Root dieback of Picea abies (L) Karst., Norway spruce, seedlings is a serious problem in Scandinavian forest nurseries. We have chosen spruce seedlings infected with a pathogenic Pythium sp. isolate as an experimental system to study the interaction between the roots of a gymnosperm and a pathogen at the protein level. In this infection system, necroses on the hypocotyl and browning of the upper part of the roots appear within 2 days. Within 10 days the seedlings are completely wilted. Low pH soluble PR-proteins from infected and uninfected roots have been resolved on isoelectrofocusing (IEF) gels. Chitinases, chitosanases and β-1,3-glucanases have been detected enzymatically after IEF. Our results show that more than 30 different pathogenesis-related (PR) proteins accumulated in roots after pathogen infection. PR proteins of low and high isoelectric points appeared within 2 days after infection. In uninfected plants, only one acidic protein was detected. Eight different isoforms of chitinases accumulated after pathogen infection. Two acidic chitinases were constitutively expressed, and one of these strongly accumulated following pathogen infection. Three chitosanase activities were observed in infected plants, while no chitosanase activity was detected in uninfected plants. Also, no β-1,3-glucanase activity was observed in uninfected plants. One acidic β-1,3-glucanase was detected in infected roots after the second day of infection. A second acidic β-1,3-glucanase of relatively higher pI was detected on the fourth day. To our knowledge, this is the first report describing the response of gymnosperm roots to pathogen infection at the protein level. For the first time, we show that PR proteins accumulating after pathogen infection also include chitosanases. It can be concluded from our results that although gymnosperms are evolutionarily very distant from angiosperms, in terms of the production of PR proteins the response of gymnosperm roots resembles that observed in angiosperms. © 1993 Academic Press, Inc.
