Abstract
To determine the location of specific molecules within tissues or cells, immunological techniques are frequently used. However, immunolocalization of small molecules, such as jasmonic acid (JA) and its bioactive amino acid conjugate, JA-isoleucine, requires proper fixation and embedding methods as well as specific antibodies. In this chapter, we present a method to prepare plant tissues for the detection of jasmonates, including the chemical fixation to immobilize JA within the tissue, the subsequent embedding in a suitable medium, and the immunolabeling procedure itself.
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
Similar content being viewed by others
References
Dewitte W, Van Onckelen H (2001) Probing the distribution of plant hormones by immunocytochemistry. Plant Growth Regul 33:67–74
Mielke K, Forner S, Kramell R, Conrad U, Hause B (2011) Cell-specific visualization of jasmonates in wounded tomato and Arabidopsis leaves using jasmonate-specific antibodies. New Phytol 190:1069–1080
Aloni R, Schwalm K, Langhans M, Ullrich CI (2003) Gradual shifts in sites of free-auxin production during leaf-primordium development and their role in vasculat differentiation and leaf morphogenesis in Arabidopsis. Planta 216:841–853
Hause B, Frugier F, Crespi M (2006) Immunolocalization. In: Mathesius U, Journet E-P, Sumner LW (eds) The Medicago truncatula handbook. The Samuel Noble Foundation, Ardmore, pp 1–11. ISBN 0-9754303-1-9 (http://www.noble.org/MedicagoHandbook/)
Pena JTG, Sohn-Lee C, Rouhanifard SH, Ludwig J, Hafner M, Mihailovic A, Lim C, Holoch D, Berninger P, Zavolan M, Tuschl T (2009) miRNA in situ hybridization in formaldehyde and EDC-fixed tissues. Nat Methods 6:139–141
Tretner C, Huth U, Hause B (2008) Mechanostimulation of Medicago truncatula leads to enhanced levels of jasmonic acid. J Exp Bot 59:2847–2856
Browse J (2009) The power of mutants for investigating jasmonate biosynthesis and signaling. Phytochemistry 70:1539–1546
Li C, Liu G, Xu C, Lee GI, Bauer P, Ling H-Q, Ganal MW, Howe GA (2003) The tomato suppressor of prosystemin-mediated response2 gene encodes a fatty acid desaturase required for the biosynthesis of jasmonic acid and the production of a systemic wound signal for defense gene expression. Plant Cell 15:1646–1661
Acknowledgments
This work was supported by a grant of the Deutsche Forschungsgemeinschaft (Project HA2655/7-2 within SPP1212) to B.H.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Hause, B., Mielke, K., Forner, S. (2013). Cell-Specific Detection of Jasmonates by Means of an Immunocytological Approach. In: Goossens, A., Pauwels, L. (eds) Jasmonate Signaling. Methods in Molecular Biology, vol 1011. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-414-2_11
Download citation
DOI: https://doi.org/10.1007/978-1-62703-414-2_11
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-413-5
Online ISBN: 978-1-62703-414-2
eBook Packages: Springer Protocols