Leaf infiltration in plant science: old method, new possibilities.

The penetration of substances from the surface to deep inside plant tissues is called infiltration. Although various plant tissues may be effectively saturated with externally applied fluid, most described infiltration strategies have been developed for leaves. The infiltration process can be spontaneous (under normal atmospheric pressure) or forced by a pressure difference generated between the lamina surface and the inside of the leaf. Spontaneous infiltration of leaf laminae is possible with the use of liquids with sufficiently low surface tension. Forced infiltration is most commonly performed using needle-less syringes or vacuum pumps.Leaf infiltration is widely used in plant sciences for both research and application purposes, usually as a starting technique to obtain plant material for advanced experimental procedures. Leaf infiltration followed by gentle centrifugation allows to obtain the apoplastic fluid for further analyses including various omics. In studies of plant-microorganism interactions, infiltration is used for the controlled introduction of bacterial suspensions into leaf tissues or for the isolation of microorganisms inhabiting apoplastic spaces of leaves. The methods based on infiltration of target tissues allow the penetration of dyes, fixatives and other substances improving the quality of microscopic imaging. Infiltration has found a special application in plant biotechnology as a method of transient transformation with the use of Agrobacterium suspension (agroinfiltration) enabling genetic modifications of mature plant leaves, including the local induction of mutations using genome editing tools. In plant nanobiotechnology, the leaves of the target plants can be infiltrated with suitably prepared nanoparticles, which can act as light sensors or increase the plant resistance to environmental stress. In addition the infiltration has been also intensively studied due to the undesirable effects of this phenomenon in some food technology sectors, such as accidental contamination of leafy greens with pathogenic bacteria during the vacuum cooling process.This review, inspired by the growing interest of the scientists from various fields of plant science in the phenomenon of infiltration, provides the description of different infiltration methods and summarizes the recent applications of this technique in plant physiology, phytopathology and plant (nano-)biotechnology.