The capacity to accumulate cadmium (Cd) and zinc (Zn) was compared in Thlaspi goesingense and four populations of Thlaspi caerulescens. Two populations of T. caerulescens were grown in hydroponics at five concentrations of Cd. In addition, plants were grown in pots containing compost in which three different concentrations of Cd and two concentrations of Zn were added. A field trial was conducted to compare Zn and Cd uptake by three populations of T. caerulescens on nine selected plots of the Woburn Market Garden Experiment (UK) which had been contaminated to different degrees with heavy metals owing to past applications of sewage sludge. Results show that the four populations of T. caerulescens had the same ability to hyperaccumulate Zn but were significantly different in terms of Cd accumulation. Two populations of T. caerulescens from Southern France accumulated much more Cd than the populations from Prayon (Belgium) and Whitesike (UK). Generally, uptake of Cd was not decreased by increased concentrations of Zn in the substrate. These results indicate that the mechanisms of Cd and Zn hyperaccumulation are not identical in this species. This is the first report of hyperaccumulation of Cd by T. goesingense, but the growth of this species was markedly reduced by the large concentrations of Zn in the substrate. Future work should focus on the differences between Cd and Zn uptake in hyperaccumulator plants at the species and population level.

Changes in the production and turnover of roots in forests and grasslands in response to rising atmospheric CO2 concentrations, elevated temperatures, altered precipitation, or nitrogen deposition could be a key link between plant responses and longer-term changes in soil organic matter and ecosystem carbon balance. Here we summarize the experimental observations, ideas, and new hypotheses developed in this area in the rest of this volume. Three central questions are posed. Do elevated atmospheric CO2, nitrogen deposition, and climatic change alter the dynamics of root production and mortality? What are the consequences of root responses to plant physiological processes? What are the implications of root dynamics to soil microbial communities and the fate of carbon in soil? Ecosystem-level observations of root production and mortality in response to global change parameters are just starting to emerge. The challenge to root biologists is to overcome the profound methodological and analytical problems and assemble a more comprehensive data set with sufficient ancillary data that differences between ecosystems can be explained. The assemblage of information reported herein on global patterns of root turnover, basic root biology that controls responses to environmental variables, and new observations of root and associated microbial responses to atmospheric and climatic change helps to sharpen our questions and stimulate new research approaches. New hypotheses have been developed to explain why responses of root turnover might differ in contrasting systems, how carbon allocation to roots is controlled, and how species differences in root chemistry might explain the ultimate fate of carbon in soil. These hypotheses and the enthusiasm for pursuing them are based on the firm belief that a deeper understanding of root dynamics is critical to describing the integrated response of ecosystems to global change.

Summary

I. INTRODUCTION – A HYPOTHESIS 374

II. EFFECTS OF CALCIUM ON PHYSIOLOGICAL PROCESSES 376

1. The chemical uniqueness of calcium 376

(a) Cytotoxicity 376

(b) Binding properties 376

(c) Stimulation/displacement potential 376

2. Calcium signaling and plant responses to environmental stress 377

(a) Control principles 377

(b) Carbohydrate metabolism 378

(c) Synthesis and function of membranes and cell walls 379

(d) Disease resistance and wound repair 380

(e) Cold tolerance 381

(f) Stomatal regulation 382

III. CALCIUM UPTAKE AND DISTRIBUTION AT THE WHOLE-PLANT LEVEL 382

1. Uptake at the root-soil interface 382

2. Transport and exchange in stems 384

3. Exchange of calcium by foliage 385

IV. ECOSYSTEM PROCESSES AND CALCIUM SUPPLY 387

1. Plant succession and soil acidification 387

2. Plant adaptations to nutrient deficiency 389

(a) Morphological adaptations 389

(b) Physiological adaptations 390

V. PLANT AND ECOSYSTEM RESPONSES TO HUMAN ALTERATIONS IN CALCIUM SUPPLY 391

1. Increased atmospheric inputs of acidity 391

(a) Reductions in soil cation pools 392

(b) Inhibition of calcium uptake and effects on root function 393

(c) Increased leaching of calcium from foliage 395

(d) Physiological indicators of altered forest function 397

(e) Wood chemistry, structure and function 402

2. Forest management 404

(a) Harvesting effects on nutrient supply 404

(b) Managing forest nutrient supply 405

VI. CONCLUSION 407

1. Whole-tree perspectives 407

2. Ecosystem perspectives 408

VII. EVALUATION OF THE HYPOTHESIS 410

Acknowledgements 411

References 411

Summary

Calcium occupies a unique position among plant nutrients both chemically and functionally. Its chemical properties allow it to exist in a wide range of binding states and to serve in both structural and messenger roles. Despite its importance in many plant processes, Ca mobility is low, making Ca uptake and distribution rate a limiting process for many key plant functions. Ca plays an essential role in regulating many physiological processes that influence both growth and responses to environmental stresses. Included among these are: water and solute movement, influenced through effects on membrane structure and stomatal function; cell division and cell wall synthesis; direct or signaling roles in systems involved in plant defense and repair of damage from biotic and abiotic stress; rates of respiratory metabolism and translocation; and structural chemistry and function of woody support tissues. Forest trees, because of their size and age capacity, have been examined for evidence of limitations imposed by the timing and level of Ca supply. Examination of Ca physiology and biogeochemical cycling for forested systems reveals many indications that Ca supply places important limitations on forest structure and function. These limitations are likely to be most significant with older trees, later successional stages, high levels of soil acidity and/or high canopy Ca leaching losses, or under conditions where plant competition is high or transpiration is limited by high humidity or low soil moisture. Evidence of structural and physiological adaptations of forests to limited Ca supply; indicators of system dysfunction at many levels under reduced Ca supply; and the positive responses of diverse indicators of forest vitality in liming experiments indicate that Ca is more important to forest function and structure than has generally been recognized. Lack of recognition of Ca limitations is due in part to that fact some important plant functions are controlled by changes in very small physiologically active pools within the cytoplasm, and whole-leaf Ca levels may not reflect these limitations. An additional aspect is the fact that Ca availability has declined significantly for many forests in just the past few decades. Additional research on the role of Ca supply in resistance of forests to disease, changes in structural integrity of woody tissues, restrictions on rooting patterns and function, and uptake of other nutrients, notably N, is needed. Increased understanding of the physiological ecology of Ca supply can be anticipated to provide important insights that will aid in future protection and management of both natural and commercial forest systems.

An appendix is available for this article which was not included in the print edition of the Journal. The appendix may be found at the rear of the full text, PDF version of the article.

A popular conceptual model asserts that shade tolerance is characterized by morphological and physiological traits that enhance the net rate of carbon capture in low light. We tested this model by quantitatively reviewing growth, leaf lifespan, CO2 exchange and morphological data from 76 studies on woody seedlings grown under conditions of low light. Data were placed into three tolerance categories (intolerant, intermediate, tolerant), two light categories (less than 4% and 4–12%) and two leaf phenology categories (broad-leaved evergreen and winter deciduous). For both evergreen and deciduous groups, intolerant species had traits conferring greater growth potential than tolerant species in both light categories. These traits included greater leaf mass ratio, leaf area ratio, specific leaf area and mass-based photosynthetic rates above light compensation. However, in 0–4% light, growth rates were similar for intolerant and tolerant species, because low light together with higher respiration rates for intolerant species limited the expression of their growth potential differences. Deciduous and evergreen intolerant species were similar in many respects. However, both intermediate and tolerant deciduous species had markedly lower leaf mass ratios and higher root mass ratios than intermediate and tolerant evergreen species. In addition, deciduous species and intolerant evergreens must cope with as much as sixfold higher leaf turnover rates than tolerant evergreen species. Thus, rather than maximizing growth rates in low light, tolerant evergreen species minimize biomass loss through long leaf lifespans and low respiration rates. Tolerant deciduous species also minimize biomass losses by minimizing whole-plant respiration rates but they accomplish low biomass turnover though low leaf mass ratio and not low leaf turnover rates. Furthermore, unlike most tropical evergreens, tolerant deciduous species can gain large fractions of their total growing season carbon during short periods when the overstory is leafless and then allocate this carbon to storage (as reflected by high root mass ratios) rather than new leaves. In conclusion, we found no support for the low-light-enhanced carbon capture model of shade tolerance as viewed strictly from the perspective of physiological growth capacity. This can be explained by the disadvantages to net growth and survival of maintaining a high growth potential at low light, because high growth potential results in greater rates of whole-plant respiration, tissue turnover, herbivory and mechanical damage and in decreased storage. Thus, shade tolerance can be characterized by traits that maximize survival and net growth, where net growth includes losses to all agents.

Summary 373

I. Introduction: Arum-types and Paris-types 374

II. Possible functional implications 375

III. Extent of the two classes in the plant kingdom 377

1. Bryophytes and Pteridophytes 377

2. Gymnosperms 379

3. Angiosperms 379

IV. Is the distinction between classes useful? 383

V. The structural basis 383

VI. The role of the fungal genome 384

VII. Physiology revisited 384

VIII. Conclusions 385

Acknowledgements 386

References 386

This review describes diversity in the structure of (vesicular)–arbuscular (VA) mycorrhizas, i.e. endomycorrhizas formed by Glomalean fungi. In particular, we consider the extent in the plant kingdom of the two classes first described by Gallaud (1905). These are: (1) the Arum-type, defined on the basis of an extensive intercellular phase of hyphal growth in the root cortex and development of terminal arbuscules on intracellular hyphal branches; (2) the Paris-type, defined by the absence of the intercellular phase and presence of extensive intracellular hyphal coils. Arbuscules are intercalary structures on the coils. However, there have been many reports that in Paris-types arbuscules are relatively few in numbers, small, or absent altogether.

A survey of the literature has revealed that Paris-types occur more frequently in the plant kingdom than Arum-types and predominate in ferns, gymnosperms and many wild angiosperms. The cultivated herbs that are the subject of much experimental work are mostly Arum-types. Although evidence is still limited, there are differences at the family level. In 41 angiosperm families there are records of only Paris-type VA mycorrhizas and in 30 families records of only Arum-types. Another 21 families have examples of both classes, or intermediates between them. Accordingly, we consider whether the original division into two classes is still useful. We conclude that it is when considering the physiology of the symbiosis and especially the issue of whether different fungus/host interfaces have specialized roles in transfer of inorganic nutrients and organic carbon between the partners. If there is no such specialization between hyphal coils and arbuscules, then the latter might not be necessary for the function of Paris-types. This would account for reports of the infrequency or absence of arbuscules in this class. The control exerted on structures by the genomes of host and fungus, and possible reasons (anatomical and physiological) for the existence of the VA mycorrhizal structures, are discussed. The presence or absence of extensive intercellular spaces and differences in the wall structure of cortical cells might be particularly important in determining which type of VA mycorrhiza is formed.

Despite the usually high abundance of iron (Fe) in soils, the low solubility of Fe-bearing minerals restricts the available Fe pools in most aerobic soils to levels that are far below those required for microbial or plant growth. To acquire the necessary amounts of Fe from the environment, organisms have evolved mechanisms that enhance the solubility and dissolution rate of Fe(iii) oxyhydroxides prevailing in aerobic soils. Chemically, these mechanisms are based on weakening of the Fe–O bond by reduction, chelation and protonation. Physiologically, two distinct and in all known cases mutually exclusive strategies can be distinguished: the excretion of siderophores capable of solubilizing external ferric Fe and subsequent uptake of the ferric siderophore complex; and reduction of Fe(iii) prior to uptake of the more soluble Fe2+ ion. With the exception of graminaceous species, in which Fe uptake is based on the former mechanism, the latter strategy is found in all cormophytes and certain algae, yeast and bacteria. In higher plants, the increase in their capacity to convert extracellular ferric to ferrous Fe is part of a series of physiological and morphological events that act in concert to achieve appropriate internal levels of Fe. It is this amalgam of features that determines the Fe efficiency of a species or cultivar that in turn affects the yield of economically important plants and the natural distribution of species. Adaptive changes to limited Fe availability have been studied at the molecular, physiological and whole-plant level. This review summarises current knowledge of the components of reduction-based Fe uptake in plants and presents an integrated view of the present understanding of mechanisms that control the rate and extent of Fe absorption by roots.

Summary 241

I. INTRODUCTION: CAESIUM IN THE ENVIRONMENT 242

II. UPTAKE OF CAESIUM BY PLANT ROOTS 243

1. Evidence for multiple mechanisms of Cs+uptake by plant roots 243

2. Caesium uptake is affected by the presence of other cations 244

3. Caesium inhibits the uptake of other cations 244

III. MOLECULAR MECHANISMS CATALYSING CAESIUM UPTAKE 245

1. ‘High-affinity’ transport mechanisms 245

2. Inward-rectifying potassium (KIR) channels 245

3. Outward-rectifying potassium (KOR) channels 248

4. Voltage-insensitive cation (VIC) channels 249

5. Ca2+-permeable channels 249

IV. MODELLING CAESIUM INFLUX TO ROOT CELLS 249

1. Predicted Cs+influx through high-affinity mechanisms 250

2. Predicted Cs+influx through cation channels 250

3. Predicted dependence of Cs+influx on [Cs+]ext 252

V. PERSPECTIVE 253

Acknowledgements 254

References 254

Caesium (Cs) is a Group I alkali metal with chemical properties similar to potassium (K). It is present in solution as the monovalent cation Cs+. Concentrations of the stable caesium isotope 133Cs in soils occur up to 25 μg g−1 dry soil. This corresponds to low micromolar Cs+ concentrations in soil solutions. There is no known role for Cs in plant nutrition, but excessive Cs can be toxic to plants. Studies of the mechanism of Cs+ uptake are important for understanding the implications arising from releases of radioisotopes of Cs, which are produced in nuclear reactors and thermonuclear explosions. Two radioisotopes of Cs (134Cs and 137Cs) are of environmental concern owing to their relatively long half-lives, emissions of β and γ radiation during decay and rapid incorporation into biological systems. The soil concentrations of these radioisotopes are six orders of magnitude lower than those of 133Cs. Early physiological studies demonstrated that K+ and Cs+ competed for influx to excised roots, suggesting that the influx of these cations to root cells is mediated by the same molecular mechanism(s). The molecular identity and/or electrophysiological signature of many K+ transporters expressed in the plasma membrane of root cells have been described. The inward-rectifying K+ (KIR), outward-rectifying K+ (KOR) and voltage-insensitive cation (VIC) channels are all permeable to Cs+ and, by analogy with their bacterial counterparts, it is likely that ‘high-affinity’ K+/H+ symporters (tentatively ascribed here to KUP genes) also transport Cs+. By modelling cation fluxes through these transporters into a stereotypical root cell, it can be predicted that VIC channels mediate most (30–90%) of the Cs+ influx under physiological conditions and that the KUP transporters mediate the bulk of the remainder. Cation influx through KIR channels is likely to be blocked by extracellular Cs+ under typical ionic conditions in the soil. Further simulations suggest that the combined Cs+ influxes through VIC channels and KUP transporters can produce the characteristic ‘dual isotherm’ relationship between Cs+ influx to excised roots and external Cs+ concentrations below 200 μM. Thus, molecular targets for modulating Cs+ influx to root cells have been identified. This information can be used to direct future genetic modification of plants, allowing them to accumulate more, or less, Cs and thereby to remediate contaminated sites.

We studied interspecific and ontogenetic relationships between the size of a leaf and the primary diameter of the internode bearing it. Although these two variables are known to be strongly correlated across species, the form of this relationship has not been studied. In a re-analysis of published data on interspecific comparisons of 69 temperate tree species, we showed the existence of a strong relationship between twig cross-sectional area (before secondary growth) and surface area of leaves borne by it, within each of three morphological groups, deciduous angiosperms, evergreen angiosperms, and gymnosperms. Within each of these groups, this relationship is isometric: across species, primary cross-sectional area of the stem increases proportionally with leaf surface area. When we consider the relationship between the cross-sectional area of a twig and the surface area of one leaf borne by it, the y-intercepts for this relation are different for the three groups. However, when total leaf surface area per first-year shoot is considered, no differences remained between gymnosperms and evergreen angiosperms, but deciduous angiosperms continued to be distinct. This difference between deciduous and evergreen groups could be due to differences in leaf volume (evergreen species have thicker leaves than deciduous) or in traits related to a trade-off between life span of leaves and their physiological behaviour.

We present results of the first quantitative study of the relationship between leaf size and primary diameter of the stem during ontogeny. Both these parameters increase during development of the plant from seedling to adult. For the four tree species examined, the relationship between primary cross-sectional area of the stem and leaf surface area is also isometric.

These results bear on a functional interpretation of the relationship between leaf and stem dimensions, suggesting that vascular supply is directly proportional to the requirements of leaves supported by the stem.

There is a growing recognition that accurate predictions of plant and ecosystem responses to global change require a better understanding of the mechanisms that control acquisition of growth-limiting resources. One such key mechanism is root physiological capacity to acquire nutrients. Changes in kinetics of root nitrogen (N) uptake might influence the extent to which terrestrial ecosystems will be able to sequester excesses in carbon (C) and N loads. Despite its significant role in determining plant and ecosystem cycling of C and N, there is little information on whether, or how, root nutrient uptake responds to global change. In this review various components of global change, namely increased CO2 concentration, increased soil temperature and increased atmospheric N deposition and their effects on kinetics of root nutrient uptake are examined. The response of root nutrient uptake kinetics to high CO2 is highly variable. Most of this variability might be attributable to differences in experimental protocols, but more recent evidence suggests that kinetic responses to high CO2 are also species-specific. This raises the possibility that elevated CO2 might alter community composition by shifting the competitive interaction of co-occurring species. Uptake of NH4+ and NO3− seem to be differentially sensitive to high CO2, which could influence ecosystem trajectory toward N saturation. Increased soil temperature might increase N and P uptake capacity to a greater extent in species from warm and fluctuating soil habitats than in species from cold and stable soil environments. The few available data also indicate that increased soil temperature elicits a differential effect on uptake of NH4+ versus NO3−. Root uptake kinetics are generally down-regulated in response to long-term exposure to atmospheric N deposition. The extent of this down-regulation might, however, vary among species, stages of succession, land-use history and plant demand. Nonetheless, it is suggested that root N uptake kinetics might be an accurate biological indicator of the ecosystem capacity to retain N. The results reviewed here clearly highlight the scanty nature of the literature in the area of root nutrient absorption responses to global change. It is also clear that effects of one component of global change on root nutrient absorption capacity might be counterbalanced by another. Therefore, the generalizations offered here must be viewed with caution and more effort should be directed to rigorously test these initial observations in future research.

Laboratory experiments have shown a large difference in specific leaf area (SLA, leaf area: leaf mass) between species from nutrient-poor and nutrient-rich habitats, but no systematic difference in the construction costs (the amount of glucose required to construct 1 g biomass). We examined how far these patterns are congruent with those from field-grown plants. An analysis was made of the vegetation in a range of grasslands and heathlands differing in productivity. The SLA of the dominant species in 15 different habitats was determined, as well as chemical composition and construction costs of bulk samples of leaves. SLA in the field was generally lower than in the laboratory, but showed consistency in that the ranking across species remained the same. Species from highly productive habitats had higher SLA than those from sites of low productivity, although individual species sometimes deviated substantially from the general trend. Construction costs were similar for plants from different habitats. This was mainly due to the positive correlation between an expensive class of compounds (proteins) and a cheap one (minerals).

Colonization of mycorrhizal fungi was studied in a Pinus muricata forest on the coast in California, USA, burned by a stand-replacing wildfire in October 1995. Naturally established field seedlings of P. muricata were harvested 1 yr after the fire. The species composition of the mycorrhizal fungi on these field seedlings was dominated by Rhizopogon species, Wilcoxina mikolae and Tomentella sublilacina. Bioassays, set up with soil collected immediately after the fire, were used to determine which mycorrhizal species had colonized the burned area from resistant propagules. The P. muricata seedlings in these bioassays were dominated by suilloid and ascomycetous fungi, the same fungi which dominated the mycorrhizal flora of seedlings in pre-fire bioassays derived from the same forest site, suggesting that resistant propagules were the primary inoculum source for naturally establishing seedlings. Drying of post-fire soil for 1 month raised the number of bioassay seedlings associated with Rhizopogon olivaceotinctus, while the number of bioassay seedlings associated with Rhizopogon ochraceorubens was reduced. Fire appeared to have either stimulated or provided a competitive advantage to R. olivaceotinctus, which increased in abundance on the post-fire bioassay and field seedlings. Soil collected from the burned area was diluted with sterile soil in three different concentrations, and the number and frequency of mycorrhizal taxa on bioassay seedlings decreased with increased dilution. Although precise quantification was not possible, propagules of the Rhizopogon species were much more abundant than those of Tomentella or Wilcoxina species. Differences between the mycorrhizal associates of bioassay seedlings, naturally regenerated seedlings, and different inoculum sources are discussed.

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.

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.

As a result of increasing anthropogenic nitrogen deposition, N availability in many forest ecosystems, which are normally N-limited, has been enhanced. We discuss the impacts of this increased N availability on the ectomycorrhizal (ECM) symbiosis which is generally regarded as an adaptation to nutrient limited conditions. Nitrogen deposition can influence fruit-body formation by ECM fungi, the production and distribution of the extraradical mycelium in the soil and the formation of ectomycorrhizas.

Available data from long-term N deposition studies indicate that the most prominent effects might be discernible above-ground (i.e. on the formation of fruit bodies). ‘Generalist’ species, forming a symbiosis with a wide range of tree species, seem to be less affected by increased N availability than ‘specialist’ species, especially those living in symbiosis with conifers. However, the importance of below-ground investigations to determine the impacts of N deposition on the ECM symbiosis must not be underestimated. Culture experiments show an optimum N concentration for the formation of extraradical mycelium and mycorrhizas. Often, negative effects only become visible at comparatively high N concentrations, but the use of a few easily cultivated species of ECM fungi, which are adapted to higher N concentrations, undermines our ability to generalize.

So far, N deposition experiments in the field have only shown minor changes in the below-ground mycorrhizal population, as estimated from the investigation of mycorrhizal root tips. However, effects on the ECM mycelium, which is the main fungal component in terms of nutrient uptake, cannot be excluded and need further consideration.

Because the photoassimilate supply from the plant to the fungal partner is crucial for the maintenance of the ECM symbiosis, we discuss the possible physiological implications of increasing N inputs on the allocation of C to the fungus. Together with ultrastructural changes, physiological effects might precede obvious visible changes and might therefore be useful early indicators of negative impacts of increasing N inputs on the ECM symbiosis.

Defining root death in studies of root dynamics is problematic because cell death occurs gradually and the resulting effects on root function are not well understood. In this study, metabolic activity of grape roots of different ages was assessed by excised root respiration and tetrazolium chloride reduction. We investigated changes in metabolic activity and patterns of cell death occurring with root age and changes in root pigmentation. Tetrazolium chloride reduction of roots of different ages was strongly correlated to respiration (R2 = 0.786). As roots aged, respiration and tetrazolium chloride reduction declined similarly, with minimum metabolic activity reached at six weeks. Tetrazolium chloride reduction indicated that the onset of root browning corresponded to a 77% reduction in metabolic activity (P < 0.001). Anatomical examination of roots at each pigmentation stage showed that even though some cells in brown roots were still alive, these roots were functionally dead. The effect of using different definitions of root death in relation to root survivorship was determined in a study of ‘Concord’ grapes with two pruning treatments, using three criteria for root death: browning, blackening or shriveling, and disappearance. There was no effect of vine pruning on root life span when life span was defined as the time from first appearance to the onset of browning. However, if death was judged as the point when roots either became black or shriveled or disappeared, vine pruning decreased root life span by 34% and 40%, respectively (P < 0.001), and also increased the decay constant for root decomposition by about 45% (P < 0.001). We conclude that the discrepancy among determinations of root life span assessed with different definitions of death might be partly caused by the latter evaluations of root life span incorporating a portion of root decomposition in definitions of root death.

This review describes the life cycle of Mesembryantheum crystallinum L. (the common ice plant, Aizoaceae, Caryophyllales), a halophyte with a developmentally programmed switch from C3 photosynthesis to Crassulacean acid metabolism (CAM) which is accelerated by salinity and drought. Since there has been controversy regarding the interplay between genes and environmental stimuli during the development of M. crystallinum, it is timely to summarize the life cycle for a defined set of conditions. We seek to establish the framework whereby five stages of development can be described in terms of morphology, physiology, and molecular biology. Stages 1 and 2, representing germination and growth of a juvenile form, show a determinate pattern of growth. Although specific genes for salt tolerance can be induced at these stages, stress early in development prevents progression to the mature form (stages 3–5) in which the plants advance to mature growth, flowering, and seed development. Growth in stage 3 is indeterminate in the absence of stress, but development and flowering are accelerated by environmental stresses, and CAM is constitutively expressed. Depending on the severity of the stress, plants start to flower (stage 4) and then die from the roots, ultimately with only seed capsules remaining viable, with salt sequestered into large epidermal bladder cells (stage 5). We highlight responses to salinity leading to compartmentation of ions and compatible solutes, turgor maintenance, and CAM. Finally, the molecular genetics of the ice plant are characterized, emphasizing selected genes and their products. We conclude with an analysis of the multiple stages of growth as an ecological adaptation to progressive stress. The initial determinate and inflexible juvenile phase provides a critical mass of plant material which supports the indeterminate, mature phase. Depending on the degree of stress, the mature form is then propelled towards flowering and seedset.

We have explored leaf-level plastic response to light and nutrients of Quercus ilex and Q. coccifera, two closely related Mediterranean evergreen sclerophylls, in a factorial experiment with seedlings. Leaf phenotypic plasticity, assessed by a relative index (PI = (maximum value - minimum)/maximum) in combination with the significance of the difference among means, was studied in 37 morphological and physiological variables. Light had significant effects on most variables relating to photosynthetic pigments, chlorophyll fluorescence and gas exchange, whereas nutrient treatment had a significant effect in only 10% of the variables. Chlorophyll content was higher in the shade whereas carotenoid content and nonphotochemical quenching increased with light. Nutrient limitations increased the xanthophyll-cycle pool but only at high light intensities, and the same interaction between light and nutrients was observed for lutein. Predawn photochemical efficiency of PSII was not affected by either light or nutrients, although midday photochemical efficiency of PSII was lower at high light intensities. Photosynthetic light compensation point and dark respiration on an area basis decreased with light, but photosynthetic capacity on a dry mass basis and photochemical quenching were higher in low light, which translated into a higher nitrogen use efficiency in the shade. We expected Q. ilex, the species of the widest ecological distribution, to be more plastic than Q. coccifera, but differences were minor: Q. ilex exhibited a significant response to light in 13% more of the variables than Q. coccifera, but mean PI was very similar in the two species. Both species tolerated full sunlight and moderate shade, but exhibited a reduced capacity to enhance photosynthetic utilization of high irradiance. When compared with evergreen shrubs from the tropical rainforest, leaf responsiveness of the two evergreen oaks was low. We suggest that the low leaf-level responsiveness found here is part of a conservative resource use strategy, which seems to be adaptive for evergreen woody plants in Mediterranean-type ecosystems.

We review four hypotheses for the control of carbon acquisition by roots, and conclude that the functional equilibrium hypothesis can offer a good description of C acquisition by roots relative to shoots, but is deficient mechanistically. The hypothesis that import into roots is solely dependent on export from the shoot, itself determined by features of the shoot alone (the ‘push’ hypothesis), is supported by some but not all the evidence. Similarly, the idea that root demand, a function of the root alone, determines import into it (the ‘pull’ hypothesis), is consonant with some of the evidence. The fourth, general, hypothesis (the ‘shared control’ hypothesis) – that acquisition of C by roots is controlled by a range of variables distributed between root and shoot – accords with both experiment and theory. Top-down metabolic control analysis quantifies the control of C flux attributable to root relative to source leaf. We demonstrate that two levels of mechanistic control, short-term regulation of phloem transport and control of gene expression by compounds such as sugars, underlie distributed control. Implications for the impact of climate change variables are briefly discussed.

The mechanical behaviour of large foliage leaves in response to static and dynamic mechanical forces is reviewed in the context of a few basic engineering principles and illustrated in terms of species drawn from a variety of vascular plant lineages. When loaded under their own weight or subjected to externally applied forces, petioles simultaneously bend and twist, and thus mechanically operate as cantilevered beams. The stresses that develop in petioles reach their maximum intensities either at their surface or very near their centroid axes, where they are accommodated either by living and hydrostatic tissues (parenchyma and collenchyma) or dead and stiff tissues (sclerenchyma and vascular fibres) depending on the size of the leaf and the species from which it is drawn. Allometric analyses of diverse species indicate size-dependent variations in petiole length, transverse shape, geometry and stiffness that accord well with those required to maintain a uniform tip-deflection for leaves with laminae differing in mass. When dynamically loaded, the laminae of many broad-leaved species fold and curl into streamlined objects, thereby reducing the drag forces that they experience and transmit to their subtending petioles and stems. From a mechanical perspective, the laminae of these species operate as stress-skin panels that distribute point loads more or less equally over their entire surface. Although comparatively little is known about the mechanical structure and behaviour of foliage leaves, new advances in engineering theory and computer analyses reveal these organs to be far more complex than previously thought. For example, finite-element analyses of the base of palm leaves reveal that stresses are decreased when these structures are composed of anisotropic as opposed to isotropic materials (tissues).

Nutrient conservation plays an important role in plants adapted to infertile environments. Nutrients can be conserved mainly by extending the life span of plant parts and/or by minimizing the nutrient content of those parts that are abscissed. Together these two parameters (life span and resorption) define the mean residence time (MRT) of a nutrient. In this review we summarize available information on nitrogen resorption and life span, and evaluate their relationship to the MRT of nitrogen, both between and within species. Abundant information with respect to nitrogen resorption efficiency and life span is available at the leaf level. By definition, woody evergreen plants have a much longer leaf life span than species of other life-forms. Conversely, differences in resorption efficiency among life-forms or among plants in habitats differing in soil fertility appear to be small. Inter-specific variation in leaf life span is much larger than intra-specific variation (factor of >200 compared with 2, respectively), while resorption efficiency varies by about the same magnitude at both levels (factor of 3.8 compared with 2.7, respectively). The importance of resorption efficiency in determining leaf-level MRT increases exponentially towards and above the maximum resorption efficiency observed in nature. This effect is independent of leaf life span, which may explain the lack of life-form related differences in resorption efficiency. When scaling up from the leaf to the whole-plant level, fundamental differences in turnover rate among different plant organs must be considered. Woody species invest c. 50% of their net productivity into their low-turnover stems, while in herbaceous species the life span of stems is only slightly longer than that of leaves. As a result, nutrient turnover of woody (evergreen and deciduous) plants is generally lower than that of herbaceous species (herbs and graminoids) on a whole-plant basis. At the intra-specific level empirical data show that both biomass life span (i.e. the inverse of biomass loss rate) and resorption efficiency are important sources of variation in MRT. However, we argue that the relative importance of resorption efficiency in explaining variation in MRT is lower at the inter-specific level, whereas the reverse is true for life span. This is because variation in MRT and life span is much larger at the inter-specific level compared with variation in resorption efficiency. Plant traits related to nutrient conservation are discussed with respect to their implications for leaf structure, plant growth, competition, succession and ecosystem nutrient cycling.