Isolation of Apoplast

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1511; p. 233
• Main Authors: Andreasson, Erik, Abreha, Kibrom B, Resjö, Svante
• Format: Journal Article
• Language: English
• Published: United States 2017
• Subjects: Buffers; Cell Fractionation - instrumentation; Cell Fractionation - methods; Culture Media - chemistry; Extracellular Space - chemistry; Phosphates - chemistry; Plant Leaves - chemistry; Plant Proteins - genetics; Plant Proteins - isolation & purification; Plant Proteins - metabolism; Protoplasts - chemistry; Solanum tuberosum - chemistry; Vacuum; Secretome; Apoplast; Apoplastic protein; Phosphate buffer; Vacuum infiltration; Extracellular space; Protease inhibitor; Soluble fraction
• ISSN: 1940-6029
• DOI: 10.1007/978-1-4939-6533-5_18

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.