Biotechnology of Plant Secondary Metabolism pp 35-42

Part of the Methods in Molecular Biology book series (MIMB, volume 1405) | Cite as

Collection of Apoplastic Fluids from Arabidopsis thaliana Leaves

  • Svend Roesen Madsen
  • Hussam Hassan Nour-Eldin
  • Barbara Ann Halkier
Protocol

Abstract

The leaf apoplast comprises the extracellular continuum outside cell membranes. A broad range of processes take place in the apoplast, including intercellular signaling, metabolite transport, and plant-microbe interactions. To study these processes, it is essential to analyze the metabolite content in apoplastic fluids. Due to the fragile nature of leaf tissues, it is a challenge to obtain apoplastic fluids from leaves. Here, methods to collect apoplastic washing fluid and guttation fluid from Arabidopsis thaliana leaves are described.

Key words

Apoplastic fluid Guttation fluid Vacuum infiltration Glucosinolates 

References

  1. 1.
    Sattelmacher B (2001) Tansley review no. 22 - the apoplast and its significance for plant mineral nutrition. New Phytol 149:167–192CrossRefGoogle Scholar
  2. 2.
    Baker CJ, Kovalskaya NY, Mock NM, Owens RA, Deahl KL, Whitaker BD, Roberts DP, Hammond RW, Aver’yanov AA (2012) An internal standard technique for improved quantitative analysis of apoplastic metabolites in tomato leaves. Physiol Mol Plant Pathol 78:31–37CrossRefGoogle Scholar
  3. 3.
    Robinson JM, Bunce JA (2000) Influence of drought-induced water stress on soybean and spinach leaf ascorbate-dehydroascorbate level and redox status. Int J Plant Sci 161:271–279CrossRefPubMedGoogle Scholar
  4. 4.
    Lee YW, Jin S, Sim WS, Nester EW (1995) Genetic evidence for direct sensing of phenolic compounds by the VirA protein of Agrobacterium tumefaciens. Proc Natl Acad Sci U S A 92:12245–12249PubMedCentralCrossRefPubMedGoogle Scholar
  5. 5.
    Foyer CH, Descourvières P, Kunert KJ (1994) Protection against oxygen radicals: an important defence mechanism studied in transgenic plants. Plant Cell Environ 17:507–523CrossRefGoogle Scholar
  6. 6.
    Halkier BA, Gershenzon J (2006) Biology and biochemistry of glucosinolates. Annu Rev Plant Biol 57:303–333CrossRefPubMedGoogle Scholar
  7. 7.
    Fan J, Crooks C, Creissen G, Hill L, Fairhurst S, Doerner P, Lamb C (2011) Pseudomonas sax genes overcome aliphatic isothiocyanate-mediated non-host resistance in Arabidopsis. Science 331:1185–1188CrossRefPubMedGoogle Scholar
  8. 8.
    Madsen SR, Olsen CE, Nour-Eldin HH, Halkier BA (2014) Elucidating the role of transport processes in leaf glucosinolate distribution. Plant Physiol 166:1450–1462. doi:10.1104/pp.114.246249 PubMedCentralCrossRefPubMedGoogle Scholar
  9. 9.
    Andersen TG, Nour-Eldin HH, Fuller VL, Olsen CE, Burow M, Halkier BA (2013) Integration of biosynthesis and long-distance transport establish organ-specific glucosinolate profiles in vegetative Arabidopsis. Plant Cell 25:3133–3145PubMedCentralCrossRefPubMedGoogle Scholar
  10. 10.
    Nour-Eldin HH, Andersen TG, Burow M, Madsen SR, Jorgensen ME, Olsen CE, Dreyer I, Hedrich R, Geiger D, Halkier BA (2012) NRT/PTR transporters are essential for translocation of glucosinolate defence compounds to seeds. Nature 488:531–534CrossRefPubMedGoogle Scholar
  11. 11.
    Pilot G, Stransky H, Bushey DF, Pratelli R, Ludewig U, Wingate VPM, Frommer WB (2004) Overexpression of glutamine dumper1 leads to hypersecretion of glutamine from hydathodes of Arabidopsis leaves. Plant Cell 16:1827–1840PubMedCentralCrossRefPubMedGoogle Scholar
  12. 12.
    Boudart G, Jamet E, Rossignol M, Lafitte C, Borderies G, Jauneau A, Esquerre-Tugaye MT, Pont-Lezica R (2005) Cell wall proteins in apoplastic fluids of Arabidopsis thaliana rosettes: identification by mass spectrometry and bioinformatics. Proteomics 5:212–221CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Svend Roesen Madsen
    • 1
  • Hussam Hassan Nour-Eldin
    • 1
  • Barbara Ann Halkier
    • 1
  1. 1.DynaMo Center of Excellence, Department of Plant and Environmental Sciences, Faculty of ScienceUniversity of CopenhagenFrederiksberg CDenmark

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