title: Isolation of Apoplast.
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.
authors.affiliation: Department of Plant Protection Biology, Swedish University of Agricultural Sciences, Sundsvägen 14, SE-230 53, Alnarp, Sweden.
authors.forename: Erik
authors.initials: E
authors.lastname: Andreasson
authors.affiliation: Department of Plant Protection Biology, Swedish University of Agricultural Sciences, Sundsvägen 14, SE-230 53, Alnarp, Sweden.
authors.forename: Kibrom B
authors.initials: KB
authors.lastname: Abreha
authors.affiliation: Department of Plant Protection Biology, Swedish University of Agricultural Sciences, Sundsvägen 14, SE-230 53, Alnarp, Sweden. svante.resjo@slu.se.
authors.forename: Svante
authors.initials: S
authors.lastname: Resjö
mesh_terms: D002021:Buffers; D002458:Cell Fractionation / Q000295:instrumentation / Q000379:methods*; D003470:Culture Media / Q000737:chemistry; D005110:Extracellular Space / Q000737:chemistry*; D010710:Phosphates / Q000737:chemistry; D018515:Plant Leaves / Q000737:chemistry*; D010940:Plant Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D011523:Protoplasts / Q000737:chemistry; D011198:Solanum tuberosum / Q000737:chemistry*; D014618:Vacuum
chemical_list: D002021:Buffers; D003470:Culture Media; D010710:Phosphates; D010940:Plant Proteins
keywords: Apoplast; Apoplastic protein; Extracellular space; Phosphate buffer; Protease inhibitor; Secretome; Soluble fraction; Vacuum infiltration