Abstract
The apoplast can be described as the soluble fraction of the extracellular space of plant tissue, and it plays an important role in signaling, nutrient transport, and plant–pathogen interactions. In this protocol, we describe a method where leaves are infiltrated with phosphate buffer under vacuum. The apoplast can then be extracted by centrifugation and simultaneously collected in a protease inhibitor solution. Using this protocol, typically 3 μg of apoplastic proteins can be obtained in a volume of 300 μL from five potato leaflets, with minimal contamination by non-apoplastic proteins.
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References
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Acknowledgements
We would like to thank Ashfaq Ali and Erik Alexandersson (among other current and former lab members) for their work in developing and testing this protocol. We would also like to thank Mia Mogren for the SDS-PAGE analysis shown in Fig. 2.
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Andreasson, E., Abreha, K.B., Resjö, S. (2017). Isolation of Apoplast. In: Taylor, N., Millar, A. (eds) Isolation of Plant Organelles and Structures. Methods in Molecular Biology, vol 1511. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6533-5_18
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DOI: https://doi.org/10.1007/978-1-4939-6533-5_18
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6531-1
Online ISBN: 978-1-4939-6533-5
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