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.

Patterns of molecular genetic variation were examined in seed collections of Plantago major and Plantago intermedia, used previously to investigate the variations in ozone (O3) resistance of these species across Europe. Total genomic DNA was amplified with random primers (random amplied polymorphic DNA (RAPD) and inter- single sequence repeats (SSR)) to produce 73 genetic markers. In addition, allozyme and chloroplast variations were surveyed. Genetic markers were examined for association with O3 resistance in 18 British populations of P. major as well as 27 continental European populations of P. major and P. intermedia. Two populations that exhibited increased resistance to O3 following several years' exposure to high O3 concentrations in the field showed decreased genetic variation over time. In addition, their genetic composition showed no drastic change, which suggests that the change in resistance to O3 was probably the result of selection on genotypes already present in local populations (selection in situ). It appears that selection for O3 resistance may occur in independent populations, and also may involve a number of genetically determined traits. Consequently the finding that plants with similar degrees of O3 resistance are not closely related was not unexpected. However, the finding of an association of several genetic markers with O3 resistance merits further investigation.

Variation in stomatal development and physiology of mature leaves from Alnus glutinosa plants grown under reference (current ambient, 360 μmol mol−1 CO2) and double ambient (720 μmol mol−1 CO2) carbon dioxide (CO2) mole fractions is assessed in terms of relative plant growth, stomatal characters (i.e. stomatal index and density) and leaf photosynthetic characters. This is the first study to consider the effects of elevated CO2 concentration on the distribution of stomata and epidermal cells across the whole leaf and to try to ascertain the cause of intraleaf variation. In general, a doubling of the atmospheric CO2 concentration enhanced plant growth and significantly increased stomatal index. However, there was no significant change in relative stomatal density. Under elevated CO2 concentration there was a significant decrease in stomatal conductance and an increase in assimilation rate. However, no significant differences were found for the maximum rate of carboxylation (Vcmax) and the light saturated rate of electron transport (Jmax) between the control and elevated CO2 treatment.

The inter- and intracellular distribution of phosphate in non-mycorrhizal and ectomycorrhizal roots of Pinus sylvestris was analysed by energy-dispersive X-ray spectroscopy after cryofixation, freeze-drying, pressure infiltration with resin and dry cutting of the material. The results show (1) that in non-mycorrhizal roots the distribution of phosphate across the root cross section is relatively homogenous, (2) that the intracellular distribution of phosphate depends on the external supply, and (3) that the phosphate absorption can be increased by a supply of NH4. Under phosphate starvation, the phosphate is mainly localized in the cell cytoplasm, whereas under high external phosphate this element accumulates in the vacuoles of cortical cells. It can be inferred that at low external conditions phosphate is mainly located in the cytoplasm of cells to protect metabolism from deficiency, that phosphate is translocated to the shoot via symplastic and apoplastic pathways, and that especially the vacuolar pool size is regulated by the external phosphate supply. The nutrition of the host plant under phosphate starvation was improved by a mycorrhizal infection with Suillus bovinus. However, this effect might not be the same for all ectomycorrhizal infections and supply conditions. With high external phosphate an ectomycorrhizal infection with S. bovinus had no effect on intracellular phosphate within the roots, the shoot contents of mycorrhizal plants were less than those of non-mycorrhizal seedlings and translocation rate across the mycorrhizal interface appeared to be independent of the availability of phosphate. Intracellular phosphate in fungal cells of mycorrhizal roots was independent of the external supply and seemed to be regulated by the formation of polyphosphates. A supply of (NH4)2HPO4, which led to an increase in cytoplasmic phosphate levels in the hyphae of the Hartig net of the Paxillus involutus mycorrhiza also led to higher phosphate in the plant cell compartments. Therefore, it can be inferred that the translocation of phosphate from fungus to host plant across the mycorrhizal interface is regulated by the phosphate concentration in the cytoplasm of the Hartig net and by the efflux rate into the interfacial apoplast.

To study the relationships between groups of organisms and the degree to which these relationships are consistent across major climatic gradients, we analysed the testate amoeba (Protozoa) communities, vegetation and water chemistry of one peatland in five countries: Switzerland, The Netherlands, Great Britain, Sweden and Finland, as part of the BERI (Bog Ecosystem Research Initiative) project. The relationships between the different data sets and subsets were investigated by means of detrended correspondence analysis, canonical correspondence analysis and Mantel permutation tests. The comparison of data on vegetation and testate amoebae showed that inter-site differences are more pronounced for the vegetation than for the testate amoebae species assemblage. Testate amoebae are a useful tool in multi-site studies and in environmental monitoring of peatlands because: (1) the number of species in Sphagnum-dominated peatlands is much higher than for mosses or vascular plants; (2) most peatland species are cosmopolitan in their distributions and therefore less affected than plants by biogeographical distribution patterns, thus differences in testate amoeba assemblages can be interpreted primarily in terms of ecology; (3) they are closely related to the ecological characteristics of the exact spot where they live, therefore they can be used to analyse small-scale gradients that play a major role in the functioning of peatland ecosystems. This study revealed the existence of small-scale vertical gradients within the vegetation and life-form niche separation in response to water chemistry. The deep-rooted plants such as Carex spp. and Eriophorum spp. are related to the chemistry of water sampled at or near the ground water table, whereas the mosses are not. Testate amoebae were shown to be ecologically more closely related to the chemistry of water sampled at or near the water table level and to the mosses than to the deep-rooted plants.

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.

The effect of temperature on germination rate was assessed for seeds of 31 wild plant species and four cultivated species growing in the UK. The temperature at which seed first germinated ranged from 2 °C (Brassica rapa) to 11 °C (several species). As the temperature progressively increased, the percentage of seed that germinated rapidly increased to near 100%, whereas the duration for germination progressively decreased up to the thermal optimum. Above the thermal optimum, the duration for germination initially remained constant, and then rapidly increased until the thermal maximum was exceeded when no seed germinated. To assess the effect of temperature on germination rates, the reciprocal of the duration (1/duration) for 50% germination was calculated and regressed against temperature. For most species a linear regression accounted for >90% of the variation in germination rates for temperatures up to the thermal optimum. From the regressions, the base temperatures (Tb, below which no germination is expected) and the thermal constant (S, expressed in degree days) for 50% germination were calculated. The extremes were the alpine species Dryas octopetala, which had a high Tb (c. 12.0 °C) and low S (26 degree days), and Saxifraga tridactylites which had a low estimated Tb (c. −1.8 °C) and a high S (136 degree days). The estimated value of S increased with increasing percentage germination, and although S varied considerably between species with similar values of Tb, a weak inverse relationship between Tb and S was indicated. A thermal time approach was shown to be useful when analysing and comparing thermal requirements for germination of the seed of wild plant species. It showed, for example, that percentage germination often decreased as temperatures approached Tb, and that for some seed germination ceased at temperatures well above Tb.

The inter- and intracellular distribution of the elements calcium, potassium and sodium in non-mycorrhizal and mycorrhizal roots of Pinus sylvestris dependent on different external nutrient supply conditions was detected by energy-dispersive X-ray microanalysis after cryofixation, freeze-drying and pressure infiltration of the material. In non-mycorrhizal and mycorrhizal roots, calcium was mainly detectable in the apoplastic regions. The levels in vacuoles and cytoplasm were near the limits of detection by X-ray microanalysis. Incubation with high concentrations of potassium and sodium, or mycorrhizal infection with Suillus bovinus and Pisolithus tinctorius reduced the amounts of calcium detectable in the roots, especially in the apoplast of cortical cells. The studies revealed that: potassium is mainly localized in cytoplasm and cell walls; the cytoplasmic content is regulated over a wide range of external potassium concentrations; potassium levels in the inner parts of roots are higher than in the outer parts. Mycorrhizal infection with Suillus bovinus had no effect on the inter- and intracellular distribution of potassium in roots but, if the external supply was low, the potassium content in shoots was reduced. In non-mycorrhizal pine roots and those infected with Paxillus involutus an increase in the sodium content of all cell compartments was observed after treatment with high external concentrations of NaH2PO4. However, an increase in sodium content in mycorrhizas of S. bovinus was not detected. The X-ray microanalytical results are discussed in relation to the apoplastic movement of nutrients in non-mycorrhizal and mycorrhizal fine roots of pine and to the demand for these nutrients in different intracellular compartments.

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 apoplasmic permeability of ectomycorrhizal roots of intact Eucalyptus pilularis seedlings infected with Pisolithus tinctorius on aseptic agar plates was examined using the nonbinding fluorochrome 8-hydroxypyrene-1,3,6-trisulphonate and lanthanum ions in conjunction with anhydrous freeze substitution and dry sectioning. Most mycorrhizas formed in the air above the agar surface, and in these the sheath rapidly became nonwettable and impermeable to the fluorochrome but was nevertheless permeable to lanthanum ions. In a few mycorrhizas which developed in contact with the agar the sheath remained permeable to both tracers when fully developed. This increased hydrophobicity of the sheath in mycorrhizas in the air above the agar surface might be explained by deposition of hydrophobins, but nevertheless it still allows an apoplasmic pathway for radial movement of ions. Regardless of their sheath permeation both apoplasmic tracers were always found throughout the Hartig net and were arrested at the Casparian bands and suberin lamellae of the exodermis. It is concluded that the fluorochrome must have moved longitudinally along the Hartig net which is a region of higher permeability than the sheath. Casparian bands in the exodermis of ectomycorrhizal roots have similar properties to those in nonmycorrhizal roots in excluding solutes and their exclusion of lanthanum ions indicates that they are not permeable to ions. The data do not support the concept of a totally sealed apoplasmic exchange compartment, but the differential permeability suggests that the sheath might allow radial transfer of ions but block loss of sugars and organic molecules of similar size.

Cuticular hydrocarbon composition was determined for 18 populations of Avicennia germinans (Avicenniaceae) collected from West Africa, Florida, the Pacific coast of Mexico and French Guyana. Variation in covariance structure among the hydrocarbons was evaluated from correlations among the traits for each of the populations and from a hierarchical common principal components analysis. Principal components ordinations of populations based on the 171 z* transformed correlation coefficients, onto which a minimal spanning tree was superimposed, suggested a network in which French Guyanan populations formed a nexus from which African populations differentiated biochemically from Floridan and Mexican groups. Common principal components analysis provided further support for the differentiation in covariance structure among these biochemical traits of African populations. Pacific coast Mexican populations appeared to have diverged less in covariation among traits from Atlantic American sources than did the northern (Floridan) and southern (French Guyanan) Atlantic American populations from one another. The complete isolation of Pacific coast sources after emergence of the Central American isthmus during the Pliocene provides a reference for comparison of levels of differentiation among regions that suggests an underlying pattern of divergence in this species. Partial Mantel matrix tests revealed significant geographic effects, with no significant climatic effects. Our results provide support for recognition of a major differentiation between Old and New World sources within the taxon and indicate the need for a full revision of this species.

A mechanistic model was developed, to simulate the main facets of photoinhibition in phytoplankton. Photoinhibition is modelled as a time dependent decrease in the initial slope of a photosynthesis versus irradiance curve, related to D1 (photosystem II reaction centre protein) damage and non-photochemical quenching. The photoinhibition model was incorporated into an existing ammonium-nitrate nutrition interaction model capable of simulating photoacclimation and aspects of nitrogen uptake and utilization. Hence the current model can simulate the effects of irradiance on photosynthesis from sub-saturating to inhibitory photon flux densities, during growth on different nitrogen sources and under nutrient stress. Model output conforms well to experimental data, allowing the extent of photoinhibition to be predicted under a range of nutrient and light regimes. The ability of the model to recreate the afternoon depression of photosynthesis and the enhancement of photosynthesis during fluctuating light suggests that these two processes are related to photoinhibition. The model may be used to predict changes in biomass and/or carbon fixation under a wide range of oceanographic situations, and it may also help to explain the progression to dominance of certain algal species, and bloom formation under defined irradiance and nutrient conditions.

The mycobionts of Piceirhiza bicolorata, a distinct ectomycorrhizal morphotype of conifers and hardwoods, have been identified by internal transcribed spacer 1 (ITS1) nuclear ribosomal DNA (rDNA) sequence comparison of the fungi involved. Samples of Piceirhiza bicolorata were obtained from seedlings of Picea abies, Pinus sylvestris, Betula pubescens, Populus tremula, Quercus robur and Salix phylicifolia. In an initial screening, the fungus amplified with universal ITS primers from ectomycorrhizal root samples of P. bicolorata shared approx. 95% ITS1 sequence identity with the ericoid mycorrhizal fungus Hymenoscyphus ericae. A total of 77 out of 88 (= 87.5%) DNA samples (i.e. 52/56 root samples and 25/32 axenic culture isolates) of P. bicolorata were successfully amplified with a taxon-selective primer designed for exclusive amplification of H. ericae-like strains. Forty-seven amplicons were sequenced, yielding 15 different ITS1 genotypes that differed by 1–14 nucleotide character state changes. An inferred ITS1 phylogeny (maximum parsimony) showed that a single major evolutionary lineage of P. bicolorata embraced the historically important H. ericae isolates in a 100% bootstrap-supported clade. The 15 P. bicolorata genotypes were positioned in four subclades, roughly corresponding to morphological groups of P. bicolorata isolates observed in axenic culture. Culture isolates of H. ericae and P. bicolorata share some common morphological features including slow, dense growth and formation of short aerial hyphal aggregates. Our results suggest that members of the H. ericae aggregate participate in the formation of the distinct ectomycorrhizal morphotype P. bicolorata. This opposes the widely accepted discrimination of ericoid and ectomycorrhizal mycobionts of the boreal forest ecosystem. The high prevalence of the P. bicolorata morphotype on pioneer seedlings of P. sylvestris, B. pubescens and S. phylicifolia at a copper mine spoil was remarkable. Hypotheses of possible nutrient mobilization and detoxification potentials of the fungal associates of P. bicolorata are discussed. We hypothesize that ericoid and ectomycorrhizal plants may share mycobionts of the H. ericae aggregate.

A new group of darkly pigmented root-infecting fungi was isolated from cereal roots obtained from six different locations in northeastern Germany. Similar random amplified polymorphic DNA(RAPD) patterns and restriction profiles of amplified rDNA were used as a basis for classifying the isolates in a separate group. The isolates demonstrating mycelial and infection characteristics typical of Gaeumannomyces graminis could be differentiated from the varieties of G. graminis as well as from Gaeumannomyces cylindrosporus/Phialophora graminicola using RAPD Polymerase chain reaction (PCR) and rDNA Restriction-fragment length polymorphism (RFLP) analysis. Phylogenetic analysis of the Internal transcribed spacer (ITS) regions suggests that the isolates form a distinct group (named group ‘E’) situated within the Gaeumannomyces–Phialophora complex between the branch of the G. graminis varieties and Gaeumannomyces incrustans/Magnaporthe poae. Isolates of group E produced lobed hyphopodia and were shown in biotests to be non-pathogenic to wheat, oats, Italian Ryegrass and Chewings Fescue, suggesting it is a benign parasite which colonizes cereals or grasses without destroying vascular tissue. Furthermore, curved phialospores could be found. Summarizing the results presented, this new group could be classified as a new species of Phialophora. Although isolates of group E were found at only six of the 32 investigated locations, they composed up to 50% of total isolates of the Gaeumannomyces–Phialophora complex at these sites. Because of the non-pathogenic behaviour, the new group may be of value as biological control agents for pathogenic fungi.

On the basis of the evidence that insect fungivory has the potential to affect fungal reproductive fitness, we investigated the effects of two specialist ciid beetles (Octotemnus glabriculus and Cis boleti) on the reproductive potential of their host fungus, Coriolus versicolor. We found, from field data, a negative correlation between the number of individuals of O. glabriculus inhabiting C. versicolor fruit bodies and the percentage of the fungal spore-producing surface (hymenium) that was functional. By contrast, the number of C. boleti inhabiting C. versicolor fruit bodies did not correlate with the percentage of functional hymenium. Experimentally, O. glabriculus and C. boleti reduced the reproductive potential of C. versicolor by 58% and 30%, respectively, whereas the combined trophic activity of both beetles caused a reduction of 64%. This latter effect was not significantly different from that caused by O. glabriculus alone. These findings disagree with previous assertions that insect fungivory on fruit bodies has only neutral effects on fungal fitness. We conclude that in the short-term, fungivory by ciids significantly decreases the area of functional hymenium of C. versicolor and is likely to reduce fungal reproductive fitness. Within this perspective the evolution of certain fungal characteristics (i.e. chemical composition, consistency and phenology) can be interpreted as being driven by fungivory.

The effects of mycorrhiza formation in combination with elevated CO2 concentrations on carbon metabolism of Norway spruce (Picea abies) seedlings and aspen (Populus tremula × Populus tremuloides) plantlets were analysed. Plants were inoculated for 6 wk with the ectomycorrhizal fungi Amanita muscaria and Paxillus involutus (aspen only) in an axenic Petri-dish culture at 350 and 700 μl l−1 CO2 partial pressure. After mycorrhiza formation, a stimulation of net assimilation rate was accompanied by decreased activities of sucrose synthase, an increased activation state of sucrose-phosphate synthase, decreased fructose-2,6-bisphosphate and starch, and slightly elevated glucose-6-phosphate contents in source leaves of both host species, independent of CO2 concentration. Exposure to elevated CO2 generally resulted in higher net assimilation rates, increased starch as well as decreased fructose-2,6-bisphosphate (aspen only) content in source leaves of both mycorrhizal and nonmycorrhizal plants. Our data indicate only slightly improved carbon utilization by mycorrhizal plants at elevated CO2. They demonstrate however, that both factors which modulate the sink-source properties of plants increase the capacity for sucrose synthesis in source leaves mainly by allosteric enzyme regulation.

New Phytologist 144 (1999), 195–202

In the October 1999 issue of New Phytologist, we published the research paper entitled ‘Identification and expression analysis of two fungal cDNAs regulated by ectomycorrhiza and fruit body formation’ by Uwe Nehls et al. (New Phytol. (1999) 144, 195–202). Since its publication, the authors have identified an important error in Fig. 4: the labels SC13 and SC25 are in incorrect positions and should be transposed.

We apologise to our readers for this mistake.

Most work on root proliferation to a localized nutrient supply has ignored the possible role of mycorrhizal fungi, despite their key role in nutrient acquisition. Interactions between roots of Plantago lanceolata, an added arbuscular mycorrhiza (AM) inoculum and nitrogen capture from an organic patch (Lolium perenne shoot material) dual-labelled with 15N and 13C were investigated, to determine whether root proliferation and nitrogen (N) capture was affected by the presence of AM fungi. Decomposition of the organic patch in the presence and absence of roots peaked in all treatments at day 3, as shown by the amounts of 13CO2 detected in the soil atmosphere. Plant N concentrations were higher in the treatments with added inoculum 10 d after patch addition, but thereafter did not differ among treatments. Plant phosphorus concentrations at the end of the experiment were depressed by the addition of the organic residue in the absence of mycorrhizal inoculum. Although uninoculated plants were also colonized by mycorrhizal fungi, colonization was enhanced at all times by the added inoculum. Addition of the AM inoculum increased root production, observed in situ by the use of minirhizotron tubes, most pronouncedly within the organic patch zone. Patch N capture by the end of the experiment was c. 7.5% and was not significantly different as a result of adding an AM inoculum. Furthermore, no 13C enrichments were detected in the plant material in any of the treatments showing that intact organic compounds were not taken up. Thus, although the added AM fungal inoculum benefited P. lanceolata seedlings in terms of P concentrations of tissues it did not increase total N capture or affect the form in which N was captured by P. lanceolata roots.

The effect of soil flooding on arbuscular-mycorrhizal (AM) fungal colonization of wetland plants was investigated using Panicum hemitomon and Leersia hexandra, two semi-aquatic grasses (Graminaceae) that grow along a wide hydrologic gradient in Carolina bay wetlands of the southeastern US coastal plain. Three related investigations were conducted along the dry-to-wet gradient in these wetlands; a field survey of AM fungal root colonization in eight wetlands, monthly monitoring of colonization patterns in P. hemitomon over a growing season, and an inoculum potential bioassay of soils collected along the gradient. The field survey showed that AM fungal colonization was strongly negatively correlated with water depth, but colonization was present in most root samples. The monthly assessment indicated that AM fungal colonization was lowest in plots that were consistently wet but rose as some plots underwent seasonal drying. The inoculum potential assay of dry, intermediate, and wet soils performed under both dry and saturated conditions showed that soils that were wet for >1 yr had the same ability to form mycorrhizas in bait plants as those that had remained dry. These findings suggested that the lower degree of colonization in wet areas observed in the field survey was because of the presence of surface water rather than low numbers of mycorrhizal propagules in the soil. Overall, the results of these investigations show that flooding is partially but not totally inhibitory to AM fungal colonization of wetland grasses.