Assessment of recombinants that arise from the use of a TMV-based transient expression vector

A potential use of virus-based transient expression vectors is the large-scale production of commercial specialty products, which would require the inoculation of many acres of plants with the viral vector. However, there are several concerns about the widespread use of virus-based vectors. Among these are the spread of the engineered virus to susceptible plants and the generation and persistence of recombinant viruses in the environment. Using a Tobacco mosaic virus (TMV)-based transient gene expression vector, 30B, which expresses the jellyfish green fluorescent protein (30B-GFP), we describe the predominant types of hybrid viruses that developed in plants. In general, the recombinants deleted the foreign gene and repeated sequences, retaining only those sequences required for optimal replication and movement. In pathogenicity studies and challenge experiments designed to make a comparative assessment of the competitiveness of the recombinants with the parent virus, the recombinants had reduced vigor and were less competitive and pathogenic than TMV, a virus which is already present in the areas where tobacco is grown.