Ca2+ is Required by Clubroot Resistant Turnip Cells for Transient Increases in PAL Activity that Follow Inoculation with Plasmodiophora brassicae

Phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) activity in clubroot disease-resistant turnip calli was transiently increased by 20h after the inoculation with Plasmodiophora brassicae spores. The magnitude of the increase in PAL activity was four to six times higher than constitutive PAL activity. There was no transient increase in PAL activity in susceptible calli. Preincubation of calli in Ca2+-free medium or the removal of Ca2+ from cell surfaces by ethylene glycol bis (2-aminoethyl ether)-N,N,N¢,N¢-tetraacetic acid-chelation, completely inhibited induced PAL activity. The influx of exogenous Ca2+ into cells appears necessary for this pathogen induced PAL activity. Verapamil and the calmodulin inhibitor W7 almost completely inhibited induced PAL activity at 1 and 0.1 mm, respectively. Neomycin, ruthenium red and (1-(6-[(17b- 3-Methoxyestra-1,3,5-(10)-trien-17-yl)amino]hexyl)-1Hpyrrole- 2,5-dione) did not inhibit induced PAL activity. Thus, verapamil and N-(6-aminohexyl)-5-chloro- 1-naphthalenesulphonamide hydrochloride-sensitive Ca2+-mediated signalling process appear necessary for P. brassicae induced PAL activity. As the protein synthesis inhibitor cycloheximide (CHX) blocked the induced increasing PAL activity, de novo synthesis of PAL appears to be required for turnip cell defence reactions against P. brassicae