Applying the branch-in-bag method, naturally seeded Scots pine (Pinus sylvestris L.) trees, 25–30 yr old, were subjected to two CO2 concentrations (350 and 700 μmol mol−1) and two soil-nitrogen-supply regimes for three growing seasons (1994–96). Gas exchange and chlorophyll a fluorescence in detached shoots were measured simultaneously in a diffuse radiation field. Elevated CO2 did not lead to a significant ‘downward regulation’ in the light-saturated rate of net photosynthesis (Pn.max), the maximum apparent quantum yield (αA.max) or the maximum photochemical efficiency (Fv/Fm) of photosystem II (PS II). However, the elevated CO2 significantly decreased the light-saturated stomatal conductance and increased the sensitivity of stomatal conductance to change in low photon-flux densities. The high soil–nitrogen supply significantly increased photosynthetic capacity, as manifested by increases in Pn.max, αA.max, Fv/Fm, and the effective photochemical efficiency (ΔF/F′m) at low photon-flux densities, did not, on the other hand, enhance the magnitude of photosynthetic response to elevated CO2 concentration. In addition, the treatment-induced modifications in fluorescence parameters are discussed in detail.

After cooling the roots and shoot apex of barley, there is a reduction in sucrose export from, and an increased total accumulation of carbohydrate and induction of fructan synthesis in, source leaves. We investigated carbohydrate accumulation in individual leaf cells. Using the microsampling technique and microfluorometric enzymatic assay, the amounts of various carbohydrates were measured in mesophyll, bundle sheath and epidermal cells. Epidermal cells take no part in carbohydrate partitioning and have very low concentrations of sugars even when leaf export is reduced. Even without cooling, however, there are remarkable changes in sucrose concentration (up to 200 mM) in mesophyll and bundle sheath cells during the photoperiod. The activity of acid invertase in samples from individual cells was very low, c. 3 μmol sucrose (g f. wt)−1 h−1. Interrelationships between these cell types and the control of fructan metabolism are discussed.

Mineral nutrient concentrations were determined in leaves of 83 mostly herbaceous species collected from central England. Most samples were analysed for N, P, K, Ca, Mg, Na, Fe, Al, Mn, Cu and Zn. Concentrations of K, N and P showed similar levels of interspecific variability, with the highest concentrations being 6–9 times the lowest. Mg and (especially) Ca were much more variable, with the highest concentrations being 24 and 49 times the lowest respectively. Only in the case of P concentration was the majority of the variance in the data found at or below the species level. Most of the variance in Ca and Mg concentrations was between monocots and dicots. Concentrations of N and P were strongly positively correlated with each other. Only Ca and Mn were consistently associated with soil pH, positively and negatively respectively. Dicots tended to accumulate more Ca and Mn from high soil concentrations than did monocots. Concentration of P was significantly positively correlated with maximum potential relative growth rate. Plants of woodland and arable habitats contained high concentrations of P, and those of pasture and skeletal habitats contained low concentrations of P. The P[ratio ]N ratio was higher in plants of arable habitats. Species with P-rich leaves tended to be currently increasing in abundance. The results suggest that plants with nutrient-rich foliage grow quickly, dominate nutrient-rich ecosystems and are generally increasing as a result of the eutrophication and disturbance arising from human exploitation.

Spontaneous hybrids between the two diploids Poa infirma Kunth and P. supina Schrader were obtained at the rate of 0·2%. No reciprocal maternal effect was observed. Almost all hybrids survived, grew normally but were sterile. Hybrids were diploid, and resembled in morphology the tetraploid P. annua L. (annual meadow grass). In their isozymes, they showed additivity of the parental patterns, and were very similar to those found in P. annua populations. The ease with which hybrids were obtained and the similarity of their morphological and isozyme pattern to P. annua suggest that the two diploid species are closely related to the allotetraploid species, and are perhaps its parent species.

The effects of u.v.-B radiation (280–315 nm) on the fungi occurring on the lammas leaves of pedunculate oak (Quercus robur L.) were examined using saplings that were exposed at an outdoor facility to supplemental levels of u.v.-B radiation under treatment arrays of cellulose diacetate-filtered fluorescent lamps, which also produce u.v.-A radiation (315–400 nm). Saplings were also exposed to u.v.-A radiation alone under control arrays of polyester-filtered lamps, and to ambient levels of solar radiation under arrays of unenergized lamps. The u.v.-B treatment corresponded to a 30% elevation above the ambient level of erythemally-weighted u.v.-B radiation. The fungi were examined weekly over a 4-month-period in summer and autumn 1995 using two techniques, the spore fall and leaf impression methods, which differentiated between those fungi occurring on the upper (adaxial) and lower (abaxial) surfaces of the leaves. The abundances of Aureobasidium pullulans (De Bary) Arnaud and Sporobolomyces roseus Kluy. et van Niel, two leaf yeasts which had adaxial[ratio ]abaxial ratios of <1 under ambient levels of u.v.-B radiation, were negatively correlated with increasing ambient levels of u.v.-B radiation and were significantly reduced on adaxial leaf surfaces by supplemental levels of u.v.-B. There were few effects of supplemental u.v.-B radiation on the abundances of these yeasts on abaxial leaf surfaces. The abundances of the dematiaceous hyphomycetes, Cladosporium spp. and Epicoccum nigrum Link., species with adaxial[ratio ]abaxial ratios of [ges ]1 under ambient levels of u.v.-B radiation, were not correlated with ambient levels of u.v.-B radiation, nor were they usually affected on either leaf surface by supplemental u.v.-B radiation. Alternaria spp. and Microdochium nivale (Fr.) Samuels & Hallet showed consistent responses on adaxial leaf surfaces to u.v.-A radiation applied under control and treatment arrays. Our results suggest that current levels of shortwave radiation already influence the distribution of fungi on leaf surfaces and that future increases in u.v.-B radiation will directly affect the abundances of specific phylloplane fungi.

The accumulation of fructo-oligosaccharides and the activities of fructosyltransferase (sucrose[ratio ]sucrose 1F-fructosyltransferase (SST), 1F-fructosyltransferase (1F-FT) and 6G-fructosyltransferase (6G-FT)) in the bulbs of three onion cultivars were investigated from June to September 1993. The total fructo-oligosaccharide content increased from June until August, then decreased in September, except in one cultivar. The levels of neokestose and its related tetrasaccharides (1F, 6G-di-β-D-fructofuranosyl sucrose and 6G (1-β-D-fructofuranosyl)2sucrose) were higher than those of 1-kestose and nystose throughout growth. The activities of 6G-FT, 1F -FT and SST were high in June and July, then decreased; SST activity was very low in September. The activity ratios of 6G-FT to 1F-FT varied between 1·86 and 2·65 over the growth period. Two trisaccharides, three tetrasaccharides and four pentasaccharides were identified, together with a mixture of hexa- and heptasaccharides, all of which were synthesized in vitro from 0·1 M sucrose by an enzyme preparation of onion bulbs harvested in August. Octa- and nonasaccharides other than the saccharides formed from sucrose were also synthesized from 0·1 M 1-kestose or 0·1 M neokestose. All the saccharides produced from sucrose, 1-kestose or neokestose by the crude enzyme prepared from onion bulbs were identical to the saccharides occurring naturally in onion bulbs.

Measurements of the chitin content of the rooting horizons of a typical mor-humus heathland soil, indicate that chitin can contain in excess of 20% of the total nitrogen in the litter (L) horizon and 30% in the fermentation (F) horizon. Much of this chitin-nitrogen is thought to be contained in the mycelial walls of soil fungi.

Experiments were therefore designed to test the hypothesis that such sources of N could be rendered accessible to the ericaceous plants by their fungal endophytes. Mycelium of the ericoid endophyte Hymenoscyphus ericae (Read) Korf & Kernan and of the ectomycorrhizal fungus Suillus bovinus (Fr.) O. Krantze were grown in liquid culture before being killed and added either in the intact condition, or after fractionation, as sole sources of N to sterile media upon which were grown H. ericae in pure culture, or mycorrhizal and non-mycorrhizal plants of Vaccinium macrocarpon Ait. and Calluna vulgaris L. The abilities of the test organisms to utilize the nitrogen contained in the intact mycelial necromass, or in its fractions, were assessed by determining their yields and nitrogen concentrations of their tissues.

It was revealed that H. ericae was able to produce significantly higher yield when grown on intact fungal necromass than when provided with equivalent concentrations of N in the form of ammonium. Its yields on mycelial fractions were lower, but still significantly greater than those obtained in the controls lacking N. Significantly greater yields and N contents were also found in the ericaceous plants grown with these nitrogenous substrates in the mycorrhizal condition. Without H. ericae they had no access to the substrates. The possible ecological implications of these results are discussed.

The relative ozone resistance of 20 European and two American populations of Plantago major was examined, and relationships with climatic factors at the source of the plant material were explored using data provided by participants in the ICP-Crops initiative (International Co-operative Programme to Investigate the Effects of Air Pollutants and Other Stresses on Agricultural and Semi-Natural Vegetation). Plants grown from seed were exposed to either charcoal/Purafil® filtered air (CF<5 nmol mol−1 O3) or CF+ozone (70 nmol mol−1 O37 h d−1) over a 2-wk period in controlled environment chambers, and effects on mean plant relative growth rate (R) and allometric root/shoot growth (K) determined. Ozone resistance (R%) was calculated from (R03/RCF)×100.

Populations exhibited contrasting sensitivities to ozone, without the development of typical visible symptoms of injury. A positive relationship was found between relative ozone resistance and descriptors of the ozone-climate at the site of seed collection for the year of, and the 2 yr before, seed collection. The best predictors of inherent ozone resistance were shown to be cumulative ozone exposure indices calculated according to current United Nations Economic Commission for Europe (UN-ECE) critical level guidelines for the pollutant (i.e. the accumulated hourly average ozone exposure over a threshold level of 40 nmol mol−1 (AOT40) or 30 nmol mol−1 (AOT30) calculated during daylight hours for the consecutive 3-month period of the year experiencing the highest ozone concentrations). No relationships were found between ozone resistance and climatic factors (temperature, precipitation, sunshine hours, humidity) or the concentrations of other air pollutants (SO2, NO2, NO).

These findings support the view that current ambient levels of ozone in many regions of Europe are high enough to promote evolution of resistance to the pollutant in native plant populations. The significance of these findings to the debate over the establishment of separate critical levels for the protection of natural and semi-natural vegetation is discussed.

Possible limitations of sink strength for fructan storage in tubers of Jerusalem artichoke (Helianthus tuberosus L. cv. Waldspindel) were studied in 2-yr field experiments. Treatments included the removing of side branches and the breaking of apical dominance in unbranched plants. Data from both treatments and the comparison of results obtained for 2 yr, which differed in temperature and total irradiance, indicated that fructan storage in tubers during the storage phase was not limited by supply of assimilate (sucrose). Moreover, fructan concentrations were very constant despite large variations in tuber yields and yield components (e.g. number of tubers per plant, tuber length). However, stimulation of cell expansion and diameter of tubers increased fructan concentrations, probably owing to a larger vacuolar volume in the storage cells. We concluded that fructan storage in tubers of Jerusalem artichoke is not source-limited as far as carbon import into tubers is concerned. However, a strong shoot control, most probably regulated via phytohormones, limits sink strength. Fructan concentrations in tubers might not only be increased by sink activity (fructan-synthesizing enzymes) but also by sink capacity (vacuolar volume). In this respect the shoot might play a decisive role not only as a temporary storage pool for carbohydrates but also as a source of native growth-regulators which control sink capacity.

Cell walls were isolated and purified from Ri T-DNA-transformed carrot roots to determine whether arbuscular mycorrhizal (AM) fungi need a signal from the host root in order to form appressoria. Large cell-wall pieces were generated by the Parr nitrogen bomb technique. Purified cell walls were sterilized and inoculated with single spores of Gigaspora gigantea (Nicol. & Gerd.) Gerdemann & Trappe and Gigaspora margarita Becker & Hall using an in vitro culture technique. Appressoria only formed on epidermal cell walls isolated from host roots (Daucus carota L.) but did not form on epidermal cell walls isolated from non-host roots (Beta vulgaris L.). Although appressoria formed on host-cell walls, penetration hyphae did not fully develop. The results of this study indicate that appressorium formation by these AM fungi is a contact recognition event that does not require a signal secreted from the host root or the presence of intact host cells.

Hydrogen sulphide emission in lichens as a response to low concentration SO2 fumigation was investigated. In an open flow-through system several lichen species were fumigated with 36 ppb SO2. Two species were also fumigated with higher concentrations (72, 119, 122 and 198 ppb SO2). Hydrogen sulphide emission was monitored concurrently by cryogenic trapping and analysis with gas chromatography. All tested species increased H2S emission significantly in response to fumigation with 36 ppb SO2. Parmelina tiliacea (L.) Hale and Cladina rangiferina (L.) Wigg. released significantly more H2S (0·098±0·015 and 0·073±0·013 pmol H2S g−1 d. wt s−1, respectively) than Parmelina quercina (Ach.) Hale, Ramalina menziesii Tayl. and Parmelia sulcata Tayl. (0·028±0·01, 0·025±0·014 and 0·023±0·013 pmol H2S g−1 d. wt s−1, respectively). Release of H2S in Hypogymnia physodes was enhanced by increasing SO2 concentrations up to 72 ppb SO2. No significant difference in H2S emission in the dark vs. in the light was found. Generally, no correlation was found between photosynthetic activity and H2S emission for the tested species. Uptake of SO2 was similar for all species, at 24·7±5·6 pmol SO2 g−1 d. wt s−1 in 36 ppb SO2 and increasing at greater SO2 concentrations. Therefore, H2S-S release represents only 0·11–0·74% of SO2-S uptake.

The interaction between the race 1 of the cowpea rust fungus (Uromyces vignae Barclay) and the resistant cowpea (Vigna unguiculata (L.) Walp.) cv. Queen Anne is characterized by the deposition of callose around intracellular fungal structures. Ultrastructural examination of the early stages of infection by basidiospores of the fungus revealed two types of deposits induced by intracellular hyphae: a non-callose collar at the penetration site and a callose encasement around the invasion hypha. The callose encasement was developed from the site where the fungus encountered the inside of the plant cell wall and was separated from the fungus by the extrahyphal membrane and an extension of plant plasma membrane. The incidence of encasements was reduced in plants treated with inhibitors of transcription (actinomycin D), protein synthesis (cycloheximide), protein glycosylation (tunicamycin) and microfilament polymerization (cytochalasin E). Inhibitors of Golgi-associated vesicle transfer (monensin, brefeldin A) and anti-microtubule agents (colchicine, oryzalin) had no effect. When the fungus was killed by heat treatment in either the resistant or the susceptible cultivar, callose was deposited at various locations along the fungus, mostly in the extrahyphal matrix. The data suggest that unlike the extrahaustorial membrane surrounding D-haustoria of this fungus, the extrahyphal membrane is capable of generating callose. Since this process does not normally occur in the susceptible or the resistant cv. even when callose is deposited in the latter by regions of the plasma membrane not associated with the fungus, we conclude that the deposition of callose by the extrahyphal membrane is inhibited by the living fungus.

Two species of Bromus adapted to different climatic conditions were studied to determine their responses to chilling and water stress. Enzymes of sucrose and fructan metabolism as well as fructoside content were measured in roots and shoots. The results indicate that Bromus pictus (adapted to a cold desert area) has a constitutive fructan synthesis, whilst B. auleticus (adapted to a warmer northern climate) only accumulated fructan under cold stress. Microscopic observations of roots exposed to fluorescent probes ascertained cell viability and showed that the apex of roots in the northern species were more affected by the water-deficit treatment than the southern species. The presence of fructans in B. pictus and their absence in B. auleticus support the hypothesis that fructans play a role in plant tolerance to environmental stress.

The 15N natural abundance and N concentrations of fruit bodies from 70 species (23 genera) of ectomycorrhizal fungi found in boreal forests are presented. Large intraspecific and intrageneric differences were found, e.g. 8·3‰ 15N in the species Dermocybe crocea and 12·6‰ in the genus Cortinarius. In addition, significant differences in both δ15N and %N were found between different parts of fruit bodies, with cap material giving consistently higher values. Proteins and amino acids were enriched by 9·7±0·4‰ (mean±1 SE) relative to chitin, irrespective of the part of the fruit body examined. Chitin had δ15N values similar to that of plant hosts. The higher δ15N and %N values of the caps than of the stipes probably reflect a higher portion of proteins and amino acids in the caps. The δ15N of mycorrhizal fungi can be a function of the N species used (organic N, NH4+, NO3−), the depth of soil at which the mycelium occurs, and metabolic fractionations. The metabolic fractionations, e.g. potential transaminations during the flux of N from the soil through the fungus to the plant, make it difficult, at present, to make inferences about sources of N based on δ15N values alone. No effect of sample drying temperature on δ15N values of fungal material was detected.

We compared the plasma membrane, tonoplast and mitochondrial Mg-ATPase activities of microsomes isolated from mycorrhizal and non-mycorrhizal sunflower (Helianthus annuus L.) and onion (Allium cepa L.) roots during a time-course experiment (21–60 d). No significant difference was observed between the protein levels of root microsomes from mycorrhizal and non-mycorrhizal sunflower. However, the protein content of microsomes obtained from mycorrhizal onion roots increased when the symbiosis was well established (more than 50% of root length colonized by the arbuscular mycorrhizal fungus). Kinetic studies revealed that vanadate-sensitive (plasma membrane-associated) ATPase activity of sunflower root microsomes increased with mycorrhizal colonization. However, the vanadate-sensitive ATPase activity of microsomes from mycorrhizal onion roots decreased compared with those from non-mycorrhizal controls. Increased activity of nitrate (tonoplast-associated) and azide (mitochondrial membrane-related)-sensitive ATPase activities was detected in extracts obtained from 45- and 60-d-old mycorrhizal onion plants. A slight increase in azide-sensitive ATPase activity was detected in microsomes of mycorrhizal sunflower. The existence of different physiological and biochemical strategies in sunflower and onion in relation to the establishment of this mutualistic symbiosis is discussed.

The hypaphorine concentration in Pisolithus tinctorius Coker & Couch hyphae colonizing Eucalyptus roots was 3 to 5 times higher than in adjacent parts of the fungal colony. This phenomenon, observed 24 h after inoculation, was also recorded in several-month-old, well-established ectomycorrhizas. Accumulation was controlled by specific root-derived diffusible molecules: it can be induced through a membrane, but not by non-host plants. In pure culture, high hypaphorine concentration was found only in the youngest mycelium, i.e. the outer 2 mm of the colony. Fungal hypaphorine had no IAA-like activity on Eucalyptus root development and therefore could not be considered as an auxin analogue; instead, a strong reduction of root hair elongation was recorded.

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.

Barley (Hordeum vulgare L. cv. Agneta) was cultivated hydroponically under continuous light, constant temperature and relative humidity. The relative growth rate (RGR) was kept at 25% by an exponential supply of complete medium for 2 wk. The cultures were then transferred to phosphorus-free media and the amounts of total P, fructans, starch, sucrose, glucose and fructose in sink and source leaves were measured at 0, 12, 24, 48, 72, 120 and 156 h. The capacity of net fructan synthesis (i.e. total activity of sucrose[ratio ]sucrose 1-fructosyltransferase (1-SST) and sucrose[ratio ]fructan 6-fructosyltransferase (6-SFT)), the activity of fructan exohydrolase (FEH), as well as the activity of acid invertase were also measured in the two types of leaves. Omission of P from the growth medium caused a decrease in the P content in both sink and source leaves, whereas the RGR was not affected during the 156 h of the experiment. The fructan concentration increased in low-P plants whilst there was little or no effect on the concentrations of starch, sucrose, glucose and fructose. There was a slight increase in the capacity of net fructan synthesis in both sink and source leaves at 156 h after the onset of P-starvation. There was no change in the activity of FEH or acid invertase. The results indicate that in barley leaves the mechanism for carbon partitioning into fructans is more sensitive towards low P conditions than the mechanism for carbon partitioning into starch.

Interspecific and intraspecific variation in the internal transcribed spacer (ITS) region of rDNA of ectomycorrhizal fungi of 44 species in 17 genera were examined using the restriction fragment length polymorphism (RFLP) method. For each species, two to five herbarium vouchers (mainly basidiocarps), collected throughout Fennoscandia, were examined. The ITS region was amplified using the polymerase chain reaction (PCR) and the universal primers ITS1 and ITS4, and subjected to RFLP analysis with three endonucleases. Intraspecific polymorphisms in the ITS region were found in seven species (in nine of the 132 herbarium vouchers). Polymorphisms were due to length mutations, ranging from 5 to 15 bp in four of the seven polymorphic species and mutations in endonuclease restriction sites in six species, mostly affecting only one endonuclease, but in two species two endonucleases. Using a single endonuclease, a unique RFLP pattern could be obtained for more than half the investigated species. By combining different endonucleases, 34 (77%) of the species could be distinguished from another. The remaining RFLP types occurred in one genus. On the basis of the low intra- but high interspecific variation in the ITS region, it is concluded that most ectomycorrhizas formed by the 44 investigated species should be recognized by comparison with this dataset, if the mycorrhizas are sampled from a site located in Fennoscandia. However, in datasets from even larger geographical areas encompassing a higher degree of intraspecific variation in the ITS region, or when mycorrhizas from several sites distant from each other are compared, it might be necessary to include local reference species.

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.