title: A cytometry microparticle platform approach for screening tobacco microRNA changes after agrobacterium delivery.
abstract: MicroRNAs are a class of non-coding regulatory RNAs that can modulate development as well as alter innate antiviral defenses in plants. In this study we explored changes in Nicotiana benthamiana tobacco microRNA expression as it relates to expression of a recombinant anti-Ebola GP1 antibody. The antibody was delivered to tobacco leaves through a bacterial Agrobacterium tumefaciens "agroinfiltration" expression strategy. A multiplex microparticle-based cytometry assay tracked the expression changes of 53 host tobacco microRNAs. Our results revealed that the most abundant microRNAs in actively growing leaves corresponded to nanoparticle probes specific to nta-mir-6149 and nta-miR-168b. After agroinfiltration, probes specific for nta-mir-398, and nta-mir-482d were significantly altered in their respective expression levels, however changes were partially attributed to the infiltration broth medium used in the antibody delivery process. Confirmation of nta-mir-398 and nta-mir-482d expression changes was also verified through RT-qPCR. To our knowledge this study is the first to profile medium and Agrobacterium injection at the microRNA level through a multiplex microparticle approach.
authors.affiliation: Chemical and Biological Signature Sciences Group, Pacific Northwest National Laboratory, Richland, WA 99354, USA. Electronic address: joshua.powell@pnnl.gov.
authors.forename: Joshua D
authors.initials: JD
authors.lastname: Powell
authors.affiliation: The Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Chen
authors.affiliation: The Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Hugh S
authors.initials: HS
authors.lastname: Mason
mesh_terms: D016960:Agrobacterium tumefaciens / Q000378:metabolism; D000914:Antibodies, Viral / Q000235:genetics; D029043:Ebolavirus / Q000276:immunology; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods; D015870:Gene Expression; D005822:Genetic Vectors; D035683:MicroRNAs / Q000235:genetics*; D018515:Plant Leaves / Q000235:genetics; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D018749:RNA, Plant / Q000235:genetics*; D012680:Sensitivity and Specificity; D014026:Nicotiana / Q000235:genetics* / Q000382:microbiology
chemical_list: D000914:Antibodies, Viral; D035683:MicroRNAs; D018749:RNA, Plant
keywords: Agrobacterium tumefaciens; Ebola; Nicotiana benthamiana; Tobacco