Flavonoid-induced calcium signalling in Rhizobium leguminosarum bv. viciae.

• Legume-rhizobium symbiosis requires a complex dialogue based on the exchange of diffusible signals between the partners. Compatible rhizobia express key nodulation (nod) genes in response to plant signals - flavonoids - before infection. Host plants sense counterpart rhizobial signalling molecules - Nod factors - through transient changes in intracellular free-calcium. Here we investigate the potential involvement of Ca(2+) in the symbiotic signalling pathway activated by flavonoids in Rhizobium leguminosarum bv. viciae. • By using aequorin-expressing rhizobial strains, we monitored intracellular Ca(2+) dynamics and the Ca(2+) dependence of nod gene transcriptional activation. • Flavonoid inducers triggered, in R. leguminosarum, transient increases in the concentration of intracellular Ca(2+) that were essential for the induction of nod genes. Signalling molecules not specifically related to rhizobia, such as strigolactones, were not perceived by rhizobia through Ca(2+) variations. A Rhizobium strain cured of the symbiotic plasmid responded to inducers with an unchanged Ca(2+) signature, showing that the transcriptional regulator NodD is not directly involved in this stage of flavonoid perception and plays its role downstream of the Ca(2+) signalling event. • These findings demonstrate a key role played by Ca(2+) in sensing and transducing plant-specific flavonoid signals in rhizobia and open up a new perspective in the flavonoid-NodD paradigm of nod gene regulation.