Systemic Agrobacterium tumefaciens-mediated transfection of viral replicons for efficient transient expression in plants.
Plant biotechnology relies on two approaches for delivery and expression of heterologous genes in plants: stable genetic transformation and transient expression using viral vectors. Although much faster, the transient route is limited by low infectivity of viral vectors carrying average-sized or large genes. We have developed constructs for the efficient delivery of RNA viral vectors as DNA precursors and show here that Agrobacterium-mediated delivery of these constructs results in gene amplification in all mature leaves of a plant simultaneously (systemic transfection). This process, called "magnifection", can be performed on a large scale and with different plant species. This technology combines advantages of three biological systems (the transfection efficiency of A. tumefaciens, the high expression yield obtained with viral vectors, and the post-translational capabilities of a plant), does not require genetic modification of plants and is faster than other existing methods.
Marillonnet
Sylvestre
S
Icon Genetics, Biozentrum Halle, Weinbergweg 22, D-06120 Halle (Saale), Germany.
Thoeringer
Carola
C
Kandzia
Romy
R
Klimyuk
Victor
V
Gleba
Yuri
Y
D016960:Agrobacterium tumefaciens / Q000502:physiology*; D027461:Beta vulgaris / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors; D016679:Genome, Viral; D030821:Plants, Genetically Modified; D012367:RNA, Viral / Q000235:genetics*; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism*; D014027:Tobacco Mosaic Virus / Q000235:genetics*; D014162:Transfection / Q000379:methods*; D014779:Virus Replication
D012367:RNA, Viral