A tobamovirus expression vector for agroinfection of legumes and Nicotiana

The highest recombinant protein expression levels in plants have been achieved using tobacco mosaic virus (TMV) vectors via agroinoculation of the tobacco, Nicotiana benthamiana. These vectors have been utilized for pharmaceutical protein production and also can serve as rapid gene expression screens for proteonomics. We have constructed a similar vector based on the legume-infecting tobamovirus, sunn hemp mosaic virus (SHMV), by deleting the coat protein gene (SHMV eliminate coat protein gene or SHEC). SHEC/GFP co-agroinoculated with a 35S/p19 binary yielded 600 microg GFP/gfw (25% TSP) in N. benthamiana. In the absence of p19, SHEC/GFP expression was nearly eliminated. SHEC also yielded strong GUS production in agroinoculated Medicago trunculata, Pinto bean, cowpea, pea and lentil even without the aid of systemic infection. A full-length version (SHAC, SHMV alternate coat protein) was created by adding to SHEC the coat protein subgenomic promoter and ORF from the tobamovirus, tobacco mild green mottle virus (TMGMV). SHAC induced a slowly developing, symptomless infection of N. benthamiana and may be of use as a virus induced gene silencing (VIGS) vector.