Agroinfection as a rapid method for propagating Cowpea mosaic virus-based constructs.
To increase the efficiency of infections with Cowpea mosaic virus (CPMV)-based constructs, clones suitable for agroinfection were constructed. Full-length copies of RNA-1 and RNA-2 were inserted between the sequence of a Cauliflower mosaic virus (CaMV) 35S promoter and a nos terminator and were introduced into the Agrobacterium tumefaciens plasmid, pBINPLUS. Infiltration of leaves of either Nicotiana benthamiana or cowpea (Vigna unguiculata) with a bacterial suspension containing a mixture of the RNA-1- and RNA-2-based plasmids resulted in the plants developing typical CPMV symptoms. To confirm the utility of this approach for use with CPMV-based vectors, a GFP construct based on RNA-2 was adapted for agroinfection. Infiltration of N. benthamiana leaves with a mixture of Agrobacteria containing this construct and the RNA-1 plasmid resulted in high levels of GFP expression. The results demonstrate that agroinfection is a suitable method for the propagation of CPMV-based derivatives.
Liu
Li
L
Department of Metabolic Biology, John Innes Centre, Colney Lane, Norwich NR47UH, UK.
Lomonossoff
George
G
D016960:Agrobacterium tumefaciens / Q000821:virology; D017798:Comovirus / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology*; D007887:Fabaceae / Q000821:virology; D010935:Plant Diseases / Q000821:virology*; D010957:Plasmids; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D015183:Restriction Mapping / Q000379:methods; D014026:Nicotiana / Q000821:virology; D014773:Virology / Q000379:methods; D014779:Virus Replication / Q000502:physiology*
D012367:RNA, Viral