A sensitive and rapid RNA silencing suppressor activity assay based on alfalfa mosaic virus expression vector. 2019

Mireya Martínez-Pérez, and José A Navarro, and Vicente Pallás, and Jesús A Sánchez-Navarro
Department of Molecular and Evolutionary Plant Virology, Instituto de Biología Molecular y Celular de Plantas (IBMCP) (UPV-CSIC), Universitat Politècnica de Valencia, Ingeniero Fausto Elio s/n, 46022 Valencia, Spain. Electronic address: mimarpe6@etsid.upv.es.

Plant viruses express RNA silencing suppressor (RSS) proteins to counteract plant defence mechanisms. Here, we describe a method to assess the RSS activity based on an alfalfa mosaic virus (AMV) RNA 3 expression vector and transgenic Nicotiana tabacum plants that express the P1 and P2 subunits of the AMV replicase (P12 plants). Inoculation of P12 plants with different AMV RNA 3 constructs expressing different HC-Pro mutants that differ in their RSS capabilities, revealed a perfect correlation between necrotic lesions on inoculated leaves and RSS activity. Protoplast analysis showed that the RSS activity correlated with the accumulation of the AMV RNAs. A direct comparison between three RSS activity assays and the AMV-P12 system revealed that the coat protein of carnation mottle virus displays RSS activity with the four assays and reduced the accumulation of the siRNAs.

UI MeSH Term Description Entries
D010641 Phenotype The outward appearance of the individual. It is the product of interactions between genes, and between the GENOTYPE and the environment. Phenotypes
D010935 Plant Diseases Diseases of plants. Disease, Plant,Diseases, Plant,Plant Disease
D010942 Plant Viruses Viruses parasitic on plants. Phytophagineae,Plant Virus,Virus, Plant,Viruses, Plant
D005822 Genetic Vectors DNA molecules capable of autonomous replication within a host cell and into which other DNA sequences can be inserted and thus amplified. Many are derived from PLASMIDS; BACTERIOPHAGES; or VIRUSES. They are used for transporting foreign genes into recipient cells. Genetic vectors possess a functional replicator site and contain GENETIC MARKERS to facilitate their selective recognition. Cloning Vectors,Shuttle Vectors,Vectors, Genetic,Cloning Vector,Genetic Vector,Shuttle Vector,Vector, Cloning,Vector, Genetic,Vector, Shuttle,Vectors, Cloning,Vectors, Shuttle
D012367 RNA, Viral Ribonucleic acid that makes up the genetic material of viruses. Viral RNA
D012680 Sensitivity and Specificity Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed) Specificity,Sensitivity,Specificity and Sensitivity
D015870 Gene Expression The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION. Expression, Gene,Expressions, Gene,Gene Expressions
D017794 Alfalfa mosaic virus The type species of the genus ALFAMOVIRUS that is non-persistently transmitted by aphids. Alfalfa mosaic viruses
D020868 Gene Silencing Interruption or suppression of the expression of a gene at transcriptional or translational levels. Gene Inactivation,Inactivation, Gene,Silencing, Gene
D023061 Gene Order The sequential location of genes on a chromosome. Gene Arrangement,Gene Position,Arrangement, Gene,Arrangements, Gene,Gene Arrangements,Gene Positions,Order, Gene,Position, Gene,Positions, Gene

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