title: Delivery and expression of functional viral RNA genomes in planta by agroinfiltration.
abstract: Agroinfiltration is a simple, efficient, and powerful approach for transient expression of viral genes as well as DNA-based expression of full-length RNA genomes of plant viruses for studies leading to understanding of replication, movement, and assembly. Most importantly, it results in synchronous delivery of Agrobacterium transformants to a majority of cells encompassing the infiltrated area and is therefore ideal for examining the biological activities of viruses having multipartite genomes. Because of the high transformation rate and efficient accumulation of mRNAs, the method is also ideal for analyzing biological activities of viral genomes with defective replication and cell-to-cell movement characteristics.
authors.affiliation: University of California, Riverside, Riverside, California.
authors.forename: Padmanaban
authors.initials: P
authors.lastname: Annamalai
authors.affiliation: University of California, Riverside, Riverside, California.
authors.forename: A L N
authors.initials: ALN
authors.lastname: Rao
mesh_terms: D018014:Gene Transfer Techniques; D016679:Genome, Viral / Q000502:physiology*; D018515:Plant Leaves / Q000235:genetics / Q000382:microbiology*; D012328:RNA Viruses / Q000235:genetics* / Q000502:physiology; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D012231:Rhizobium / Q000235:genetics* / Q000472:pathogenicity; D014170:Transformation, Genetic; D019065:Virus Assembly; D014779:Virus Replication
chemical_list: D012367:RNA, Viral
references.citation: Annamalai, P. and Rao, A.L.N. 2005. Agrobacterium-mediated DNA expression of functional genome components of brome mosaic virus in planta: Transient expression of viral capsid protein and RNA packaging independent of replication. Virology 338:96-111.
references.citation: Balucombe, D.C. 1999. Fast forward genetics based on virus induced gene silencing. Curr. Opin. Plant Biol. 2:109-113.
references.citation: Been, M.D. 1994. Cis-and trans-acting ribozymes from human pathogen, hepatitis delta virus. Trends Biochem. Sci. 19:251-256.
references.citation: Bendahmane, A., Querci, M., Kanyuka, K., and Baulcombe, D.C. 2000. Agrobacterium transient expression system as a tool for the isolation of disease resistance genes: Application to the R×2 locus in potato. Plant J. 21:73-81.
references.citation: Boyer, J.C. and Haenni, A.L. 1994. Infectious transcripts and cDNA clones of RNA viruses. Virology 198:415-426.
references.citation: Brigneti, G., Voinnet, O., Li, W.X., Ji, L.H., Ding, S.W., and Baulcombe, D.C. 1998. Viral pathogenicity determinants are suppressors of transgene silencing in Nicotiana benthamiana. EMBO J. 17:6739-6746.
references.citation: Bruening, G., Buzayan, J., Gerlach, W., and Hampel, A. 1988. Small RNA autolytic cleavage and ligation. In From Proteins to Ribosomes, Vol. 1 (R.H. Sarma and M.H. Sarma, eds.) pp. 239-248. Adenine Press, Schenectady, New York.
references.citation: Daros, J.A. and Flores, R. 2004. Arabidopsis thaliana has the enzymatic machinery for replicating representative viroid species of the family pospiviroidae. Proc. Natl. Acad. Sci. U.S.A. 101:6792-6797.
references.citation: Engebrecht, J., Brent, R., and Kaderbhai, M.A. 1991. Minipreps of plasmid DNA. In Current Protocols in Molecular Biology (F.M. Ausubel, R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, J.A. Smith, and K. Struhl, eds.) pp. 1.6.1-1.6.10. John Wiley & Sons, Hoboken, N.J.
references.citation: Grimsley, N., Hohn, T., Davies, J.W., and Hohn, B. 1987. Agrobacterium mediated delivery of infectious maize streak virus into maize plants. Nature 325:177-179.
references.citation: Hellens, R., Mullineaux, P., and Klee, H. 2000. A guide to Agrobacterium binary Ti vectors. Trends Plant Sci. 5:446-451.
references.citation: Ishikawa, M., Janda, M., Krol, M.A., and Ahlquist, P. 1997. In vivo DNA expression of functional brome mosaic virus RNA replicons in Sacchromyces cerevisiae. J. Virol. 71:7781-7790.
references.citation: Johansen, L.K. and Carrington, J.C. 2001. Silencing on the spot. Induction and suppression of RNA silencing in Agrobacterium-mediated transient expression system. Plant Physiol. 126:930-938.
references.citation: Kapila, J., De Rycke, R., van Montagu, M., and Angenon, G. 1997. An Agrobacterium-mediated transient gene expression system for intact leaves. Plant Sci. 122:101-108.
references.citation: Leiser, R.M., Ziegler-Graff, V., Reutenauer, A., Herrbach, E., Lemaire, O., Guilley, H., Richards, K., and Jonard, G. 1992. Agroinfection as an alternative to insects for infecting with beet western yellows luteovirus. Proc. Natl. Acad. Sci. U.S.A. 89:9136-9140.
references.citation: Li, H.W., Lucy, A.P., Guo, H.S., Li, W.X., Ji, L.H., Wong, S.M., and Ding, S.W. 1999. Strong host resistance targeted against a viral suppressor of the plant gene silencing defense mechanism. EMBO J. 18:2683-2691.
references.citation: Liu, L. and Lomonossoff, G.P. 2002. Agroinfection as a rapid method for propagating cowpea mosaic virus based constructs. J. Virol. Methods 105:343-348.
references.citation: Lu, R., Malcuit, I., Moffett, P., Ruiz, M.T., Peart, J., Wu, A.J., Rathjen, J.P., Bendahmane, A., Day, L., and Baulcombe, D.C. 2003. High-throughput virus induced gene silencing implicates heat shock protein 90 in plant disease resistance. EMBO J. 22:5690-5699.
references.citation: Palanichelvam, K., Cole, A.B., Shababi, M., and Schoelz, J.E. 2000. Agroinfiltration of cauliflower mosaic virus gene VI elicits hypersensitive response in Nicotiana species. Mol. Plant. Microbe Interact. 13:1275-1279.
references.citation: Sambrook, J. and Russell, D.W. 2001. Molecular Cloning: A Laboratory Manual, 3rd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.
references.citation: Vlot, C.A., Neelman, L., Linthorst, H.J.M., and Bol, J.F. 2001. Role of the 3′-untranslated regions of alfalfa mosaic virus RNAs in the formation of a transiently expressed replicase in plants and in the assembly of virions. J. Virol. 75:6440-6449.
references.citation: Voinnet, O., Rivas, S., Mestre, P., and Baulcombe, D.C. 2003. An enhanced transient expression system in plants based on suppression of gene silencing by the p19 protein of tomato bushy stunt virus. Plant J. 33:949-956.
references.citation: Voytas, D. 2000. Agarose gel electrophoresis. In Current Protocols in Molecular Biology (F.M. Ausubel, R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, J.A. Smith, and K. Struhl, eds.) pp. 2.5A.1-2.5A.9. John Wiley & Sons, Hoboken, N.J.
references.citation: Weigel, D. and Glazebrook, J. 2002. Arabidopsis: A laboratory manual, pp. 119-141. Cold Spring Harbor Laboratory Press, New York.
references.citation: Yang, Y., Li, R., and Qi, M. 2000. In vivo analysis of plant promoters and transcription factors by agroinfiltration of tobacco leaves. Plant J. 22:543-551.