Seed Dormancy pp 329-339 | Cite as

Tissue-Printing Methods for Localization of RNA and Proteins that Control Seed Dormancy and Germination

  • Wioletta E. Pluskota
  • Kent J. Bradford
  • Hiro Nonogaki
Part of the Methods in Molecular Biology book series (MIMB, volume 773)


A number of genes and proteins are expressed in a tissue- or cell layer-specific manner. Spatial patterns of gene expression are critical to understanding gene function. Tissue printing provides a simple and rapid method to analyze localization of mRNA and protein at the tissue and cellular levels. This is especially convenient for gene expression analysis in hard tissues, such as seeds that are often difficult to section. Seed RNA or protein can be transferred onto a suitable membrane by printing the cut surface of a bisected seed. This method has been used successfully to determine mRNA and protein localization in seed research. The resolution of printed seed images and RNA and protein signals in tissue printing is sufficient to identify embryo- or endosperm-specific expression of various genes and proteins. In some cases, these studies have contributed to elucidating the spatial characteristics of hydrolytic enzymes putatively involved in the completion of germination and/or early postgerminative growth. By the same principle, tissue-printing methods could also be valuable for elucidating the spatial characteristics of genes/proteins that control the inception, maintenance, and termination of seed dormancy.

Key words

Tissue printing Seed mRNA Protein Localization Gene expression Dormancy Germination 



This work was supported by National Science Foundation grant IBN-0237562 (to H. Nonogaki).


  1. 1.
    Ogawa, M., Hanada, A., Yamauchi, Y., Kuwahara, A., Kamiya, Y., and Yamaguchi, S. (2003) Gibberellin biosynthesis and response during Arabidopsis seed germination. Plant Cell 15, 1591–604.Google Scholar
  2. 2.
    Yamauchi, Y., Ogawa, M., Kuwahara, A., Hanada, A., Kamiya, Y., and Yamaguchi, S. (2004) Activation of gibberellin biosynthesis and response pathways by low temperature during imbibition of Arabidopsis thaliana seeds. Plant Cell 16, 367–78.Google Scholar
  3. 3.
    Nakabayashi, K., Okamoto, M., Koshiba, T., Kamiya, Y., and Nambara, E. (2005) Genome-wide profiling of stored mRNA in Arabidopsis thaliana seed germination: epigenetic and genetic regulation of transcription in seed. Plant J 41, 697–709.Google Scholar
  4. 4.
    Varner, J. E., and Ye, Z. (1994) Tissue printing. FASEB J 8, 378–84.Google Scholar
  5. 5.
    Nonogaki, H., Gee, O. H., and Bradford, K. J. (2000) A Germination-specific endo-β-mannanase gene is expressed in the micropylar endosperm cap of tomato seeds. Plant Physiol 123, 1235–46.Google Scholar
  6. 6.
    Chen, F., and Bradford, K. J. (2000) Expression of an expansin is associated with endosperm weakening during tomato seed germination. Plant Physiol 124, 1265–74.Google Scholar
  7. 7.
    Wu, C. T., Leubner-Metzger, G., Meins, F. Jr., and Bradford, K. J. (2001) Class I β-1,3-glucanase and chitinase are expressed in the micropylar endosperm of tomato seeds prior to radicle emergence. Plant Physiol 126, 1299–313.Google Scholar
  8. 8.
    Yamaguchi, S., Kamiya, Y., and Sun, T.-p. (2001) Distinct cell-specific expression patterns of early and late gibberellin biosynthetic genes during Arabidopsis seed germination. Plant J 28, 443–53.Google Scholar
  9. 9.
    Lefebvre, V., North, H., Frey, A., Sotta, B., Seo, M., Okamoto, M., Nambara, E., and Marion-Poll, A. (2006) Functional analysis of Arabidopsis NCED6 and NCED9 genes indicates that ABA synthesised in the endosperm is involved in the induction of seed dormancy. Plant J 45, 309–19.Google Scholar
  10. 10.
    Sawada, Y., Aoki, M., Nakaminami, K., Mitsuhashi, W., Tatematsu, K., Kushiro, T., Koshiba, T., Kamiya, Y., Inoue, Y., Nambara, E., and Toyomasus, T. (2008) Phytochrome- and gibberellin-mediated regulation of abscisic acid metabolism during germination of photoblastic lettuce seeds. Plant Physiol 146, 1386–96.Google Scholar
  11. 11.
    Sawada, Y., Katsumata, T., Kitamura, J., Kawaide, H., Nakajima, M., Asami, T., Nakaminami, K., Kurahashi, T., Mitsuhashi, W., Inoue, Y., and Toyomasu, T. (2008) Germination of photoblastic lettuce seeds is regulated via the control of endogenous physiologically active gibberellin content, rather than of gibberellin responsiveness. J Exp Bot 59, 3383–93.Google Scholar
  12. 12.
    Wu, C.-T., and Bradford, K. J. (2003) Class I chitinase and β-1,3-glucanase are differentially regulated by wounding, methyl jasmonate, ethylene, and gibberellin in tomato seeds and leaves. Plant Physiol 133, 263–73.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Wioletta E. Pluskota
    • 1
    • 2
  • Kent J. Bradford
    • 3
  • Hiro Nonogaki
    • 2
  1. 1.Department of Plant Physiology and BiotechnologyUniversity of Warmia and MazuryOlsztynPoland
  2. 2.Department of HorticultureOregon State UniversityCorvallisUSA
  3. 3.Department of Plant SciencesUniversity of CaliforniaDavisUSA

Personalised recommendations