In the field, pathogens exist in a diverse and dynamic ecological environment with other organisms (Blakeman, 1993; Preece & Dickinson, 1971). We tend to assume that these other organisms are ‘benign’ or perhaps in competition with the pathogen to occupy a particular niche. However, bacteria are often observed to be in close association with pathogens, as witnessed by the difficulty of eliminating them during isolation of pathogens involved in plant diseases. We now have clear evidence (Dewey et al., pp. 489–497 in this issue) that they are sometimes neither benign, nor in competition, but actually helping the pathogen successfully infect its host even when the bacteria themselves are not pathogens of that host.

Chlorophyll fluorescence measurements were used to test whether externally applied abscisic acid (ABA) has an effect on the desiccation tolerance of lichens and their sensitivity to constant water saturation. A surplus of ABA did not decrease the time required to regain full photosynthetic capacity after prolonged dry periods. However, an effect of ABA could be observed when lichens were permanently hydrated at moderately high temperatures. Lichens suffered less from constant saturation if ABA was added to the incubation medium. These results suggest a positive effect of ABA on membrane function.

Podetia of the terricolous lichen Cladina stellaris (Opiz) Brodo, collected in an unpolluted rural area in N. Finland, were wetted with either H2O, diluted acidic solutions of H2SO4, HNO3 and NaHSO3, simulating acidic rain, or the following salts: K2SO4, KCl, CuSO4, CuCl2, Cu(NO3)3, ZnSO4, ZnCl2, Zn(NO3)2, FeSO4, FeCl2, Fe2(SO4)3, FeCl3 and Fe(NO3)3. The samples were further exposed to combined treatments in both acidic solutions, simulating acidic rain, and heavy metal salts in solution. Lichen samples wetted with H2O at pH 6·8, diluted solutions of H2SO4, HNO3 or a mixture of these two acids produced low concentrations of endogenous ethylene. The application of NaHSO3 greatly increased the production of ethylene. The application of KCl induced a higher rate of ethylene production than the application of K2SO4 solutions. The application of Cu-containing solutions enhanced the production of ethylene. The influence of Zn was smaller than that of Cu. Iron was the most effective heavy metal to promote the production of ethylene: very high ethylene concentrations were detected upon the application of FeCl2. Combined treatments in H2SO4 or H2SO4+HNO3 followed by either FeCl2 or FeSO4, yielded higher concentrations of ethylene than the same treatments in a reversed order. The role of Fe ions in the production of ethylene is discussed in reference to previous works dealing with ethylene production in higher plants, fungi and algae.

Colonization of roots by arbuscular mycorrhizal (AM) fungi in several annual crops in two consecutive seasons was compared with, in the second season, the density of fungal propagules in the soil with the use of a bioassay. Root density decreased down the soil profile in both years in all crops, and a high proportion of roots were mycorrhizal throughout the profile. AM colonization decreased down the profile in cotton and lablab in the second season only. The bioassay indicated that most propagules of AM fungi in soils under cotton were located near the surface, with virtually no propagules at 1 m. The absence of propagules at depth indicates a lack of mycelium deep in the soil, and suggests that mycorrhizas are primarily initiated in the surface soil and that the fungi colonize the root system mostly through secondary spread down the profile. The use of AM colonization in the field as an indicator of propagule density and symbiotic function should be qualified by an understanding of the depth in the soil from which roots were extracted.

Of the three component organs of the grass leaf – blade, sheath and ligule – the ligule is the least studied and the least understood. Traditionally, it has been assumed to act in a passive way in protecting the culm and leaves that it encloses from the entry of water, dust and harmful spores. However, ultrastructural and cytochemical studies of the membranous ligules of several taxa, particularly Lolium species, have challenged that rather simplistic view and suggest that these ligules play a more active role in the life of the grass plant as a secretory tissue. This review summarizes the evidence for the latter notion, assesses the validity of both the passive and active hypotheses of membranous ligule function and notes similarities between membranous grass ligules, root caps and lycopsid ligules.

A novel extracellular fructanhydrolase was isolated from the culture filtrate of Lactobacillus paracasei ssp. paracasei P 4134 grown on a mineral medium supplemented with fructan extracted from Timothy (Phleum pratense L.) as the only carbon source. The enzyme was purified by a combination of ammonium sulphate precipitation, affinity chromatography, preparative isoelectric focusing and anion-exchange chromatography. As a result of these procedures, the specific enzyme activity increased 93-fold, with a final yield of 28·4%. The substrate-specific activities against different fructan types were determined by incubating the enzyme fractions with fructan extracted from Timothy (predominantly β-2,6 fructosyl-fructose linkages), inulin from Dahlia tubers (mostly β-2,1 fructosyl-fructose linkages) and sucrose. The purified enzyme catalysed the hydrolysis of β-2,6-linked fructan more rapidly than the β-2,1 linkages of inulin. Additionally, the enzyme showed low ability to hydrolyse sucrose. Fructose was the main product of the degradation of Timothy fructan and inulin, indicating a high exohydrolytic activity of the enzyme. It is proposed that the fructan-degrading enzyme from L. paracasei ssp. paracasei P 4134 is a β-D-fructan-fructohydrolase (EC 3.2.1.80). The enzyme preparation showed a single protein band in sodium dodecyl sulphate-polyacrylamide gel electrophoresis with a mobility corresponding to molecular weight of c. 42 kDa. It was concluded that only one molecular weight of fructan-degrading enzyme exists in L. paracasei ssp. paracasei P 4134.

Fructan: fructan fructosyltransferase (FFT) activity was purified about 300-fold from leaves of Lolium rigidum Gaudin by a combination of affinity chromatography, gel filtration, anion exchange and isoelectric focusing. The FFT activity was free of sucrose: sucrose fructosyltransferase and invertase activities. It had an apparent pI of 4·7 as determined by isoelectric focusing, and a molecular mass of about 50000 (gel filtration). The FFT activity utilized the trisaccharides 1-kestose and 6G-kestose as sole substrates, but was not able to use 6-kestose as sole substrate. The FFT activity was not saturated when assayed at concentrations of 1-kestose, 6G-kestose or (1,1)-kestotetraose of up to 400 mM. The rate of reaction of the FFT activity was most rapid when assayed with 1-kestose and was less rapid when assayed with 6G-kestose, (1,1)-kestotetraose or (1,1,1)-kestopentaose. The FFT activity when assayed at a relatively high concentration of enzyme activity (approximately equivalent to about half the activity in crude extracts per gram fresh mass) did not synthesize fructan of degree of polymerization > 6, even during extended assays of up to 10 h. When assayed with a combination of 1-kestose and uniformly labelled [14C]sucrose as substrates, the major reaction was the transfer of a fructosyl residue from 1-kestose to sucrose resulting in the re-synthesis of 1-kestose. Tetrasaccharide and 6G-kestose were also synthesized. When assayed with 6G-kestose and [14C]sucrose as substrates, the major reaction of the FFT activity was the synthesis of tetrasaccharide. However, some synthesis of 1-kestose and re-synthesis of 6G-kestose also occurred. When 6G-kestose was the sole substrate for the FFT activity, synthesis of tetrasaccharide was 2·7 to 3·4-fold slower than when 1-kestose was used as the sole substrate. Owing to differences in the fructan [ratio ] sucrose fructosyltransferase activity of the FFT with each of the trisaccharides, net synthesis of tetrasaccharide by the FFT was altered significantly in the presence of sucrose. The magnitude of this effect depended on the concentration of the trisaccharides. In the presence of sucrose, 6G-kestose could be a substrate of equivalent importance to 1-kestose for synthesis of tetrasaccharide.

The abundance and cellular location of Fe-containing superoxide dismutase (Fe-SOD) in trichomes of Nodularia, Aphanizomenon and Anabaena collected from various depths in the Baltic Sea, and in trichomes of a cultured Nodularia strain, BC Nod-9427, isolated from the Baltic Sea, was examined by immunogold labelling. For trichomes collected from natural populations the areal concentration of Fe-SOD labelling decreased with depth: trichomes collected from surface accumulations had between 8 and 11 gold particles μm−2 whereas trichomes collected from a depth of 18 m were unlabelled. When trichomes collected from a depth of 10 m (mean areal labelling density 0·5 gold particles μm−2) were exposed to the higher irradiances present at 1 m, the areal concentration of Fe-SOD increased to 3·5–4 gold particles μm−2 within 4 h. When cultures of Nodularia strain BC Nod-9427, adapted to low light (10 μmol m−2 s−1), were transferred to an incident irradiance of 1350 μmol m−2 s−1, a doubling of the areal concentration of Fe-SOD gold label was observed within 1 h. Addition of 3-(3,4-dichlorophenyl)-1,1′-dimethylurea (DCMU) to cultures immediately before their transfer to increased illumination resulted in a decrease in areal Fe-SOD concentrations whereas addition of CdCl2 caused an increase over and above that induced by the elevated irradiance. These results suggest that Baltic Sea cyanobacteria are able to modulate their Fe-SOD content but that this might be in response to oxidative stress rather than to light per se.

The photosynthetic properties of cyanobiont lichens from contrasting habitats were measured to identify whether the increased assimilation rates which characterized Peltigera membranacea (Ach.) Nyl. from an exposed habitat were correlated with increased carbon-concentrating mechanism (CCM) activity. The results were contrasted with data obtained from two populations of Peltigera praetextata (Flörke ex Sommerf.) Zopf collected from dry and damp microhabitats within a shaded woodland and Peltigera leucophlebia (Nyl.) Gyelnik, which has been shown to lack a carbon-concentrating mechanism. The differences in assimilation rates between the cyanobiont lichens were not accounted for by differences in chlorophyll content. Peltigera membranacea from the exposed habitat which had the highest assimilation rates had the lowest Gamma; and K0·5 values and accumulated the greatest Ci-pool indicating that increased Ci accumulation contributed towards the higher assimilation rates shown by these species. The convexity of the light response curve for the cyanobiont lichens decreased with increasing assimilation rates. This might have indicated a diversion of electron transport to energize the carbon-concentrating mechanism. The apparent quantum efficiency of CO2 assimilation (ΦCO2) was correlated with the genus of lichen photobiont. All cyanobiont lichens had comparable values for ΦCO2 which were greater than that of the tripartite Peltigera leucophlebia. Light compensation points reflected the exposure of the habitats with higher compensation points characterizing the cyanobiont population from the exposed crag and the tri-partite population from the open grassland. Carbon isotope discrimination values for organic matter and measured instantaneously were the same for all cyanobiont lichens and were comparable with values recorded for species with a carbon-concentrating mechanism. Carbon isotope measurements for P. leucophlebia were typical of those recorded for species without a carbon-concentrating mechanism. Variation in source isotope signature and refixation of respiratory CO2 were considered to be significant factors in determining organic matter and instantaneous carbon-isotope discrimination. These factors might have masked any subtle variation in carbon-isotope discrimination which resulted from variable CCM activity. The functional significance of increased carbon-concentrating mechanism activity in cyanobiont lichens occupying exposed habitats is discussed.

Ice-core records of the concentration of atmospheric CO2 and its stable isotope ratio (δ13Ca) have shown that the global C cycle has not remained in steady-state over the past 11000 yr, implying a possible change in vegetation activity over this period. Here we evaluated the ecophysiological responses of the dwarf willow (Salix herbacea) over the past 9000 yr by measuring the stable carbon isotope composition and stomatal characters of a unique, well dated, high-latitude (68 °N) sub-fossil leaf sequence. After correction for corresponding changes in δ13Ca, a 9000-yr record of variations in the ratio of intercellular (ci) to atmospheric (ca) CO2 concentration was established. Intercellular[ratio ]atmospheric CO2 concentration ratios provide a time-integrated indicator of the set-point of leaf gas exchange, and the historical variations revealed in this record have been interpreted as an impact of environmental changes on leaf gas exchange. The sequence shows a progressive fall in ci/ca 9000–3000 yr BP as well as the climatic effects of the Medieval Warm Period, the Little Ice Age and the post-industrial CO2 rise. Leaf stomatal index (proportion of epidermal cells as stomata), but not stomatal density, was significantly (P<0.01) correlated with Holocene atmospheric CO2 variations. A process-based interpretation of the changes in ci/ca was made using two different coupled photosynthesis-stomatal conductance models. Calculated in this way, S. herbacea photosynthetic rates were relatively stable throughout the Holocene whilst stomatal conductance progressively declined. Both, however, showed the marked effects of the Medieval Warm Period and the Little Ice Age. Overall, the results demonstrate that S. herbacea leaf metabolism, like the global C cycle, has not remained in steady state during the Holocene but has responded to changes in atmospheric CO2 concentration and short-term climatic oscillations.

Light, fluorescence and electron microscopy were employed to follow the development of the endodermis in cluster roots and lateral roots of Grevillea robusta A. Cunn. ex R. Br. Endodermal cells had three different origins: rootlet endodermis arose from the rootlet meristem; endodermis covering the primordium shortly after initiation came from division of parental endodermis; cells at the junction between parent and rootlet endodermis developed from re-differentiated rootlet cortical cells. In the cluster root, the Casparian band formed in three ways, and was not initially present opposite the two sets of single xylem elements in the rootlet stele. A new clearing technique was developed that allowed visualization of xylem, suberized endodermis, Casparian band formation and phenolic compounds. In lateral roots, endodermal differentiation was asynchronous, but was related to position relative to protoxylem poles. However, the observed delay began before these poles had differentiated. At the tip of mature rootlets, which are determinate, the endodermis terminates in a ‘dome’ of cells, with the initial cell differentiating as an endodermal cell. Results are discussed in terms of determinate development in roots and the spatial and temporal contexts within which this development takes place.

Pollen counting is an important element of research in pollination, and the majority of current methods involve manual counting. An image-processing technique has been developed for accurate counting and sizing of small particles, and this was tested on almond pollen grains. Our automatic process was found to be several times faster than manual counting and more accurate, with an average error of only 3%. Because of its simplicity, it is an attractive tool for research on such topics as pollen viability or germination. It was found that in the methods using vials there were ‘residues’ of c. 17% on the vial walls and on the handling tools. In addition, the first drop from each vial contained, on average, 1·65 times more grains than the average number of grains in all drops, whereas the last drop from each vial contained fewer grains than the average. This phenomenon was correlated with the non-uniform pollen distribution in the suspension. An automatic counting system is therefore, recommended, to avoid the problem, otherwise a calibration procedure should be used that accounts for both the residues on the tools and the non-uniformity of contents of sample drops.

Because nitrogen is the mineral nutrient needed in largest amounts by plants, it is usually also the limiting factor for plant growth in terrestrial ecosystems (Vitousek & Howarth, 1991). Consequently, the deposition of oxidized and reduced N compounds will almost invariably have large effects in these systems, and because N availability not only regulates plant growth but also that of organisms at other trophic levels, disturbances of several ecosystem processes might occur. The alternations introduced by deposition of atmospheric N compounds are both of a quantitative and of a qualitative nature. Moreover, N deposition can have phytotoxic as well as growth-stimulating effects.

This short commentary gives a personal view of some of the possible consequences of N deposition on plants. It refers particularly to the oral presentations given by Professor Heinz Rennenberg and Dr Marta Peréz-Soba, and to the discussions held after their talks where appropriate. Separate attention is given to four different consequence of anthropogenic N-deposition: N-availability; N-form, N-uptake by the shoot, and the period of N-uptake. Finally, I have tried to adopt an ecosystem perspective and discuss briefly the concept of critical loads of N.

The amounts of cyclic AMP (cAMP), fructose-2,6-bisphosphate (F26BP), trehalose and glycogen were determined in cell suspension cultures of the ectomycorrhiza-forming fungus Amanita muscaria (L. ex Fr.) Hooker. For the assay of cAMP a protocol was developed that enabled the detection of as little as 50 fmol of this secondary messenger by an enzyme-linked immuno assay (EIA). Values varied from <1 and up to 5 pmol cAMP mg−1 d. wt according to the age of the fungal culture. Typically, a transient increase in cAMP occurred after c. 4 d of culture of the fungus on glucose-containing medium. This increase (up to 100%) was followed by the start of the logarithmic growth phase, and by a more persistent increase in F26BP. In parallel, glucose in the medium started to decrease, whilst the amounts of fungal carbohydrates, especially the disaccharide trehalose, increased. From these data we assume that a high initial rate of glucose uptake caused an increase in the fungal pools of storage carbohydrates and, via activation of an adenylate cyclase, of cAMP. According to data reported for yeast cells this should enhance the formation of F26BP by phosphorylation of relevant enzymes. In animal and yeast cells an increase in the concentration of F26BP stimulates glycolysis by activation of the ATP-dependent phosphofructokinase (PFK). A. muscaria also possesses an F26BP activated PFK and, under conditions of symbiosis, host-derived carbohydrates are supplied mainly in the form of glucose. The implications of these findings to the regulation of carbohydrate metabolism of symbiotic plant root/fungus structures (ectomycorrhiza) are discussed.

Pathozone behaviour of soil-borne plant pathogenic fungi is characterized by curves for the change in probability of infection with distance of inoculum from a host. We identify three major components that give rise to the pathozone profile for infection efficiency: the germinability of inoculum, given that it occurs in the pathozone (P1), the growth of the resulting fungal colony outwards to make contact with the host (P2), and the infectivity of mycelium once contact is made (P3). The probability of infection (P) is then given by the product, P1 × P2 × P3. Using Raphanus sativus and two contrasting types of inoculum of Rhizoctonia solani as a model experimental system, we measured each of the three components by quantifying germinability, colony architecture (to determine the chance of contact by one or more hyphae) and the change in susceptibility over time as a measure of infectivity as mycelium arrives from different distances away. The germinability of inoculum is uniform across the pathozone whereas the probability of contact declines with distance, and the net effect of host susceptibility and infectivity increases. The characteristics of these components result in pathozone profiles that vary in shape. Infection efficiency can decline in an exponential or sigmoidal fashion with increasing distance from the host or follow a curve that rises close to the host and then falls asymptotically to zero with increasing distance. Colony architecture was summarized by the distribution of the furthest extent of hyphal growth amongst colonies growing out from single inoculum units (described, because of its asymmetrical pattern, by a gamma distribution) and by the radial density of hyphae on concentric circles at different distances from the centre of the colony (described by a negative binomial distribution with parameters changing with distance).

The mechanistic model for the components of pathozone behaviour is tested by comparing the magnitude and shape of pathozone profiles predicted by the model with independent estimates for P obtained by placing inoculum at fixed distances from the host, and measuring the proportion of successful infections. The shape depends on the relative magnitudes and change with distance of the three components, P1, P2 and P3, which differed between the two types of inoculum. The profiles could be reproduced accurately when the probability of contact was based on the distribution and density of hyphae reaching the host rather than on the furthest extent of hyphal growth amongst colonies. The model is used to analyse the effects of replication and stochastic variability in the components of pathozone behaviour on the comparison of treatments for disease control.

The tissue-specific and developmental expression of histone 2A was studied in onion (Allium cepa ‘Robusta’), using northern blots. Histone 2A expression was enriched in basal tissues, particularly in the inner, meristematically active parts of bulbs. The expression was assessed during a time course of bulb development, dormancy onset and post-harvest sprouting in field-grown material. During bulb development histone 2A expression in the inner bulb declined rapidly during bulb ripening, reaching a minimum with the onset of dormancy. During post-harvest storage, expression increased slowly, reaching a peak in the spring, coinciding with the first observed sprout emergence. It was concluded that in onion, as in other plant systems, histone 2A expression is linked to cell division and dormancy level, the peak in expression during post-harvest storage indicating the time of dormancy breakage. In cultivars where post-harvest sprouting occurred much earlier or much later than in ‘Robusta’, this expression peak occurred at about the same time of year, regardless of sprouting time. It was concluded that differences in storage longevity between cultivars were not due to differing times of dormancy breakage. Factors controlling the rate of sprout emergence post-dormancy are likely to be major determinants of storage capacity.

The study characterized the influence of cucumber mosaic virus (CMV) on the growth response of two annual weeds to nitrogen. Plants were grown individually along a N gradient from 4 to 32 mmol l−1 and data were interpreted using growth analysis. Plant biomass increased with N concentration and was significantly higher for healthy than infected plants at the two highest N concentrations. Healthy plants of Portulaca oleracea L. were characterized by lower biomass allocation to leaves and higher biomass allocation to roots than infected plants; no change in biomass allocation was recorded for Stellaria media Vill. Relative growth rate (rgr), net assimilation rate (nar) and specific leaf area (sla) of plants increased with increasing N concentration. Healthy plants of P. oleracea were characterized by a higher rgr and nar and a lower sla than infected plants, whereas healthy S. media had a higher rgr but a similar nar and sla or leaf weight ratio (lwr) compared with infected plants. The consequences of these results on the population dynamics of weeds and virus spread are discussed.

The purpose of this work was to establish how the distribution of local curvatures changed during the mushroom stem gravitropic reaction and to suggest a suitable mathematical model based on these new data.

The gravitropic bending of base- and apex-pinned Coprinus cinereus (Fries) S. F. Gray stems was recorded on videotapes. The images were captured from the tapes after each 10 min, rotated by 45° and transformed into tables of changing co-ordinates of points for each stem. The non-linear regression of these points was performed using Legendre polynomials. From the resulting equations the patterns of changing local curvature for 50 subsections per stem during 400 min of gravitropic reaction were calculated.

It was observed that base-pinned stems first bent from the apex, but later the curvature of this part decreased, and in the late stages the apex became nearly completely straight again. Subsections, located about one third of stem length from the base determined the main part of the final curvature. The free basal part of the apex-pinned stems bent upward and after a certain bending time also began to straighten. However, this process started significantly later and was weaker. Bending of the subsections close to the pinned apex did not stop when they reached the vertical position, and the final angle of gravitropic curvature could exceed 180°.

Plotting various functions of local bending speed and its derivatives against each other and against local angle indicated that, if the hypothetical signal about reorientation arises in the apex, its propagation towards the base did not follow simple wave or simple diffusion laws. The importance of the local angle of all subsections both for signal origin and transmission was established and a signal transmission equation, involving local angle of each subsection, was derived. After creating a suitable program this partial differential equation was solved numerically. The generated shapes of the bending stem coincided in high degree with experimentally observed images.

The effects of Cd and Zn on cross-colonization by Paxillus involutus of Scots pine seedlings was examined by using pairs of ectomycorrhizal (ECM) and non-mycorrhizal (NM) seedlings grown in the same vessel. This was done to assess, first, the ability of P. involutus to colonize NM Scots pine seedlings by growth from colonized roots of other Scots pine seedlings in the presence of Cd or Zn, and, second whether ECM colonization of Scots pine by P. involutus provided a competitive advantage over NM seedlings. Ectomycorrhizal colonization of Scots pine was shown to be more sensitive than Scots pine itself to Cd and Zn, but prior colonization did provide a competitive advantage with respect to biomass production. This beneficial effect over NM seedlings was, however, equal in the control, Cd and Zn treatments, and was due simply to growth stimulation in the presence of ECM colonization. Cross-colonization from an ECM to a NM seedling was reduced but not prevented by Cd and Zn. Cd had a more negative effect on cross-colonization than on initial colonization of seedlings, whereas Zn had an equally inhibitory effect on both parameters. These results have important implications for plant establishment on metal-contaminated sites. If cross-colonization between plants is reduced by toxic metals, plant establishment on contaminated sites might be retarded.

For wood mycologists, the work of Savory (1954a,b) marked an important step in the understanding of decay processes by lignolytic fungi. His description of decay by ascomycete and deuteromycete fungi in wood from industrial water cooling towers revealed a particular pattern observed previously by Bailey & Vestal (1937), Tamblyn (1937) and Barghoorn & Linder (1944) in wood from other situations. This type of decay was termed ‘soft rot’ because of the spongy texture of the wood surface. Soft rot, characterized by hyphal penetration of the wood cell walls in a fashion similar to sap-stain fungi, results in the formation of decay cavities in the central zone of wood cell walls, which leads to their ultimate destruction. Soft rot was therefore described as distinct from the brown and white rot forms of wood decay normally attributed to lignolytic basidiomycetes and some of the larger ascomycetes, such as members of the Xylariaceae. Indeed, members of the genera Daldinia, Hypoxylon and Xylaria have usually been considered to be white rot fungi. However, the boundaries between these three types of fungal decay have since become less clear-cut and a report in this issue, of a study into hyphal penetration of the reaction zone in beechwood (Schwarze and Baum, pp. 129–140), identifies different penetration mechanisms.