N-butyryl-homoserine lactone, a bacterial quorum-sensing signaling molecule, induces intracellular calcium elevation in Arabidopsis root cells

N-acyl-l-homoserine lactones (AHLs) are quorum sensing (QS) signal molecules that are commonly used in gram-negative bacteria. Recently, it has become evident that AHLs can influence the behavior of plant cells. However, little is known about the mechanism of the plants’ response to these bacterial signals. Calcium ions (Ca²⁺), ubiquitous intracellular second messengers, play an essential role in numerous signal transduction pathways in plants. In this study, the cytosolic free Ca²⁺ concentration (Ca²⁺]_cyt) was measured by a luminometric method in the excised root cells of Arabidopsis plants that were treated with N-butyryl-homoserine lactone (C4-HSL). There was a transient and immediate increase in Ca²⁺]_cyt levels, and the highest level (0.4 μM), approximately 2-fold higher than the basal level, was observed at the 6th second after the addition of 10 μM C4-HSL. Pretreatments with La³⁺, verapamil or ethylene glycol tetraacetic acid (EGTA) inhibited the increase in Ca²⁺]_cyt caused by C4-HSL, whereas it remained unaffected by pretreatment with Li⁺, indicating that the Ca²⁺ contributing to the increase in Ca²⁺]_cyt was mobilized from the extracellular medium via the plasma membrane Ca²⁺ channels but not from the intracellular Ca²⁺ stores. Furthermore, electrophysiological approaches showed that the transmembrane Ca²⁺ current was significantly increased with the addition of C4-HSL. Taken together, our observations suggest that C4-HSL may act as an elicitor from bacteria to plants and that Ca²⁺ signaling participates in the ability of plant cells to sense the bacterial QS signals.