title: Knock-down of heat-shock protein 90 and isocitrate lyase gene expression reduced root-knot nematode reproduction.
abstract: Crop losses caused by nematode infections are estimated to be valued at USD 157 billion per year. Meloidogyne incognita, a root-knot nematode (RKN), is considered to be one of the most important plant pathogens due to its worldwide distribution and the austere damage it can cause to a large variety of agronomically important crops. RNA interference (RNAi), a gene silencing process, has proven to be a valuable biotechnology alternative method for RKN control. In this study, the RNAi approach was applied, using fragments of M. incognita genes that encode for two essential molecules, heat-shock protein 90 (HSP90) and isocitrate lyase (ICL). Plant-mediated RNAi of these genes led to a significant level of resistance against M. incognita in the transgenic Nicotiana tabacum plants. Bioassays of plants expressing HSP90 dsRNA demonstrated a delay in gall formation and up to 46% reduction in eggs compared with wild-type plants. A reduction in the level of HSP90 transcripts was observed in recovered eggs from plants expressing dsRNA, indicating that gene silencing persisted and was passed along to first progeny. The ICL knock-down had no clear effect on gall formation but resulted in up to 77% reduction in egg oviposition compared with wild-type plants. Our data suggest that both genes may be involved in RKN development and reproduction. Thus, in this paper, we describe essential candidate genes that could be applied to generate genetically modified crops, using the RNAi strategy to control RKN parasitism.
authors.affiliation: First, second, third, fourth, fifth, sixth, eighth, ninth, and eleventh authors: Embrapa Genetic Resources and Biotechnology, Laboratory of Molecular Plant-Pest Interaction, Brasília, DF, Brazil; first, second, and third authors: University of Brasília, Department of Cell Biology, Graduate Program in Molecular Biology, Brasília, DF, Brazil; seventh author: Institut National de la Recherche Agronomique, Sophia-Antipolis, France; eighth author: Federal University of Minas Gerais, Botany Department, Belo Horizonte, MG, Brazil; tenth author: Embrapa Cerrados, Laboratory of Phytopathology, Planaltina, DF, Brazil; and eleventh author: Catholic University of Brasília, Graduate Program in Genomic Sciences and Biotechnology, Brasília, DF, Brazil.
authors.forename: Isabela Tristan
authors.initials: IT
authors.lastname: Lourenço-Tessutti
authors.affiliation: First, second, third, fourth, fifth, sixth, eighth, ninth, and eleventh authors: Embrapa Genetic Resources and Biotechnology, Laboratory of Molecular Plant-Pest Interaction, Brasília, DF, Brazil; first, second, and third authors: University of Brasília, Department of Cell Biology, Graduate Program in Molecular Biology, Brasília, DF, Brazil; seventh author: Institut National de la Recherche Agronomique, Sophia-Antipolis, France; eighth author: Federal University of Minas Gerais, Botany Department, Belo Horizonte, MG, Brazil; tenth author: Embrapa Cerrados, Laboratory of Phytopathology, Planaltina, DF, Brazil; and eleventh author: Catholic University of Brasília, Graduate Program in Genomic Sciences and Biotechnology, Brasília, DF, Brazil.
authors.forename: José Dijair Antonino
authors.initials: JD
authors.lastname: Souza Junior
authors.affiliation: First, second, third, fourth, fifth, sixth, eighth, ninth, and eleventh authors: Embrapa Genetic Resources and Biotechnology, Laboratory of Molecular Plant-Pest Interaction, Brasília, DF, Brazil; first, second, and third authors: University of Brasília, Department of Cell Biology, Graduate Program in Molecular Biology, Brasília, DF, Brazil; seventh author: Institut National de la Recherche Agronomique, Sophia-Antipolis, France; eighth author: Federal University of Minas Gerais, Botany Department, Belo Horizonte, MG, Brazil; tenth author: Embrapa Cerrados, Laboratory of Phytopathology, Planaltina, DF, Brazil; and eleventh author: Catholic University of Brasília, Graduate Program in Genomic Sciences and Biotechnology, Brasília, DF, Brazil.
authors.forename: Diogo
authors.initials: D
authors.lastname: Martins-de-Sa
authors.affiliation: First, second, third, fourth, fifth, sixth, eighth, ninth, and eleventh authors: Embrapa Genetic Resources and Biotechnology, Laboratory of Molecular Plant-Pest Interaction, Brasília, DF, Brazil; first, second, and third authors: University of Brasília, Department of Cell Biology, Graduate Program in Molecular Biology, Brasília, DF, Brazil; seventh author: Institut National de la Recherche Agronomique, Sophia-Antipolis, France; eighth author: Federal University of Minas Gerais, Botany Department, Belo Horizonte, MG, Brazil; tenth author: Embrapa Cerrados, Laboratory of Phytopathology, Planaltina, DF, Brazil; and eleventh author: Catholic University of Brasília, Graduate Program in Genomic Sciences and Biotechnology, Brasília, DF, Brazil.
authors.forename: Antônio Américo Barbosa
authors.initials: AA
authors.lastname: Viana
authors.affiliation: First, second, third, fourth, fifth, sixth, eighth, ninth, and eleventh authors: Embrapa Genetic Resources and Biotechnology, Laboratory of Molecular Plant-Pest Interaction, Brasília, DF, Brazil; first, second, and third authors: University of Brasília, Department of Cell Biology, Graduate Program in Molecular Biology, Brasília, DF, Brazil; seventh author: Institut National de la Recherche Agronomique, Sophia-Antipolis, France; eighth author: Federal University of Minas Gerais, Botany Department, Belo Horizonte, MG, Brazil; tenth author: Embrapa Cerrados, Laboratory of Phytopathology, Planaltina, DF, Brazil; and eleventh author: Catholic University of Brasília, Graduate Program in Genomic Sciences and Biotechnology, Brasília, DF, Brazil.
authors.forename: Regina Maria Dechechi Gomes
authors.initials: RM
authors.lastname: Carneiro
authors.affiliation: First, second, third, fourth, fifth, sixth, eighth, ninth, and eleventh authors: Embrapa Genetic Resources and Biotechnology, Laboratory of Molecular Plant-Pest Interaction, Brasília, DF, Brazil; first, second, and third authors: University of Brasília, Department of Cell Biology, Graduate Program in Molecular Biology, Brasília, DF, Brazil; seventh author: Institut National de la Recherche Agronomique, Sophia-Antipolis, France; eighth author: Federal University of Minas Gerais, Botany Department, Belo Horizonte, MG, Brazil; tenth author: Embrapa Cerrados, Laboratory of Phytopathology, Planaltina, DF, Brazil; and eleventh author: Catholic University of Brasília, Graduate Program in Genomic Sciences and Biotechnology, Brasília, DF, Brazil.
authors.forename: Roberto Coiti
authors.initials: RC
authors.lastname: Togawa
authors.affiliation: First, second, third, fourth, fifth, sixth, eighth, ninth, and eleventh authors: Embrapa Genetic Resources and Biotechnology, Laboratory of Molecular Plant-Pest Interaction, Brasília, DF, Brazil; first, second, and third authors: University of Brasília, Department of Cell Biology, Graduate Program in Molecular Biology, Brasília, DF, Brazil; seventh author: Institut National de la Recherche Agronomique, Sophia-Antipolis, France; eighth author: Federal University of Minas Gerais, Botany Department, Belo Horizonte, MG, Brazil; tenth author: Embrapa Cerrados, Laboratory of Phytopathology, Planaltina, DF, Brazil; and eleventh author: Catholic University of Brasília, Graduate Program in Genomic Sciences and Biotechnology, Brasília, DF, Brazil.
authors.forename: Janice
authors.initials: J
authors.lastname: de Almeida-Engler
authors.affiliation: First, second, third, fourth, fifth, sixth, eighth, ninth, and eleventh authors: Embrapa Genetic Resources and Biotechnology, Laboratory of Molecular Plant-Pest Interaction, Brasília, DF, Brazil; first, second, and third authors: University of Brasília, Department of Cell Biology, Graduate Program in Molecular Biology, Brasília, DF, Brazil; seventh author: Institut National de la Recherche Agronomique, Sophia-Antipolis, France; eighth author: Federal University of Minas Gerais, Botany Department, Belo Horizonte, MG, Brazil; tenth author: Embrapa Cerrados, Laboratory of Phytopathology, Planaltina, DF, Brazil; and eleventh author: Catholic University of Brasília, Graduate Program in Genomic Sciences and Biotechnology, Brasília, DF, Brazil.
authors.forename: João Aguiar Nogueira
authors.initials: JA
authors.lastname: Batista
authors.affiliation: First, second, third, fourth, fifth, sixth, eighth, ninth, and eleventh authors: Embrapa Genetic Resources and Biotechnology, Laboratory of Molecular Plant-Pest Interaction, Brasília, DF, Brazil; first, second, and third authors: University of Brasília, Department of Cell Biology, Graduate Program in Molecular Biology, Brasília, DF, Brazil; seventh author: Institut National de la Recherche Agronomique, Sophia-Antipolis, France; eighth author: Federal University of Minas Gerais, Botany Department, Belo Horizonte, MG, Brazil; tenth author: Embrapa Cerrados, Laboratory of Phytopathology, Planaltina, DF, Brazil; and eleventh author: Catholic University of Brasília, Graduate Program in Genomic Sciences and Biotechnology, Brasília, DF, Brazil.
authors.forename: Maria Cristina Mattar
authors.initials: MC
authors.lastname: Silva
authors.affiliation: First, second, third, fourth, fifth, sixth, eighth, ninth, and eleventh authors: Embrapa Genetic Resources and Biotechnology, Laboratory of Molecular Plant-Pest Interaction, Brasília, DF, Brazil; first, second, and third authors: University of Brasília, Department of Cell Biology, Graduate Program in Molecular Biology, Brasília, DF, Brazil; seventh author: Institut National de la Recherche Agronomique, Sophia-Antipolis, France; eighth author: Federal University of Minas Gerais, Botany Department, Belo Horizonte, MG, Brazil; tenth author: Embrapa Cerrados, Laboratory of Phytopathology, Planaltina, DF, Brazil; and eleventh author: Catholic University of Brasília, Graduate Program in Genomic Sciences and Biotechnology, Brasília, DF, Brazil.
authors.forename: Rodrigo Rocha
authors.initials: RR
authors.lastname: Fragoso
authors.affiliation: First, second, third, fourth, fifth, sixth, eighth, ninth, and eleventh authors: Embrapa Genetic Resources and Biotechnology, Laboratory of Molecular Plant-Pest Interaction, Brasília, DF, Brazil; first, second, and third authors: University of Brasília, Department of Cell Biology, Graduate Program in Molecular Biology, Brasília, DF, Brazil; seventh author: Institut National de la Recherche Agronomique, Sophia-Antipolis, France; eighth author: Federal University of Minas Gerais, Botany Department, Belo Horizonte, MG, Brazil; tenth author: Embrapa Cerrados, Laboratory of Phytopathology, Planaltina, DF, Brazil; and eleventh author: Catholic University of Brasília, Graduate Program in Genomic Sciences and Biotechnology, Brasília, DF, Brazil.
authors.forename: Maria Fatima
authors.initials: MF
authors.lastname: Grossi-de-Sa
mesh_terms: D000818:Animals; D005260:Female; D015870:Gene Expression; D006360:Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism; D015801:Helminth Proteins / Q000235:genetics / Q000378:metabolism; D007522:Isocitrate Lyase / Q000235:genetics* / Q000378:metabolism; D010935:Plant Diseases / Q000276:immunology* / Q000469:parasitology; D018517:Plant Roots / Q000166:cytology / Q000235:genetics / Q000276:immunology / Q000469:parasitology; D030821:Plants, Genetically Modified; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics; D012098:Reproduction; D014026:Nicotiana / Q000166:cytology / Q000235:genetics / Q000276:immunology* / Q000469:parasitology; D014431:Tylenchoidea / Q000145:classification / Q000235:genetics* / Q000472:pathogenicity / Q000502:physiology
chemical_list: D006360:Heat-Shock Proteins; D015801:Helminth Proteins; D012330:RNA, Double-Stranded; D007522:Isocitrate Lyase