Abstract
Plants are constantly facing abiotic and biotic stresses. To continue to thrive in their environment, they have developed many sophisticated mechanisms to perceive these stresses and provide an appropriate response. There are many ways to study these stress signals in plant, and among them, protoplasts appear to provide a unique experimental system. As plant cells devoid of cell wall, protoplasts allow observations at the individual cell level. They also offer a prime access to the plasma membrane and an original view on the inside of the cell. In this regard, protoplasts are particularly useful to address essential biological questions regarding stress response, such as protein signaling, ion fluxes, ROS production, and plasma membrane dynamics. Here, the tools associated with protoplasts to comprehend plant stress signaling are overviewed and their potential to decipher plant defense mechanisms is discussed.
Highlights
Plants are exposed to myriads of potential stresses that can be harmful to their development
transient expression assays (TEA) are performed by isolating protoplasts from plant tissues, transfecting them in the presence of polyethylene glycol and calcium (Yoo et al, 2007; Lin et al, 2014) or through electroporation (Miao and Jiang, 2007) and incubating them for 2–24 h
Di-4ANEPPDHQ has notably been used on protoplasts from rice transgenic plants that lack fatty acid hydroxylase 1 and 2 (FAH1/2), enzymes responsible for the formation of 2-hydroxy sphingolipids (2-OH-SL), precursors of glycosylinositol phosphorylceramides (GIPC), that are both located at the plasma membrane (PM) in Arabidopsis
Summary
Plants are exposed to myriads of potential stresses that can be harmful to their development. Through cell surface and intracellular protein receptors, the plant is capable of sensing multiple molecular stress factors, such as MAMPs (microbe-associated molecular patterns), PAMPs (pathogen-associated molecular patterns), and DAMPs (damageassociated molecular patterns), initiating a cascade of signal transduction leading to a rapid and effective response from the plant (Cook et al, 2015; Couto and Zipfel, 2016) Both biotic and abiotic stresses share some early signaling events such as the production of reactive oxygen species (ROS) by NADPH oxidases, activation of protein kinases, receptors, or co-receptors through phosphorylation (Kadota et al, 2015; Yu et al, 2017; Zipfel and Oldroyd, 2017; Bigeard and Hirt, 2018), and rapid and transient change of ion fluxes (Jones and Dangl, 2006; Bigeard et al, 2015; Lamers et al, 2020). We will enlighten and discuss the advantages and the limitations of protoplasts as a proxy for whole tissues or plants
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