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 mu g of apoplastic proteins can be obtained in a volume of 300 mu L from five potato leaflets, with minimal contamination by non-apoplastic proteins.

Keywords

Apoplast; Apoplastic protein; Secretome; Soluble fraction; Protease inhibitor; Phosphate buffer; Vacuum infiltration; Extracellular space

Published in

Methods in Molecular Biology
2017, volume: 1511, number: 1511, pages: 233-240
Book title: Isolation of Plant Organelles and Structures
ISBN: 978-1-4939-6531-1
Publisher: Springer

Authors' information