Abstract
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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
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.
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.
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.
Varner, J. E., and Ye, Z. (1994) Tissue printing. FASEB J 8, 378–84.
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.
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.
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.
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.
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.
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.
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.
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.
Acknowledgment
This work was supported by National Science Foundation grant IBN-0237562 (to H. Nonogaki).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Pluskota, W.E., Bradford, K.J., Nonogaki, H. (2011). Tissue-Printing Methods for Localization of RNA and Proteins that Control Seed Dormancy and Germination. In: Kermode, A. (eds) Seed Dormancy. Methods in Molecular Biology, vol 773. Humana Press. https://doi.org/10.1007/978-1-61779-231-1_19
Download citation
DOI: https://doi.org/10.1007/978-1-61779-231-1_19
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-230-4
Online ISBN: 978-1-61779-231-1
eBook Packages: Springer Protocols