Skip to main content
SpringerLink
Log in

Tracking of the CaMV-35S promoter performance in GFP transgenic tobacco, with a special emphasis on flowers and reproductive organs, confirmed its predominant activity in vascular tissues

  • Original Paper
  • Published:
Plant Cell, Tissue and Organ Culture Aims and scope Submit manuscript

Abstract

Constitutive promoters are the most common promoters used to drive the expression of various genes in monocots and dicots. Therefore, it is of intense interest to ascertain their expression patterns in various plant species, organs and during their ontogenic development. In this study, the activity of the CaMV 35S promoter in transgenic tobacco plants was assessed. In contrast to other studies, performed rather on the primary transformants (T0 generation), here, individuals of T1 and T2 generations were used. The expression profiles of the CaMV 35S promoter were tracked within various plant organs and tissues using the GFP marker. Special attention was given to floral tissues for which the original data regarding the CaMV 35S expression were obtained. As expected, distinct developmental and organ/tissue specific expression patterns in a plant body were observed. CaMV 35S activity was detected in most of the plant tissues and during different developmental stages. The GFP signal was not visible in dry seeds only, but it became clearly apparent within 24–48 h after sowing onto the medium, what, among other things, enables the discrimination of transgenic and non-transgenic seeds/seedlings. Afterwards, the most pronounced GFP fluorescence intensity was usually visible in various vascular tissues of both, T1 and T2 plants, indicating the high promoter activity. A stable manifestation of the promoter was retained in the next T2 generation without any evident changes or losses of activity, showing the expression stability of the CaMV 35S.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

CaMV 35S:

Cauliflower mosaic virus promoter

GFP:

Green fluorescent protein

GUS:

β-Glucuronidase

Kn:

kanamycin

RFP:

Red fluorescent protein

RT-PCR:

Reverse transcription polymerase chain reaction

T-DNA:

Transferred DNA

YFP:

Yellow fluorescent protein

gfp :

Green fluorescent protein gene

mgfp5-ER :

Modified gene for Green fluorescent protein

nptII :

Neomycin Phosphotransferase II gene

nos (promoter):

Nopaline Synthase promoter

uidA :

β-Glucuronidase gene

References

  • Battraw MJ, Hall TC (1990) Histochemical analysis of CaMV 35S promoter-β-glucuronidase gene expression in transgenic rice plants. Plant Mol Biol 15:527–538. doi: 10.1007/BF00017828

    Article  PubMed  CAS  Google Scholar 

  • Benfey PN, Chua N-H (1989) Regulated genes in transgenic plants. Science 244:174–181

    Article  PubMed  CAS  Google Scholar 

  • Benfey PN, Ren L, Chua N-H (1990a) Tissue specific expression from CaMV 35S enhancer subdomains in early stages of plant development. EMBO J 9:1677–1684

    PubMed  CAS  Google Scholar 

  • Benfey PN, Ren L, Chua N-H (1990b) Combinatorial and synergistic properties of CaMV 35S enhancer subdomains. EMBO J 9:1685–1696

    PubMed  CAS  Google Scholar 

  • Cornejo MJ, Luth D, Balkenship KM, Andreson OD, Blechl AE (1993) Activity of a maize ubiquitin promoter in transgenic rice. Plant Mol Biol 23:567–581. doi: 10.1007/BF00019304

    Article  PubMed  CAS  Google Scholar 

  • Custers JBM, Snepvangers SCHJ, Jansen HJ, Zhang L, van Lookeren C (1999) The 35S-CaMV promoter is silent during early embryogenesis but is activated during non-embryogenic sporophytic development in microspore culture. Protoplasma 208:257–264. doi: 10.1007/BF01279097

    Article  CAS  Google Scholar 

  • Haseloff J, Siemering KR, Prasher DC, Hodge S (1997) Removal of a cryptic intron and subcellular localisation of green fluorescent protein are required to mark transgenic Arabidopsis plants brightly. PNAS USA 94:2122–2127. doi: 10.1073/pnas.94.6.2122

    Article  PubMed  CAS  Google Scholar 

  • Horsch RB, Fry JE, Hoffmann NL, Eicholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231

    Article  CAS  Google Scholar 

  • Hraška M, Rakouský S, Kocábek T (2005) Use of simple semiquantitative method for appraisal of green fluorescent protein gene expression in transgenic tobacco plants. Biol Plant 49:313–316. doi: 10.1007/s10535-005-3316-z

    Article  Google Scholar 

  • Hraška M, Rakouský S, Čurn V (2006) Green fluorescent protein as a vital marker for non-destructive detection of transformation events in transgenic plants. Plant Cell Tiss Organ Cult 86:303–318. doi: 10.1007/s11240-006-9131-1

    Article  CAS  Google Scholar 

  • Jefferson RA, Ravanagh TA, Bevan MW (1987) GUS fusion: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907

    PubMed  CAS  Google Scholar 

  • Malik K, Wu K, Li X-Q, Martin-Heller T, Hu M, Foster E, Tian L, Wang C, Ward K, Jordan M, Brown D, Gleddie S, Simmonds D, Zheng S, Simmonds J, Miki B (2002) A constitutive gene expression system derived from the tCUP cryptic promoter elements. Theor Appl Genet 105:505–514. doi: 10.1007/s00122-002-0926-0

    Article  PubMed  CAS  Google Scholar 

  • Mascarenhas JP, Hamilton DA (1992) Artifacts in the localization of GUS activity in the anthers of petunia transformed with a CaMV 35S-GUS construct. Plant J 2:405–408. doi: 10.1111/j.1365-313X.1992.00405.x

    Article  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497. doi: 10.1111/j.1399-3054.1962.tb08052.x

    Article  CAS  Google Scholar 

  • Odell JT, Nagy F, Chua N-H (1985) Identification of DNA sequence required for activity of cauliflower mosaic virus 35S promoter. Nature 313:810–812

    Article  PubMed  CAS  Google Scholar 

  • Ooms G, Hooykaas PJJ, van Veen RJM, van Beelen P, Regensburg-Tuink TJG, Schilperoot RA (1982) Octopine Ti-plasmid deletion mutants of Agrobacterium tumefaciens with emphasis on right side of T-region. Plasmid 7:15–29

    Article  PubMed  CAS  Google Scholar 

  • Prasher DC, Eckenrode VK, Ward WW, Prendergast FG, Cormier MJ (1992) Primary structure of the Aequorea victoria green-fluorescent protein. Gene 111:229–233

    Article  PubMed  CAS  Google Scholar 

  • Preťová A, Obert B, Wetzstein HY (2001) Leaf developmental stage and tissue location affect the detection of β-glucuronidase in transgenic tobacco plants. Biotech Lett 23:555–558. doi: 10.1023/A:1010311524048

    Article  Google Scholar 

  • Rooke L, Byrne D, Salgueiro S (2000) Marker gene expression driven by the maize ubiquitin promoter in transgenic wheat. Ann Appl Biol 136:167–172. doi: 10.1111/j.1744-7348.2000.tb00022.x

    Article  CAS  Google Scholar 

  • Stewart CN Jr (2001) The utility of green fluorescent protein in transgenic plants. Plant Cell Rep 20:376–382. doi: 10.1007/s002990100346

    Article  PubMed  CAS  Google Scholar 

  • Sunilkumar G, Mohr L-A, Lopata-Finch E, Emani Ch, Rathore KS (2002) Developmental and tissue specific expression of CaMV 35S promoter in cotton as revealed by GFP. Plant Mol Biol 50:463–474. doi: 10.1023/A:1019832123444

    Article  PubMed  CAS  Google Scholar 

  • Terada R, Shimamoto K (1990) Expression of CaMV 35S-GUS gene in transgenic rice plants. Mol Gen Genet 220:389–392

    Article  CAS  Google Scholar 

  • Tsien RY (1998) The green fluorescent protein. Ann Rev Biochem 67:509–544. doi: 10.1146/annurev.biochem.67.1.509

    Article  PubMed  CAS  Google Scholar 

  • Williamson JD, Hirsch-Wyncott ME, Larkins BA, Gelvin SB (1989) Differential accumulation of a transcript driven by the CaMV 35S promoter in transgenic tobacco. Plant Physiol 90:1570–1576

    Article  PubMed  CAS  Google Scholar 

  • Yang N-S, Christou P (1990) Cell type specific expression of a CaMV 35S-GUS gene in transgenic soybean plants. Develop Genet 11:289–293

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors are grateful to Stanislav Vitha (Microscopy and Imaging Center Texas A&M University) for his valuable critical comments and revisions. The authors would like also gratefully acknowledge the financial support received from Ministry of Education, Youth and Sport of the Czech Republic (grants 1M06030, 1PO5ME800 and MSM 60076658-06). Marek Hraška received the support from grant GA ČR 31/H160 provided by the Grant Agency of the Czech Republic. Linguistic revision was kindly performed by John McAvoy.

Author information

Authors and Affiliations

  1. Faculty of Health and Social Studies, University of South Bohemia, Jírovcova 24, 370 01, Ceske Budejovice, Czech Republic

    Marek Hraška & Slavomír Rakouský

  2. Department of Genetics, Faculty of Science, University of South Bohemia, Branišovská 31, 370 05, Ceske Budejovice, Czech Republic

    Marek Hraška & Slavomír Rakouský

  3. Biotechnological Centre, Faculty of Agriculture, University of South Bohemia, Studentská 13, 370 05, Ceske Budejovice, Czech Republic

    Vladislav Čurn

Authors
  1. Marek Hraška
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Slavomír Rakouský
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Vladislav Čurn
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Slavomír Rakouský.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hraška, M., Rakouský, S. & Čurn, V. Tracking of the CaMV-35S promoter performance in GFP transgenic tobacco, with a special emphasis on flowers and reproductive organs, confirmed its predominant activity in vascular tissues. Plant Cell Tiss Organ Cult 94, 239–251 (2008). https://doi.org/10.1007/s11240-007-9312-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-007-9312-6

Keywords

Search

Navigation