Positive and negative sense coat protein gene-mediated protection against poplar mosaic carlavirus in Nicotiana benthamiana

Plants of Nicotiana benthamiana were transformed with four constructs based on the coat protein gene of a poplar mosaic carlavirus (PMV) isolate from the UK. The four constructs were: the capsid protein coding sequence plus a portion of the adjacent sequence encoding a protein with a molecular mass of 14 kDa (CP14k); the capsid protein coding sequence in the positive sense (CPP); a mutated capsid protein coding sequence (CPM) and the capsid protein coding sequence in the negative sense (CPN). Forty-one regenerated plants, after selection for their kanamycin resistance, were confirmed by PCR to contain the appropriate sequences. Virus coat protein was detected in small amounts in 50% of the plants transformed with the CP14k or CPP constructs. Primary transformants showed a range of reactions to challenge with two isolates of PMV. These varied from apparently no infection in inoculated or in later-formed young leaves, as assessed by ELISA, to typical systemic symptoms associated with large amounts of serologically detected virus. There was no correlation between the level of protection against virus infection and the observed accumulation of transgene protein product. Plants were protected whether transformed with the coat protein coding sequence in the positive or negative sense.