The photosensitive endoplasmic reticulum-chloroplast contact site.

Abstract

The endoplasmic reticulum (ER) forms contact sites with the chloroplast. Exposing contact sites that contain both the chloroplast and the ER to localised high-fluence, wavelength specific, 405 nm violet light, hereinafter referred to as photostimulation, induces multiple, potentially interacting intra- and intercellular responses. The responses vary depending on the tissue type of the cell and the chloroplast. Photostimulating the ER-chloroplast contact sites in growing epidermal cells of the hypocotyl of Arabidopsis thaliana, produces a wave of cytoplasmic ionic calcium that traverses the cell, spreading radially to other cells around the circumference of the hypocotyl. A transient ER stress accompanies the calcium wave. These responses occur in older epidermal cells (5-8 days post-germination) with nonmotile chloroplasts tethered to the ER and the cell cortex but do not occur with motile or dividing chloroplasts. Dividing chloroplasts show a markedly different association with the ER, which forms a ring around the fission plane, similar to that of dividing mitochondria. Inhibition of calcium channels with lanthanum has no effect. Photostimulation of only the ER results in no ER stress and a calcium wave with a different spatiotemporal signature: delayed release and lower magnitude, with no accompanying ER stress response. Likewise, photostimulation of the chloroplast only, without the ER, produces no calcium wave or ER stress. General chloroplast photobleaching or restructuring caused by photostimulation is not the cause of this response; photostimulation with 488 nm of the same intensity and power as 405 nm photostimulation produces no change in cytosolic calcium levels. The pH of the ER decreases, indicating the involvement of ER ion transporters in the response. A wave of increased reactive oxygen species (ROS) in mitochondria and nuclei accompanies photostimulation. Together, these data support a model by which tethered ER-chloroplast contact sites constitute a unique subcellular photosensitive region and are part of an ER-mediated signalling network. Lay Abstract: The endoplasmic reticulum (ER) forms contact sites with the chloroplast. Shining violet (405 nm) light on the chloroplast with its associated ER produces a calcium wave through the cell that is communicated to other cells. This is correlated with a wave of transient denaturation of the luminal proteins of the ER (ER stress) and increased reactive oxygen species (ROS) in mitochondria. The wavelength dependence and precise cellular location of the light stimulation implies a novel way for plants to sense light. The movement of the response through the cell is consistent with the mediation of the response by a subcellular network, such as that formed by the ER.