title: RNA interference as a gene silencing tool to controlTuta absolutain tomato (Solanum lycopersicum).
abstract: RNA interference (RNAi), a gene-silencing mechanism that involves providing double-stranded 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. We selected two target genes (Vacuolar ATPase-AandArginine kinase) based on the RNAi response reported for these genes in other pest species. In view of the lack of an artificial diet forT. 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 deliverin planta-transcribed dsRNA to target insect genes.Tuta absolutalarvae 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 byT. absolutain these plants. Our results demonstrate the feasibility of RNAi as an alternative method for controlling this critical tomato pest.
authors.affiliation: Centro de Energia Nuclear na Agricultura, Universidade de São Paulo, Piracicaba, São Paulo, Brazil; Escola Superior de Agricultura "Luiz de Queiroz" (ESALQ), Universidade de São Paulo, Piracicaba, São Paulo, Brazil.
authors.forename: Roberto A
authors.initials: RA
authors.lastname: Camargo
authors.affiliation: Departamento de Bioquímica e Biologia Tecidual, Universidade Estadual de Campinas , Campinas , São Paulo , Brazil.
authors.forename: Guilherme O
authors.initials: GO
authors.lastname: Barbosa
authors.affiliation: Centro de Energia Nuclear na Agricultura, Universidade de São Paulo, Piracicaba, São Paulo, Brazil; Escola Superior de Agricultura "Luiz de Queiroz" (ESALQ), Universidade de São Paulo, Piracicaba, São Paulo, Brazil.
authors.forename: Isabella Presotto
authors.initials: IP
authors.lastname: Possignolo
authors.affiliation: Escola Superior de Agricultura "Luiz de Queiroz" (ESALQ), Universidade de São Paulo , Piracicaba , São Paulo , Brazil.
authors.forename: Lazaro E P
authors.initials: LE
authors.lastname: Peres
authors.affiliation: Department of Plant Biology & Pathology, Rutgers, The State University of New Jersey , New Brunswick , NJ , United States.
authors.forename: Eric
authors.initials: E
authors.lastname: Lam
authors.affiliation: Centro de Energia Nuclear na Agricultura, Universidade de São Paulo, Piracicaba, São Paulo, Brazil; Departamento de Botânica, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Joni E
authors.initials: JE
authors.lastname: Lima
authors.affiliation: Centro de Energia Nuclear na Agricultura, Universidade de São Paulo , Piracicaba , São Paulo , Brazil.
authors.forename: Antonio
authors.initials: A
authors.lastname: Figueira
authors.affiliation: Departamento de Bioquímica e Biologia Tecidual, Universidade Estadual de Campinas , Campinas , São Paulo , Brazil.
authors.forename: Henrique
authors.initials: H
authors.lastname: Marques-Souza
keywords: Agro-infiltration; Gene silencing; Micro-tom; Pest control; RNAi; Transgenic; Tuta absoluta; dsRNA delivery; dsRNA uptake