Nicotiana benthamiana : Its History and Future as a Model for Plant–Pathogen Interactions

Department of Plant Pathology, University of Kentucky, Lexington 40546, USA.
Molecular Plant-Microbe Interactions (Impact Factor: 3.94). 09/2008; 21(8):1015-26. DOI: 10.1094/MPMI-21-8-1015
Source: PubMed

ABSTRACT Nicotiana benthamiana is the most widely used experimental host in plant virology, due mainly to the large number of diverse plant viruses that can successfully infect it. Additionally, N. benthamiana is susceptible to a wide variety of other plant-pathogenic agents (such as bacteria, oomycetes, fungi, and so on), making this species a cornerstone of host-pathogen research, particularly in the context of innate immunity and defense signaling. Moreover, because it can be genetically transformed and regenerated with good efficiency and is amenable to facile methods for virus-induced gene silencing or transient protein expression, N. benthamiana is rapidly gaining popularity in plant biology, particularly in studies requiring protein localization, interaction, or plant-based systems for protein expression and purification. Paradoxically, despite being an indispensable research model, little is known about the origins, genetic variation, or ecology of the N. benthamiana accessions currently used by the research community. In addition to addressing these latter topics, the purpose of this review is to provide information regarding sources for tools and reagents that can be used to support research in N. benthamiana. Finally, we propose that N. benthamiana is well situated to become a premier plant cell biology model, particularly for the virology community, who as a group were the first to recognize the potential of this unique Australian native.

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Available from: Naidu Rayapati, Aug 30, 2014
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    • "Agrobacterium transfection and plant viruses (Goodin et al., 2008), as well as its growth pattern and high leaf:plant biomass ratio. The facility has the capacity to grow over 4 million N. benthamiana plants at a single time in a highly automated multilevel growing environment under proprietary LED lighting fixtures. "
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    ABSTRACT: Rapid, large-scale manufacture of medical countermeasures can be uniquely met by the plant-made-pharmaceutical platform technology. As a participant in the Defense Advanced Research Projects Agency (DARPA) Blue Angel project, the Caliber Biotherapeutics facility was designed, constructed, commissioned and released a therapeutic target (H1N1 influenza subunit vaccine) in <18 months from groundbreaking. As of 2015, this facility was one of the world's largest plant-based manufacturing facilities, with the capacity to process over 3500 kg of plant biomass per week in an automated multilevel growing environment using proprietary LED lighting. The facility can commission additional plant grow rooms that are already built to double this capacity. In addition to the commercial-scale manufacturing facility, a pilot production facility was designed based on the large-scale manufacturing specifications as a way to integrate product development and technology transfer. The primary research, development and manufacturing system employs vacuum-infiltrated Nicotiana benthamiana plants grown in a fully contained, hydroponic system for transient expression of recombinant proteins. This expression platform has been linked to a downstream process system, analytical characterization, and assessment of biological activity. This integrated approach has demonstrated rapid, high-quality production of therapeutic monoclonal antibody targets, including a panel of rituximab biosimilar/biobetter molecules and antiviral antibodies against influenza and dengue fever.
    Plant Biotechnology Journal 09/2015; 13(8):1191-1198. DOI:10.1111/pbi.12469 · 5.75 Impact Factor
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    • "Indeed, expression can be detected as little as 2–3 days after gene transfer, avoiding the lengthy process of stable transformation, and allowing large-scale genetic analyses. In the last 20 years, transient expression assays enabled the validation of many plant gene functions, as well as promoter activity and transgene functionality, especially in model species like N. benthamiana (Goodin et al., 2008). Recently, as described above, transient expression assays have also become a key technology for better understanding grapevine biology. "
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    ABSTRACT: In the past few years, the usefulness of transient expression assays has continuously increased for the characterization of unknown gene function and metabolic pathways. In grapevine (Vitis vinifera L.), one of the most economically important fruit crops in the world, recent systematic sequencing projects produced many gene data sets that require detailed analysis. Due to their rapid nature, transient expression assays are well suited for large-scale genetic studies. Although genes and metabolic pathways of any species can be analysed by transient expression in model plants, a need for homologous systems has emerged to avoid the misinterpretation of results due to a foreign genetic background. Over the last 10 years, various protocols have thus been developed to apply this powerful technology to grapevine. Using cell suspension cultures, somatic embryos, leaves or whole plantlets, transient expression assays enabled the study of the function, regulation and subcellular localization of genes involved in specific metabolic pathways such as the biosynthesis of phenylpropanoids. Disease resistance genes that could be used for marker-assisted selection in conventional breeding or for stable transformation of elite cultivars have also been characterized. Additionally, transient expression assays have proved useful for shaping new tools for grapevine genetic improvement: synthetic promoters, silencing constructs, minimal linear cassettes or viral vectors. This review provides an update on the different tools (DNA constructs, reporter genes, vectors) and methods (Agrobacterium-mediated and direct gene transfer methods) available for transient gene expression in grapevine. The most representative results published thus far are then described.
    Plant Biotechnology Journal 12/2014; 12(9):1231–1245. DOI:10.1111/pbi.12294 · 5.75 Impact Factor
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    • "Data obtained here demonstrate that the ability of CLBVbased vectors to trigger VIGS can be different depending on the host used and the results obtained in a host cannot be extrapolated to the other. N. benthamiana has been one of the model plants for VIGS studies (Goodin et al., 2008, Kumagai et al., 1995, Thomas et al., 2001) and many viral vectors have been assayed for VIGS in it, in addition to their natural hosts (Muruganantham et al., 2009). However, there must be aware that the results obtained may be conditioned by its lack of a functional RDR1. "
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    ABSTRACT: Virus induced gene silencing (VIGS) is an effective technology for gene function analysis in plants. We assessed the VIGS effectiveness in Nicotiana benthamiana and citrus plants of different Citrus leaf blotch virus (CLBV)-based vectors, using inserts of the phytoene desaturase (pds) gene. While in N. benthamiana the silencing phenotype was induced only by the construct carrying a 58-nt pds hairpin, in citrus plants all the constructs induced the silencing phenotype. Differences in the generation of secondary small interfering RNAs in both species are believed to be responsible for differential host-species effects. The ability of CLBV-based vectors to silence different endogenous citrus genes was further confirmed. Since CLBV-based vectors are known to be stable and induce VIGS in successive flushes for several months, these vectors provide an important genomic tool and it is expected that they will be useful to analyze gene function by reverse genetics in the long-lived citrus plants.
    Virology 04/2014; DOI:10.1016/j.virol.2014.04.017 · 3.32 Impact Factor
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