SUMMARYThe paper-sandwich technique for simultaneous inoculation of populations of root apices, was used to produce numbers of synchronously developing ectomycorrhizas, and to permit direct time-sequencing of stages in the infection process in eucalypts. After one day there was evidence of chemical interaction hetween fungus and host. By two days, sheath formation had started in the cap region, cap cells were invaded hy fungus, and apical root tissues commenced to swell. By four days all the anatomical features of an ectomycorrhiza were present, including the Hartig net. Thereafter, mycorrhizal features were merely enhanced and perpetuated hy subsequent acrogenous development of the fungus-root dual organ. The process of mycorrhizal perpetuation involved the same sequence of events (sheath development at the apex associated with invasion of cap cells, modification of other root tissues, then finally formation of Hartig net) as did the initial infection process.

SummaryDevelopment of ectomycorrhizas involves multiple genes that are implicated in a complex series of interdependent, sequential steps. Current research into ectomycorrhiza development and functioning is aimed at understanding this plant-microbe interaction in a framework of the developmental and physiological processes that underlie colonization and morphogenesis. After a brief introduction to the ectomycorrhizal symbiosis, the present article highlights recent work on the early signal exchange taking place between symbionts, and sketches the way functional genomics is altering our thinking about changes in gene expression during the early steps of the ectomycorrhiza development.© New Phytologist (2001) 151 : 145 -154

Signals leading to mycorrhizal differentiation are largely unknown. We have studied the sensitivity of the root system from plant model Arabidopsis thaliana to hypaphorine, the major indolic compound isolated from the basidiomycetous fungus Pisolithus tinctorius. This fungi establishes ectomycorrhizas with Eucalyptus globulus. Hypaphorine controls root hair elongation and counteracts the activity of indole-3-acetic acid on root elongation on A. thaliana, as previously reported for the host plant. In addition, we show that hypaphorine counteracts the rapid upregulation by indole-3-acetic acid and 1-naphthalenic-acetic acid of the primary auxin-responsive gene IAA1 and induces a rapid, transient membrane depolarization in root hairs and suspension cells, due to the modulation of anion and K+ currents. These early responses indicate that components necessary for symbiosis-related differentiation events are present in the nonhost plant A. thaliana and provide tools for the dissection of the hypaphorine-auxin interaction.

An easy to use method is presented here to compare and study the mineral phosphate-solubilizing activity of ectomycorrhizal fungi. This technique can discriminate between strains with differing phosphate-solubilizing activities. Synthetic mineral phosphates, crystalline or amorphous, were differentially solubilized by 10 ectomycorrhizal fungi. Natural crystalline phosphates studied do not seem to be solubilized by fungi under similar experimental conditions. Paxillus involutus 1 appears to be able to solubilize calcium phosphates using either ammonium or nitrate nitrogen, but the other isolates were able to effectively solubilize phosphate only in the presence of ammonium. This has implications regarding the possible mechanism used to solubilize phosphate by these isolates. Recrystallization can be seen in the culture medium if calcium ions are present. The type of crystals depends on the phosphate source and on the fungal strain. This technique is suitable for screening a large number of ectomycorrhizal strains. The significance of phosphate-solubilizing activity to plant growth stimulation needs to be determined by field trials. Key words: ectomycorrhizal fungus, phosphate, solubilization.

SUMMARYVesicular-arbuscular (VA) mycorrhizal fungi and ectomycorrhizal fungi were observed together in the same root system, and even within the same root apices, of seedlings of Eucalyptus dumosa A. Cunn. ex Schau. On a population basis, there was a succession of two overlapping mycorrhizal epidemics-VA mycorrhizas followed by ectomycorrhizas.This succession was interpreted as follows. Early build-up of VA mycorrhizas was related to initial higher inoculum potential of that fungus and to rapidity of colonization of individual roots. The ectomycorrhizal fungus was more successful later in achieving secondary infections by hyphal spread and in colonization of lateral root branches. The ectomycorrhizal fungus had no difficulty infecting pre-existing VA mycorrhizas, but the ectomycorrhizal sheath provided a barrier to subsequent VA mycorrhizal infections. While the proportion of VA mycorrhizas to ectomycorrhizas changed between two and five months, the overall proportion of mycorrhizal roots remained constant, suggesting that there could be competition between the different fungi for limiting substrate.

Summary Seedlings of Eucalyptus dumosa A. Cunn. ex Schau., derived from a parent tree growing in a patch of calcareous soil, grew very poorly or not at all in that same soil unless they had formed abundant mycorrhizas. Other seedlings of that tree, planted in non‐calcareous soil from the same locality, grew well with negligible mycorrhizas. Similar results were obtained using synthetic calcareous and non‐calcareous potting mixtures. E. dumosa appears therefore to be primarily a calcifuge species, which becomes secondarily or facultatively calcicole as a consequence of mycorrhiza formation. Inflow of calcium into the seedlings with many mycorrhizas was the same as that into seedlings with few, but phosphorus inflow was greater into the mycorrhizal system. Mycorrhizas were presumably more effective at mobilizing and accumulating phosphate from insoluble calcium phosphates in the calcareous substrates than were uninfected roots. Early infection of the seedlings was almost entirely by VA endomycorrhizal fungi, in contrast to the ectomycorrhizal fungi which are usual in eucalypts. Most of the growth stimulus seemed to be associated with these early endomycorrhizas. Ectomycorrhizas formed later and tended to replace the endomycorrhizas. The ectomycorrhizas which succeeded the endomycorrhizas in inoculated pots were of a different type (hyaline) from those (black) which developed spontaneously, presumably from airborne spores, on uninfected roots in uninoculated pots.

SUMMARYOxalic acid produced by an isolate of the mycorrhizal fungus Paxillus involutus (Batsch. ex Fr.) Fr. in still broth culture was assayed using high-performance liquid chromatography. Production of oxalic acid per unit of mycelium was influenced by nitrogen source and concentrations of exogenous calcium and bicarbonate ions. Nitrate-grown mycelia produced large quantities of oxalic acid; ammonium-grown mycelia produced small quantities regardless of what other ions were present. Calcium concentrations between 10 and 50 meq T^ slightly enhanced oxalic acid production in the presence of nitrate; concentrations between 250 and 500 meq 1"^ depressed production. Small additions of bicarbonate ions substantially increased oxalic acids production when nitrate was present. When calcium was present, most of the oxalic acid was associated with the mycelium, probably as calcium oxalate. In the absence of calcium, most of the oxalic acid occurred free in the culture medium. It is concluded that in calcareous soils, bicarbonate and nitrate are more important than calcium in stimulating oxalate production. The significance and underlying mechanisms of oxalate production are discussed.

Summary• Ectomycorrhizal hyphal growth is shown to be stimulated by a phenol compound isolated from Eucalyptus globulus ssp. bicostata root exudates, highlighting the importance of phenolics in host-fungal interaction.• HPLC analysis allowed separation and identification of phenolic compounds from Eucalyptus seedling tissues and root exudates. The activity of the flavonol, rutin, was tested on a range of mycorrhizal and saprophytic fungi.• Rutin stimulated Pisolithus hyphal growth by more than twofold, and the fungus responded significantly to concentrations as low as 1 pM; only a few strains responded.• Rutin from Eucalyptus globulus ssp. bicostata root exudates is a flavonoid signal for Pisolithus , and is the first such flavonoid signal identified. A rutin gradient could contribute to orientating hyphal elongation toward the root tip thereby favouring mycorrhizal infection, and might also influence the interaction between fungi in the rhizosphere.