Abstract
Plant virus infectious clones are important tools with wide‐ranging applications in different areas of biology and medicine. Their uses in plant pathology include the study of plant–virus interactions, and screening of germplasm as part of prebreeding programmes for virus resistance. They can also be modified to induce transient plant gene silencing (Virus Induced Gene Silencing – VIGS) and as expression vectors for plant or exogenous proteins, with applications in both plant pathology and more generally for the study of plant gene function. Plant viruses are also increasingly being investigated as expression vectors for in planta production of pharmaceutical products, known as molecular farming. However, plant virus infectious clones may pose a risk to the environment due to their ability to reconstitute fully functional, transmissible viruses. These risks arise from both their inherent pathogenicity and the effect of any introduced genetic modifications. Effective containment measures are therefore required. There has been no single comprehensive review of the biosafety considerations for the contained use of genetically modified plant viruses, despite their increasing importance across many biological fields. This review therefore explores the biosafety considerations for working with genetically modified plant viruses in contained environments, with focus on plant growth facilities. It includes regulatory frameworks, risk assessment, assignment of biosafety levels, facility features and working practices. The review is based on international guidance together with information provided by plant virus researchers.
Highlights
Plant viruses are important because of the threat they pose to global crop production, and due to their use as research tools in molecular plant pathology and other areas of biotechnology. (Rybicki 2015; Scholthof et al 2011)
We review individual risk assessments and protocols provided by researchers currently working with plant virus infectious clones (ICs) in Europe and the USA
Plant virus ICs are important molecular tools in many areas of biology. Their status as both genetically modified microorganisms (GMMs) and plant pathogens necessitates their containment by a combination of biological, physical and operational measures to prevent harm to the environment and allay concerns regarding perceived or potential risks to human health
Summary
In the case of molecular farming, researchers use genetic modific tio to ‘di ble’ vi uecig exogenous proteins, for example by removing genes required for dissemination or transmission (Gleba et al 2007, Hefferon 2014) Their evaluation of risk relies on the assumption that this modification is stable and would persist should the infectious clone be released into the environment, without acknowledging that the virus could be rendered fully infective via viral recombination, whilst maintaining the genes for exogenous protein production (Phillipson & Weekes 2005). It is important to consider the possibility of dual use during a risk assessment, to liaise with the appropriate competent authorities and to instigate proportionate containment and security measures to guard against misappropriation
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