Seedlings of the myco-heterotrophic orchid Corallorhiza trifida which had been germinated in the field in mesh bags developed hyphal links and mycorrhizas with Betula pendula and Salix repens, but not with Pinus sylvestris, when transplanted into soil microcosms. The fungus connecting the myco-heterotroph to Betula and Salix formed endomycorrhiza in the orchid with typical pelotons, but formed ectomycorrhizas with the autotrophs. The orchid plants, when linked to Betula and Salix by fungal hyphae, gained 6–14% in weight over 25–28 wk. In microcosms supporting P. sylvestris, and in control microcosms which lacked autotrophs, the Corallorhiza plants lost 13% of their weight over the same period. In the course of the 28-wk experimental period new Corallorhiza seedlings, in addition to those added as part of the experiment, appeared in the microcosms containing Salix and Betula but not in the Pinus microcosms. Shoots of Betula and Salix plants grown in association with Corallorhiza were fed with 14CO2, and the movement of the isotope was subsequently traced by a combination of digital autoradiography and tissue oxidation. Direct transfer of C from both autotrophs to the myco-heterotroph occurred in all cases where the associates had become connected by a shared fungal symbiont. Orchid seedlings lacking these hyphal connections, introduced to the microcosms as controls immediately before isotope feeding, failed to assimilate significant amounts of C. The results provide the first experimental confirmation that growth of Corallorhiza trifida can be sustained by supply of C received directly from an autotrophic partner through linked fungal mycelia.

The effects of increased nitrogen influx on Sphagnum growth and on interspecific competition between Sphagnum species were studied in a 3-yr experiment in mires situated in two areas with different rates of airborne N deposition. Sphagnum growth was recorded after various supplementary N influxes (0, 1, 3, 5 and 10 g m −2 yr−1) in hummocks and lawn communities. Sphagnum biomass production decreased with increasing N influx in both areas. After the first season at the low-deposition site, Sphagnum showed an increased growth in length with the intermediate N treatment, but in the second and third seasons the control treatment had the highest growth in length. Capitulum dry mass increased with increasing N influx. Sphagnum N concentration and N/P quotient were higher at the high- than at the low-deposition site. The low quotient at the low-deposition site, together with the initial growth increase with intermediate N supplements, indicates that growth was N-limited at this site, but our lowest N supplement was sufficient to reduce growth. The N treatments had no effect on interspecific competition between the Sphagnum species. This indicates that the species have similar responses to N. The species studied all occur naturally on ombrotrophic, N-poor sites and show low tolerances to increased N influx. Reduced Sphagnum production may affect the carbon balance, changing the mires from C sinks to sources.

The function of a leaf is photosynthesis, which requires the interception of light and access to atmospheric CO2 while controlling water loss. This paper examines the influence of leaf anatomy on both light capture and CO2 diffusion. As photosynthetic metabolism is spread between many chloroplasts, a leaf faces the challenge of matching light capture by a given chloroplast with the metabolic capacity of that chloroplast. Chloroplasts nearest the leaf surface receive the greatest irradiance and therefore absorb more light per unit chlorophyll than chloroplasts in the centre of a leaf. Electron transport and carbon fixation capacities per unit of chlorophyll decline with increasing depth in the leaf, to compensate for the decline in light absorbed per unit chlorophyll. Many key photosynthetic protein complexes in chloroplasts have nuclear encoded genetic information. Consequently, all chloroplasts within a given cell have a similar metabolic complement, which limits the potential gradient of photosynthetic capacity per unit chlorophyll across the leaf. A simple model couples light absorption through the leaf (based on the Beer–Lambert law) with the profile of chlorophyll through a leaf and the gradient in photosynthetic capacity. It is validated by comparison with 14CO2 fixation profiles through spinach leaves obtained in various studies. The model can account for published 14C fixation profiles obtained with blue, red and green light of different irradiances and white light applied in different combinations to the adaxial and abaxial surfaces of spinach leaves. The model confirms that spongy mesophyll increases the apparent extinction coefficient of chlorophyll compared to palisade tissue. The palisade tissue nearest the surface which receives light facilitates the penetration of light to a greater depth, while spongy mesophyll promotes scattering to enhance light absorption, thus reducing the gradient in light absorbed per unit chlorophyll through a leaf. CO2 fixation faces a diffusional limitation, which necessitates Rubisco to be spread evenly across the cell walls exposed to intercellular airspace. Mesophyll cell structure reflects the need to have a large cell surface per unit volume exposed to airspaces. The regular array of columnar cells in palisade tissue, or cell lobing in monocot leaves, results in greater exposed surface per unit tissue volume than spongy mesophyll. The exposed surface area per unit leaf area scales with photosynthetic capacity such that the difference in CO2 partial pressure between substomatal cavities and the sites of carboxylation within chloroplasts is, on average, independent of photosynthetic capacity of the leaf. However, Rubisco specific activity declines as the Rubisco content per unit leaf area increases due to greater internal diffusional limitations.

We examined the importance of seed size in the early establishment of seedlings of four cotyledonous species restricted to nutrient-impoverished soils in south-western Australia. Experimentally removing cotyledons from seedlings increased mortality and reduced root-penetration, growth rate and final weight of plants after 12 wk, in proportion to seed size. The impact of removal of the cotyledons was least in the smallest-seeded, Eucalyptus loxophleba Benth., grown in the more fertile of two soils, and greatest in the largest-seeded, Hakea psilorrhyncha R. M. Barker, whose growth was otherwise unaffected by soil type. In the smallest-seeded species, cotyledon content of most mineral nutrients increased over time, especially in the more fertile soil, while chlorophyll content showed some decrease by the end of the experiment. In the other species, most nutrients (especially N, P, K and Cu in the less fertile soil) were translocated from the cotyledons to the growing plant, and chlorophyll content declined markedly. The translocated N, P, K, Mg and Cu in these species made up a substantial part of the total plant content by 12 wk after germination, while total Ca and Fe tended to increase in the cotyledons via substantial soil uptake. The results indicate, contrary to reports for some other species, that the cotyledons have an important nutritional function in the early establishment of seedlings in these species. The results also show that, in the larger-seeded species, young seedlings rely to a greater extent on nutrient support from the cotyledons than from the soil. We propose that large seeds might be an adaptation for enhancing establishment in nutrient-poor soils.

The ability of the mycorrhizal fungus Paxillus involutus to mobilize nitrogen and phosphorus from discrete patches of beech (Fagus sylvatica), birch (Betula pendula) and pine (Pinus sylvestris) litter collected from the fermentation horizon of three forest soils, and to transfer the nutrients to colonized B. pendula Roth seedlings, was investigated in transparent observation chambers. The mycelium of P. involutus foraged intensively in all three types of litter, leading to a significant decline in their phosphorus contents after 90 d. Over the same period only one of the litter types, beech, showed more than a 10% loss of its N contents. Exploitation of the litter led to invigoration of the vegetative mycelium of the fungus throughout the chambers as well as to significant increases of biomass production and leaf area in seedlings grown in the plus litter (+L) relative to those in minus litter (−L) systems. The yield increases were associated with gains in whole plant tissue content and concentration of P, but in content only in the case of N. Calculations suggest that a major proportion of the phosphorus lost from litter originated in its organic fraction. The possible basis of the discrepancy between values of N loss from litter and gain by the plant is discussed and the extent to which the distinctive pattern of nutrient mobilization is a feature peculiar to this fungus-plant combination is considered. It is concluded that nutrient mobilization from natural organic substrates in the fermentation horizon of forest soils may be a key function of the vegetative mycelium of mycorrhizal systems. The need for experimental analyses of a greater range of fungus-plant partnerships is stressed.

Summary 421

I. INTRODUCTION 421

II. EFFECTS OF OZONE ON REPRODUCTION 423

1. Pollen germination and pollen tube growth 424

2. Floral initiation and development 428

3. Effects on seed and fruit yield and yield components 433

4. Effects of ozone on seed and fruit quality, germination and seedling growth 437

III. INFLUENCE OF REPRODUCTIVE HABIT AND IMPLICATIONS FOR FIELD-GROWN PLANTS 438

IV. CONCLUSIONS AND FUTURE RESEARCH 441

Acknowledgements 442

References 442

Sexual reproductive development is a crucial stage in the life cycle of higher plants as any impairment of the processes involved might have significant implications for the productivity of crop plants and the survival of native species. There is considerable evidence that exposure to ozone, even at current ambient levels in many industrialized countries, reduces grain and fruit yields and adversely affects yield quality. It is also well established that sensitivity to ozone may differ not only between species, but also between cultivars and populations of individual species, and that the impact of exposure is highly dependent on ozone concentration and the duration and timing of exposure. However, few studies have attempted to distinguish between the direct effects of air pollutants on reproductive development, and indirect effects mediated by injury to the vegetative organs and associated changes in the supply of assimilates and other essential resources to support reproductive growth, or the levels of endogenous growth regulators. This review considers the impact of ozone on the reproductive biology of agricultural and native species, and examines its direct effects on specific reproductive processes. The extent to which compensatory responses redress the adverse effects of exposure is also explored, with particular reference to recent studies of Brassica napus (oilseed rape), Brassica campestris (Wisconsin Fast Plants), Plantago major (greater plantain) and Triticum aestivum (wheat).

Evolution of 14CO2 from onion roots and the intraradical hyphae of Gigaspora margarita Becker & Hall was examined by radiorespirometry after addition of 14C-labelled glucose or sucrose to mycorrhizal or non-mycorrhizal roots. In mycorrhizas, the respiration rate from glucose was about twice that from sucrose. The respiration rate from glucose in the mycorrhizas was much higher than that in the non-mycorrhizal roots, but no differences between mycorrhizal and non-mycorrhizal roots were found in the respiration from sucrose. Intraradical hyphae were isolated from mycorrhizas by enzyme digestion and homogenization followed by Percoll® gradient centrifugation. The 14C-labelled glucose, fructose or sucrose was added to the isolated hyphae and the subsequent evolution of 14CO2 was measured. The hyphae mainly used glucose as a substrate for respiration. Although sucrose or fructose was utilized to some degree, the respiration rate from glucose was much higher than that from sucrose and fructose. This is the first direct evidence of use of glucose by the intraradical hyphae of arbuscular mycorrhizal fungi in the symbiotic state.

The causes and consequences of biodiversity are central themes in ecology. Perhaps one reason for much of the current interest in biodiversity is the belief that the loss of species (by extinction) or their gain (by invasion) will significantly influence ecosystem function. Arbuscular mycorrhizal (AM) fungi are components of most terrestrial ecosystems and, while many research programs have shown that variability among species or isolates of AM fungi does occur (Giovannetti & Gianinazzi-Pearson, 1994), the basis for this variability and its consequences to the function of communities and ecosystems remains largely unexplored. Smith et al. (pp. 357–366 in this issue) now show clearly that ecologically significant functional diversity exists among AM fungal species in the regions of the soil from which they absorb phosphate, and their results suggest that such diversity may have significant ecological consequences.

Observations were made on the vertical distribution of colonies of Aphanizomenon flos-aquae for 9 d at a drift-station east of Bornholm Island in the Baltic Sea. The buoyant colonies were dispersed in the upper layers of the water column during periods of wind-induced mixing but floated up during calm periods. From measurements of the vertical light extinction, surface irradiance and the photosynthesis versus irradiance curve, calculations were made of the changes in the daily integral of photosynthesis with respect to time and depth throughout the water column. From these calculations it is demonstrated that net photosynthesis by the population of Aphanizomenon flos-aquae increased nearly threefold by floating up after a deep mixing event. It is estimated that, averaged over alternating periods of calm and mixing, the buoyancy provided by gas vesicles in this organism will result in a nearly twofold increase in photosynthesis. A quantitative analysis has been made of the relationship of the daily integral of photosynthesis by the Aphanizomenon population with the mean depth of the population in the water column and the daily insolation. The analysis shows that the integral decreases linearly with respect to mean depth.

The present work describes the morphogenesis and cytological characteristics of ‘branched absorbing structures’ (BAS, formely named arbuscule-like structures, ALS), small groups of dichotomous hyphae formed by the extraradical mycelium of arbuscular mycorrhizal (AM) fungi. Monoxenic cultures of the AM fungus Glomus intraradices Smith & Schenck and tomato (Lycopersicum esculentum Mill.) roots allowed the continuous, non-destructive study of BAS development. These structures were not observed in axenic cultures of the fungus under different nutritional conditions or in unsuccessful (asymbiotic) monoxenic cultures. However, extraradical mycelium of G. intraradices formed BAS immediately after fungal penetration of the host root and establishment of the symbiosis. The average BAS development time was 7 d under our culture conditions, after which they degenerated, becoming empty septate structures. Certain BAS were closely associated with spore formation, appearing at the spore's substending hypha. Branches of these spore-associated BAS (spore-BAS) usually formed spores. Electron microscopy studies revealed that BAS and arbuscules show several ultrastructural similarities. The possible role of BAS in nutrient uptake by the mycorrhizal plant is discussed.

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.

Seedlings and old trees of Scots pine in self-regenerating, old, virgin boreal forest in northern Sweden were compared in terms of the species composition of their associated ectomycorrhizal (EM) fungi. The natural regeneration of seedlings was mimicked by annual seeding for 10 yr (1986–1995) in intact field vegetation at three sites. In 1995, all seedlings were harvested, and 171 were examined for mycorrhizas. Twenty-five soil cores were taken in order to study the mycorrhizas on the established trees. Most short roots were mycorrhizal. Using mycorrhizal morphology and restriction fragment length polymorphisms (RFLP) analysis, we were able to distinguish 43 ectomycorrhizal taxa. Fourteen of these taxa were identified using a sporocarp and mycelial culture based internal transcribed spacer (ITS)-RFLP reference database, and another three were identified by mycorrhizal morphology. Cenococcum geophilum, Piloderma croceum and Suillus variegatus were present on the seedlings, irrespective of age, as well as on the old trees. At one forest, 23 ectomycorrhizal taxa were found as mycorrhizas, of which 10 occurred on both seedlings and old trees. These 10 species accounted for 92 and 73% of the mycorrhizas on the seedlings and old trees, respectively. A 3-yr survey of ectomycorrhizal sporocarps at the same site revealed 62 EM taxa. The degree of similarity between the composition of EM species reflected by sporocarps and by mycorrhizas, was low. Ectomycorrhizal species present in our ITS-RFLP database constituted 7.5–19% of the mycorrhizas on seedlings and old trees, whereas they constituted 80–95% of the total production of epigeous sporocarps. The unidentified taxa comprised 70–87% of the mycorrhizas. Our findings support the view that the species composition of mycorrhizas colonizing naturally regenerated seedlings in forests is similar to that of mycorrhizas colonizing surrounding trees. We suggest that the concept of the mycelial network be expanded in order to embrace both the significance of interconnections between different trees as well as the continuity or perpetuation of EM fungal communities.

Vulnerability of stem xylem to cavitation was measured in 10 species of conifers using high pressure air to induce xylem embolism. Mean values of air pressure required to induce a 50% loss in hydraulic conductivity (φ50) varied enormously between species, ranging from a maximum of 14.2±0.6 MPa (corresponding to a xylem water potential of −14.2 MPa) in the semi-arid species Actinostrobus acuminatus to a minimum of 2.3±0.2 MPa in the rainforest species Dacrycarpus dacrydioides. Mean φ50 was significantly correlated with the mean rainfall of the driest quarter within the distribution of each species. The value of φ50 was also compared with leaf drought tolerance data for these species in order to determine whether xylem dysfunction during drought dictated drought response at the leaf level. Previous data describing the maximum depletion of internal CO2 concentration (ci) in the leaves of these species during artificial drought was strongly correlated with φ50 suggesting a primary role of xylem in effecting leaf drought response. The possibility of a trade-off between xylem conductivity and xylem vulnerability was tested in a sub-sample of four species, but no evidence of an inverse relationship between φ50 and either stem-area specific (Ka) or leaf-area specific conductivity (K1) was found.

Axenic cultures of lichen photobionts isolated from bark-inhabiting lichen thalli of the Physcietum adscendentis Ochsner were identified by light microscopy and sequence comparisons of internal transcribed spacer rDNAs to investigate principles of lichenization within a defined lichen sociological unit. The photobiont identity of eight lichen species is reported for the first time (photobiont species in square brackets): Lecania cyrtella (Ach.) Th. Fr. [Trebouxia arboricola Puym.], Lecania naegelii (Hepp) Diederich & v. d. Boom [Dictyochloropsis symbiontica Tscherm.-Woess], Candelaria concolor (Dicks) B. Stein [Trebouxia jamesii (Hildreth & Ahmadjian) Gärtner], Candelariella cf. reflexa (Nyl.) Lettau [T. jamesii], Lecanora spec. [T. arboricola], Phaeophyscia orbicularis (Neck.) Moberg [T. impressa Ahmadjian], Physcia adscendens (Fr.) H. Olivier [T. impressa] and Lecidella elaeochroma (Ach.) M. Choisy [T. arboricola] and could be confirmed for another two species, Physcia stellaris (L.) Nyl. [Trebouxia impressa] and Xanthoria parietina (L.) Th. Fr. [Trebouxia arboricola]. The observation that pioneer lichens without vegetative propagules, growing on smooth bark, had Trebouxia arboricola as photobiont can be explained by the assumption of a free-living population of Trebouxia arboricola. Species of photobionts from Xanthoria parietina were morphologically and genetically different from those of Physcia adscendens and Phaeophyscia orbicularis, respectively; a finding that does not support the previous assumption that Xanthoria parietina takes over its algal partner from a Physcia species, at least at the sites investigated.

Elevated CO2 increases root growth and fine (diam. [les ]2 mm) root growth across a range of species and experimental conditions. However, there is no clear evidence that elevated CO2 changes the proportion of C allocated to root biomass, measured as either the root[ratio ]shoot ratio or the fine root[ratio ]needle ratio. Elevated CO2 tends to increase mycorrhizal infection, colonization and the amount of extramatrical hyphae, supporting their key role in aiding the plant to more intensively exploit soil resources, providing a route for increased C sequestration. Only two studies have determined the effects of elevated CO2 on conifer fine-root life span, and there is no clear trend. Elevated CO2 increases the absolute fine-root turnover rates; however, the standing crop root biomass is also greater, and the effect of elevated CO2 on relative turnover rates (turnover[ratio ]biomass) ranges from an increase to a decrease. At the ecosystem level these changes could lead to increased C storage in roots. Increased fine-root production coupled with increased absolute turnover rates could also lead to increases in soil organic C as greater amounts of fine roots die and decompose. Although CO2 can stimulate fine-root growth, it is not known if this stimulation persists over time. Modeling studies suggest that a doubling of the atmospheric CO2 concentration initially increases biomass, but this stimulation declines with the response to elevated CO2 because increases in assimilation are not matched by increases in nutrient supply.

Models have been formulated for monospecific stands in which canopy photosynthesis is determined by the vertical distribution of leaf area, nitrogen and light. In such stands, resident plants can maximize canopy photosynthesis by distributing their nitrogen parallel to the light gradient, with high contents per unit leaf area at the top of the vegetation and low contents at the bottom. Using principles from game theory, we expanded these models by introducing a second species into the vegetation, with the same vertical distribution of biomass and nitrogen as the resident plants but with the ability to adjust its specific leaf area (SLA, leaf area[ratio ]leaf mass). The rule of the game is that invaders replace the resident plants if they have a higher plant carbon gain than those of the resident plants. We showed that such invaders induce major changes in the vegetation. By increasing their SLA, invading plants could increase their light interception as well as their photosynthetic nitrogen-use efficiency (PNUE, the rate of photosynthesis per unit organic nitrogen). By comparison with stands in which canopy photosynthesis is maximized, those invaded by species of high SLA have the following characteristics: (1) the leaf area index is higher; (2) the vertical distribution of nitrogen is skewed less; (3) as a result of the supra-optimal leaf area index and the more uniform distribution of nitrogen, total canopy photosynthesis is lower. Thus, in dense canopies we face a classical tragedy of the commons: plants that have a strategy to maximize canopy carbon gain cannot compete with those that maximize their own carbon gain. However, because of this strategy, individual as well as total canopy carbon gain are eventually lower. We showed that it is an evolutionarily stable strategy to increase SLA up to the point where the PNUE of each leaf is maximized.

We investigated interactions between plant roots, protozoa and nematodes after addition of patches containing inorganic or organic nitrogen in order to determine whether root proliferation could explain the capture of N by the plant from the patch. Decomposition of a 15N/13C, dual-labelled, organic patch in the absence of plant roots was also examined. In the decomposing patch the amounts of 13C and 15N remaining co-varied and both declined with time. Nematode numbers increased. However, protozoan biomass and inorganic N (NO3− and NH4+) availability did not significantly alter as decomposition of the patch progressed. Addition of inorganic N patches, as NH4NO3 solutions, to the first lateral to emerge from the main seminal root axis of Lolium perenne L. seedlings had no effect on root growth compared with controls 16 d after addition. Protozoan biomass increased. Furthermore, log protozoan biomass and NO3− concentrations of the growth medium were significantly (P<0·05) and positively related. Plant response (i.e. biomass production, N capture and root length) to an added organic patch was examined using five different grass species (Festuca arundinacea L., Phleum pratense L., Poa pratensis L., Dactylis glomerata L. and L. perenne). Total plant biomass was significantly (P<0·05) repressed by an organic patch. Plant N content was reduced when an organic patch was present but N concentrations were greater. Roots were generally slow to proliferate within the patch but there was a significant (P<0·05) species×patch interaction for root length within the patch at harvest and in the 2-cm band below it. However, 15N capture by the plants was not related to mean root length duration. All species captured similar amounts of 15N (c. 3–5%) at harvest as a percentage of the initial 15N added in the organic patch. Similarly, the percentage of the total N captured from the patch was not related to the proportion of the root weight within the patch. The fraction of the captured N from the organic patch as a percentage of the plants' total N, however, did differ among species. Substantial amounts (>62%) of the 15N initially added remained in the patch at harvest. Much less (c. 13–21%) 13C remained in the patch. Protozoan biomass and nematode numbers increased significantly (P<0·05) in the organic patch, although the relationship between the two groups was not significant. As in the inorganic N study, the relationship between log protozoan biomass and NO3− concentrations in the soil was significantly positive. We conclude that, when grown in monoculture, plants' N capture from an organic patch is not a simple function of root proliferation. External factors, not plant attributes, are more important in controlling patch exploitation.

The processes of symbiotic germination and seedling development were analysed in the myco-heterotrophic orchid Corallorhiza trifida, seeds of which were buried in ‘packets’ either adjacent to or at varying distances from adult plants in defined communities of ectomycorrhizal tree species. Germination occurred within eight months of burial under Betula–Alnus and within seven months under Salix repens. It was always associated with penetration of the suspensor by a clamp-forming mycorrhizal fungus. Four distinct developmental stages were defined and the rates of transition through these stages were plotted. There was no evidence of a relationship between extent of germination or rate of development and the presence of naturally distributed plants of C. trifida at the spatial scale of 1 m. The best germination and the most rapid rate of development of C. trifida seedlings occurred in a Salix repens community located at a considerable distance from any extant C. trifida population. Determination of internal transcribed spacer (ITS) RFLPs and of gene sequences of the fungi involved in symbiotic germination and growth of C. trifida, revealed them to belong exclusively to the Thelephora–Tomentella complex of the Thelephoraceae. These fungi are known also to be ectomycorrhizal associates of trees. It is hypothesized that the rate of growth of the C. trifida seedlings is determined by the ability of the fungal symbionts to transfer carbon from their ectomycorrhizal co-associates.

An experiment was performed to find out whether ectomycorrhizal (ECM) fungi alter the nitrogen (N) isotope composition, δ15N, of N during the transport of N from the soil through the fungus into the plant. Non- mycorrhizal seedlings of Pinus sylvestris were compared with seedlings inoculated with either of three ECM fungi, Paxillus involutus, Suillus bovinus and S. variegatus. Plants were raised in sand in pots supplied with a nutrient solution with N given as either NH4+ or NO3−. Fractionation against 15N was observed with both N sources; it decreased with increasing plant N uptake, and was larger when NH4+ was the source. At high ratios of Nuptake/Nsupplied there was no (NO3−), or little (NH4+), fractionation. There seemed to be no difference in fractionation between ECM and non-mycorrhizal plants, but fungal rhizomorphs were sometimes enriched in 15N (up to 5‰ at most) relative to plant material; they were also enriched relative to the N source. However, this enrichment of the fungal material was calculated to cause only a marginal decrease (−0.1‰ in P. involutus) in δ15N of the N passing from the substrate through the fungus to the host, which is explained by the small size of the fungal N pool relative to the total N of the plant, i.e. the high efficiency of transfer. We conclude that the relatively high 15N abundance observed in ECM fungal species should be a function of fungal physiology in the ECM symbiosis, rather than a reflection of the isotopic signature of the N source(s) used. This experiment also shows that the δ15N of plant N is a good approximation of δ15N of the available N source(s), provided that N is limiting growth.

Interactions between saprotrophic and ectomycorrhizal fungi have been largely ignored, although their mycelia often share the same microsites. The mycelial systems show general similarity to each other and, although the enzymatic potential of the saprotrophic fungi is generally considered to be higher, the importance of organic nutrient sources to ectomycorrhizal fungi is now widely accepted. In the experiments described here, nutritional interactions involving transfer of elements from one mycelium to the other have been monitored dynamically using radioactive tracers and a non-destructive electronic autoradiography system. Microcosms were used in which mycelial systems of the ectomycorrhizal fungi Suillus variegatus and Paxillus involutus, extending from Pinus sylvestris host plants, were confronted with mycelia of the saprotroph Hypholoma fasciculare extending from wood blocks. The fungi showed a clear morphological confrontation response. The mycorrhizal mycelium often formed dense patches over the Hypholoma mycelia. Up to 25% of the 32P present in the Hypholoma mycelium was captured by the mycorrhizal fungi and translocated to the plant host within 30 d. The transfer of 32P to the saprotroph from labelled mycorrhizal mycelium was one to two orders of magnitude lower. The significance of this transfer as a ‘short cut’ in nutrient cycling is discussed.