Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 102, Issue 2, pp 245–250 | Cite as

Transient gene expression in rose petals via Agrobacterium infiltration

  • Aneela Yasmin
  • Thomas DebenerEmail author
Original Paper


The study of gene function in roses is hampered by the low efficiency of transformation systems and the long time span needed for the generation of transgenic plants. For some functional analyses, the transient expression of genes would be an efficient alternative. Based on current protocols for the transient expression of genes via the infiltration of Agrobacterium into plant tissues, we developed a transient expression system for rose petals. We used β-glucuronidase (GUS) as a marker gene to optimize several parameters with effects on GUS expression. The efficiency of expression was found to be dependent on the rose genotype, flower age, position of petals within a flower, Agrobacterium strain and temperature of co-cultivation. The highest GUS expression was recorded in petals of the middle whirls of half-bloomed flowers from cultivars of ‘Pariser Charme’ and ‘Marvel’.


Rosa Rose petals Agrobacterium mediated transient expression 



Transfer deoxyribonucleic acid


RNA interference




Optical density



The first author is thankful to Deutscher Akademischer Austausch Dienst (DAAD), Higher Education Commission of Pakistan (HEC) and Sindh Agriculture University, Tandojam for the award of scholarship and study leave.


  1. Batoko H, Zheng HQ, Hawes C, Moore I (2000) A Rab1 GTPase is required for transport between the endoplasmic reticulum and golgi apparatus and for normal golgi movement in plants. Plant Cell 12:2201–2217CrossRefPubMedGoogle Scholar
  2. Bevan MW (1984) Binary Agrobacterium vectors for plant transformation. Nuceic Acids Res 22:8711–8721CrossRefGoogle Scholar
  3. Clemente T (2006) Nicotiana (Nicotiana tobaccum, Nicotiana benthamiana). In: Wang K (ed) Agrobacterium protocols, 2nd edn. Humana press, New Jersey, pp 143–154Google Scholar
  4. Dillen W, De Clercq J, Kapila J, Zambre M, Van Montagu M, Angenon G (1997) Effect of temperature on Agrobacterium tumefaciens-mediated gene transfer to plants. Plant J 6:1459–1463CrossRefGoogle Scholar
  5. Dohm A, Ludwig C, Schiling D, Debener T (2001) Transformation of roses with genes for antifungal proteins. Acta Hortic 547:27–33Google Scholar
  6. Hellens R, Mullineaux P, Klee H (2000) Technical focus: a guide to Agrobacterium binary Ti vectors. Trends Plant Sci 5(10):446–451CrossRefPubMedGoogle Scholar
  7. Janssen BJ, Gardner RC (1989) Localized transient expression of GUS in leaf discs following cocultivation with Agrobacterium. Plant Mol Biol 14:61–72CrossRefGoogle Scholar
  8. Jefferson RA, Kavanagh TA, Bevan MV (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907PubMedGoogle Scholar
  9. Joh LD, Wroblewski T, Ewing NN, VanderGheynst JS (2005) High-level transient expression of recombinant protein in lettuce. Biotechnol Bioeng 91(7):861–871CrossRefPubMedGoogle Scholar
  10. Kapila J, Rycke RD, Montagu MV, Angenon G (1997) An Agrobacterium-mediated transient gene expression system for intact leaves. Plant Sci 122:101–108CrossRefGoogle Scholar
  11. Kim MJ, Baek K, Park CM (2009) Optimization of conditions for transient Agrobacterium-mediated gene expression assays in Arabidopsis. Plant Cell Rep 28:1159–1167CrossRefPubMedGoogle Scholar
  12. Marchant R, Power JB, Lucas JA, Davey MR (1998) Biolistic transformation of rose (Rosa hybrida L.). Ann Bot 81:109–114CrossRefGoogle Scholar
  13. McCullen CA, Binns AN (2006) Agrobacterium tumefaciens and plant cell interactions and activities required for interkingdom macromolecular transfer. Annu Rev Cell Dev Biol 22:101–127. doi: 10.1146/annurev.cellbio.22.011105.102022 CrossRefPubMedGoogle Scholar
  14. McIntosh KB, Hulm JL, Young LW, Bonham-Smith PC (2004) A rapid Agrobacterium-mediated Arabidopsis thaliana transient assay system. Plant Mol Biol Report 22:53–61CrossRefGoogle Scholar
  15. R Development Core Team (2009) R: a language and environment for statistical computing. ISBN 3-9000051-07-0, URL:
  16. Santos-Rosa M, Poutaraud A, Merdinoglu D, Mestre P (2008) Development of a transient expression system in grapevine via agro-infiltration. Plant Cell Rep 27:1053–1063. doi: 10.1007/s00299-008-0531-z CrossRefPubMedGoogle Scholar
  17. Schöb H, Kunz C, Meins F Jr (1997) Silencing of transgenes introduced into leaves by agroinfiltration: a simple, rapid method for investigation of sequence requirements for gene silencing. Mol Gen Genet 256:581–585CrossRefPubMedGoogle Scholar
  18. Schweizer P, Pokorny J, Abderhalden O, Dudler R (1999) A transient assay system for the functional assessment of defense-related genes in wheat. Mol Plant-Microbe Interact 12:647–654CrossRefGoogle Scholar
  19. Sheludko YV, Sindarovska YR, Gerasymenko IM, Bannikova MA, Kuchuk NV (2007) Comparison of several Nicotiana species as hosts for high-scale Agrobacterium-mediated transient expression. Biotechnol Bioeng 3:608–614CrossRefGoogle Scholar
  20. Van der Hoorn JAL, Laurent F, Roth R, De Wit PJGM (2000) Agroinfiltration is a versatile tool that facilitates comparative analyses of Avr9/cf-9-induced and Avr4/Cf-4-induced necrosis. Mol Plant Microbe Interact 13:439–446CrossRefPubMedGoogle Scholar
  21. Van Engelen FA, Molthoff JW, Conner AJ, Nap JP, Pereira A, Stiekema WJ (1995) pBINPLUS: an improved plant transformation vector based on pBIN19. Transgenic Res 4:288–290CrossRefPubMedGoogle Scholar
  22. Wroblewski T, Tomczak A, Michelmore R (2005) Optimization of Agrobacterium mediated transient expression assays for lettuce, tomato and Arabidopsis. Plant Biotech J 3:259–273CrossRefGoogle Scholar
  23. Zhang X, Henriques R, Lin S, Niu Q, Chua N (2006) Agrobacterium-mediated transformation of Arabidopsis Thaliana using the floral dip. Nat Protoc 1(2):1–6CrossRefGoogle Scholar
  24. Zottini M, Barizza E, Costa A, Formentin E, Ruberti C, Carimi F, Schiavo FL (2008) Agroinfiltration of grapevine leaves for fast transient assays of gene expression and for long-term production of stable transformed cells. Plant Cell Rep 27:845–853. doi: 10.1007/s00299-008-0510-4 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  1. 1.Institute of Plant Genetics, Department of Molecular BreedingLeibniz University of HannoverHannoverGermany

Personalised recommendations