Optimization of transient Agrobacterium-mediated gene expression system in leaves of Nicotiana benthamiana

• Autorzy: Wydro M. , Kozubek E. , Lehmann P.
• Języki publikacji: EN

Abstrakty

EN

Here we report on a simple and reproducible system of Agrobacterium-mediated transient gene expression assay that utilizes infiltration of young Nicotiana benthamiana leaves. Although some of the phenomena described in this paper have been already reported by other researchers, here we have further developed them. The highest level of transient gfp gene expression was detected in the youngest leaves of N. benthamiana infiltrated with A. tumefaciens strains AGL0 and EHA105 precultured in the presence of 450-600 µM acetosyringone. Although the maximum level of transient gfp gene expression was restricted presumably by RNA silencing, it was completely suppressed in the presence of the viral protein HC-Pro. The transient expression system described here can be used to identify new viral suppressors of RNA silencing, for detailed analysis of unidentified genes and for industrial production of proteins in plants as well.

Słowa kluczowe

EN

green fluorescent protein; HC-Pro protein; Nicotiana benthamiana; viral suppressor; gene expression; Agrobacterium; RNA silencing; leaf

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2006
• Tom: 53
• Numer: 2
• Opis fizyczny: p.289-298,fig.,ref.

Twórcy

autor

Wydro M.

• Polish Academy of Sciences, Poznan, Poland

autor

Kozubek E.

autor

Lehmann P.

Bibliografia

• Al-Kaff NS, Covey SN, Kreike MM, Page AM, Pinder R, Dale PJ (1998) Transcriptional and posttranscriptional plant gene silencing in response to a pathogen. Science 279: 2113–2115.
• Anandalakshmi R, Pruss GJ, Ge X, Marathe R, Mallory AC, Smith TH, Vance VB (1998) A viral suppressor of gene silencing in plants. Proc Natl Acad Sci USA 95: 13079–13084.
• Baulcombe DC (1996) RNA as a target and an initiator of post-transcriptional gene silencing in transgenic plants. Plant Mol Biol 32: 79–88.
• Baulcombe D (2004) RNA silencing in plants. Nature 431: 356–363.
• Baulcombe DC, Malnár A (2004) Crystal structure of p19 — a universal suppressor of RNA silencing. Trends Biochem Sci 29: 279–281.
• Dougherty WG, Parks TD (1995) Transgenes and gene suppression: telling us something new? Curr Opin Cell Biol 7: 399–405.
• English JJ, Mueller E, Baulcombe DC (1996) Suppression of virus accumulation in transgenic plants exhibiting silencing of nuclear genes. Plant Cell 8: 179–188.
• Fischer R, Vaquero-Martin C, Sack M, Drossard J, Emans N, Commandeur U (1999) Towards molecular farming in the future: transient protein expression in plants. Biotechnol Appl Biochem 30: 113–116.
• Fischer R, Stoger E, Schillberg S, Christou P, Twyman R (2004) Plant-based production of biopharmaceuticals. Curr Opin Plant Biol 7: 152–158.
• Halfhill MD, Millwood RJ, Rufty TW, Weissinger AK, Stewart CN (2003) Spatial and temporal patterns of green fluorescent protein (GFP) fluorescence during leaf canopy development in transgenic oilseed rape, Brassica napus L. Plant Cell Rep 22: 338–343.
• Hamilton A, Voinnet O, Chappell L, Baulcombe D (2002) Two classes of short interfering RNA in RNA silencing. EMBO J 21: 4671–4679.
• Haseloff J, Siemering KR, Prasher DC, Hodge S (1997) Removal of a cryptic intron and subcellular localization of green fluorescent protein are required to mark transgenic Arabidopsis plants brightly. Proc Natl Acad Sci USA 94: 2122–2127.
• Hiei Y, Ohta S, Komari T, Kumashiro T (1994) Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J 6: 271–282.
• Horn ME, Woodard SL, Howard JA (2004) Plant molecular farming: systems and products. Plant Cell Rep 22: 711–720.
• Jeoung JM, Krishnaveni S, Muthukrishnan S, Trick HN, Liang GH (2002) Optimization of sorghum transformation parameters using genes for green fluorescent protein and β-glucuronidase as visual markers. Hereditas 137: 20–28.
• Johansen LK, Carrington JC (2001) Silencing on the spot. Induction and suppression of RNA silencing in the Agrobacterium-mediated transient expression system. Plant Physiol 126: 930–938.
• Kapila J, DeRycke R, VanMontagu M, Angenon G (1997) An Agrobacterium-mediated transient gene expression system for intact leaves. Plant Sci 122: 101–108.
• Kasschau KD, Carrington JC (1998) A counterdefensive strategy of plant viruses: suppression of posttranscriptional gene silencing. Cell 95: 461–470.
• Kozak M (1984) Point mutations close to the AUG initiator codons affect the efficiency of translation of rat preproinsulin in vivo. Nature 308: 241–246.
• Lakatos L, Szittya G, Silhavy D, Burgyan J (2004) Molecular mechanism of RNA silencing suppression mediated by p19 protein of tombusviruses. EMBO J 23: 876–884.
• Lindbo JA, Silva-Rosales L, Proebsting WM, Dougherty WG (1993) Induction of a highly specific antiviral state in transgenic plants: implications for regulation of gene expression and virus resistance. Plant Cell 5: 1749–1759.
• Mallory AC, Reinhart BJ, Bartel D, Vance VB, Bowman LH (2002) From the cover: a viral suppressor of RNA silencing differentially regulates the accumulation of short interfering RNAs and mikro-RNAs in tobacco. Proc Natl Acad Sci USA 99: 15228–15233.
• Maximova SN, Dandekar AM, Guiltinan MJ (1998) Investigation of Agrobacterium-mediated transformation of apple using green fluorescent protein: high transient expression and low stable transformation suggest that factors other than T-DNA transfer are rate-limiting. Plant Mol Biol 37: 549–559.
• Pruss G, Ge X, Shi XM, Carrington JC, Vance VB (1997) Plant viral synergism: the potyviral genome encodes a broad-range pathogenicity enhancer that transactivates replication of heterologous viruses. Plant Cell 9: 858–868.
• Qu F, Ren T, Morris TJ (2003) The coat protein of turnip crincle virus suppresses posttranscriptional gene silencing at an early initiation step. J Virol 77: 511–522.
• Reed JC, Kasschau KD, Prokhnevsky AI, Gopinath K, Pogue GP, Carrington JC, Dolja VV (2003) Suppressor of RNA silencing encoded by Beet yellows virus. Virology 306: 203–209.
• Roth BM, Pruss GJ, Vance VB (2004) Plant viral suppressors of RNA silencing. Virus Res 102: 97–108.
• Sambrook J, Russell DW (2001) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
• Siemering KR, Golbik R, Sever R, Haseloff J (1996) Mutations that suppress the thermosensitivity of green fluorescent protein. Curr Biol 6: 1653–1663.
• Smith HA, Swaney SL, Parks TD, Wernsman EA, Dougherty WG (1994) Transgenic plant virus resistance mediated by untranslatable sense RNAs: expression, regulation, and fate of nonessential RNAs. Plant Cell 6: 1441–1453.
• Tenllado F, Llave C, Diaz-Ruiz JR (2004) RNA interference as a new biotechnological tool for the control of virus diseases in pants. Virus Res 102: 85–96.
• Thomas CL, Leh V, Lederer C, Maule AJ (2003) Turnip crincle virus coat protein mediates suppression of RNA silencing in Nicotiana benthamiana. Virology 306: 33–41.
• Vaquero C, Sack M, Chandler J, Drossard J, Schuster F, Monecke M, Schillberg S, Fischer R (1999) Transient expression of a tumor-specific single-chain fragment and a chimeric antibody in tobacco leaves. Proc Natl Acad Sci USA 96: 11128–11133.
• Voinnet O, Lederer C, Baulcombe DC (2000) A viral movement protein prevents systemic spread of the gene silencing signal in Nicotiana benthamiana. Cell 103: 157–167.
• Voinnet O, Rivas S, Mestre P, Baulcombe D (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.