those in the hemolymph of Heliothis virescens. We evaluated six housekeeping genes (18S rRNA, EF1-α, Actin, Ubiquitin, 60S RP and β-Tubulin) across dsRNA treatments (injection and feeding) in nymphs and adults of BMSB and identified 18S rRNA and 60S RP as the best genes to use as a reference in reverse-transcriptase quantitative PCR (RT-qPCR). Homologs of 13 genes that were shown to function as effective RNAi targets in other insects were identified and evaluated by injecting dsRNA targeting these homologs into BMSB adults. Five out of 13 dsRNAs tested caused more than 70% mortality by seven days after injection of dsRNA. Feeding dsRNA targeting five of these genes (IAP, ATPase, SNF7, GPCR, and PPI) to nymphs caused more than 70% mortality by three of the five dsRNAs tested. These data suggest that feeding dsRNA causes target gene knockdown and mortality in BMSB.

Titel:	Improving RNAi in the Brown Marmorated Stink Bug: Identification of target genes and reference genes for RT-qPCR.
Autor/in / Beteiligte Person:	Mogilicherla, K ; Howell, JL ; Palli, SR
Link:	Full Text Finder (Volltext)
Zeitschrift:	Scientific reports, Jg. 8 (2018-02-27), Heft 1, S. 3720
Veröffentlichung:	London : Nature Publishing Group, copyright 2011-, 2018
Medientyp:	academicJournal
ISSN:	2045-2322 (electronic)
DOI:	10.1038/s41598-018-22035-z
Schlagwort:	Animals RNA, Double-Stranded genetics RNA, Ribosomal, 18S genetics Heteroptera genetics RNA Interference physiology Reverse Transcriptase Polymerase Chain Reaction methods
Sonstiges:	Nachgewiesen in: MEDLINE Sprachen: English Publication Type: Journal Article; Research Support, U.S. Gov't, Non-P.H.S. Language: English [Sci Rep] 2018 Feb 27; Vol. 8 (1), pp. 3720. <i>Date of Electronic Publication: </i>2018 Feb 27. MeSH Terms: Heteroptera / genetics ; RNA Interference / physiology ; Reverse Transcriptase Polymerase Chain Reaction / *methods ; Animals ; RNA, Double-Stranded / genetics ; RNA, Ribosomal, 18S / genetics References: Peptides. 2014 Mar;53:307-14. (PMID: 24394433) ; J Insect Physiol. 2011 Feb;57(2):231-45. (PMID: 21078327) ; J Insect Physiol. 2010 Mar;56(3):227-35. (PMID: 19837076) ; Nature. 1998 Feb 19;391(6669):806-11. (PMID: 9486653) ; Sci Rep. 2017 Aug 7;7(1):7379. (PMID: 28784998) ; Genome Biol. 2002 Jun 18;3(7):RESEARCH0034. (PMID: 12184808) ; Methods. 2001 Dec;25(4):402-8. (PMID: 11846609) ; Insect Biochem Mol Biol. 2007 Jul;37(7):732-8. (PMID: 17550829) ; PLoS One. 2014 Mar 03;9(3):e87235. (PMID: 24595215) ; J Med Entomol. 2016 Mar;53(2):304-14. (PMID: 26659858) ; J Biosci. 2011 Mar;36(1):153-61. (PMID: 21451256) ; Apoptosis. 2011 Mar;16(3):235-48. (PMID: 21274634) ; Insect Biochem Mol Biol. 2017 Nov;90:53-60. (PMID: 28951282) ; Biotechnol Lett. 2004 Mar;26(6):509-15. (PMID: 15127793) ; Genome Biol. 2008 Jan 17;9(1):R10. (PMID: 18201385) ; Environ Entomol. 2013 Aug;42(4):627-41. (PMID: 23905725) ; Sci Rep. 2017 Dec 6;7(1):17059. (PMID: 29213068) ; PLoS One. 2016 May 04;11(5):e0152730. (PMID: 27144586) ; Pest Manag Sci. 2011 Feb;67(2):175-82. (PMID: 21061270) ; Science. 2015 Feb 27;347(6225):991-4. (PMID: 25722411) ; Plant Mol Biol. 2012 Jan 31;:null. (PMID: 22290409) ; Biochim Biophys Acta. 2001 Jan 15;1499(3):191-8. (PMID: 11341966) ; Insect Biochem Mol Biol. 2016 Nov;78:78-88. (PMID: 27687845) ; Anal Biochem. 1976 May 7;72:248-54. (PMID: 942051) ; Cancer Res. 2004 Aug 1;64(15):5245-50. (PMID: 15289330) ; PLoS One. 2017 Feb 9;12 (2):e0171861. (PMID: 28182760) ; PLoS One. 2014 Feb 26;9(2):e88483. (PMID: 24586332) ; Arch Insect Biochem Physiol. 2005 Apr;58(4):234-42. (PMID: 15756700) ; Curr Opin Insect Sci. 2014 Dec;6:1-8. (PMID: 26705514) ; J Mol Endocrinol. 2002 Aug;29(1):23-39. (PMID: 12200227) ; Nat Biotechnol. 2007 Nov;25(11):1322-6. (PMID: 17982443) ; Sci Rep. 2017 Aug 21;7(1):8913. (PMID: 28827780) ; RNA Biol. 2016 Jul 2;13(7):656-69. (PMID: 27245473) Substance Nomenclature: 0 (RNA, Double-Stranded) ; 0 (RNA, Ribosomal, 18S) Entry Date(s): Date Created: 20180301 Date Completed: 20190923 Latest Revision: 20190923 Update Code: 20240513 PubMed Central ID: PMC5829169

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Improving RNAi in the Brown Marmorated Stink Bug: Identification of target genes and reference genes for RT-qPCR.