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RESEARCH PRODUCT

Ergosterol elicits oxidative burst in tobacco cells via phospholipase A2 and protein kinase C signal pathway

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Ergosterol, a typical fungal sterol, induced in tobacco (Nicotiana tabacum L. cv. Xanthi) suspension cells the synthesis of reactive oxygen species and alkalization of the external medium that are dependent on the mobilization of calcium from internal stores. We used specific inhibitors to elucidate the signal pathway triggered by ergosterol compared with cryptogein, a proteinaceous elicitor of Phytophthora cryptogea. HerbimycinA and genistein, inhibitors of tyrosine protein kinases, had no effect on the oxidative burst and pH changes induced by bothelicitors.Similarly,H-89,aninhibitorofproteinkinaseA,hadnoeffectontheinductionofthesedefensereactions.However,theresponse to both elicitors was completely blocked by NPC-15437, a specific inhibitor of animal protein kinase C (PKC). The responses induced by cryptogein but not those induced by ergosterol were inhibited by U73122 and neomycin, inhibitors of phospholipase C (PLC). On the other hand, the activity of phospholipaseA2 (PLA2) measured using a fluorogenic substrate was stimulated by ergosterol and not by cholesterol and cryptogein.A specific inhibitor of PLA2, arachidonic acid trifluoromethyl ketone (AACOCF3), inhibited the pathway stimulated by ergosterol but not that induced by cryptogein. These results suggest that the cryptogein-induced signal pathway leading to the oxidative burst and DpH changes includes PLC and PKC, whereas this response induced by ergosterol includes PLA2 and PKC. © 2004 Elsevier SAS. All rights reserved.