Colonization of Orchis morio protocorms by a mycorrhizal fungus: effects of nitrogen nutrition and glyphosate in modifying the responses

Abstract

Effects of nitrogen nutrition and application of glyphosate (as Roundup™) on the interactions between protocorms of Orchis morio and a mycorrhizal Rhizoctonia species were investigated. Protocorms for all experiments were raised clonally in liquid culture. Split plates were used to separate the direct effects of composition of the media from effects mediated via the fungus. Mycorrhizal interactions, including coil formation and prolonged protocorm growth, were established when a relatively low nitrogen supply to the fungus (as NH4)2SO4) was combined with a high carbohydrate supply. Rejection of the fungus, associated with phenolic production, wall thickening, and lack of protocorm growth, was observed with high-carbon, high-nitrogen medium. Low carbohydrate supply, with either high or low nitrogen supply, was associated with breakaway parasitism, symptoms of soft rot, and lack of subsequent protocorm growth. Application of glyphosate at 0.5 or 1.0 mM had no effect on fungal growth and at 1.0 mM did not cause death of asymbiotic protocorms, but resulted in failure of mycorrhizal initiation. Coils were never formed and the fungus ramified through the tissues of the protocorms. The parasitism induced by glyphosate differed from breakaway parasitism, and there were no symptoms of soft rot. Assays for activity of phenylalanine ammonia lyase (PAL) and for orchinol were carried out to assess the effects of the treatments on the shikimic acid pathway. Low activity of PAL and low quantities of orchinol (together with another unidentified phenolic compound) were detected in asymbiotic protocorms. Concentrations of both were increased in the presence of the fungus, but no significant differences were observed in the various symbiotic responses in the absence of glyphosate. This is the first report of the presence of orchinol in orchid protocorms. Previous work has always been carried out on tubers, which are not usually colonized by the mycorrhizal fungi. Application of glyphosate resulted in increases in both PAL activity and orchinol production. The results are discussed in the context of orchid–fungus interactions and the mechanism of glyphosate action. Key words: orchid, mycorrhiza, parasitism, cell walls, glyphosate, phenylalanine ammonia lyase (PAL), orchinol.