Nod factors stimulate plasma membrane delimited phospholipase C activity in vitro

Abstract

Nod factors are lipo‐chito‐oligosaccharides secreted by Rhizobium to initiate deformation of root hairs and other changes in host plants. Since Nod factor‐induced changes in intracellular calcium occur in responsive root hairs, we tested if phospholipase C (PLC) activity is stimulated by Nod factors. Plasma membranes were isolated from the nodulation‐competent zone of roots of Vigna unguiculata to assay PLC activity in vitro. Nod factors isolated from Rhizobium sp. NGR234, NodNGR[S] and NodNGR[Ac] significantly increased PLC activity and this increase in activity was inhibited in the presence of the PLC inhibitors, neomycin and U‐73122. The response appears specific as PLC activity was not significantly induced neither by the 4‐sugar, N,N′,N′′,N′′′‐tetracetylchitotetraose (TACT), or the five‐sugar, penta‐N‐acetylchitopentaose (PACT), backbone of Nod factors. The G‐protein activators, GTPγS and the aluminium fluoride complex, had no effect on PLC activity in the presence or absence of NodNGR[S], suggesting that Nod factors act independently of G‐proteins in vitro. However, the combination of oleic acid and TACT mimicked the effect of Nod factors on PLC activity indicating that the presence of the lipid tail may be critical. Also this combination of compounds acted synergistically together to evoke root hair deformation in vivo. Our results indicate that Nod factors can modulate membrane delimited PLC activity and indicate that PLC is likely to be a component of the Nod factor‐signalling pathway.