It is becoming increasingly apparent that the long-distance signalling associated with many developmental processes is complex and that novel hormone-like signals may play substantial roles. The past decades have seen several substances (e.g. brassinosteroids, systemin and other polypeptides, mevalonic and jasmonic acids, polyamines, oligosaccharides, flavonoids, and quinones) vie for a place among the classical plant hormones (e.g. Spaink, 1996). Recent microinjection and grafting studies have also shown that RNA may act as a long-distance signal (Jorgensen et al., 1998; Xoconostle-Cázares et al., 1999). In this issue, Hannah et al. describe long-distance signalling and the regulation of root–shoot partitioning in dwarf lethal or dosage-dependent lethal (DL) mutants of common bean (Shii et al., 1980, 1981), and present evidence indicating that substances in addition to classical plant hormones (e.g. cytokinins) may be involved.

As in the report by Hannah et al., much of the evidence for roles of unidentified long-distance signals in the control of plant development is indirect. The possibility that a small number of long-distance signals might control a multitude of developmental processes arises through the potential for differences in tissue sensitivity, fluctuations in hormone levels and differences in the nature of responses of different tissues to the same hormone. Consequently, particular hormones may influence numerous processes seemingly simultaneously, yet independently. Even so, long-distance signalling is involved in processes as diverse as root–shoot balance, senescence, branching, flowering, nodulation, stress responses and nutrient uptake. Through comparison of even a few different developmental processes, progress can be made to reveal the true complexity of plant development. Using this approach it is also clear that many unknown signals may be involved.

32nd Annual Air Pollution Workshop

Auburn University in Auburn, AL, USA, April 2000

Air pollution has profound effects on agriculture, forests and natural ecosystems. The first Air Pollution Workshop was held over 30 years ago, and the most vital issues have always been highlighted within this forum. This year, forest health and passive sampling of air pollutants were two key areas of interest.

Summary 421

I. INTRODUCTION 421

II. EFFECTS OF OZONE ON REPRODUCTION 423

1. Pollen germination and pollen tube growth 424

2. Floral initiation and development 428

3. Effects on seed and fruit yield and yield components 433

4. Effects of ozone on seed and fruit quality, germination and seedling growth 437

III. INFLUENCE OF REPRODUCTIVE HABIT AND IMPLICATIONS FOR FIELD-GROWN PLANTS 438

IV. CONCLUSIONS AND FUTURE RESEARCH 441

Acknowledgements 442

References 442

Sexual reproductive development is a crucial stage in the life cycle of higher plants as any impairment of the processes involved might have significant implications for the productivity of crop plants and the survival of native species. There is considerable evidence that exposure to ozone, even at current ambient levels in many industrialized countries, reduces grain and fruit yields and adversely affects yield quality. It is also well established that sensitivity to ozone may differ not only between species, but also between cultivars and populations of individual species, and that the impact of exposure is highly dependent on ozone concentration and the duration and timing of exposure. However, few studies have attempted to distinguish between the direct effects of air pollutants on reproductive development, and indirect effects mediated by injury to the vegetative organs and associated changes in the supply of assimilates and other essential resources to support reproductive growth, or the levels of endogenous growth regulators. This review considers the impact of ozone on the reproductive biology of agricultural and native species, and examines its direct effects on specific reproductive processes. The extent to which compensatory responses redress the adverse effects of exposure is also explored, with particular reference to recent studies of Brassica napus (oilseed rape), Brassica campestris (Wisconsin Fast Plants), Plantago major (greater plantain) and Triticum aestivum (wheat).

Summary 449

I. INTRODUCTION 450

II. THE PARTNERS 451

1. Cyanobionts and their role 451

2. Hosts and their role 453

3. Location of cyanobionts in their hosts 455

III. INITIATION AND DEVELOPMENT OF SYMBIOSES 458

1. Initiation of symbioses 458

2. Geosiphon pyriforme 458

3. Cyanolichens 459

4. Liverworts and hornworts 460

5. Azolla 460

6. Cycads 461

7. Gunnera 461

IV. THE SYMBIOSES 462

1. Geographical distribution and ecological significance 462

2. Benefits to the partners 462

(a) Benefits to the cyanobionts 462

(b) Benefits to the hosts 463

3. Duration and stability 463

4. Mode of transmission and perpetuation 463

5. Recognition between the partners 464

6. Specificity and diversity 464

7. Symbiosis-related genes 465

8. Modifications of the cyanobiont 466

(a) Growth and morphology 466

(b) Photosynthesis and carbon metabolism 467

(c) Glutamine synthetase 467

(d) Heterocysts 469

(e) N2fixation 470

9. Nutrient exchange 471

(a) Carbon 471

(b) Nitrogen 472

V. EVOLUTIONARY ASPECTS 472

VI. ARTIFICIAL SYMBIOSES 474

VII. FUTURE OUTLOOK AND PERSPECTIVES 475

1. Cryptic symbioses 476

2. Developmental profile of symbiotic tissues 476

3. Sensing and signalling 476

4. Genetic aspects 476

5. Physiological and biochemical aspects of nutrient exchange 477

6. Microaerobiosis 477

7. Potential applications 477

Acknowledgements 477

References 477

Cyanobacteria are an ancient, morphologically diverse group of prokaryotes with an oxygenic photosynthesis. Many cyanobacteria also possess the ability to fix N2. Although well suited to an independent existence in nature, some cyanobacteria occur in symbiosis with a wide range of hosts (protists, animals and plants). Among plants, such symbioses have independently evolved in phylogenetically diverse genera belonging to the algae, fungi, bryophytes, pteridophytes, gymnosperms and angiosperms. These are N2-fixing symbioses involving heterocystous cyanobacteria, particularly Nostoc, as cyanobionts (cyanobacterial partners). A given host species associates with only a particular cyanobiont genus but such specificity does not extend to the strain level. The cyanobiont is located under a microaerobic environment in a variety of host organs and tissues (bladder, thalli and cephalodia in fungi; cavities in gametophytes of hornworts and liverworts or fronds of the Azolla sporophyte; coralloid roots in cycads; stem glands in Gunnera). Except for fungi, the hosts form these structures ahead of the cyanobiont infection. The symbiosis lasts for one generation except in Azolla and diatoms, in which it is perpetuated from generation to generation. Within each generation, multiple fresh infections occur as new symbiotic tissues and organs develop. The symbioses are stable over a wide range of environmental conditions, and sensing–signalling between partners ensures their synchronized growth and development. The cyanobiont population is kept constant in relation to the host biomass through controlled initiation and infection, nutrient supply and cell division. In most cases, the partners have remained facultative, with the cyanobiont residing extracellularly in the host. However, in the water-fern Azolla and the freshwater diatom Rhopalodia the association is obligate. The cyanobionts occur intracellularly in diatoms, the fungus Geosiphon and the angiosperm Gunner a. Close cell–cell contact and the development of special structures ensure efficient nutrient exchange between the partners. The mobile nutrients are normal products of the donor cells, although their production is increased in symbiosis. Establishment of cyanobacterial–plant symbioses differs from chloroplast evolution. In these symbioses, the cyanobiont undergoes structural–functional changes suited to its role as provider of fixed N rather than fixed C, and the level of intimacy is far less than that of an organelle. This review provides an updated account of cyanobacterial–plant symbioses, particularly concerning developments during the past 10 yr. Various aspects of these symbioses such as initiation and development, symbiont diversity, recognition and signalling, structural–functional modifications, integration, and nutrient exchange are reviewed and discussed, as are evolutionary aspects and the potential uses of cyanobacterial–plant symbioses. Finally we outline areas that require special attention for future research. Not only will these provide information of academic interest but they will also help to improve the use of Azolla as green manure, to enable us to establish artificial N2-fixing associations with cereals such as rice, and to allow the manipulation of free-living cyanobacteria for photobiological ammonia or hydrogen production or for use as biofertilizers.

Crosses between certain genotypes of common bean result in dwarfing of F1 plants and lethal dwarfing in a proportion of the F2 population. This is under the control of the semi-dominant alleles, DL1 and DL2 at two complementary loci which are expressed in the root and shoot respectively. The various DL genotypes can be simulated by grafting. The graft combination DL1DL1dl2dl2/dl1dl1DL2DL2 was found to have a significantly higher root dry matter fraction than either parent. Lethally dwarfed plants (DL1DL1DL2DL2) and the analogous lethal graft combination (dl1dl1DL2DL2/DL1DL1dl2dl2) exhibit failure of root growth and have very low root fractions. Hybrids or graft combinations with failed roots ceased growth and accumulated large amounts of starch throughout their hypocotyls. In sterile culture, both lethal dwarfs and lethal graft combinations were able to grow roots if sucrose was added to the growth medium. This indicates that a failure of sucrose translocation to the roots is probably responsible for failed root growth. Data from screening the DL genotypes of 49 cultivars could be fully explained using the DL system hypothesis, and grafting proved to be efficient for identifying DL genotype. The DL system might be of fundamental importance in root–shoot partitioning. Current evidence favours the hypothesis that failure of root growth is the outcome of excessively high sink strength of shoots compared to roots, which might arise from signalling incompatibilities between the genotypes.

In a study on the mechanism of stimulated petiole elongation in submerged plants, oxygen concentrations in petioles of the flood-tolerant plant Rumex palustris were measured with micro-electrodes. Short-term submergence lowered petiole partial oxygen pressure to c. 19 kPa whereas prolonged submergence under continuous illumination depressed oxygen levels to c. 8–12 kPa after 24 h. Oxygen levels in petioles depended on the presence of the lamina, even in submerged conditions, and on available light. In darkness, petiole oxygen levels in submerged plants dropped quickly to values as low as 0.5–4 kPa. It is hypothesized that prolonged submergence in the light is accompanied by a decrease in carbon dioxide in the petiole. Submergence-enhanced petiolar elongation rate was compared with emergent plants. Peak daily elongation rates occurred at the end of the dark period in emergent plants, but in the middle of the light period in submerged plants. We suggest that this shift in daily elongation pattern is induced by dependence of growth on photosynthetically derived oxygen in submerged plants. Implications of reduced oxygen for ethylene production are raised. Levels of 1- aminocyclopropane-1-carboxylic acid synthase and 1-aminocyclopropane-1-carboxylic acid oxidase and ethylene sensitivity are cited as potential factors in hypoxia-induced ethylene release.

Three MADS-box cDNA clones and two corresponding genomic sequences (gDNAs) have been isolated from the bryophyte Physcomitrella patens and sequenced. Our findings indicate that the genes may be expressed in a tissue- or age-specific manner, and that expression of one of them is regulated by an alternative splicing mechanism. Conceptual translation of the clones reveals that the encoded MADS-domain proteins have the typical plant-domain pattern (MIKC). Additionally, there is a high degree of conservation of intron number and positions between angiosperm MADS-box genes and the moss loci. These observations confirm the homology of moss and higher plant MADS-box genes. We conclude that the MIKC pattern evolved in MADS-box genes after the separation of the plant lineage from that of fungi and animals, and that it must have been present in the common ancestor of mosses, ferns and seed plants. Therefore it evolved at least 400 million yr ago. Phylogenetic analysis of a large subset of the sequenced plant MADS-box genes, incorporating those from P. patens, indicates that the bryophyte genes are not orthologues of spermatophyte genes belonging to any of the well recognized higher plant gene subfamilies. This conclusion accords well with reports that the known fern MADS-box genes also comprise subfamilies distinct from those of higher plants. Therefore we tentatively propose that the gene duplication and diversification events that created the MADS-box gene subfamilies, discernible in extant angiosperm and other spermatophyte groups, occurred after separation of the moss and fern lineages from the lineage which produced the higher plants.

Relationships between nitrogen (N) content and growth are routinely measured in plants. This study determined the effects of N on the separate morphological and physiological components of plant growth, to assess how N-limited growth is effected through these components. Lettuce (Lactuca sativa) plants were grown hydroponically under contrasting N-supply regimes, with the external N supply either maintained continuously throughout the period of study, or withdrawn for up to 14 d. Richards' growth functions, selected using an objective curve-fitting technique, accounted for 99.0 and 99.1% of the variation in plant dry weight for control and N-limited plants respectively. Sublinear relationships occurred between N and relative growth rates under restricted N-supply conditions, consistent with previous observations. There were effects of treatment on morphological and physiological components of growth. Leaf weight ratio increased over time in control plants and decreased in N- limited plants. Shoot:root ratio followed a similar pattern. On a whole-plant basis, assimilation of carbon decreased in N-limited plants, a response paralleled by differences in stomatal conductance between treatments. Changes in C assimilation, expressed as a function of stomatal conductance to water vapour, suggest that the effects of N limitation on growth did not result directly from a lack of photosynthetic enzymes. Relationships between plant N content and components of growth will depend on the availability of different N pools for remobilization and use within the plant.

The effects of increased nitrogen influx on Sphagnum growth and on interspecific competition between Sphagnum species were studied in a 3-yr experiment in mires situated in two areas with different rates of airborne N deposition. Sphagnum growth was recorded after various supplementary N influxes (0, 1, 3, 5 and 10 g m −2 yr−1) in hummocks and lawn communities. Sphagnum biomass production decreased with increasing N influx in both areas. After the first season at the low-deposition site, Sphagnum showed an increased growth in length with the intermediate N treatment, but in the second and third seasons the control treatment had the highest growth in length. Capitulum dry mass increased with increasing N influx. Sphagnum N concentration and N/P quotient were higher at the high- than at the low-deposition site. The low quotient at the low-deposition site, together with the initial growth increase with intermediate N supplements, indicates that growth was N-limited at this site, but our lowest N supplement was sufficient to reduce growth. The N treatments had no effect on interspecific competition between the Sphagnum species. This indicates that the species have similar responses to N. The species studied all occur naturally on ombrotrophic, N-poor sites and show low tolerances to increased N influx. Reduced Sphagnum production may affect the carbon balance, changing the mires from C sinks to sources.

The dynamic-chamber technique was used to investigate the correlation between NH3 and NO2 fluxes and different climatic and physiological parameters: air temperature; relative air humidity; photosynthetic photon fluence rate; NH3 and NO2 concentrations; transpiration rate; leaf conductance for water vapour; and photosynthetic activity. The experiments were performed with twigs from the sun crown of mature beech trees (Fagus sylvatica) at a field site (Höglwald, Germany), and with 12-wk-old beech seedlings under controlled conditions. Both sets of experiments showed that NO2 and NH3 fluxes depended linearly on NO2 and NH3 concentration, respectively, in the concentration ranges representative for the field site studied, and on water-vapour conductance as a measure for stomatal aperture. The NO2 compensation point determined in the field studies (the atmospheric NO2 concentration with no net NO2 flux) was 1.8–1.9 nmol mol−1. The NH3 compensation point varied between 3.3 and 3.5 nmol mol−1 in the field experiments, and was 3.0 nmol mol−1 in the experiments under controlled conditions. The climatic factors T and PPFR were found to influence both NO2 and NH3 fluxes indirectly, by changing stomatal conductance. Whilst NO2 flux showed a response to changing relative humidity that could be explained by altered stomatal conductance, increased NH3 flux with increasing relative humidity (>50%) depended on other factors. The exchange of NO2 between above-ground parts of beech trees and the atmosphere could be explained exclusively by uptake or emission of NO2 through the stomata, as indicated by the quotient between measured and predicted NO2 conductance of approx. 1 under all environmental conditions examined. Neither internal mesophyll resistances nor additional sinks could be observed for adult trees or for beech seedlings. By contrast, the patterns of NH3 flux could not be explained by an exclusive exchange of NH3 through the stomata. Deposition into additional sinks on the leaf surface, as indicated by an increase in the quotient between measured and predicted NH3 conductance, gained importance in high air humidity, when the stomata were closed or nearly closed and/or when atmospheric NH3 concentrations were high. Although patterns of NH3 gas exchange did not differ between different months or years at high NH3 concentrations (c. 140 nmol mol−1), it must be assumed that emission or deposition fluxes at low ambient NH3 concentration (0.8 and 4.5 nmol mol−1) might vary significantly with time because of variation in the NH3 compensation point.

Fruiting and deblossomed plants of strawberry (Fragaria × ananassa) were exposed to 92 ppb ozone or filtered air in open-top chambers for 69 d. Flower and fruit production, relative growth rate of leaf area, leaf gas exchange and plant biomass were investigated. Ozone caused an initial acceleration in inflorescence production, which was followed by a reduction in inflorescence production, fruit set, and, later, individual fruit weight, although total fruit yield was not affected before the end of the fumigation period. Ozone accelerated leaf senescence and had a greater negative effect on the rate of photosynthesis in older than in younger leaves in fruiting and deblossomed plants, but the response of net photosynthesis to ozone did not differ between the two groups of plants. Relative growth rate of leaf area was the first parameter to be reduced by ozone fumigation, with the effect being significant in fruiting, but not in deblossomed, plants. Final above-ground biomass was also significantly decreased by ozone in fruiting plants, but not in deblossomed plants. Root and crown biomass were not significantly affected by ozone fumigation in either fruiting or deblossomed plants.

The activities of nitrate reductase and glutamine synthetase were evaluated in young plants of Faidherbia albida, a tropical woody legume, fed with different N sources under hydroponic conditions. Results showed that assimilation of both NO3− and NH4+ preferentially took place in shoots. A basal amount of nitrate reductase activity was detected in shoots of plants grown with an NO3−-free solution or placed under N2-fixing conditions, and also in nodules of N2-fixing plants. This strongly suggests that constitutive nitrate reductase activity is present in these organs. Analyses of the soluble nitrogenous content showed that the major form of N in the different organs was α-amino acids (particularly amides), irrespective of the N status of the culture conditions. The same result was obtained for nodulated plants grown in local sandy soil. In this case, amide-N generally accounted for more than 40% of the total soluble N. This was especially true in nodules. Ureide-N never exceeded 9% of the total soluble N and did not appear to increase with increasing nodule nitrogenase activity. Amides were also predominant in three N2-fixing Sahelian acacias (Acacia seyal, A. nilotica and A. tortilis), showing that F. albida does not differ from Sahelian Acacia in terms of the metabolism of fixed N. However, like another Sahelian acacia growing preferentially near water (A. nilotica), F. albida can be distinguished from acacias growing strictly in arid zones (A. seyal and A. tortilis) in terms of initial growth, water and nitrate management.

The photobiont ultrastructure of the epiphytic lichens Bryoria fuscescens and Bryoria fremontii was studied along the pollution gradient from two Cu-Ni smelters in Nikel and Monchegorsk in northern Finland and north-western Russia. The relationship between ultrastructural characteristics of B. fuscescens and environmental factors (i.e. climate, atmospheric SO2 and bark element concentrations) was studied by using a principal component analysis (PCA) aiming to assess the air quality in a northern environment. Based on PCA, increased plasmolysis and mitochondrial changes in the Trebouxia photobiont were significantly correlated with elevated pollutant concentrations. Degenerated cells, showing altered chloroplasts and electron-translucent pyrenoglobuli, occurred in lichens growing 35–50 km from the Monchegorsk smelter. Cell wall and cytoplasmic lipid volumes, and size of pyrenoglobuli, positively correlated with the distance from the Monchegorsk smelter. Vacuoles and electron-opaque vacuolar deposits were significantly increased at the Finnish site in the vicinity of a pulp mill. Swelling of mitochondrial cristae and thylakoids showed little correlation with environmental factors, but indicated of initial stage of injuries and were observed at several slightly polluted sites in northern Finland and north-western Russia. The results suggest that the severe photobiont injuries of lichens are strongly associated with poor air quality.

Accumulation of lead in the crustose lichen Acarospora smaragdula sensu lato is reported in the vicinity of an ore- processing plant where it is subjected to acidification and metal particulate fallout. A combination of light microscopy, X-ray element mapping, field emission scanning electron microscopy (FESEM) and other analytical techniques identifies Pb accumulation within specific fungal tissues derived from smelter particles (PM10s). No Pb was detected within the photobiont layer. Our studies suggest that Pb is highly mobile under the prevailing acidic conditions, and is fixed within the lichen cortex and melanized apothecia. Lead is also accumulated within the medulla and at the rock–lichen interface where it may precipitate as amorphous botryoidal encrustations on medullary hyphae and iron-rich particles. Modern FESEMs and microprobes enable analysis of minute quantities of material, and are important tools in understanding the fate of metals within lichens necessary to develop their use as predictive and sensitive bioindicators of aerial particulate contaminants. We suggest that crustose lichens, hitherto largely ignored in metal pollution studies, may be useful bioindicators of aerial particulate contaminants in polluted areas where macrolichens are absent.

Plant species that persist during succession, from the colonization to the stabilization stages, face major environmental changes. Such changes are believed to have significant effects on species performance. In subarctic coastal dune systems, Leymus mollis colonizes the embryo dunes, on the upper limit of the beach. It reaches its maximum density on the foredune, but also grows on older, stabilized ridges. This paper reports on the phenotypic variations of some ecophysiological traits associated with the persistence of L. mollis on a dune system on the east coast of Hudson Bay (northern Quebec). Leymus mollis ramets tend to have a lower net carbon assimilation rate and water use efficiency, and a higher substomatal CO2 concentration on the stabilized dune than on the foredune. However, these physiological differences cannot be explained by differences in leaf morphology or nitrogen content. Under controlled conditions, ecophysiological differences observed in the field disappear, suggesting that these are not genetic but determined by environmental changes along the foredune-stabilized dune gradient. We propose that higher net carbon assimilation rate on the foredune might be related to higher sink strength in relation to the growth-stimulating effect of sand burial.

Very little is known about the physiological interactions between plant hosts and symptomless endophytic fungi despite their widespread occurrence. We investigated the impact of two such fungi, Colletotrichum musae and Fusarium moniliforme, upon the photosynthetic capacity of two crop plants, banana and maize, respectively. Endophyte-free plants were obtained first and then infected with the fungi. Measurements of total chlorophyll content revealed very little difference between endophyte-free and infected plants of banana, whereas in maize they showed 50% reductions in the endophyte-infected plants. The maximum photochemical capacity (Fv/Fm ) was measured in order to determine if the plants had any photoinhibitory effect caused by biotic or abiotic factors. After 45 d of growth, endophyte-free banana plants had similar values of Fv/Fm to plants typical of nonstressed conditions, whereas the endophyte-infected plants showed a reduction of approx. 15%. Unlike banana, infected maize plants displayed values of Fv/Fm similar to those of control and endophyte-free plants, indicating that the maximum photochemical capacity was not affected by infection. The light response curves of both species showed that the photosynthethic capacity was severely reduced in endophyte-infected plants, reaching saturation at c. 400 μmol m−2s−1 whereas the control and endophyte-free plants were saturated at much higher photon flux densities. In banana the effect seemed to be due to an impairment of electron transport in the thylakoid membranes. By contrast, reduction of the photosynthetic capacity in maize was due to a reduction in chlorophyll content, leading to a decrease in the electron transport components and a consequent reduction in carbohydrate synthesis. It is possible that the reduction in the maximum yield of photosynthesis in both crops was caused by toxins produced by the fungi. Nevertheless there were no major macroscopic effects on the plants to indicate disease symptoms.

A survey of the endophytic fungi in fronds of Livistona chinensis was carried out in Hong Kong. The endophyte assemblages identified using morphological characters consisted of 16 named species and 19 ‘morphospecies’, the latter grouped based on cultural morphology and growth rates. Arrangement of taxa into morphospecies does not reflect species phylogeny, and therefore selected morphospecies were further identified based on ribosomal DNA (rDNA) sequence analysis. The 5.8S gene and flanking internal transcribed spacers (ITS1 and ITS2) regions of rDNA from 19 representative morphospecies were amplified by the polymerase chain reaction and sequenced. Phylogenetic analysis based on 5.8S gene sequences showed that these morphospecies were filamentous Ascomycota, belonging in the Loculoascomycetes and Pyrenomycetes. Further identification was conducted by means of sequence comparison and phylogenetic analysis of both the ITS and 5.8S regions. Results showed that MS704 belonged to the genus Diaporthe and its anamorph Phomopsis of the Valsaceae. MS594 was inferred to be Mycosphaerella and its anamorph Cladosporium of the Mycosphaerellaceae. MS339, MS366, MS370, MS395, MS1033, MS1083 and MS1092 were placed in the genus Xylaria of the Xylariaceae. MS194, MS375 and MS1028 were close to the Clypeosphaeriaceae. MS191 and MS316 were closely related to the Pleosporaceae within the Dothideales. The other 5 morphospecies, MS786, MS1043, MS1065, MS1076 and MS1095, probably belong in the Xylariales. The value of using DNA sequence analysis in the identification of endophytes is discussed.

Measurements of the electric potential difference across the hyphal wall and the cell membrane were made on external hyphae of three species of arbuscular mycorrhizal fungus Gigaspora margarita, Scutellospora calospora and Glomus coronatum and on germ tubes of Gi. margarita. The values of transmembrane electric potential difference recorded (∼ –40 mV) are less negative than those previously reported from hyphae of arbuscular mycorrhizal fungi closely associated with roots and from filamentous fungi. The external hyphae of arbuscular mycorrhizal fungi grown in soil had similar values of electric potential difference to those grown in soil-less culture, and to germ tubes. Thermodynamic calculations showed that despite these low values of electric potential difference, efficient high-affinity uptake of phosphate is possible. The transmembrane electric potential difference of germ tubes of Gi. margarita became more negative when plant root extract was added to the medium, showing for the first time that the early stages of interaction between plant and fungus occur via direct effects on the plasma membrane rather than via effects on gene expression. Addition of K+ reversibly depolarized the transmembrane electric potential difference of germ tubes of Gi. margarita, indicating that despite the low electric potential difference the fungus has control over the permeability of the plasmamembrane to K+.