Abstract
The GUS gene of E. coli, encoding β-glucuronidase, has been widely used as a reporter gene in plant transformation. However, β-glucuronidase activity in transgenic wheat leaf or root tissue is rarely observed or reported. To address this question, we investigated three wheat lines transformed with the GUS reporter gene. We found all three lines expressed GUS mRNA as well as β-glucuronidase protein in their leaf and root tissues as detected by RNA gel blot, ELISA, and immunoblot analyses. However, β-glucuronidase enzyme activity was only detected in pollen grains from the transgenic plants. Fluorometric and histochemical assays performed in the presence of wheat tissue extracts indicated that wheat leaf and root tissues contain inhibitor(s) of β-glucuronidase activity, but pollen grains contain much lower concentrations. Further characterizations indicated that the inhibitor(s) is of low molecular weight (<10 kDa) and is non-proteinaceous.
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Abbreviations
- GUS:
-
β-glucuronidase
References
D Becker R Brettschneider H Lörz (1994) ArticleTitleFertile transgenic wheat from microprojectile bombardment of scutellar tissue Plant J. 5 299–307
AH Christensen P Quail (1996) ArticleTitleUbiquitin promoter-based vectors for high level expression of selectable and/or screenable marker genes in monocotyledonous plants Transgenic Res. 5 213–218
M De Block J Botterman M Vandewiele J Dockx C Thoen V Gossele NR Movva C Thompsen M Van-Montagu J Leemans (1987) ArticleTitleEngineering herbicide resistance in plants by expression of a detoxifying enzyme EMBO J. 6 2513–2518
DB Duncan (1955) ArticleTitleMultiple range and multiple F test Biometrics 11 1–42
ML Edwards JI Gooper (1985) ArticleTitlePlant virus detection using a new form of indirect ELISA J. Virol. Methods 11 309–319
L Folling A Olesen (2001) ArticleTitleTransformation of wheat (Triticum aestivum L.) microspore-derived callus and microspores by particle bombardment Plant Cell Rep. 20 629–636
WJ Gordon-Kamm TM Spencer ML Mangano et al. (1990) ArticleTitleTransformation of maize cells and regeneration of fertile transgenic plants Plant Cell 2 603–618
DG He A Mouradov YM Yang E Mouradova KJ Scott (1994) ArticleTitleTransformation of wheat (Triticum aestivum L.) through electroporation of protoplasts Plant Cell Rep. 14 192–196
L Hodal A Bochardt JE Nielsen O Mattsson FT Okkels (1992) ArticleTitleDetection, expression and specific elimination of endogenous β-glucuronidase activity in transgenic plants Plant Sci. 87 115–122
RA Jefferson TA Kavanagh MW Bevan (1987) ArticleTitleGUS␣fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants EMBO J. 6 3901–3907
SP Lanning LE Talbert FH McNeal WL Alexander CF McGuire H Bowman G Carlson G Jackson J Eckhoff G Kushnak V Stewart G Stallknecht (1992) ArticleTitleRegistration of “Hi-Line” wheat Crop Sci. 32 283–284
L Li R Qu A Kochko Particlede C Fauquet RN Beachy (1993) ArticleTitleAn improved rice transformation system using the biolistic method Plant Cell Rep. 12 250–255
RM Lister J Hammond DL Clement (1983) ArticleTitleComparison of interdermal and intermuscular injection for raising plant virus antisera for use in ELISA J.Virol. Methods 6 179–182
D McElroy AD Blowers B Jenes R Wu (1991) ArticleTitleConstruction of expression vector based on the rice actin 1 (Act 1) 5′ region for use in monocot transformation Mol. Gen. Genet. 231 150–160
MJ Muhitch (1998) ArticleTitleCharacterization of pedicel β-glucuronidase activity in developing maize (Zea mays) kernels Physiol. Plant. 104 423–430
NS Nehra RN Chibbar N Leung K Caswell C Mallard L Steinhauer M Baga KK Kartha (1994) ArticleTitleSelf-fertile transgenic wheat plants regenerated from isolated scutellar tissues following microprojectile bombardment with two distinct gene constructs Plant J. 5 285–297
E Sivamani A Bahieldin JM Wraith T Al-Niemi WE Dyer T-HD Ho R Qu (2000) ArticleTitleImproved biomass productivity and water use efficiency under water deficit conditions in transgenic wheat constitutively expressing the barley HVA1 gene Plant Sci. 155 1–9
DA Somers HW Rhines W Gu HF Kaeppler WR Bushnell (1992) ArticleTitleFertile, transgenic oat plants Bio/Technol. 10 1589–1594
AP Sorokin X-Y Ke DF Chen MC Elliot (2000) ArticleTitleProduction of fertile wheat plants via tissue electroporation Plant Sci. 156 227–233
TM Spencer WJ Gordon-Kamm RJ Daines WG Start PG Lemaux (1990) ArticleTitleBialaphos selection of stable transformation from maize cell culture Theor. Appl. Genet. 79 625–631
V Vasil V Srivastava AM Castillo ME Fromm IK Vasil (1993) ArticleTitleRapid production of transgenic wheat plants by direct bombardment of cultured immature embryos Bio/Technol. 11 1553–1558
Y Wan PG Lemaux (1994) ArticleTitleGeneration of large numbers of independently transformed fertile barley plants Plant Physiol. 104 37–48
JT Weeks OD Anderson AE Blechl (1993) ArticleTitleRapid production of multiple independent lines of fertile transgenic wheat (Triticum aestivum L.). Plant Physiol 102 1077–1084
SD Zvereva GA Romanov (2000) ArticleTitleReporter gene for plant genetic engineering: characteristics and detection Russ. J. Plant Physiol. 47 424–432
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Bahieldin, A., Eissa, H.F., Mahfouz, H.T. et al. Evidence for non-proteinaceous inhibitor(s) of β-glucuronidase in wheat (Triticum aestivumL.) leaf and root tissues. Plant Cell Tiss Organ Cult 82, 11–17 (2005). https://doi.org/10.1007/s11240-004-5890-8
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DOI: https://doi.org/10.1007/s11240-004-5890-8