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Effects and Effectiveness of Two RNAi Constructs for Resistance to Pepper golden mosaic virus in Nicotiana benthamiana Plants
Laboratorio de Fitopatología, Centro de Investigaciones Biológicas del Noroeste, Instituto Politécnico Nacional 195, Col. Playa Palo de Santa Rita, La Paz, Baja California Sur, 23096, Mexico
Departamento de Ingeniería Genética, Centro de Investigación y de Estudios Avanzados del IPN, Unidad Irapuato, Km. 9.6 Libramiento Norte, Irapuato, Guanajuato, 36821, Mexico
Departamento de Biología Medioambiental, Centro de Investigaciones Biológicas, Ramiro de Maeztu 9, Madrid, 28040, Spain
* Author to whom correspondence should be addressed.
Received: 17 October 2013; in revised form: 14 November 2013 / Accepted: 20 November 2013 / Published: 28 November 2013
Abstract: ToChLPV and PepGMV are Begomoviruses that have adapted to a wide host range and are able to cause major diseases in agronomic crops. We analyzed the efficacy of induced resistance to PepGMV in Nicotiana benthamiana plants with two constructs: one construct with homologous sequences derived from PepGMV, and the other construct with heterologous sequences derived from ToChLPV. Plants protected with the heterologous construct showed an efficacy to decrease the severity of symptoms of 45%, while plants protected with the homologous construct showed an efficacy of 80%. Plants protected with the heterologous construct showed a reduction of incidence of 42.86%, while the reduction of incidence in plants protected with the homologous construct was 57.15%. The efficacy to decrease viral load was 95.6% in plants protected with the heterologous construct, and 99.56% in plants protected with the homologous construct. We found, in both constructs, up-regulated key components of the RNAi pathway. This demonstrates that the efficacy of the constructs was due to the activation of the gene silencing mechanism, and is reflected in the decrease of viral genome copies, as well as in recovery phenotype. We present evidence that both constructs are functional and can efficiently induce transient resistance against PepGMV infections. This observation guarantees a further exploration as a strategy to control complex Begomovirus diseases in the field.
Keywords: Hairpin RNA; Mixed infections; PepGMV; RNAi constructs; ToChLPV; Viral load
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Medina-Hernández, D.; Rivera-Bustamante, R.F.; Tenllado, F.; Holguín-Peña, R.J. Effects and Effectiveness of Two RNAi Constructs for Resistance to Pepper golden mosaic virus in Nicotiana benthamiana Plants. Viruses 2013, 5, 2931-2945.
Medina-Hernández D, Rivera-Bustamante RF, Tenllado F, Holguín-Peña RJ. Effects and Effectiveness of Two RNAi Constructs for Resistance to Pepper golden mosaic virus in Nicotiana benthamiana Plants. Viruses. 2013; 5(12):2931-2945.
Medina-Hernández, Diana; Rivera-Bustamante, Rafael F.; Tenllado, Francisco; Holguín-Peña, Ramón J. 2013. "Effects and Effectiveness of Two RNAi Constructs for Resistance to Pepper golden mosaic virus in Nicotiana benthamiana Plants." Viruses 5, no. 12: 2931-2945.