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

PeerJ

article

Roberto A. Camargo¹  Guilherme O. Barbosa³  Isabella Presotto Possignolo¹  Lazaro E. P. Peres²  Eric Lam⁴  Joni E. Lima¹  Antonio Figueira¹  Henrique Marques-Souza³ 

[1] Centro de Energia Nuclear na Agricultura, Universidade de São Paulo;Escola Superior de Agricultura “Luiz de Queiroz” ,(ESALQ), Universidade de São Paulo;Departamento de Bioquímica e Biologia Tecidual, Universidade Estadual de Campinas;Department of Plant Biology & Pathology, Rutgers, The State University of New Jersey;Departamento de Botânica, Universidade Federal de Minas Gerais

关键词: Pest control;    Gene silencing;    RNAi;    dsRNA delivery;    Transgenic;    dsRNA uptake;    Micro-tom;    Tuta absoluta;    Agro-infiltration;   

DOI  :  10.7717/peerj.2673

学科分类：社会科学、人文和艺术（综合）

来源: Inra

【 摘 要 】

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-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.

【 授权许可】

CC BY