The three youngest age-classes of needles of Norway spruce (Picea abies) were collected from four sites in the Krusne Hory Mountains (Czech Republic) characterized by different levels of damage caused by environmental pollution. Histochemical methods did not reveal any differences in localization of phenolics among the needles. Mesophyll cells close to the epidermis of needles and cells around resin ducts and substomatal cavities often accumulated higher amounts of phenolics than the rest of the mesophyll cells, but this was independent of age and damage. Needles of different age- and damage-class did not show any marked changes in general lignification pattern. However, a lower intensity of histochemical detection of lignin was observed in needles from the most damaged site. This finding was confirmed by chemical analysis using thioglycolic acid. Generally, the amount of lignin in mesophyll cells was lower in damaged trees than in healthy ones. Using the Folin–Ciocalteau method, no significant differences in the total content of phenolics were observed in the needles, although HPLC revealed marked alterations in the forms of seven phenolic acids. Concentrations of conjugated forms of phenolic acids (esters and glycosides) were higher in damaged needles (255.9 μg g−1 f. wt) than in healthy needles (189.8 μg g−1 f. wt). By contrast, content of esterified phenolic acids incorporated into cell walls was higher in needles from healthy trees (101.1 μg g−1 f. wt) than in damaged needles (78.3 μg g−1 f. wt). Marked differences were also observed in the activity of soluble peroxidases, although the activity of ionically bound forms was approximately the same in healthy and damaged needles. The total amounts of chlorophylls and carotenoids decreased as environmental damage increased.

Physcomitrella patens, recently renamed Aphanoregma patens, has been transformed with the plasmid, pBI426. On selective medium approx. 30% of regenerants expressed the transformed phenotype transiently (transients). The remaining 70% (transformants) retained their transformed phenotype (GUS-positive and resistant to G418) indefinitely when subcultured repeatedly on selective medium. However, most lost this phenotype after one or two passages through nonselective medium (unstable transformants). Approximately 0.2% of transformants retained their transformed phenotype after numerous passages through nonselective medium (stable transformants). Using PCR methodology, it has been shown that loss of the transformed phenotype by unstable transformants is invariably accompanied by disappearance of the transgenic DNA. Southern blot analysis data argue strongly that unstable transformants cultured under selective conditions contain unintegrated pBI426 as circular concatenates consisting of 3–40 copies of the plasmid. Under selective conditions, it appears that replication and/or partitioning of these extrachromosomal concatemers might be growth rate-limiting. This is the first report of a transgenic, autonomously replicating extrachromosomal element in a photosynthetic plant. A single copy of pBI426 has been inserted into the moss genome in each of three stable transformants analysed.

Resistance to complete submergence was tested in three Rumex species that occur in the Dutch river forelands. The species differ in both habitat and life history characteristics. The annual or biennial R. maritimus and the biennial or short lived perennial R. palustris grow on frequently flooded mud flats of low elevation, while the perennial R. thyrsiflorus can be found on dykes and river dunes that are seldom flooded. The flooding characteristics of the habitats of the three species were determined. These data were used to design experiments to determine the survival and biomass development of the three species during submergence and the influence of plant size and light level on these parameters. It was shown in all three species that plants submerged during daytime were much more resistant to flooding than those submerged at night. This is most probably due to the generation of oxygen or carbohydrates by underwater photosynthesis. Mature plants of the three species showed higher survival after submergence than juvenile plants, which might be caused by higher carbohydrate levels in the taproots of mature plants. In addition, the three species clearly differed in survival and biomass development during submergence. Rumex thyrsiflorus, the species least subjected to flooding, is least tolerant to complete submergence. Rumex maritimus, which can avoid the floods by having a short life cycle, is less tolerant to submergence than R. palustris, which has to survive the floods as a vegetative plant. It was noted that some plants that survived the flooding period itself, still died in the following period of drained conditions, possibly due to post-anoxic injury.

The early development of indigenous and introduced Scots pine (Pinus sylvestris L.) ectomycorrhiza in natural forest humus was examined using molecular fingerprinting techniques. Non-mycorrhizal or mycorrhizal (Suillus bovinus (L. ex Fr.) O. Kuntze or Paxillus involutus (Batsch ex Fr.) Fr.) seedlings were placed in transparent two-dimensional microcosms on a thin layer of sieved humus to allow observation of ectomycorrhizal development and fungal growth. Twelve ectomycorrhizal morphotypes, mainly based on colour, gross morphology and outer mantle structure, were identified over a 3-month period. No successional trends in mycorrhiza formation were observed. For further characterization, individual ectomycorrhiza representing the different morphotypes were subjected to analyses of esterase (EST) isozyme profiles and RFLPs of the internal transcribed spacer (ITS) from fungal rRNA genes following amplification using PCR. All 10 morphotypes analysed displayed different esterase isozyme profiles, and the characteristic S. bovinus species diagnostic band (S. b. EST8) was detected in two white morphotypes. Ten Pink and Black ectomycorrhiza were all separated into one and three groups, respectively. A fast-running plant-specific polymorphic locus (Z) was also confirmed in most ectomycorrhizal morphotypes. Diagnostic species-specific (S. bovinus and Pink) EST bands were detected in intact external mycelium colonizing soil. Successful amplification of the ITS from individual ectomycorrhiza, of eight different morphotypes, was found to be mainly influenced by the DNA template concentration. Different RFLPs (Hinf I, Mbo I and Hha I) of the ITS placed the white morphotypes into two groups, one corresponding to S. bovinus, in agreement with the esterase fingerprinting. By contrast, Pink and Beige morphotypes displayed different EST profiles but nearly identical RFLP fingerprints. The ITS amplification success rate of standardized DNA template concentrations from 10 individual Pink and Black ectomycorrhiza was 90 and 50%, respectively. In all successful amplifications, the black morphotype yielded the smallest ITS fragment, similar to earlier reports for Cenococcum geophilum, that gave identical RFLPs. The ecological significance of the observed mycorrhizal diversity and combined application of these two identification methods is discussed.

Arbuscular mycorrhizal (AM) symbioses are known to play a role in increased resistance of plants against soilborne pathogens. Mechanisms involved in this phenomenon are not yet well understood. This work investigates possible roles of endoproteolytic activities in bioprotection of Pisum sativum roots by Glomus mosseae against Aphanomyces euteiches. First, it is demonstrated that bioprotection occurs only in pre-mycorrhizal plants. Second, endoproteolytic activities were analysed qualitatively and quantitatively during AM symbiosis, in plants infected with either zoospores or mycelium of A. euteiches, and in mycorrhizal plants infected with the pathogen. In mycorrhizal symbiosis a progressive increase in endoproteolytic activities was observed following root colonization by G. mosseae. By contrast, in roots inoculated with A. euteiches, a drastic increase in endoproteolytic activities was observed which was correlated with the amount of pathogen occurring in roots. Qualitative differences were seen among the endoproteolytic activities detected in roots inoculated with zoospores or mycelium. The constitutive as well as mycorrhizal and pathogen-induced activities were further characterized as ‘trypsin-like’ serine endoproteases. Interestingly, in a situation of bioprotection, only low levels of the activities normally associated with the infection by A. euteiches were detected, suggesting that the synthesis of these proteins is directly linked to the growth or virulence of the pathogen.

The ability of four species of vesicular–arbuscular mycorrhizal (VAM) fungi to increase phosphorus uptake and growth of clover plants (Trifolium subterraneum L.) at different levels of soil compaction and P application was studied in a pot experiment. Dry matter in the shoots and roots of clover plants decreased with increasing soil compaction. Colonization by Glomus intraradices Schenck & Smith and Glomus sp. City Beach WUM16 increased plant growth and P uptake up to a bulk density of 1·60 Mg m−3, although the response was smaller as soil compaction was increased. Glomus etunicatum Becker & Gerdeman and Glomus mosseae (Nicol. & Gerd.) Gerdemann & Trappe had no effect on the shoot d. wt and P uptake when the bulk density of the soil was [ges ]1·40 and [ges ]1·60 Mg m−3, respectively. Soil compaction to a bulk density of 1·60 Mg m−3 had no effect on the percentage of root length colonized by G. intraradices and Glomus sp. City Beach, but total root length colonized decreased as soil compaction was increased. Decreased P uptake and growth of clover plants colonized by G. intraradices and Glomus sp. City Beach, with increasing soil compaction up to a bulk density of 1·60 Mg m−3, was mainly attributed to a significant reduction in total root length colonized and in the hyphal biomass. Soil compaction, which increased bulk density from 1·20 to 1·75 Mg m−3, reduced the O2 content of the soil atmosphere from 0·16 to 0·05 m3 m−3. The absence of any observable mycorrhizal growth response to any of the four species of VAM fungi in highly compacted soil (bulk density = 1·75 Mg m−3) was attributed to the significant decrease in the O2 content of the soil atmosphere, change in soil pore size distribution and, presumably, to ethylene production.

Azolla filiculoides Lam. causes serious weed problems in Britain, but its long-term survival might be limited by winter death. The aim of this study was to establish the low temperature responses and limitations of A. filiculoides sporophytes.

In the laboratory, normal vegetative growth was shown to continue at 5°C. Reddening of plants was a response to low temperature and high light conditions which could be prevented by shading. Adult plants died after short (18 h) exposure to −4°C but survived sub-zero temperatures >−4°C. Evidence was found of seasonal changes in chill tolerance, but not in freeze tolerance.

In outdoor culture, plants survived encasement in ice and air temperatures to −5°C. Additional evidence suggested that natural populations can readily survive air temperatures much lower than this. Microclimatic effects are likely to be responsible for this discrepancy between laboratory and outdoor culture results.

Three phenotyes were identified; survival, colonizing and mat forms.

It is easy to dismiss bryophytes as ‘lower’ plants, mere primitive precursors long since left behind in the evolutionary race, and of only rather esoteric and incidental biological interest. But this is to let oneself be led astray by a simplistic image of a tidy evolutionary tree – an image that served Darwin well a century and a half ago (Desmond & Moore, 1992), but which we should now see as an intricately branched evolutionary bush with innumerable shoots reaching out from all depths to the growing apices that represent the present day. The earliest land plants may indeed have been at a bryophyte level of organization, but modern bryophytes, no less than vascular plants, are the product of some 450 million years’ evolution since that time (Edwards et al., 1998). Raven (1977, 1984) has emphasized the importance of the evolution of supracellular transport systems in the origin of vascular land plants. Bryophytes, on the other hand, evolved desiccation tolerance and represent an alternative strategy of adaptation to life on land, photosynthesizing and growing when water is available, and suspending metabolism when it is not. They are limited by their mode of life, but also liberated : they are prominent on hard substrates such as rock and bark, which are impenetrable to roots and untenable to vascular plants. Bryophytes (in species numbers the second biggest group of green land plants) may be seen as the mobile phones, notebook computers and diverse other rechargeable battery-powered devices of the plant world – not direct competitors for their mains-based equivalents, but a lively and sophisticated complement to them.

The number and type of isoforms of superoxide dismutase (SOD) and their activities were compared in mitochondria and peroxisomes isolated from cotyledons of three different oilseed seedlings. Mitochondrial and peroxisomal isoforms of SOD could be distinguished in nondenaturing polyacrylamide gels by their differential sensitivities to KCN and/or H2O2. The type of SOD was not the same for each organelle in each of the three oilseed species. For example, a single Mn–SOD was found in cotton and cucumber mitochondria, whereas four CuZn–SODs were present in mitochondria from sunflower. At least one CuZn–SOD isoform was found in the peroxisomes of all three species. Cucumber peroxisomes contained both a CuZn–SOD and a Mn–SOD, cotton peroxisomes contained a single CuZn–SOD, whilst four separate CuZn–SODs, but no Mn–SOD were found in sunflower peroxisomes. Using antibodies against CuZn–SOD from watermelon peroxisomes or from chloroplasts of Equisetum, a single polypeptide of c. 16·5 kDa was detected on immunoblots of peroxisomal fractions from the three species. Post-embedment, electron-microscopic double immunogold-labelling showed that CuZn–SOD, with malate synthase used as marker enzyme of peroxisomes, was localized in the matrix of these organelles of all three species. These results suggest that CuZn–SOD is a characteristic matrix enzyme of peroxisomes in oilseed cotyledons.

Plant roots enter symbiotic as well as pathogenic interactions with fungi in the rhizosphere. We studied the response of bean (Phaseolus vulgaris L. cv. Saxa) roots to infection by the arbuscular mycorrhizal fungus Glomus mosseae (Nicol. & Gerd.) Gerdemann & Trappe and the pathogen Fusarium solani (Mart.) Sacc. f. sp. phaseoli (Burkholder) Snyder & Hansen. In a time-course study of the symbiotic interaction between bean roots and G. mosseae, covering all stages of mycorrhiza development, we detected little change in the expression of the defence-related genes chitinase, β-1,3-glucanase and phenylalanine ammonia-lyase compared with non-mycorrhizal control roots. The only difference observed was a transient increase in chalcone synthase transcripts at later stages of mycorrhizal root colonization. In interactions with the pathogen, a marked induction of chitinase and phenylalanine ammonia-lyase expression was observed at the level of both the transcripts and enzyme activities. Class I β-1,3-glucanase levels strongly increased at the transcript level, but there was little change in the overall β-1,3-glucanase enzyme activity. In the non-host interaction between common bean and Fusarium solani (Mart.) Sacc. f. sp. pisi (Linford) Snyder & Hansen defence responses increased only slightly and transiently, if at all.

The outcome of dual infection of the grass Lolium perenne L. by arbuscular mycorrhizal (AM) fungi and the parasitic angiosperm Rhinanthus minor L. was investigated in a glasshouse study. Colonization of L. perenne roots by AM fungi was significantly reduced by the presence of R. minor, as was host growth which fell by 44–51%. It was concluded that these two responses were linked, with AM colonization declining in response to the reduction in availability of host carbon. Parasite growth and reproductive output rose by 58% and 47% respectively when the hosts were mycorrhizal. These trends were unrelated to the attachment success of the parasite, but were accompanied by a significant increase in the formation of secondary haustoria. The benefits afforded the parasite when the hosts were mycorrhizal were attributed to increased carbon and nutrient flux resulting from alternations in sink strength. Host responses to parasitism and mycorrhizal colonization were not affected by the interaction between the two symbionts. However, the suggestion is made that the interaction between the AM fungi and parasite might have long-term ecological implications for the host species via its impact on parasite fecundity.

Nitrogen (N) allocated to leaf growth in forage grasses and legumes following severe defoliation is predominately mobilized from the remaining root and leaf sheath tissues, since both N uptake from the soil and N2 fixation are severely down-regulated for several days. The hypothesis that a low N reserve status at the time of defoliation limits N remobilization and leaf regrowth was tested with contrasting cultivars of Lolium perenne (cvs Aberelan and Cariad) in flowing solution culture. Plants were grown under ‘high’ or ‘low’ (uptake of N decreased by 50%) regimes of N supply for 10 d before a single severe defoliation. Labelling with 15N was used to assess the importance of N reserves, including putative vegetative storage proteins, relative to N translocated from concurrent uptake, as a source of leaf N during regrowth. Leaf regrowth, N uptake and N mobilization were all affected by previous N supply. Low plant N status at the time of defoliation increased regrowth dry weight of ‘Aberelan’ by 10% and translocation of N absorbed from the medium by 23%, while mobilization of N reserves was decreased by 56%. On the contrary, regrowth dry weight of ‘Cariad’ was decreased by 23%, and translocation of N absorbed by 21% in low plant N status, compared with high plant N status. Concentrations of soluble protein in roots and remaining leaf sheaths decreased after defoliation in plants only under optimal N supply. Analysis of soluble proteins in sheath material by SDS–PAGE suggested that three polypeptides (55, 36.6 and 24 kDa) might function as vegetative storage proteins, although they were of low abundance in plants, subjected to monthly harvests, grown in controlled conditions and in the field. The apparent antagonism between uptake of NH4+ or NO3− by roots and mobilization of N reserves is discussed together with evidence for functional vegetative storage proteins in L. perenne.

Concentrations of NH4+, NO3−, PO43−, K+, Ca2+ and Mg2+ in snow meltwater resulting from summer snow showers were monitored before and after its passage through monospecific stands of the Antarctic macrolichen Usnea sphacelata R. Br. The sampling was conducted under field conditions near Casey Station in East Antarctica between January and March. Total snow deposition during the 61-d period was 44±1 mm (rainfall equivalent depth) delivering 362±10, 87±2 and 9±1 μmol m−2 of NH4+, NO3− and PO43−, respectively. Meltwater that had percolated through U. sphacelata was depleted in NH4+ and NO3− equating with a retention by the lichen of 87 and 92%, respectively, of the total wet deposition of these ions. Lichen-modified meltwater was slightly enriched in PO43−, but because the volume of the lichen percolate was smaller than that of the original snow deposition, the lichen achieved a net gain of 9% of the total P deposited. Lichen percolate was also enriched in metal cations. Potassium loss associated with the melting of the heaviest snowfall (18 mm) was equivalent to only 0·05% of the total K in the lichen suggesting that ion loss did not signal significant cellular damage. There was also a progressive increase in NH4+ concentration in unmodified meltwater from 3 to 21 nmol ml−1 over a 3-d period whereas levels in the lichen-modified meltwater remained unchanged at [les ]4 nmol ml−1. This enrichment in NH4+ might have resulted from dry deposition onto the melting snow pack of NH3 emitted from nearby penguin rookeries. During the study, tagged thalli of U. sphacelata made a 2% loss in dry mass although they appeared healthy and, at the end of the study, showed an effective quantum yield (ΔF/Fm′) comparable with field material. The results are discussed in relation to the time of year that is likely to be most suitable for lichen growth in this continental Antarctic environment and the potential growth-led demand for N in U. sphacelata.

Endophytes of the genus Epichloë (Clavicipitaceae, Ascomycota) are systemic symbionts of cool-season grasses. Their interactions with grass hosts may vary between mutualistic and pathogenic depending on the mode of endophyte reproduction. Sexual strains prevent flowering and seed set (choke disease) of the host and can be horizontally transmitted by ascospores, while asexual strains remain asymptomatic and are vertically transmitted through seeds. In Switzerland nearly all plants of Brachypodium sylvaticum (Huds.) P.B. are infected by Epichloë sylvatica Leuchtmann & Schardl, but choke symptoms are formed very rarely, and are restricted to particular locations and to a minority of plants at those locations. Earlier research has revealed that E. sylvatica is genetically differentiated into sexual and asexual subpopulations. Given the high level of infection and assuming horizontal transmission of sexual strains, multiple host infections have been predicted. In this study, 25 plants out of 63 examined by isozyme analysis were found to be infected by two or three different endophyte genotypes. In most cases endophyte genotypes appeared to be correlated with the symptom type of a particular tiller, suggesting that the fungal genome controls choke formation and that the sexual and asexual subpopulations are separated at the ramet (tiller) level rather than at the genet (plant) level. These conclusions were further supported by analyses with log-linear models of the population structure of E. sylvatica at four locations where choke symptoms were present. These analyses also revealed a geographic structure in the asexual subpopulation but not in the sexual subpopulation which could be caused by the different dispersal ranges of their propagules. The rare occurrence of sexually reproducing strains and the dominance of a single genotype in asymptomatic plant populations may be explained by the colonization history of B. sylvaticum and its endophyte in Switzerland.

Responses of leaf gas exchange and above-ground growth of beech (Fagus sylvatica L.) and Norway spruce (Picea abies Karst.) to atmospheric CO2 enrichment (374 μl l−1 vs. 590 μl l−1) and increased wet deposition of N (5 vs. 50 kg N ha−1 a−1) in combination with two natural forest soil types (‘acidic’ and ‘calcareous’) were studied in large open-top chambers. Eight juvenile beech and spruce trees from different provenances, together with a ground cover composed of five understorey species, were established in each of 32 model ecosystems. Both beech and spruce showed sustained enhancement of photosynthesis in response to atmospheric CO2 enrichment during the first 2 yr of treatment. Nevertheless, switching measurement CO2 concentrations revealed partial downward adjustment of photosynthesis in trees grown in elevated CO2, beech generally showing more pronounced downward adjustment than spruce. The responsiveness of photosynthesis to CO2 enrichment did not vary significantly among trees from different provenances. Stomatal conductance was reduced under elevated CO2 in both tree species. In spruce, the radial growth of the main stem and the annual production of wood (shoot-wood dry mass of current-year lateral shoots), needle dry mass, and assimilation area per tree were stimulated both by CO2 enrichment and increased N deposition, but were not significantly affected by soil type by year 2. In contrast, in beech, the radial growth of the stem and the total leaf number, foliage dry mass, and assimilation area per tree were all not significantly affected by elevated CO2 and increased N deposition when responses of the two soil types were pooled, but were greater on calcareous than on acidic soil by year 2. However, CO2 interacted with soil type in beech: irrespective of the N deposition rate, saplings showed growth stimulation on the calcareous soil but responded negatively to CO2 enrichment on the acidic soil (where growth was slower). Our results suggest that complex interactions between CO2, species and soil quality need to be accounted for when attempting to predict forest development in a future CO2-rich world.

Solution culture experiments were made using two subspecies of Cerastium fontanum and Cochlearia pyrenaica ssp. alpina (Bab.) Dalby and Festuca rubra L. of ultramafic and non-ultramafic origin to test: (1) whether Mg and Ni toxicities are likely to exclude non-ultramafic races of the three species; (2) whether tolerance to Mg and Ni toxicities by the ultramafic plants differed markedly among the species; and (3) to use (2) to generate a hypothesis about controlling factors of species distribution on the Meikle Kilrannoch (MK1) ultramafics. The solution cultures were based on soil solution extracts with experimentally varied concentrations of Mg, Ni and Na. The solution with high (7·5 mM) Mg and high (0·7 mg l−1) Ni concentrations simulated the ultramafic soil solution. Care was taken to use a Fe source which did not react with Ni and microelements. Of the non-ultramafic plants, the growth of Cerastium was reduced by high concentrations of Mg but not by Ni, whereas both the Cochlearia and the Festuca suffered severe reduction in biomass production in the presence of high Mg or Ni or both. Of the ultramafic plants, Festuca was tolerant of high Mg and Ni concentrations; Cerastium was less tolerant of high Mg; and Cochlearia had low shoot production in high Ni (although the effect was ameliorated by high Mg). The different interspecific responses of the ultramafic plants to Mg and Ni supported field observations that the three species might occupy microhabitats which differ in Mg and Ni.

The axial and radial refilling with water of cut dry branches (up to 80 cm tall) of the resurrection plant Myrothamnus flabellifolia was studied in both acro- and basipetal directions by using 1H-NMR imaging. NMR measurements showed that the conducting elements were not filled simultaneously. Axial water ascent occurred initially only in a cluster of a very few conducting elements. Refilling of the other conducting elements and of the living cells was mainly achieved by radial extraction of water from these initial conducting elements. With time, xylem elements in a few further regions were apparently refilled axially. Radial water spread through the tissue occurred almost linearly with time, but much faster in the acropetal than in the basipetal direction. Application of hydrostatic pressure (up to 16 kPa) produced similar temporal and spatial radial refilling patterns, except that more conducting elements were refilled axially during the first phase of water rise. The addition of raffinose to the water considerably reduced axial and radial spreading rates. The polarity of water climbing was supported by measurements of the water rise in dry branches using the ‘light refraction’ (and, sometimes, the ‘leaf recurving’) method. Basipetal refilling of the xylem conduit exhibited biphasic kinetics; the final rise height did not exceed 20–30 cm. Three-cm-long branch pieces also showed a directionality of water climbing, ruling out the possibility that changes in the conducting area from the base to the apex of the branches were responsible for this effect. The polarity of water ascent was independent of gravity and also did not change when the ambient temperature was raised to c. 40 °C. At external pressures of 50–100 kPa the polarity disappeared, with basipetal and acropetal refill times of the xylem conduit of tall branches becoming comparable. Refilling of branches dried horizontally (with a clinostat) or inverted (in the direction of gravity) showed a pronounced reduction of the acropetal water rise to or below basipetal water climbing level (which was unaffected by this treatment). Unlike water, benzene and acetone climbing showed no polarity. In the case of benzene, the rise kinetics (including the final heights) were comparable with those measured acropetally for water, whereas with acetone the rise height was less. Transmission electron microscopy of dry branches demonstrated that the inner surfaces of the conducting tracheids and vessels were lined with a continuous osmiophilic (lipid) layer, as postulated by the kinetic analysis and light microscopy studies. The thickness of the layer varied between 20 and 80 nm. The parenchymal and intervessel pits as well as numerous tracheid corners contained opaque inclusions, presumably also consisting of lipids. Electron microscopy of rehydrated plants showed that the lipid layer was either thinned or had disintegrated and that numerous vesicle-like structures and lipid bodies were formed (together with various intermediate structural elements). These, many other data and the physical–chemical literature imply that several (radial) driving forces (such as capillary condensation, Marangoni forces, capillary, osmotic and turgor pressure forces) operate when a few conducting elements become axially refilled with water. These forces apparently lead to an avalanche-like radial refilling of most of the conducting elements and living cells, and thus to the removal of the ‘internal cuticle’ and of the hydrophobic inclusions in the pits. The polarity of water movement presumably results from high resistances in the basipetal direction, which are created by local gradients in the thickness of the lipid film as a result of draining under gravity in response to drought. There are striking similarities in morphology and function between the xylem-lining lipid film and the lung surfactant film lining the pulmonary air spaces of mammals.

The effects of full-spectrum solar radiation and of solar radiation deprived of total u.v. radiation (λ<395 nm) on the endemic Mediterranean red alga Rissoella verruculosa (Bertoloni) J. Agardh were studied in situ in early summer, from sunrise to sunset. Photosynthetic performance, pigment content and the activities of enzymes related to nutrient uptake, were monitored under both radiation conditions throughout a daylight period.

The doses of solar radiation, measured on the day during which the experiments were carried out (24 June 1997), were 9228·25, 1109·70 and 13·03 kJ m−2 for PAR (λ=400–700 nm), u.v.-A (λ=315–400 nm) and u.v.-B (λ=280–315 nm), respectively. Under these conditions, a clear daily variation in photosynthetic performance was found. However, no significant differences were detected between the two radiation conditions. At noon, strong sunlight impaired O2 evolution by 75%, but complete recovery occurred during the afternoon, reaching similar values to those measured in the early morning. By contrast, photoinhibition of optimal quantum yield (Fv/Fm), and the relative electron transport rate was followed by only a slight recovery during the afternoon. The rate of photosynthesis in air (simulating emersion conditions), estimated by CO2 exchange, showed a negative balance at noon, which was accentuated in plants exposed to the full-solar spectrum.

Significant changes in the activities of nitrate reductase and carbonic anhydrase were found throughout the day under both radiation conditions. In thalli receiving solar radiation deprived of u.v., total CA activity decreased throughout the day. However, in thalli exposed to full-spectrum solar radiation, the activity of the enzyme tended to increase in the afternoon, correlating with an increase in NR activity.

Phytosiderophore (PS) release occurs under both iron and zinc deficiencies in representative Poaceae and has been speculated to be a general adaptive response to enhance the acquisition of micronutrient metals. We tested this hypothesis within an on-going study of the role of micronutrient metal nutrition for patterns of spontaneous vegetation in relation to soil pH and carbonate content. Hordelymus europaeus (L.) Harz, a negative grass species commonly found on soils rich in CaCO3 in Western Central Europe, was subjected to deficiencies of Fe, Zn, Mn and Cu using chelator-buffered nutrient solutions. PS release rates were determined at 3–5 d intervals during onset and development of deficiency symptoms. Plant dry matter yields and nutrient concentrations, measured at three time points were used to construct growth curves for calculation of PS release per unit root mass. In comparison with trace metal-sufficient control plants, dry matter production was markedly reduced in the Fe, Zn, Mn and Cu-deficiency treatments, with final relative yields of 6, 11, 15 and 31%, respectively. The phytosiderophore produced under Fe- and Cu-deficiency treatments was identified, using HPLC, as desoxymugineic acid. The highest rate of PS release (18 μmol g−1 root d. wt in 2 h) was measured in the Fe-deficiency treatment, and there was substantial release in the Cu-deficiency treatment (7·25 μmol g−1 root d. wt in 2 h). No PS release above control levels (2·6 μmol g−1 root d. wt in 2 h) was observed in the Zn- or Mn-deficiency treatments (1·5 and 2·6 μmol g−1 root d. wt in 2 h, respectively). It remains to be clarified whether PS release in response to Cu deficiency is a primary reaction to the deficiency, or is caused by a chain of events similar to that observed in Zn-deficient wheat, which involves internal Fe deficiency. Our results suggest that PS release in the native plant species H. europaeus is a specific response to Fe and Cu deficiency and is not significantly induced in response to deficiencies of Zn and Mn. Induction of the PS mechanism in different plant species might be more diverse than previously thought.

There has been conflicting debate over many years regarding the trophic status of Matsutake. Here we examined the morphology, structure and ultrastructure of Pinus densiflora roots collected from a Tricholoma matsutake Shiro within a pure Japanese red pine stand. Molecular investigations (PCR–RFLP analyses) indicated that T. matsutake was the highly dominant fungus within both the Shiro and the colonized root tips, suggesting that reported root morphology modifications can be attributed to T. matsutake infection. The external morphology of Matsutake-colonized roots is consistent with previous descriptions. The presence of extraradical mycelium, mantle, and intracortical Hartig net hyphae indicates clearly that T. matsutake forms an ectomycorrhizal association with P. densiflora in naturally occurring Shiros. The elucidation, for the first time, of the Hartig net ultrastructure at the host–fungus interface provides further and convincing evidence of a conventional ectomycorrhizal association. The progressive blackening, observed from base to tip in dominant mycorrhizal types, due to increased deposition of polyphenol and subsequent necrosis, appears to be a result of infection. However, the presence of highly nucleated vascular tissue indicates the viability of the vascular cylinder in these roots bearing necrotic cortices. Such a preponderance of black necrotic cortical tissues among colonized roots may reflect some atypical behaviour of T. matsutake.