Extracellular vesicles (EVs) are a heterogenous group of membranous vesicles comprising apoptotic bodies, microvesicles and exosomes. The distinct species are classified by their size and origin. Despite there is upcoming interest on animal derived EVs- resulting in an increasing knowledge of their function, composition, and cargo- little is known about EVs in the plant kingdom [1]. Agarose gels in concentrations between 0.5% - 2% are typically used for electrophoretic separation of nucleic acids [2]. Our investigations have shown that they are also suitable for quick and easy EV detection and purification. We isolated EVs from the apoplastic fluid (APF) of Nicotiana tabacum L., Hedera helix L., and Viscum album L. Even though intact plant derived EVs do not migrate into the gel matrix, they can be visualized inside the well. If 3,3-Dihexyloxacarbocyanine iodide (DiOC6) is added to the specimen in excess, membranous components can already be detected by eye, or with higher sensitivity, using a UV transilluminator. The detected subtype of EVs depends on the previous sample preparation. Moreover, EVs are purified from small charged contaminants and dye excess, passing through the gel.
References:
[1] Rutter B, Innes R. Plant Physiol 2017; 173: 728-741
[2] Lee P, Costumbrado J, Hsu C, Kim Y. J Vis Exp 2012; 62: 3923