This paper analyses relationships between relative growth rate (rgr), seed mass, biomass allocation, photosynthetic rate and other plant traits as well as habitat factors (rainfall and altitude) in 20 wild species of Aegilops L. and one closely related species of Amblyopyrum (Jaub. & Spach) Eig., which differ in ploidy level (diploid, tetraploid and hexaploid). The plants were grown hydroponically for 20 d in a growth chamber. The relationships between parameters were calculated either using the phylogenetic information (phylogenetically independent contrasts, PIC) or without using the phylogenetic information (trait values of taxa, TIP). The results using the two approaches were very similar, but there were a few exceptions in which the results were different (e.g. rgr vs. seed mass). Specific leaf area (sla) was positively correlated with leaf area ratio (lar) and negatively correlated with net assimilation rate (nar), which together resulted in the absence of a correlation between sla and rgr. Leaf photosynthetic rates (expressed on a mass or area basis) showed no correlation with rgr. rgr was positively correlated with the stem mass ratio and negatively with root mass ratio. Species with a lower d. wt percentage have a higher rgr. Aegilops species from locations with higher annual rainfall invested less biomass in roots and more in shoots (leaves and stems) and had a higher rgr. Diploid species had a lower seed mass and initial mass than the hybrids (tetraploid and hexaploid species), but there was no correlation of rgr with ploidy level. Polyploid species, which have higher seed mass, occur at a higher altitude than diploid species. Our results show that variation in rgr in Aegilops and Amblyopyrum spp. is associated mainly with variation in biomass allocation (proportion of biomass in stems and roots) and d. wt percentage, and not with variation in sla, leaf photosynthetic rates or seed mass.

Spring ephemerals have evolved specific growth strategies that take advantage of the high photon fluence rates of early spring. These strategies involve a sequential growth of different organs. Erythronium americanum (Ker-Gawl), a common spring ephemeral of northern maple forests, produces its roots in the autumn, although stems and leaves develop in the spring. Mycorrhizal infection of the root system occurs very rapidly and intensively. The fungi thus depend on carbohydrate reserves accumulated in the corm (underground reserve organ) for their growth and development through the winter. We found that the presence of mycorrhizas drastically decreases root growth during the cold period and is more costly of carbohydrate reserves than root growth alone. However, during the spring, the presence of mycorrhizas fully benefits the plant: we measured an annual growth rate of mycorrhizal plants twice that of fungicide-treated plants which were non-mycorrhizal. We suspect that E. americanum is highly susceptible to water stress during the growing season and might rely on its mycorrhizas for water supply to a greater extent than for nutrient supply. Nutrient uptake might occur mainly in the autumn when arbuscules are at their most abundant.

When freshly cut segments of naturally decorticated steles of horizontal roots of the Australian grass tree (Xanthorrhea preissii) are subjected to gentle suction, bubbles of gas sometimes appear, as well as liquid, in capillaries attached to the aspirated ends of the steles. We tested the hypothesis that this gas comes from vessels embolized in the intact xylem stream, and that the gas volume extruded is therefore an effective measure of the extent of this embolism. To do this, twin samples were taken from individual roots of X. preissii in the field, one was fast-frozen intact for subsequent estimation of vessel embolisms in the cryo-scanning electron microscope (CSEM), the other 15-cm segment rapidly assessed for volume of gas aspirated into a standard micropipette tube. The two measures mutually confirmed one another by showing a strong positive correlation between numbers of embolized vessels and extracted gas volume. Similar gas volumes were obtained from replicate root segments excised directly from a root when the ends of the segment were frozen before excision, and aspiration conducted after subsequent thawing of the ends under water. The pattern of changes in embolisms during unstressed conditions in early summer, shown by both CSEM and aspiration, indicated almost no embolisms before dawn, followed by a rapid rise to a peak in mid morning, than a progressive loss of embolisms in late afternoon. It was also shown that the amount of embolism did not change with time after excision of the roots up to at least 30 min. A comparison of changes in leaf transpiration with gas volumes in steles during a 24-h cycle at peak transpiration stress in mid summer showed rapid rates of transpiration in early morning and late afternoon, with an intervening period of low water loss during the rest of the day. Numbers of embolisms rose to an early morning peak, followed by apparent repair of these before noon. There was a second spate of embolisms in late afternoon, followed by complete refilling of all xylem with liquid by an hour or so after dusk. All vessels then remained fully recharged until the following dawn. We believe that aspiration is a direct and reliable technique, which offers a simple, inexpensive means of assessing the relative extent of embolism of vessels in xylem, and a means to test earlier findings by the other direct method of the CSEM. In a broad context, the technique should provide new opportunities for evaluating water relations of the xylem of whole plants.

Plantago lanceolata L. and Trifolium repens L. were grown for 16 wk in ambient (360 μmol mol−1) and elevated (610 μmol mol−1) atmospheric CO2. Plants were inoculated with the arbuscular mycorrhizal (AM) fungus Glomus mosseae (Nicol. & Gerd.) Gerdemann & Trappe and given a phosphorus supply in the form of bonemeal, which would not be immediately available to the plants. Seven sequential harvests were taken to determine whether the effect of elevated CO2 on mycorrhizal colonization was independent of the effect of CO2 on plant growth. Plant growth analysis showed that both species grew faster in elevated CO2 and that P. lanceolata had increased carbon allocation towards the roots. Elevated CO2 did not affect the percentage of root length colonized (RLC); although total colonized root length was greater, when plant size was taken into account this effect disappeared. This finding was also true for root length colonized by arbuscules. No CO2 effect was found on hyphal density (colonization intensity) in roots. The P content of plants was increased at elevated CO2, although both shoot and root tissue P concentration were unchanged. This was again as a result of bigger plants at elevated CO2. Phosphorus inflow was unaffected by CO2 concentrations. It is concluded that there is no direct permanent effect of elevated CO2 on mycorrhizal functioning, as internal mycorrhizal development and the mycorrhizal P uptake mechanism are unaffected. The importance of sequential harvests in experiments is discussed. The direction for future research is highlighted, especially in relation to C storage in the soil.

Cuttings of two clones of Norway spruce, Picea abies (L.) Karst., were exposed to three different ozone concentrations in open-top chambers during four months in the summer of 1990. The treatments were charcoal-filtered air, non-filtered air and non-filtered air with extra ozone (c. 30 ppb) added daily between 1100–1800 local time. During the last 4 wk of the exposure period, half the seedlings were drought-stressed, while the remainder were well watered. Biomass, gas exchange and water potential were measured during the drought stress period.

The ozone treatments affected the two clones very differently. High ozone reduced the rate of biomass increase in the faster-growing clone (clone M) whereas ozone generally stimulated the rate of biomass increase in the slower-growing clone (clone L). At the end of the measuring period, the high-ozone treatments reduced the rate of root biomass increase of the well watered seedlings of the M-clone, but it had no effect on the drought-stressed seedlings of the same clone, probably because the root growth was already to a large extent inhibited by the drought stress.

The treatment with the highest concentration of ozone partly protected the seedlings of the M clone against the drought stress, presumably by delaying shoot growth and thus delaying the increase in the total transpiring needle area. As a result, stomata tended to close less during the drought period in this treatment.

The results are discussed in relation to the suggested critical level for ozone effects on trees.

The impact of O3 at different stages of plant development was investigated in growth-chamber-cultivated Plantago major L. Six-d-old plants of an O3-sensitive population (‘Valsain’) were exposed to one of the following six treatments; 56 d charcoal/Purafil®-filtered air (CFA): 56 d CFA plus 70 nmol mol−1 O3 for 7 h d−1; CFA with a 14-d episode of O3 administered at days 1, 14, 28 or 42. Harvests were made every 14 d, and at the final harvest (56 d) the influence of O3 on reproductive structures was assessed.

Analysis of the effects of O3 on growth and reproductive performance confirmed the sensitivity of the population to the pollutant. In the absence of the development of typical visible symptoms of foliar damage, the total d. wt of plants maintained in O3 over a 56-d period was 35% lower than that of control plants. However, the impact of the pollutant was found to decrease as plants aged. Plant relative growth rate (R) was only affected in seedlings, suggesting that effects of O3 on seedling growth were largely responsible for the decrease in accumulated biomass; the growth rate of older plants was not affected by O3. The observed shift in O3 resistance with plant age was mediated by both ‘acclimation’ and ontogenetic changes. ‘Acclimation’ was not associated with changes in O3 uptake, and there was some evidence to support the existence of compensatory growth responses. In addition to effects on vegetative growth, plants exhibited an O3-induced decline in reproductive performance; O3 reducing the number of flower spikes and seed capsules produced per plant. Ozone episodes administered at different stages of development indicated that reproductive development was particularly sensitive to O3 during the early stages of flowering.

The findings of this study are discussed in relation to evolutionary adaptation to O3 in natural plant communities.The importance of plant age, prior exposure to the pollutant and the timing of O3 episodes in relation to plant developmental stage are highlighted.

The resurrection plant Chamaegigas intrepidus Dinter (Scrophulariaceae) grows as a typical hydrophyte in shallow rock pools on granitic outcrops in arid areas of Namibia. During the rainy season, the rock pools are temporarily filled with water. When the pools dry up, C. intrepidus desiccates and survives in an air-dry condition for at least 8 months. After rewatering, the plants regain their metabolic activity in under 2 h. The desiccation of the vegetative organs is accompanied by a dramatic accumulation of abscisic acid (ABA). Beyond this, desiccation of roots is accompanied by the occurrence of specific dehydration-related proteins, whereas the leaves of C. intrepidus show high levels of dehydrins in the dehydrated as well as in the hydrated state.

Investigations in Namibia showed drastic diurnal fluctuations in the pH of the rock pools. The pH value increased from slightly acidic or neutral conditions during the morning to alkaline conditions (up to pH 12) during late afternoon. Since compartmental ABA distribution depends strongly on pH gradients across membranes, the external pH would be expected to affect the ABA relations in the plant. According to the anion trap concept, an alkaline pH in the surrounding medium should cause a release of ABA from the roots, although C. intrepidus appeared to release less ABA than the terrestrial rosettes of Valerianella locusta.

Leaf mineral concentration of Citrus aurantium (sour orange tree) was measured at bi-monthly intervals from 30 to 85 months of exposure in a long-term study on the effects of a 300 µmol mol−1 enrichment of atmospheric CO2, under conditions of high nutrient and water supply. There were clear seasonal trends in the concentrations of most of the elements studied. There were initial decreases in the leaf concentrations of N and the xylem-mobile, phloem-immobile elements Mn, Ca and Mg, as well as a significant and sustained increase in the leaf concentration of B, and no changes in the concentrations of K, Fe, Na, P, S, Zn and Cu. Interestingly, the initial reductions in the leaf concentrations of Mn, N, Ca and Mg gradually disappeared with time.

Information on ozone uptake by various tree species has been derived mainly from controlled-exposure studies of seedlings. Parallel measurements of foliar physiological processes in large trees are rare, as are measurements of local ozone concentrations within forest environments. Thus, our objectives were to characterize foliar ozone uptake in forest trees of different size and to identify a mechanistic linkage between small and large trees. Light-saturated net photosynthesis (Pn), stomatal conductance (gH2O), and ozone uptake (gO3) in seedling (<2 m), sapling (2–5 m) and mature (9–21 m) black cherry (Prunus serotina Ehrh.), northern red oak (Quercus rubra L.) and red maple (Acer rubrum L.) trees were measured monthly over two growing seasons at sites at high (1240 m) and low (600 m) elevation in the Great Smoky Mountains National Park. Seasonal ozone dose to northern red oak and red maple in the upper crown of mature trees was approx. double that of respective understorey seedlings and saplings. Seasonal ozone dose to black cherry was up to six times greater in mature trees than in seedlings and saplings. The lower ozone dose to understorey trees was mainly a function of lower gH2O, although canopy gradients in ozone concentration were indicated. Differences in Pn and gH2O between small and large deciduous forest trees were linked to variation in nitrogen concentration expressed on a leaf area basis.

Summary 167

I. INTRODUCTION 168

II. THE IMPACT OF PROTEIN STRUCTURAL INFORMATION ON THE UNDERSTANDING OF THE PRIMARY REACTIONS IN PHOTOSYNTHESIS 169

1. The structure of the purple bacterial reaction centre 170

2. The structure of PSI 172

3. The structure of PSII 173

4. The structure of antenna complexes 176

(a) Light-harvesting complexes from purple photosynthetic bacteria 177

III. DEVELOPING OVEREXPRESSION SYSTEMS FOR THE STUDY OF PLANT PROTEIN 188

1. Expression of heterologous proteins in Escherichia coli 189

2. Protein expression in the methylotrophic yeast Pichia pastoris 189

3. Expression of recombinant protein in insect cells: the baculovirus system 190

IV. CONCLUSIONS 190

This review sets out the case that now is the time for plant science to establish the technologies required for routinely studying the structure and function of plant proteins. The impact that protein structural information can have is illustrated here with reference to photosynthesis. Our understanding of the precise molecular mechanisms of the light-reactions of photosynthesis has been transformed by the combination of high-resolution protein structural data and detailed functional studies. The past few years have been a particularly exciting time to be engaged in basic plant science research. The application of modern techniques of molecular biology has allowed many key questions to be addressed. The stage is now set for an even bigger revolution as the current plant genome sequencing projects are completed. If these advances are going to be fully exploited, plant science must get to grips with studying proteins, not just genes. Reliable methods for the overexpression of proteins in their native state coupled with routine access to structure determination must become the norm rather than the exception. In 1998 there were about 9000 protein structures deposited in the Brookhaven database. Very few of these are plant proteins. This trend will have to be reversed if research in molecular plant science is to fulfil its potential.

The effects of long-term carbon dioxide enrichment on competition for nutrients and light in a ryegrass/clover association were determined for simulated swards of perennial ryegrass (Lolium perenne L. cv. Parcour) and white clover (Trifolium repens L. cv. Karina), which were grown as monocultures and in three mixtures (25/75, 50/50, 75/25), according to the replacement design, at two levels of nitrogen (N) supply (no additional N and 200 kg N ha−1) and at season-long ambient (380 ppm) and elevated (670 ppm) CO2 concentrations, in open-top chambers. Stands were cut four times, at about monthly intervals, to a height of 5 cm. Plant material was separated into different species, fresh and dry weights were determined and the content of macroelements (N, P, K, S, Mg) in both species was measured. In addition, plant height of both species at harvest dates and during several regrowth periods was monitored.

Results indicate that both species made demand on different resources and profited from growth in a mixed sward. CO2-related yield increase amounted to 16–42% for white clover whereas the effect of high CO2 on ryegrass yield ranged between −33% and +9% depending on N supply, mixture and year. As a result the contribution of white clover to total yield in mixed swards was significantly enhanced by CO2 enrichment at many harvests in both N supply treatments. Without additional N supply, shoot competition for light was intensified by CO2 enrichment to the disadvantage of ryegrass, since clover petioles grew longer and ryegrass was shorter at elevated CO2. With N fertilization, no marked effect of CO2 enrichment on interspecific competition could be observed. Since clover and total yield were increased by CO2 enrichment, nutrient requirements were also increased and potassium deficiency and increased intraspecific competition of clover for K was observed in the mixtures under elevated CO2 which had the highest nutrient withdrawal. Although white clover profited much more from CO2 enrichment in both N fertilization treatments, the suppression of ryegrass in mixed swards could only be observed under low N conditions.

Generally, the effect of N fertilization on competitive interference between both species was much greater than the effect of CO2 enrichment and it is suggested that the effect of elevated CO2 on the balance of species and the outcome of competition in a grass/clover sward is mainly dependent on the N status.

The resurrection plant Myrothamnus flabellifolia has the ability to recover from repeated prolonged and extreme desiccation cycles. During the dry state the inner walls of the xylem vessels seemed to be covered, at least partly, by a lipid film as shown by Sudan III and Nile Red staining. The lipid film apparently functioned as an ‘internal cuticle’ which prevented the adjacent parenchyma ray cells from complete water loss. The hydrophobic nature of the inner xylem walls was supported by the finding that benzene ascended as rapidly as water in the xylem of dry Myrothamnus branches. On watering, numerous lipid bodies were found in the water-conducting vessels, presumably formed from the lipid film and/or from lipids excreted from the adjacent living cells into the vessels. The presence of lipid bodies within the vessels, as well as the hydrophobic properties of the inner xylem walls, could explain the finding that the xylem pressure of hydrated, well watered plants (measured both under laboratory and field conditions with the xylem pressure probe) never dropped below c. −0.3 MPa and that cavitation occurred frequently at low negative xylem pressure values (−0.05 to −0.15 MPa). The xylem pressure of M. flabellifolia responded rapidly and strongly to changes in relative humidity and temperature, but less obviously to changes in irradiance (which varied between 10 and c. 4000 μmol m m−2 s−1). The morphological position of the stomata in the leaves could explain the extremely weak and slow response of the xylem pressure of this resurrection plant to illumination changes. Stomata were most abundant in the furrows, and were thus protected from direct sunlight. Simultaneous measurements of the cell turgor pressure in the leaf epidermal cells (made by using the cell turgor pressure probe) revealed that the xylem and the cell turgor pressure dropped in a ratio of 1:0.7 on changes in the environmental parameters, indicating a quite close hydraulic connection and, thus, water equilibrium between the xylem and cellular compartments. An increase in irradiance of c. 700 μmol m−2 s−1 resulted in a turgor pressure decrease from 0.63 to 0.48 MPa. Correspondingly, the cell osmotic pressure increased from 1.03 to 1.22 MPa. From these values and by assuming water equilibrium, the osmotic pressure of the xylem sap was estimated to be 0.25–0.4 MPa. This value seems to be fairly high but may, however, be explained by the reduction of the water volume within the vessels due to the floating lipid bodies.

Seedlings of Pseudotsuga menziesii (Mirb.) Franco, Pinus ponderosa Dougl. ex Laws, Arbutus menziesii Pursh., and cuttings of Arctostaphylos uva-ursi (L.) Spreng were grown in monoculture and in conifer-hardwood dual-culture combinations in the glasshouse and inoculated with spore slurries of six Rhizopogon species. The primary objectives were to assess and compare the pattern of host specificity between symbionts and to study the influence of co-cultured plants on ectomycorrhiza development. The Rhizopogon spp. ranged from genus-specific to multiple-host compatible. In monoculture, four Rhizopogon sp. (R. ellenae Smith, R. occidentalis Zeller & Dodge, R. smithii Hosford and R. subcaerulescens Smith) formed ectomycorrhizas with Pinus ponderosa, and two Rhizopogon sp. (R. parksii Smith and R. vinicolor Smith) formed ectomycorrhizas with Pseudotsuga menziesii. None of the fungi tested developed ectomycorrhizas on Arbutus menziesii or Arctostaphylos uva-ursi in monoculture. In dual culture, three of the four Rhizopogon species (R. ellenae, R. occidentalis and R. subcaerulescens) that formed ectomycorrhizas on Pinus ponderosa, formed some ectomycorrhizas on Arbutus menziesii and Arctostaphylos uva-ursi. Rhizopogon parksii and R. vinicolor only formed ectomycorrhizas on Pseudotsuga menziesii.

In shallow flooded parts of rich fens Mentha aquatica might thrive in deeper water than Epilobium hirsutum but previous experiments have provided no clear indication that the flooding tolerance of these species differs. In this study we investigated, by measuring growth, biomass allocation and vegetative reproduction, whether the impact of water level on vegetative reproduction might produce different lower boundaries on water level gradients. There was a striking contrast between biomass production at high water levels and the field distribution of both species. After 18 wk, the mean biomass of E. hirsutum grown in waterlogged and flooded conditions was 82% and 54%, respectively, of the mean biomass production of drained plants. Biomass of waterlogged and flooded M. aquatica was reduced to 57% and 37% in drained conditions. Waterlogged and flooded E. hirsutum had swollen stem bases and invested a high proportion of biomass in adventitious roots. Stems of M. aquatica did not swell, formed few adventitious roots and maintained an equal proportion of below-ground roots at all water levels. The effect of water level on vegetative reproduction corresponded well with the lower hydrological boundaries of both species. When waterlogged and flooded, most rhizomes of E. hirsutum emerged from above-ground parts of the stem base and were oriented in an upward direction. Plants in flooded soil allocated less biomass to rhizomes and also reduced the number and size of rhizomes. Rhizome formation of M. aquatica on the other hand was not directly affected by water level and only depended on plant size. These differences in vegetative reproduction are discussed in relation to the different abilities of both species to oxygenate their below-ground roots. It was concluded that the mode of adaptation to soil flooding might also affect vegetative reproduction and, therefore, a species' ability for long-term persistence in soil-flooded habitats.

The aim of this study was to quantify the long-term effects of Al in solution on mineral ion uptake and associated H+ release by the roots of maize cultivated in conditions compatible with field data. Two maize (Zea mays L.) cvs of differing sensitivity to Al were compared. The culture was conducted in hydroponic conditions for culture periods of 2–5 d in the presence of 0–30 μM Al. Proton fluxes were localized along the roots using a videodensitometry method with bromocresol green on agarose gel.

Aluminium reduced the uptake of anions, particularly NO3−, whereas the uptake of K+ and NH4+ was unaffected. These effects were accompanied by an increase in net H+ release which was quantitatively comparable with the reduction in NO3− uptake. The reduction in NO3− uptake and the concomitant increase in H+ release increased with the quantity of Al accumulated in the roots, which was itself dependent on the concentration of Al in solution. The Al-sensitive cultivar bound more Al to its roots than the Al-tolerant one; the reduction in NO3− uptake and the increase in H+ release were also greater in the Al-sensitive cv. Dye indicator videodensitometry showed that the increased H+ release was general all along the roots.

These results led us to specify the relation between the Al deposition in roots, the reduction in NO3− uptake and the stimulation in rhizosphere acidification by the plants. They show that, with maize, Al induces an inhibition in NO3− uptake and an increase in net H+ release in a 1[ratio ]1 ratio. These concomitant effects suggest that inhibition in NO3−/OH− exchange unmasks active H+ excretion which is otherwise insensitive to Al. Both these effects are closely related to the Al content in roots expressed per unit root length.

Swards of cocksfoot (cvs KM2, Lutetia) and perennial ryegrass (cvs Aurora, Vigor) were grown under full irrigation or severe (80 d) drought in a field experiment in the South of France. Responses of the bases of immature leaves plus enclosed tissues were made during the drought period and after rewatering.

By the end of the drought, water content had fallen from 3·0 to 0·8 gwater g−1dm, and osmotic potential from −1·0 to −4·5 MPa in all cvs. Measured minerals and water-soluble carbohydrates contributed, respectively, c 19 and 44% to osmotic potential in droughted leaf bases. The drought-sensitive cocksfoot cv. Lutetia was characterized by a large proportion of fructans having a low degree of polymerization (DP=3, 4). As drought progressed, accumulation of dehydrin transcripts and ABA were higher in leaf bases of the sensitive cv. Lutetia than in the resistant cv. KM2.

After rewatering, the water status of immature leaf bases returned to control levels in 1–2 d, and then increased further as leaves began to grow and new tissue was produced. High-DP-fructans remained unchanged in leaf bases of ‘Lutetia’ but were depleted by over 55%, and therefore remobilized, in leaf bases of other cvs after 8 d.

It is concluded that enclosed immature leaf bases survive drought by tolerating a low water status and that changes conventionally associated with desiccation tolerance are expressed most strongly in susceptible plants least able to maintain their water supply.

The response of epicuticular wax of Scots pine (Pinus sylvestris L.) needles to dry- and wet- deposited sulphur and heavy metals was investigated at six sites located 10–110 km from the Monchegorsk Cu-Ni smelter on the Kola Peninsula, North-West Russia, and in a long-term irrigation experiment where pines were exposed over four growing seasons (1991–1994) to either acid rain treatment at pH 3·1 (H2SO4), metal treatment at pH 5·7 (Cu and Ni) or a combination of these at pH 3·1. Needle wettability exhibited a closer relationship with epistomatal wax tube distribution (WTD) than with chemical composition of epicuticular wax. Water droplet contact angles (DCA) decreased towards the smelter, and significant differences due to site were noted for 26-month-old and 38-month-old needles. Significant differences due to site were determined for secondary alcohols, dehydroabietic acid and hydroxy fatty acids, the proportions of which ranged from 22·5 to 48·9%, 6·2 to 22·4% and 0·6 to 2·6% respectively, depending on site and needle age class. The proportion of dehydroabietic acid increased towards the smelter, but no gradient was observed in the proportion of secondary alcohols or hydroxy fatty acids. No major effect of experimentally applied pollutants on the chemical composition or structure of the epicuticular wax was observed. The effect of treatment on DCA was significant in 1993 and 1994 due to a 6·5–13·2 degree greater wettability of the 37–49-month-old acid-treated needles relative to the irrigated or dry controls. Sulphuric acid at pH 3·1 did not increase needle wettability when combined with copper and nickel sulphate in similar concentrations. These data indicate that S deposition, especially H2SO4, plays a more important role in needle surface deterioration than Cu and Ni. Pollutant-induced changes in epicuticular wax structure and needle wettability mimic natural wax ageing, but at an accelerated rate. Changes in wax chemical composition might also be caused by pollutant-induced metabolic changes in elongating needles.

Development of mycelial cord systems of Phanerochaete velutina (DC.: Pers.) Parmasto from 4-cm3 inocula on a nutrient-depleted non-sterile soil was studied in laboratory microcosms using image analysis techniques. Cord systems were ‘baited’ after 13 d growth with either fresh, non-sterile 4-cm3 wood baits or control Perspex® blocks of the same contact area placed behind the foraging mycelial front. After 26 d growth, mycelial ‘patches’ arose by dedifferentiation of consolidated mycelial cords in both wood- and Perspex-baited cord systems. ‘Patches’ comprised fine, highly branched separate hyphae extending radially from points of aggregated hyphae in cords. ‘Patches’ and cords could be readily distinguished by image analysis and the areas covered by patches and cords could be measured and compared. Whilst the total hyphal cover of Perspex- and wood-baited systems did not differ significantly (P>0·05), patch cover in wood-baited systems was up to 10 times greater than in Perspex-baited systems. Patches were temporary structures, regressing more rapidly with age than mycelial cords. Patch development ceased after application of a nutrient solution which replenished phosphate levels in the soil. Wood-baited mycelial systems displayed significant developmental polarity (P[les ]0·05) of both total hyphal cover (patches plus cords) and hyphae in patches towards the ‘baited’ sector of cord systems after 42 d, which corresponded with peak patch development. However, significant (P[les ]0·05) developmental polarity of the mycelial systems along the bait-inoculum line could be detected 8 d before patch formation when assessed by fractal geometry. Radiotracer studies showed that mycelial patches were not sinks for supplied 32P, but that they were sites of increased nutrient uptake capacity compared with that of mycelial cords. We discuss the need for mycelial cord systems to balance allocation of mycelial biomass between the two essential processes of colonizing wood resource units, and the acquisition of soluble inorganic nutrients from soil.

The role of carbonic anhydrase in the carbon-concentrating-mechanism of bryophytes of the class Anthocerotae was investigated by comparing the gas-exchange characteristics of material which had been incubated in the membrane-permeable Carbonic Anhydrase inhibitor ethoxyzolamide, with those of untreated material and material which had been incubated in buffer solution. In Phaeoceros laevis (Anthocerotae), incubation in ethoxyzolamide caused a depression in the rate of gross assimilation and a decrease in CO2 affinity beyond that which could be attributed to increased diffusion limitation. A range of liverworts and mosses, in which a carbon-concentrating-mechanism is absent, were also investigated. These showed no depression of rates of gross assimilation after incubation in ethoxyzolamide relative to those of untreated material. The CO2 compensation point and CO2 uptake characteristics of Phaeoceros laevis were significantly affected by incubation in ethoxyzolamide. Values of CO2 compensation point for Phaeoceros laevis rose from 2.5 Pa, after incubation in buffer, to 20 Pa after incubation in ethoxyzolamide. The CO2 compensation point for the liverworts Pellia epiphylla and Marchantia polymorpha was not significantly affected by incubation in ethoxyzolamide. Measurements of the release of CO2 at the end of a short (15 min) period of illumination revealed that, after suppression of carbonic anhydrase activity, the rapid release of a CO2 pool occurred in Phaeoceros laevis but not in the liverworts. There were also significant differences between values for fractionation measured in units per mil (‰), measured instantaneously, for Phaeoceros laevis incubated in ethoxyzolamide, compared with fractionation values for this species after incubation in buffer. Incubation in ethoxyzolamide caused fractionation values to rise from 12.4–22.7‰, indicating that the carbon-concentrating-mechanism of this species had been inactivated. Incubation in ethoxyzolamide had no effect on fractionation values for the liverworts. The convexity of the light saturation curves of liverworts and Phaeoceros laevis was also investigated, but there were no differences between groups before or after the two treatments. The data indicate an important role for carbonic anhydrase in the functioning of the carbon-concentrating-mechanism in the Anthocerotae.

To assess adverse effects of ambient ozone on injury and yield, EDU (ethylenediurea) and non-EDU-treated plants of bean (Phaseolus vulgaris L. cv. Lit) were exposed to ambient air at four rural sites in the Netherlands in 1994. After 6 wk of exposure, the proportion of leaves injured in non-EDU-treated plants was higher than that in plants treated with the antioxidant. A maximal degree of 35% injury was observed. The highest ozone levels accumulated at each site for three consecutive days before injury clearly exceeded the proposed short-term critical level for injury development. Both injury in untreated plants and the protective effect of EDU differed between sites and were not related to the measured ozone levels. EDU increased the d. wt of dry (mature) pods after 11 wk of exposure but not of green (marketable) pods after 6 wk. The effect of EDU in enhancing dry pod yield did not differ between sites and was 20% on average. The proportional yield increases in EDU-treated plants compared with those of non-EDU-treated plants at the sites were not correlated with the measured ozone levels known to adversely affect bean yield. The available evidence, however, suggests that ambient ozone was involved in the observed yield increase by EDU. When the dry pods were harvested, EDU-treated plants had more leaves than non-EDU-treated ones. The effect of EDU in delaying senescence appears to be an important factor to consider when attempting to evaluate the effect of ambient ozone on pod weight in beans using the EDU approach.