Skip to main content
SpringerLink
Log in

In Situ Hybridization to Plant Tissue Sections

  • Protocol
Plant Gene Transfer and Expression Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 49))

  • 2964 Accesses

Abstract

An important step in the analysis of gene action is to determine the spatial and temporal regulation of gene expression. The patterns of gene expression can provide clues to the function of genes and facilitate the mapping of qualitative changes in steady state mRNA levels. In situ hybridization is a cytochemical procedure first used to map sequences on chromosomes (1,2). It is also used to identify viral RNA in and around infection sites (3,4) and is commonly used for locating specific mRNA transcripts in tissue sections (5–9). The technique can also be adapted for ultrastructural resolution particularly with biotin labeled probes (10,11).

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Spiral binding cover Book
USD 169.99
Price excludes VAT (USA)
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Gall, J. G. and Pardue, M. L. (1969) Formation and detection of RNA-DNA hybrid molecules in cytological preparations. Proc. Natl. Acad. Sci. USA 63, 378–383.

    Article  PubMed  CAS  Google Scholar 

  2. Pardue, M. L. and Gall, J. G. (1969) Molecular hybridisation of radioactive DNA to the DNA of cytological preparations. Proc. Natl. Acad. Sci. USA 64, 600–604.

    Article  PubMed  CAS  Google Scholar 

  3. Brahic, M. and Haase, A. T. (1978) Detection of viral sequences of low reiteration frequency by in situ hybridisation. Proc. Natl. Acad. Sci. USA 75, 6125–6129.

    Article  PubMed  CAS  Google Scholar 

  4. Wells, M. (1990) The demonstration of viral DNA in human tissues by in situ DNA hybridisation, in In situ Hybridisation: Application to Developmental Biology and Medicine (Harris, N. and Wilkinson, D. G., eds.), Cambridge University Press, pp. 271–272.

    Google Scholar 

  5. Angerer, L. M., Cox, K. H., and Angerer, R. C. (1987) Demonstration of tissue-specific gene expression by in situ hybridisation. Methods Enzymol. 152, 649–661.

    Article  PubMed  CAS  Google Scholar 

  6. Barker, S. J., Harada, J. J., and Goldberg, R. B. (1988) Cellular localization of soybean storage protein mRNA in transformed tobacco seeds. Proc. Natl. Acad. Sci. USA 85, 458–462.

    Article  PubMed  CAS  Google Scholar 

  7. Cox, K. H., DeLeon, D. V., Angerer, L. M., and Angerer, R. C. (1984) Detection of mRNAs in sea urchin embryos, in in situ hybridisation using asymetric probes. Dev. Biol. 101, 485–502.

    Article  PubMed  CAS  Google Scholar 

  8. Langdale, J. A., Rothermel, B. A., and Nelson, T. (1988) Cellular pattern of photosynthetic gene expression in developing maize leaves. Genes Dev. 2, 106–115.

    Article  PubMed  CAS  Google Scholar 

  9. Ruiz-Avila, L., Burgess, S. R., Stiefel, V., Dolors-Ludevid, M., and Puigdomenech, P. (1992) Accumulation of cell wall hydroxyproline-rich glycoprotein mRNA is an early event in maize embryo cell differentiation. Proc. Natl. Acad. Sci. USA 89, 2414–2418.

    Article  PubMed  CAS  Google Scholar 

  10. McFadden, G. I. (1989) In situ hybridisation in plants. from macroscopic to ultra-structural resolution. Cell Biol. Int. Repts. 13, 3–21.

    Article  Google Scholar 

  11. Binder, M., Tourmente, S., Roth, J., Renaud, M., and Gehring, W. J. (1986) In situ hybridisation at the electron microscope level: localization of transcripts on ultrathin sections of Lowicryl K4M-embedded tissue using biotinylated probes and protein A-gold complexes. J. Cell. Biol. 102, 1646–1653.

    Article  PubMed  CAS  Google Scholar 

  12. Harris, N., Grindley, H., Mulchrone, J., and Croy, R. R. D. (1989) Correlated in situ hybridisation and immunocytochemical studies of legumin storage protein deposition in pea (Pisum sativum L.). Cell Biol. Int. Repts. 13, 23–35.

    Article  Google Scholar 

  13. VanDer Loss, C. M., Volkers, H. H., Rook, R., VanDer Berg, F. M., and Houthoff, H. J. (1989) Simultaneous application of in situ hybridisation and immunohistochemistry on one tissue section. Histochem. J. 21, 27–284.

    Google Scholar 

  14. Wetmur, J. G. and Davidson, N. (1968) Kinetics of renaturation of DNA. J. Mol. Biol. 31, 349–370.

    Article  PubMed  CAS  Google Scholar 

  15. Berger, C. N. (1986) In situ hybridisation of immunoglobulin-specific RNA in single cells of the B lymphocyte lineage with radiolabeled DNA probes. EMBO J. 5, 85–93.

    PubMed  CAS  Google Scholar 

  16. Lewis, M. E., Sherman, T. G., and Watson, S. J. (1985) In situ hybridisation histochemistry with synthetic oligonucleotides. Strategies and methods. Peptides 6(Suppl. 2), 75–87.

    Article  PubMed  CAS  Google Scholar 

  17. Taneja, K. and Singer, R. H. (1987) Use of oligodeoxynucleotide probes for quantitative in situ hybridisation to actin mRNA. Anal. Biochem. 166, 389–398.

    Article  PubMed  CAS  Google Scholar 

  18. Witkiewicz, H., Bolander, M. E., and Edwards, D. R. (1993) Improved design of riboprobes from pBluescript and related vectors for in situ hybridisation. Biotechniques 14, 458–463.

    PubMed  CAS  Google Scholar 

  19. Moench, T. R., Gendelman, H. F., Clements, J. E., Narayan, O., and Griffin, D. E. (1985) Efficiency of in situ hybridisation technique as a function of probe size and fixation technique. J. Virol. Methods 11, 119–130.

    Article  PubMed  CAS  Google Scholar 

  20. Smith, L. M., Handley, J., Li, Y., Martin, H., Donovan, L., and Bowles, D. J. (1992) Temporal and spatial regulation of a novel gene in barley embryos. Plant Mol. Biol. 20, 255–266.

    Article  PubMed  CAS  Google Scholar 

  21. Casey, J. and Davidson, N. (1977) Rates of formation and thermal stabilities of RNA:DNA and DNA:DNA duplexes at high concentration of formamide. Nucleic Acids Res. 4, 1539–1552.

    Article  PubMed  CAS  Google Scholar 

  22. Hemmati-Brivanlou, A., Frank, D., Bolce, M. E., Brown, B. D., Sive, H. L., and Harland, R. M. (1990) Localisation of specific mRNAs in Xenopus embryos by whole mount in situ hybridisation. Development 110, 325–330.

    PubMed  CAS  Google Scholar 

  23. Martin, R., Hoover, C., Grimme, S., Grogan, C., Holtke, J., and Kessler, C. (1990) A highly sensitive, nonradioactive DNA labeling and detection system. Biotechniques 9, 762–768.

    PubMed  CAS  Google Scholar 

  24. Leitch, I. J., Leitch, A. R., and Heslop-Harrison, J. S. (1991) Physical mapping of plant DNA sequences by simultaneous in situ hybridisation of two differently labeled fluorescent probes. Genome 34, 329–333.

    Article  Google Scholar 

  25. Mullink, H., Walboomers, J. M. M., Raap, A. K., and Meyer, C. J. L. M. (1989) Two colour DNA in situ hybridisation for the detection of two viral genomes using nonradioactive probes. Histochemistry 91, 195–198.

    Article  PubMed  CAS  Google Scholar 

  26. Singer, R. H., Lawrence, J. B., and Villnave, C. (1986) Optimisation of in situ hybridisation using isotopic and nonisotopic detection methods. Biotechniques 4, 230–250.

    CAS  Google Scholar 

  27. Glauert, A. M. (1975) The Fixation and Embedding of Biological Specimens, North-Holland Publishers, Amsterdam.

    Google Scholar 

  28. McFadden, G. I., Bonig, I., Cornish, E. C., and Clarke, A. E. (1988) A simple fixation and embedding method for use in hybridisation histochemistry of plant tissues. Histochem. J. 20, 575–586.

    Article  PubMed  CAS  Google Scholar 

  29. Von Bertalanffy, L. and Bickis, I. (1956) Identification of cytoplasmic basophilia (ribonucleic acid) by fluorescence microscopy. J. Histochem. Cytochem. 4, 481–493.

    Article  CAS  Google Scholar 

  30. Harris, J. R. (1991) Electron Microscopy in Biology—A Practical Approach. Oxford University Press, Oxford.

    Google Scholar 

  31. Royal Microscopical Society Handbooks Series. Oxford University Press, Royal Microscopical Society.

    Google Scholar 

  32. Coghlan, J. P., Aldred, P., Haralambidis, J., Niall, H. D., Penschow, J. D., and Tregear, G. W. (1985) Hybridisation histochemistry. Anal. Biochem. 149, 1–28.

    Article  PubMed  CAS  Google Scholar 

  33. Gee, C. E. and Roberts, J. L. (1983) In situ hybridisation histochemistry: a technique for the study of gene expression in single cells. DNA 2, 155–161.

    Article  Google Scholar 

  34. Meyerowitz, E. M. (1987) In situ hybridisation to RNA in plant tissue. Plant Mol. Biol. Reptr. 5, 242–250.

    Article  CAS  Google Scholar 

  35. Tecott, L. H., Eberwine, J. H., Barchas, J. D., and Valentino, K. L. (1987) Methodological considerations in the utilization of in situ hybridisation, in In Situ Hybridisation (Valentino, K., Eberwine, J., and Barchas, J. D., eds.), Oxford University Press, pp. 3–24.

    Google Scholar 

  36. Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning, A Laboratory Manual 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Agricultural Sciences, University of Bristol, UK

    Shirley R. Burgess

Authors
  1. Shirley R. Burgess
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. University of Hertfordshire, Hatfield, UK

    Heddwyn Jones

Rights and permissions

Reprints and permissions

Copyright information

© 1995 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Burgess, S.R. (1995). In Situ Hybridization to Plant Tissue Sections. In: Jones, H. (eds) Plant Gene Transfer and Expression Protocols. Methods in Molecular Biology™, vol 49. Springer, Totowa, NJ. https://doi.org/10.1385/0-89603-321-X:301

Download citation

  • DOI: https://doi.org/10.1385/0-89603-321-X:301

  • Publisher Name: Springer, Totowa, NJ

  • Print ISBN: 978-0-89603-321-4

  • Online ISBN: 978-1-59259-536-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation