Abstract
Plant cells are heavily compartmentalized, and metabolite concentrations in the various compartments differ significantly. Thus, determination of metabolite abundance in whole-cell extracts may be misleading, when the role of a compound in plant freezing tolerance shall be evaluated. Here, we describe a method for the separation of the largest compartments of plant cells, the vacuole, plastid, and cytosol. With more elaborate analysis, this method can be expanded to also resolve mitochondria and other compartments.
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
Hoermiller II, Naegele T, Augustin H et al (2017) Subcellular reprogramming of metabolism during cold acclimation in Arabidopsis thaliana. Plant Cell Environ 40:602–610
Hoermiller II, Ruschhaupt M, Heyer AG (2018) Mechanisms of frost resistance in Arabidopsis thaliana. Planta 248:827–835
Wormit A, Trentmann O, Feifer I et al (2006) Molecular identification and physiological characterization of a novel monosaccharide transporter from Arabidopsis involved in vacuolar sugar transport. Plant Cell 18:3476–3490
Schneider T, Keller F (2009) Raffinose in chloroplasts is synthesized in the cytosol and transported across the chloroplast envelope. Plant Cell Physiol 50:2174–2182
Knaupp M, Mishra KB, Nedbal L et al (2011) Evidence for a role of raffinose in stabilizing photosystem II during freeze-thaw cycles. Planta 234:477–486
Gardeström P, Wigge B (1988) Influence of photorespiration on ATP/ADP ratios in the chloroplasts, mitochondria, and cytosol, studied by rapid fractionation of barley (Hordeum vulgare) protoplasts. Plant Physiol 88:69–76
Gerhardt R, Heldt HW (1984) Measurement of subcellular metabolite levels in leaves by fractionation of freeze-stopped material in nonaqueous media. Plant Physiol 75:542–547
Farré EM, Tiessen A, Roessner U et al (2001) Analysis of the compartmentation of glycolytic intermediates, nucleotides, sugars, organic acids, amino acids, and sugar alcohols in potato tubers using a nonaqueous fractionation method. Plant Physiol 127:685–700
Fürtauer L, Weckwerth W, Nägele T (2016) A benchtop fractionation procedure for subcellular analysis of the plant metabolome. Front Plant Sci 7:1912
Boller T, Kende H (1979) Hydrolytic enzymes in the central vacuole of plant cells. Plant Physiol 63:1123–1132
Zrenner R, Willmitzer L, Sonnewald U (1993) Analysis of the expression of potato uridinediphosphate-glucose pyrophosphorylase and its inhibition by antisense RNA. Planta 190:247–252
Jelitto T, Sonnewald U, Willmitzer L et al (1992) Inorganic pyrophosphate content and metabolites in potato and tobacco plants expressing E. coli pyrophosphatase in their cytosol. Planta 188:238–244
Acknowledgments
This work was supported by grant HE 3087/10-2 from the German Science Foundation to AGH.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Hoermiller, I.I., Nägele, T., Heyer, A.G. (2020). Subcellular Compartmentation of Metabolites Involved in Cold Acclimation. In: Hincha, D., Zuther, E. (eds) Plant Cold Acclimation. Methods in Molecular Biology, vol 2156. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0660-5_18
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0660-5_18
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0659-9
Online ISBN: 978-1-0716-0660-5
eBook Packages: Springer Protocols