RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum)

Roberto A. Camargo, Guilherme O. Barbosa, Isabella Presotto Possignolo, Lazaro E.P. Peres, Eric Lam, Joni E. Lima, Antonio Figueira, Henrique Marques-Souza

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

RNA interference (RNAi), a gene-silencing mechanism that involves providing doublestranded RNA molecules that match a specific target gene sequence, is now widely used in functional genetic studies. The potential application of RNAi-mediated control of agricultural insect pests has rapidly become evident. The production of transgenic plants expressing dsRNA molecules that target essential insect genes could provide a means of specific gene silencing in larvae that feed on these plants, resulting in larval phenotypes that range from loss of appetite to death. In this report, we show that the tomato leafminer (Tuta absoluta), a major threat to commercial tomato production, can be targeted by RNAi. Weselected two target genes (Vacuolar ATPase-A and Arginine kinase) based on the RNAi response reported for these genes in other pest species. In view of the lack of an artificial diet for T. absoluta, we used two approaches to deliver dsRNA into tomato leaflets. The first approach was based on the uptake of dsRNA by leaflets and the second was based on "in planta-induced transient gene silencing" (PITGS), a well-established method for silencing plant genes, used here for the first time to deliver in planta-transcribed dsRNA to target insect genes. Tuta absoluta larvae that fed on leaves containing dsRNA of the target genes showed an ~60% reduction in target gene transcript accumulation, an increase in larval mortality and less leaf damage. We then generated transgenic 'Micro-Tom' tomato plants that expressed hairpin sequences for both genes and observed a reduction in foliar damage by T. absoluta in these plants. Our results demonstrate the feasibility of RNAi as an alternative method for controlling this critical tomato pest.

Original languageEnglish (US)
Article numbere2673
JournalPeerJ
Volume2016
Issue number12
DOIs
StatePublished - Jan 1 2016

Fingerprint

Tuta absoluta
Solanum lycopersicum
Gene Silencing
gene silencing
Lycopersicon esculentum
RNA Interference
RNA interference
double-stranded RNA
Genes
tomatoes
RNA
Insect Genes
genes
Larva
Plantae
Arginine Kinase
Insect Control
Vacuolar Proton-Translocating ATPases
Plant Genes
pests

All Science Journal Classification (ASJC) codes

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Neuroscience(all)

Cite this

Camargo, R. A., Barbosa, G. O., Possignolo, I. P., Peres, L. E. P., Lam, E., Lima, J. E., ... Marques-Souza, H. (2016). RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum). PeerJ, 2016(12), [e2673]. https://doi.org/10.7717/peerj.2673
Camargo, Roberto A. ; Barbosa, Guilherme O. ; Possignolo, Isabella Presotto ; Peres, Lazaro E.P. ; Lam, Eric ; Lima, Joni E. ; Figueira, Antonio ; Marques-Souza, Henrique. / RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum). In: PeerJ. 2016 ; Vol. 2016, No. 12.
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Camargo, RA, Barbosa, GO, Possignolo, IP, Peres, LEP, Lam, E, Lima, JE, Figueira, A & Marques-Souza, H 2016, 'RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum)', PeerJ, vol. 2016, no. 12, e2673. https://doi.org/10.7717/peerj.2673

RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum). / Camargo, Roberto A.; Barbosa, Guilherme O.; Possignolo, Isabella Presotto; Peres, Lazaro E.P.; Lam, Eric; Lima, Joni E.; Figueira, Antonio; Marques-Souza, Henrique.

In: PeerJ, Vol. 2016, No. 12, e2673, 01.01.2016.

Research output: Contribution to journalArticle

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AU - Camargo, Roberto A.

AU - Barbosa, Guilherme O.

AU - Possignolo, Isabella Presotto

AU - Peres, Lazaro E.P.

AU - Lam, Eric

AU - Lima, Joni E.

AU - Figueira, Antonio

AU - Marques-Souza, Henrique

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N2 - RNA interference (RNAi), a gene-silencing mechanism that involves providing doublestranded RNA molecules that match a specific target gene sequence, is now widely used in functional genetic studies. The potential application of RNAi-mediated control of agricultural insect pests has rapidly become evident. The production of transgenic plants expressing dsRNA molecules that target essential insect genes could provide a means of specific gene silencing in larvae that feed on these plants, resulting in larval phenotypes that range from loss of appetite to death. In this report, we show that the tomato leafminer (Tuta absoluta), a major threat to commercial tomato production, can be targeted by RNAi. Weselected two target genes (Vacuolar ATPase-A and Arginine kinase) based on the RNAi response reported for these genes in other pest species. In view of the lack of an artificial diet for T. absoluta, we used two approaches to deliver dsRNA into tomato leaflets. The first approach was based on the uptake of dsRNA by leaflets and the second was based on "in planta-induced transient gene silencing" (PITGS), a well-established method for silencing plant genes, used here for the first time to deliver in planta-transcribed dsRNA to target insect genes. Tuta absoluta larvae that fed on leaves containing dsRNA of the target genes showed an ~60% reduction in target gene transcript accumulation, an increase in larval mortality and less leaf damage. We then generated transgenic 'Micro-Tom' tomato plants that expressed hairpin sequences for both genes and observed a reduction in foliar damage by T. absoluta in these plants. Our results demonstrate the feasibility of RNAi as an alternative method for controlling this critical tomato pest.

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Camargo RA, Barbosa GO, Possignolo IP, Peres LEP, Lam E, Lima JE et al. RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum). PeerJ. 2016 Jan 1;2016(12). e2673. https://doi.org/10.7717/peerj.2673