The dynamics of early life-history stages are important for understanding regional diversity patterns in tropical trees. In this study, patterns of seed removal, seedling survival and ectomycorrhizal (ECM) colonization in monodominant and mixed tropical rain forests in Guyana were investigated during a masting event of an ECM monodominant tree, Dicymbe corymbosa. Two seed densities (1 m−2 and 6 m−2) were tested in two forest types (monodominant versus mixed) with 10 plots per treatment (40 plots total). Germination success, seed and seedling survival after 1 y were measured for the monodominant tree, and four non-dominant canopy tree species found in both forest types (Eschweilera sagotiana, Licania laxiflora, Chrysophyllum sanguinolentum and Carapa guianensis). A significant effect of forest type was detected for seed removal and seedling survival. In the mixed forest, seed removal was higher for all species, with 27% average removal compared with 7% average seed removal in the monodominant forest. Germination success was significantly lower for all but one species in the mixed forest, with average germination of 41% compared with 78% germination in the monodominant forest. Seed and seedling survival of the monodominant tree was greatest within the monodominant forest with 93% of seeds germinating and 85% of seedlings surviving after 1 y compared with 65% germination and 15% seedling survival in the mixed forest. Per cent ectomycorrhizal colonization of Dicymbe corymbosa was near 100% in the monodominant forest, but significantly lower (14%) in the mixed forest. These results suggest that seedling survival patterns, rather than seed survival, are more important for recruitment success in this system and that ectomycorrhizal inoculum may limit establishment of Dicymbe corymbosa in the mixed forest.

The extraradical mycelium of ectomycorrhizas is comprised of a network of hyphae that may initiate rhizomorphs, sclerotia and sexual reproductive structures. The development of these structures requires photosynthates produced by host trees. In this study, the initiation and early development of Laccaria bicolor (Maire) Orton fruitbodies (basidiocarps) were studied. Seedlings of Pinus resinosa Ait. and Pinus sylvestris L. were colonized by L. bicolor, a broad host epigeous ectomycorrhizal basidiomycetous fungal species, in growth pouches. Ectomycorrhizas with an extensive extraradical mycelium formed on short roots within 7-12 days after fungal inoculum was introduced. Numerous sites of hyphal aggregation, many of which subsequently developed into basidiocarp primordia, were initiated in the extraradical mycelium. Initial changes in aggregating hyphae included swelling and branching followed by growth of hyphae perpendicular to the paper wick in growth pouches. A stipe and a pileus primordium developed but none of these basidiocarp primordia matured. Distinct regions were evident in the stipe and pileus when sections were stained by various methods. Loose hyphae at the apex and periphery of the pileus were separated by mucilage. In Petri dishes with vermiculite as the substrate, basidiocarps of Laccaria laccata formed in ectomycorrhizal associations with Pinus sylvestris, developed a mature pileus with an hymenium and mature basidiospores.

Three ectomycorrhizal legume trees, Microberlinia bisulcata, Tetraberlinia bifoliolata and T. moreliana, form discrete groves in the southern part of Korup National Park, in southwest Cameroon and contribute c. 45–70% of stand basal area locally in a matrix of otherwise species-rich arbuscular mycorrhizal forest. A transplant experiment was performed to assess the importance of ectomycorrhizal infection associated with proximity to parents in seedling establishment of the grove-forming species. Nonectomycorrhizal seedlings of the three species were transplanted into plots of two forest types, one of high (HEM, within-grove) and one of very low (LEM, outside the grove) abundance of all three species as adult trees. For two species (T. moreliana and M. bisulcata) there was no difference in survival over 16 months, but for the third (T. bifoliolata) survival was best in HEM forest, and correlated with the basal area of adult trees of ectomycorrhizal species. Only one species (T. moreliana) increased in biomass over the experimental period; the others declined. There was no effect of forest type on overall growth of any species, but the survivors of two (T. moreliana and M. bisulcata) had heavier stems in the HEM forest. Differences in survival and growth of transplants between the three species were in accord with the ecology of the species as inferred from the frequency distributions of adult tree size in the forest. Seedlings became infected with ectomycorrhizas in both forest types; where there was a difference in extent of infection (T. moreliana) this was not related to survival or growth; and where there was a difference in survival (T. bifoliolata) this was not related to extent of infection. These results confirm that mycorrhizal inoculum associated with conspecific adults is neither a prerequisite nor a guarantee of seedling establishment, but indicates that in some circumstances there might be benefits of being close to parents. Further research is required to unravel the complexities of ectomycorrhizal community structure in this spatially and temporally heterogeneous forest, and to clarify the extent to which the various hosts share ectomycorrhizal partners.

Eucalyptus species are considered to have ectomycorrhizas (ECM), but many also have vesicular–arbuscular mycorrhizas (VAM) and their relative importance is unclear. Interactions between ECM and VAM fungi colonizing roots of Eucalyptus species were examined in a glasshouse experiment. This experiment investigated competition between these two types of fungi and compared benefits provided to Eucalyptus globulus and E. urophylla. Eucalyptus seedlings were inoculated with spores of the ECM fungus Laccaria lateritia and/or pot- culture soil for a VAM fungus (species of Glomus, Acaulospora or Scutellospora). Initial inoculum levels were important, as VAM fungi became established much more rapidly than Laccaria. In plants with both types of mycorrhizas, Laccaria mycorrhizas substantially increased after 2 or 3 months and the proportion of roots with VAM declined. However, the proportion of roots with VAM also decreased significantly in plants without ECM after 2 months. Thus, the relative susceptibility of eucalypt roots to these mycorrhizal associations changed. Substantial growth responses to mycorrhizal inoculation occurred when a low concentration of phosphorus fertilization was used (5 mg kg−1), but not at a higher concentration (10 mg kg−1). Treatments where Laccaria was applied, alone or in combination with a VAM fungus, resulted in the largest growth increases relative to nonmycorrhizal plants. An Acaulospora isolate was the most effective VAM fungus for E. urophylla although other VAM fungi also increased growth relative to the control. VAM fungi alone had little effect on E. globulus growth, but plants inoculated with both ECM and VAM fungi were larger than plants only inoculated with Laccaria. ECM fungi had a major impact on root system form by reducing the proportion of fine roots (specific root length) relative to nonmycorrhizal plants or those with VAM. Changes in root colonization patterns over time must be considered in studies of ECM/VAM interactions, as there can be substantial changes in the relative importance of the two associations.

Previous work in the Colorado alpine ecosystem has shown that amino acids are a potentially important N source for the sedge, Kobresia myosuroides. This plant is the only known sedge to harbour associations with ectomycorrhizal fungi. The aim of the present work was to test the hypothesis that these ectomycorrhizas transfer N from amino acids in the soil solution to the host plant, and thereby have an important role in the N nutrition of this species. We used a two-chamber system (rhizoboxes) in which K. myosuroides plants were separated from a soil chamber by nylon mesh that allowed fungal hyphae, but not plant roots, to cross it. Injections of [15N, 2-13C]glycine were made into the soil chamber. The hyphal crossings on half of the rhizoboxes were regularly disrupted to control for leakage of label across the barrier. Plants in the intact rhizoboxes showed significantly higher 15N enrichment than those in controls, and mycorrhizal root tips were significantly more enriched than bulk roots. The mycorrhizas transferred an average of 1.3% of the added 15N label to plants, a figure comparable to those obtained in previous studies in which plant roots were directly exposed to label. We conclude that fungal associations have an important role in the N nutrition of K. myosuroides by transferring N from amino acids to their hosts.

Beech nuts (Fagus sylvatica L.) were germinated and grown in soil inoculated with the ectomycorrhizal fungus Laccaria laccata or Paxillus involutus for 18–20 wk. The success of mycorrhizal infection was monitored by measuring the ergosterol contents of the mycorrhizas. Ergosterol levels ranged from 122±23 μg g−1 d. wt (Laccaria mycorrhizas) to 94±36 μg g−1 d. wt (Paxillus mycorrhizas), indicating that ectomycorrhizal symbiosis was established. In root incubation chambers, rates of sulphate uptake and the xylem loading of sulphate of excised mycorrhizas were investigated. Both types of mycorrhizas showed saturation kinetics in external sulphate concentrations from 2·5–1000 μmol l−1. Linearization of these kinetics revealed two phases with low apparent Km (Laccaria mycorrhizas: 15±3 μmol l−1; Paxillus mycorrhizas: 13±3 μmol l−1) and Vmax (Laccaria mycorrhizas: 19±3 nmol h−1 g−1 f. wt; Paxillus mycorrhizas: 25±4 nmol h−1 g−1 f. wt) at low external sulphate concentrations and significantly higher kinetic constants at higher sulphate supplies. Relative xylem loading, i.e. the portion of sulphate loaded into the xylem that was taken up, remained constant over the entire concentration range investigated (c. 4–7% of the sulphate taken up). If trees were supplied for 72 h with different N and sulphur concentrations, both uptake of sulphate and relative xylem loading were unaffected by sulphur availability, but modulated by N supply. Nitrogen depletion diminished the rates of sulphate uptake in Laccaria and Paxillus mycorrhizas. In response to higher N availability combined with sulphur depletion, sulphate uptake of Laccaria mycorrhizas, but not of Paxillus mycorrhizas, increased. Organic compounds considered to be possible signals for the regulation of sulphate uptake were fed to excised mycorrhizas. l-Cysteine but not l-methionine and glutathione (γ-Glu-Cys-Gly) inhibited sulphate uptake of the two mycorrhizas and xylem loading of sulphate was stimulated rather than inhibited by l-Cys in both types. In Paxillus mycorrhizas glutathione had a similar effect. O-Acetyl-l-serine (OAS), a precursor of l-cysteine, stimulated sulphate uptake, but did not affect xylem loading. Apparently, OAS, generated in N metabolism, and l-cysteine, a product of assimilatory sulphate reduction, act as antagonists, together mediating regulation of sulphate uptake.

We tested how nitrogen availability affects suitability of ectomycorrhizal (ECM) and non-mycorrhizal Pinus sylvestris (L.) for a polyphagous insect herbivore. Seedlings grown on sterile agar at two N levels, were infected or not with the ECM fungus Cenococcum geophilum (Sow.) Ferd. & Winge. Ultrastructural observations showed that C. geophilum formed well-developed ectomycorrhizas in pine roots. After mycorrhizal establishment, oviposition preference of the polyphagous tarnished plant bug (Lygus rugulipennis Popp.) was determined in three different experiments. The duration of these experiments and the amount of irradiance were different. Nitrogen level or mycorrhizal treatment did not have significant effects on oviposition preference. However, mycorrhizal seedlings were favoured as an oviposition site at the lower N level, but only in expt 1. Total or individual terpene, total resin acid and total or individual sugar concentrations in pine shoots were not altered by treatment. Total phenolics concentration in pine shoots and roots was also the same in different treatments. Of individual resin acids, the concentration of dehydroabietic acid was increased after mycorrhizal treatment. Total resin acid, abietic acid and sandaraco pimaric acid concentrations correlated negatively with the number of Lygus eggs, but the correlations were only marginally significant. Of individual terpenes, β-pinene and bornylacetate concentrations had a significant positive correlation with the number of Lygus eggs. Greater amounts of N predictably increased total N concentration in pine seedlings in expts 1 and 2, but mycorrhizal infection increased total nitrogen concentration only in expt 3. Total N concentration did not correlate with the number of Lygus eggs in expts 1 and 2. An early stage of mycorrhizal development did not provide mycorrhizal benefits to the host plant as a higher nutrient concentration or as increased growth. These results suggest that host–plant quality might be involved in selection of oviposition site by L. rugulipennis, but ectomycorrhizal infection at this level does not seem to be an important mediator.

The frequent co-occurrence of ectomycorrhizas and arbuscular mycorrhizas in the same ecosystem raises the question about how two functionally different mycorrhizal associations exploit a spatially heterogeneous habitat. Horizontal (in plots along a transect of 3750 m crossing forest types of low and high abundance of ectomycorrhizal (ECM) host species) and vertical (across profiles of litter and 0–5 cm, 10–15 cm, 25–35 cm soil layers) distribution of ectomycorrhizas and arbuscular mycorrhizas in relation to edaphic parameters (pH, moisture, total carbon, total nitrogen and extractable-phosphorus concentration) was compared in Korup National Park rain forest, to test the hypothesis that the two types of mycorrhiza exploit different niches. No pattern was observed in either ECM or arbuscular mycorrhizal (AM) fractional colonization with depth in the profile even though most of the variation in soil parameters was vertical. Only local (between monoliths within plots) horizontal differences were found in soil parameters. By contrast, horizontal differences in fractional colonization were found between forest types, and between monoliths, for ectomycorrhizas but only between monoliths for arbuscular mycorrhizas. The lack of a clear relationship between both horizontal and vertical distribution of ECM and AM fractional colonization and soil parameters suggests that both fungal groups are equally able to colonize the same niche, at the scale of our experiment. By contrast, a negative relationship was found between AM and ECM fractional colonization in the litter and 0–5 cm and 10–15 cm soil layers. This, in addition to the more heterogeneous horizontal distribution of arbuscular mycorrhizas in forest areas dominated by ectomycorrhizas and a patchy vertical distribution of the former as opposed to a more coherent distribution of the latter, suggests that both types of mycorrhizal fungi compete for the same niche. The co-occurrence of ectomycorrhizas and arbuscular mycorrhizas might be explained by differences between ECM and AM hosts and fungi in their colonizing abilities.

Sequence-based markers were developed to study the genetic structure and reproductive biology of the ectomycorrhizal fungus Suillus pungens Thiers & Smith in a Bishop pine (Pinus muricata D. Don) forest. Six different basidiome genotypes were found in a 1200 m2 area. Five of the six genotypes were represented by single basidiomes. The remaining genotype comprised 13 basidiomes and covered an area of at least 300 m2, with maximum measured dimensions of 40 m and 14 m. This is the largest genet of an ectomycorrhizal fungus described to date, and is likely the result of vegetative growth, because analysis of single spore isolates eliminates the possibility of genetic identity resulting from either apomixis or fortuitously indistinguishable recombinant genotypes. Genetic analysis also shows that although out-crossing appears to predominate in the population, at least a low percentage (1·4%) of spores are secondarily homothallic.

The combination of extensive vegetative growth and abundant fruiting suggests S. pungens utilizes more carbon than might be expected for a species which accounts for <3% of the total ectomycorrhizal abundance at the site. Additional carbon might come from either more efficient host–fungus transfer, pooling of carbon from the roots of different host plants, or saprophytism.

Longleaf pine (Pinus palustris Mill.) seedlings were exposed to two concentrations of atmospheric CO2 (365 or 720 μmol mol−1) and two levels of N (0·02 or 0·20 mg N g−1 soil yr−1) within open-top chambers for 20 months. Seedlings were adequately watered for 19 wk to ensure seedling establishment, after which two water-stress treatments (target values −0·5 or −1·5 MPa xylem pressure potential) were implemented. Fine-root samples were collected in July and November 1993, and in March and November 1994. Ectomycorrhizal and non-mycorrhizal short roots per unit length of fine root were quantified. The percentage of ectomycorrhizal short roots and numbers of ectomycorrhizas per unit root length were higher for seedlings grown with elevated CO2, low N and adequate water. Interactions among main treatment variables demonstrated higher percentages of ectomycorrhizal short roots, fine root length per seedling, and total numbers of ectomycorrhizas per seedling for plants grown with high CO2 (compared with ambient) or adequate water (compared with water stress) only under high N conditions. Increased fine-root length and ectomycorrhizal colonization under elevated CO2 resulted in higher (almost double) numbers of ectomycorrhizas per seedling at each sampling.

We investigated the effect of elevated CO2 on the growth and mycorrhizal colonization of three tree species native to north-eastern American forests (Betula papyrifera Marsh., Pinus strobus L. and Tsuga canadensis L. Carr). Saplings of the tree species were collected from Harvard Forest, Massachusetts, and grown in forest soil under ambient (c. 375 ppm) and elevated (700 ppm) atmospheric CO2 concentrations for 27–35 wk.

In all three species there was a trend to increasing whole-plant, total-root and fine-root biomass in elevated CO2, and a significant increase in the degree of ectomycorrhizal colonization in B. papyrifera and P. strobus, but not in T. canadensis. However, in T. canadensis the degree of colonization with arbuscular mycorrhizas increased significantly. In both the ambient and elevated environments, on the roots of B. papyrifera and P. strobus 12 distinct ectomycorrhizal morphotypes were identified. Distinct changes in the ectomycorrhizal morphotype assemblage of B. papyrifera were observed under CO2 enrichment. This change resulted in an increase in the frequency of ectomycorrhizas with a higher incidence of emanating hyphae and rhizomorphs, and resulted in a higher density of fungal hyphae in a root exclusion chamber.

Seedlings of Pseudotsuga menziesii (Mirb.) Franco were grown for six–16 months in untrenched and trenched treatments in three 90–120-yr-old mixed forests dominated by Betula papyrifera Marsh. and P. menziesii in the southern interior of British Columbia. Each forest was characterized by mesic conditions and low light intensity (PAR<200 μmol m−2 s−1) in the understorey. The objective of the study was to evaluate the influence of overstorey tree roots on (i) ectomycorrhizal fungal composition, richness and diversity, and (ii) photosynthesis and growth of understorey P. menziesii seedlings. Seventeen ectomycorrhizal morphotypes were recognized on seedlings in the untrenched treatment, and nine in the trenched treatment over the three sites. Six types occurred in both treatments, of which on average Rhizopogon vinicolor Smith type was 20 times more abundant and Thelephora type six times less abundant in the untrenched as in the trenched treatment. Of types that formed strands or rhizomorphs, eight occurred in the untrenched treatment, where they occupied on average 23% of root tips, and only four occurred in the trenched treatment over 4% of the root tips. Mean richness, diversity, and evenness of ectomycorrhizal associates per seedling were approx. twice as great in the untrenched as in the trenched treatment.

Net photosynthetic rate of P. menziesii seedlings was greater in the untrenched than in the trenched treatment in July and August, but not in September 1994. Height, diameter and biomass of seedlings did not differ between treatments, but height[ratio ]diameter ratio was greater in the untrenched treatment at time of harvest. The effect of trenching on seedling performance was attributed mainly to differences in ectomycorrhizal colonization patterns because trenching had no significant effect on soil nutrient concentrations (total C, total N, NH4-N, available N, exchangeable Ca, exchangeable Mg, exchangeable K), C[ratio ]N ratio, soil pH or light availability. Nor was there a significant difference in soil water in August, when seedlings in untrenched plots had higher net photosynthetic rates than those in trenched plots. Results suggest that influence of overstorey trees and pattern of ectomycorrhizal formation are important to P. menziesii seedling performance in deeply shaded forest environments.