Abstract
The green fluorescent protein (GFP) holds promise as a field-level transgene marker. One obstacle to the use of GFP is fluorescence variability observed within leaf canopies. In growth chamber and field experiments, GFP fluorescence in transgenic oilseed rape (Brassica napus) was shown to be variable at each leaf position over time and among different leaves on the same plant. A leaf had its highest GFP fluorescence after emergence and, subsequently, its fluorescence intensity decreased. GFP fluorescence intensity was directly correlated with the concentration of soluble protein. The concentration of the genetically linked recombinant Bacillus thuringiensis (Bt) cry1Ac endotoxin protein also was examined, and GFP fluorescence was positively correlated with Bt throughout development. The results show that GFP can be used as an accurate transgene marker but that aspects of plant developmental should be taken into account when interpreting fluorescence measurements.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Beckie HJ, Warwick SI, Nair H, Seguin-Swartz G (2003) Gene flow in commercial fields of herbicide-resistant oilseed rape (Brassica napus). Ecol Appl (in press)
Benfey PN, Chua N (1990) The cauliflower mosaic virus 35S promoter: combinatorial regulation of transcription in plants. Science 250:959–966
Blumenthal A, Kuznetzova L, Edelbaum O, Raskin V, Levy M, Sela I (1999) Measurement of green fluorescent protein in plants: quantification, correlation to expression, rapid screening and differential gene expression. Plant Sci 142:93–99
Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC (1994) Green fluorescent protein as a marker for gene expression. Science 263: 802–805
Chiu WL, Niwa Y, Zeng W, Hirano T, Kobayashi H, Sheen J (1996) Engineered GFP as a vital reporter in plants. Curr Biol 6:325–330
Downs RJ, Thomas JF (1991) Phytotron procedural manual. N C Agric Res Serv Tech Bull 244
Friedrich JW, Huffaker RC (1980) Photosynthesis, leaf resistances, and ribulose-1.5-bisphosphate carboxylase degradation in senescing barley leaves. Plant Physiol 65:1103–1107
Gan S, Amasino RW (1995) Inhibition of leaf senescence by autoregulated production of cytokinin. Science 270:1986–1988
Halfhill MD, Richards HA, Mabon SA, Stewart CN Jr (2001) Expression of GFP and Bt transgenes in Brassica napus and hybridization and introgression with Brassica rapa. Theor Appl Genet 103:362–368
Hall L, Topinka K, Huffman J, Davis L, Good A (2000) Pollen flow between herbicide-resistant Brassica napus is the cause of multiple-resistant B. napus volunteers. Weed Sci 48:688–694
Harper BK, Stewart CN Jr (2000) Patterns of green fluorescent protein expression in transgenic plants. Plant Mol Biol Rep 18:1–9
Harper BK, Mabon SA, Leffel SM, Halfhill MD, Richards HA, Moyer KA, Stewart CN Jr (1999) Green fluorescent protein as a marker for expression of a second gene in transgenic plants. Nat Biotechnol 17:1125–1129
Haseloff J, Siemering KR, Prasher D, 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
Légère A, Simard M-J, Thomas AG, Pageau D, Lajeunesse J, Warwick SI, Derksen DA (2001) Presence and persistence of volunteer oilseed rape in Canadian cropping systems. In: Proc Brighton Crop Protection Conf—weeds. British Crop Protection Council, Farnham, Surrey, UK, pp 143-148
Millwood RJ, Halfhill MD, Harkins D, Russotti R, Stewart CN Jr (2003) Instrumentation and methodology of GFP quantification in intact plant organs. Biotechniques 34:638–643
Odell J, Nagy F, Chua N (1985) Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter. Nature 313:810–812
Prasher DC, Eckenrode VK, Ward WW, Pendergast FG, Cormier MJ (1992) Primary structure of the Aequorea victoria green fluorescent protein. Gene 111:229–233
Siemering KR, Golbik R, Sever R, Haseloff J (1996) Mutations that suppress the thermosensitivity of green fluorescent protein. Curr Biol 6:1653–1663
Simard MJ, Légère A, Pageau D, Lajeunnesse J, Warwick S (2002) The frequency and persistence of oilseed rape (Brassica napus) volunteers in Québec cropping systems. Weed Technol 16:433–439
Stewart CN Jr (1996) Monitoring transgenic plants using in vivo markers. Nat Biotechnol 14:682
Thomas AG, Breve MA, Raymer PL (1991) Influence of timing and method of harvest on rapeseed yield. J Prod Agric 4:266–272
Warwick SI, Simard MJ, Légère A, Beckie HJ, Braun L, Zhu B, Mason P, Seguin-Swartz G, Stewart CN Jr (2003) Hybridization between transgenic Brassica napus L. and its wild relatives: B. rapa L., Raphanus raphanistrum L., Sinapis arvensis L., and Erucastrum gallicum (Willd.) O.E. Schulz. Theor Appl Genet DOI 10.1007/s00122-003-1278-0
Woolhouse HW (1967) The nature of senescence in plants. Sym Soc Exp Biol 21:179–213
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by M.C. Jordan
Rights and permissions
About this article
Cite this article
Halfhill, M.D., Millwood, R.J., Rufty, T.W. et al. 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 (2003). https://doi.org/10.1007/s00299-003-0696-4
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00299-003-0696-4