Plants known as hyperaccumulators take up and sequester high concentrations of potentially toxic elements from metalliferous soils. We tested the hypothesis that zinc hyperaccumulation benefits plants by deterring herbivory. In laboratory feeding trials, three species of herbivores were allowed to choose between Thlaspi caerulescens (Brassicaceae) plants grown in low-Zn and Zn-amended culture solution. Locusts (Schistocerca gregaria), slugs (Deroceras caruanae), and caterpillars (Pieris brassicae) all showed significant preferences for plants with lower foliar Zn concentrations. Such differential feeding could result in selection pressures favouring the evolution of hyperaccumulation. The findings are also relevant to current proposals to exploit hyperaccumulation as a means of remediating metal-contaminated soils.

Clones of ectomycorrhizal fungi can colonize new areas through production of vegetative mycelium or spore dispersal, but the relative importance of these processes in nature is not known. In this study, sporocarps of an ectomycorrhizal fungus, Suillus grevillei, were mapped and sampled from a Larix kaempferi stand at the foot of Mt Fuji. DNA was extracted directly from each sporocarp, and DNA polymorphism was analysed by polymerase chain reaction (PCR) amplification of inter-simple sequence repeat (ISSR) regions primed by (GTG)5, (GCC)5 and (GACA)4. Different sensitivities to detect polymorphism were found among the three primers, with (GACA)4 showing the highest sensitivity. Forty seven sporocarps were analysed by the three ISSR primers and divided into 34 genets based on combination of PCR fingerprints. In the population 28 genets were represented by individual sporocarps. In most cases, sporocarps grown in aggregation (within a circle of 50 cm diameter) showed some different ISSR band patterns. These results suggest that genets of S. grevillei at the test site are relatively small. The genetic similarities between the 34 genets were also calculated and similarity groups were determined by the criterion that all similarity F values of genets within a group were not <80%. In general, the genets within a similarity group located close to each other. The results of multiple different but highly related genets in a small area suggest that the population of S. grevillei in this stand is not spread and maintained by clonal mycelium extension but is reproduced by spore dispersal.

The effect of soil flooding on arbuscular-mycorrhizal (AM) fungal colonization of wetland plants was investigated using Panicum hemitomon and Leersia hexandra, two semi-aquatic grasses (Graminaceae) that grow along a wide hydrologic gradient in Carolina bay wetlands of the southeastern US coastal plain. Three related investigations were conducted along the dry-to-wet gradient in these wetlands; a field survey of AM fungal root colonization in eight wetlands, monthly monitoring of colonization patterns in P. hemitomon over a growing season, and an inoculum potential bioassay of soils collected along the gradient. The field survey showed that AM fungal colonization was strongly negatively correlated with water depth, but colonization was present in most root samples. The monthly assessment indicated that AM fungal colonization was lowest in plots that were consistently wet but rose as some plots underwent seasonal drying. The inoculum potential assay of dry, intermediate, and wet soils performed under both dry and saturated conditions showed that soils that were wet for >1 yr had the same ability to form mycorrhizas in bait plants as those that had remained dry. These findings suggested that the lower degree of colonization in wet areas observed in the field survey was because of the presence of surface water rather than low numbers of mycorrhizal propagules in the soil. Overall, the results of these investigations show that flooding is partially but not totally inhibitory to AM fungal colonization of wetland grasses.

We studied the transport of 15N from a soil compartment separated from a plant root compartment by a hydrophobic polytetrafluoroethylene (PTFE) membrane to plants in the presence and absence of arbuscular mycorrhizal fungi (AMF). We have previously shown that this type of membrane efficiently inhibits mass flow and diffusion of mobile ions in the soil solution in an abiotic system, but can be penetrated by the hyphae of mycorrhizal fungi. Mycorrhizal tomatoes (Lycopersicon esculentum) colonized by Glomus mosseae were grown at two N fertilizer concentrations in a root compartment. A PTFE membrane was placed between the root compartment and an adjoining soil compartment that was inaccessible to the roots but accessible to the AMF hyphae (hyphal compartment). Additional N was applied to the hyphal compartment using uniformly 15N-labelled NH4NO3. There was a flux of 15N from the hyphal compartment to the plants even in the absence of mycorrhizal fungi. However, this flux was much higher in mycorrhizal plants, which had much higher N concentrations in their shoots and roots than did the non-mycorrhizal control plants. This was particularly apparent when the root compartment had a low N fertilizer concentration. Of the total N content of mycorrhizal plants, c. 42 and 24% at the low and high N fertilizer concentrations, respectively, were estimated to originate from the hyphal compartment by transport through AMF hyphae. In the presence of mycorrhizal fungi, the flux of 15N was about three times higher than in their absence. The results show that AMF can access a soil compartment separated by a PTFE membrane, and can contribute substantially to N uptake by plants.

A systematic survey of the endophytic assemblages of Quercus ilex in central Spain has been performed, with the goal of evaluating the importance of geographical and seasonal factors on these fungal communities. Four sampling sites were selected; one of them was sampled twice, in the spring and the autumn. The collected plant material consisted of bark, twigs and leaves from eight trees per site. Fungal strains were isolated with the use of a surface-sterilization method with sodium hypochlorite. A total of 2921 fungal strains grouped into 149 ‘species’ or morphological types were recovered. The 10 dominant species, with isolation frequencies >1.5%, were Pyrenochaeta sp., Periconiella anamorph of Biscogniauxia mediterranea (De Not.) Kuntze, Pseudonectria sp., Cryptosporiopsis quercina Petrak, Alternaria alternata (Fr:) Keissl., two undetermined coelomycetes, Penicillium funiculosum Thom, Diplodia mutila Fr. apud Mont. and Ascochyta sp. Medians of fungal species per tree were significantly different among the sampled sites. The isolation frequencies of the dominant species, as well as other less frequent species, were significantly dependent on the sampling site. The degree of endophytic infection and the diversity of fungal species were significantly higher in the spring. The frequencies of all dominant species at one of the sites depended significantly on the season, except for C. quercina, Acremonium sclerotigenum (F & V Moreau ex Valenta) Gams. and D. mutila. Cluster analysis of the whole endophytic mycoflora of the sampled trees suggested that the geographical factor affects the endophytic distribution patterns more significantly than the seasonal factor.

A pathogenic strain of Fusarium oxysporum f. sp. lycopersici transformed with the glucuronidase (GUS) reporter gene was used to study the colonization process of tomato (Lycopersicon esculentum) roots in hydroponic culture. The plants were treated exactly as those used in a different study with a non-pathogenic strain of F. oxysporum in order to compare the two types of colonization process. The pathogenic strain rapidly colonized the root surface, forming a dense network of hyphae – shown by the GUS staining assay – as early as 48 h after inoculation. The first images of the pathogen penetrating into the epidermis of the root were observed 24 h after inoculation. The plant showed defence reactions mainly in the hypodermis, but also in the cortex. Generally these barriers failed to prevent the centripetal growth of the pathogen towards the stele. The GUS activity moved with the actively growing hyphae into the tissues; the stele appeared intensely stained 7 d after inoculation, whereas by this time the hyphae at the root surface did not show any more staining. The hyphae formed a dense network at the apex of the root, but direct contact between hyphae and living cells was prevented by several layers of sloughed cap cells. Although rarely observed, penetration of the pathogen into the apex was possible and led to a rapid destruction of apical cells. The only differences between this pattern of root colonization and that observed for a non-pathogenic strain of F. oxysporum appear to concern the frequency of dead apices and the intensity of fungal colonization in the cortex. When the pathogen passed around the barrier formed in the hypodermis it always reached the xylem, although this barrier and other defence reactions induced at different levels in the cortex always prevented the non-pathogenic strain from reaching the stele. These observations suggest that the main differences between the two types of interaction – between the plant and either the pathogenic or the non-pathogenic strain – are quantitative rather than qualitative.

The arbuscular mycorrhizas of bluebell (Hyacinthoides non-scripta (L.) Chouard ex Rothm.) involve several symbiotic fungi of the order Glomales. We have previously simplified the system by ignoring the taxonomic diversity of the fungi, but it is unlikely that all fungal species contribute in the same manner or to the same extent to the functioning of the symbiosis. To discover how many and which fungi take part in the bluebell mycorrhiza we sought to identify the range of arbuscular mycorrhizal (AM) fungi found in bluebell roots sampled during a complete growing season, September to June.

Although the taxonomy of the Glomales by their spores is not yet fully understood, identification is, to a large extent, possible. Arbuscular mycorrhizal communities are usually characterized by their spores but, since spores can rarely be directly associated with individual plants or plant species, a more satisfactory approach would be to identify fungal symbionts where they interact with the host plant, in the roots. Unfortunately, the intraradical mycelia of the fungi are less easily distinguished than the spores and, as yet, classification is possible only to the family level.

We have developed a method whereby different AM fungal taxa in the roots of bluebell can be distinguished by objective assessment. A large suite of morphological characteristics of the fungi in roots was recorded in samples taken at monthly intervals. Hierarchical cluster analysis of the resulting data separated six distinct AM fungal morphotypes (Scutellospora type, Acaulospora type, three Glomus types and fine endophytes) and a classification system created by which identification by eye was possible.

We compared the fungi in roots with glomalean spores in soil from the root zone of the same bluebell plants. Two species occurred in most samples, Scutellospora dipurpurescens Morton & Koske (emend. Walker, 1993) and Acaulospora koskei Blaskowski. A further six occurred sporadically, five Acaulospora spp. and Glomus rubiforme Gerdemann & Trappe) Almeida & Schenck (=Sclerocystis rubiformis). The presence of a single species of Scutellospora was consistent with a Scutellospora root morphotype which varied little. By contrast, the diversity of Acaulospora in the spore assemblage was reflected by variation within the Acaulospora morphotype. Glomus spores were very rarely found in field collections, yet Glomus morphotypes were found to be an important component of the bluebell mycorrhiza.

Because some important species are not represented in spore assemblages in the field and those that are found can only be associated with vegetational groups, not individual plants or single species, glomalean spore populations provide only a partial account of the fungi which contribute to arbuscular mycorrhizas. Although it is still not possible to identify AM fungi in roots with the same precision as their spores, the method reported here permits assessment of diversity in the roots of individual plant species, which may be applied to the investigation of mycorrhizal function and demography in natural ecosystems.

The mycobionts of Piceirhiza bicolorata, a distinct ectomycorrhizal morphotype of conifers and hardwoods, have been identified by internal transcribed spacer 1 (ITS1) nuclear ribosomal DNA (rDNA) sequence comparison of the fungi involved. Samples of Piceirhiza bicolorata were obtained from seedlings of Picea abies, Pinus sylvestris, Betula pubescens, Populus tremula, Quercus robur and Salix phylicifolia. In an initial screening, the fungus amplified with universal ITS primers from ectomycorrhizal root samples of P. bicolorata shared approx. 95% ITS1 sequence identity with the ericoid mycorrhizal fungus Hymenoscyphus ericae. A total of 77 out of 88 (= 87.5%) DNA samples (i.e. 52/56 root samples and 25/32 axenic culture isolates) of P. bicolorata were successfully amplified with a taxon-selective primer designed for exclusive amplification of H. ericae-like strains. Forty-seven amplicons were sequenced, yielding 15 different ITS1 genotypes that differed by 1–14 nucleotide character state changes. An inferred ITS1 phylogeny (maximum parsimony) showed that a single major evolutionary lineage of P. bicolorata embraced the historically important H. ericae isolates in a 100% bootstrap-supported clade. The 15 P. bicolorata genotypes were positioned in four subclades, roughly corresponding to morphological groups of P. bicolorata isolates observed in axenic culture. Culture isolates of H. ericae and P. bicolorata share some common morphological features including slow, dense growth and formation of short aerial hyphal aggregates. Our results suggest that members of the H. ericae aggregate participate in the formation of the distinct ectomycorrhizal morphotype P. bicolorata. This opposes the widely accepted discrimination of ericoid and ectomycorrhizal mycobionts of the boreal forest ecosystem. The high prevalence of the P. bicolorata morphotype on pioneer seedlings of P. sylvestris, B. pubescens and S. phylicifolia at a copper mine spoil was remarkable. Hypotheses of possible nutrient mobilization and detoxification potentials of the fungal associates of P. bicolorata are discussed. We hypothesize that ericoid and ectomycorrhizal plants may share mycobionts of the H. ericae aggregate.

Eucalyptus species are considered to have ectomycorrhizas (ECM), but many also have vesicular–arbuscular mycorrhizas (VAM) and their relative importance is unclear. Interactions between ECM and VAM fungi colonizing roots of Eucalyptus species were examined in a glasshouse experiment. This experiment investigated competition between these two types of fungi and compared benefits provided to Eucalyptus globulus and E. urophylla. Eucalyptus seedlings were inoculated with spores of the ECM fungus Laccaria lateritia and/or pot- culture soil for a VAM fungus (species of Glomus, Acaulospora or Scutellospora). Initial inoculum levels were important, as VAM fungi became established much more rapidly than Laccaria. In plants with both types of mycorrhizas, Laccaria mycorrhizas substantially increased after 2 or 3 months and the proportion of roots with VAM declined. However, the proportion of roots with VAM also decreased significantly in plants without ECM after 2 months. Thus, the relative susceptibility of eucalypt roots to these mycorrhizal associations changed. Substantial growth responses to mycorrhizal inoculation occurred when a low concentration of phosphorus fertilization was used (5 mg kg−1), but not at a higher concentration (10 mg kg−1). Treatments where Laccaria was applied, alone or in combination with a VAM fungus, resulted in the largest growth increases relative to nonmycorrhizal plants. An Acaulospora isolate was the most effective VAM fungus for E. urophylla although other VAM fungi also increased growth relative to the control. VAM fungi alone had little effect on E. globulus growth, but plants inoculated with both ECM and VAM fungi were larger than plants only inoculated with Laccaria. ECM fungi had a major impact on root system form by reducing the proportion of fine roots (specific root length) relative to nonmycorrhizal plants or those with VAM. Changes in root colonization patterns over time must be considered in studies of ECM/VAM interactions, as there can be substantial changes in the relative importance of the two associations.

Several physiological processes controlling tree phenology remain poorly understood and in particular bud dormancy. Many studies have emphasised the action of chilling temperatures in breaking dormancy. However, the effect of the preceding summer temperatures has rarely been investigated although there is some evidence that they may be involved in the settlement and intensity of dormancy as well as cold acclimation. In this paper, thermal time to budburst in relation to the duration of chilling outdoors, preceding summer temperatures and forcing temperatures was studied by outdoors experiments in seedlings of Platanus acerifolia, Vitis vinifera, Quercus pubescens and Castanea sativa. Results showed that temperatures of the preceding summer had no significant effect on the timing of budburst, P. acerifolia and Q. pubescens showed a very weak response to the duration of chilling, and the phenological characteristics of each species were found to be adapted to the climate conditions of its own geographical area. The phenological model used in this study explained 82–100% of the variance of the data without taking into account summer temperatures. Thus, although summer temperatures may be well involved in the intensity of dormancy and cold hardiness, they do not significantly affect budburst and therefore may not need to be considered in phenological models for predicting budburst.

Fungal endophytes associated with the palm, Trachycarpus fortunei, within and outside its natural geographic range were investigated. Endophytes were relatively common with colonization rates of 23–57% at the four sites sampled. The endophyte assemblages at the different sites were diverse with 75 fertile species of ascomycetes and mitosporic fungi. The assemblage composition at each site was similar and between seven and 13 species comprised 81–89% of the taxa present in relative frequencies of >1%. Glomerella cingulata and Phomopsis spp., were consistently dominant, and a large number of rare species were recorded. The diversity at each site was similar in number, but the abundance of isolates varied. The results obtained were comparable to those of previous studies of palm endophyte assemblages, but the assemblages showed more affinity with unrelated temperate hosts than with tropical palm hosts. Quantitative and qualitative differences in endophyte assemblages from old and young tissues were observed, and more isolates were recovered from old tissues independent of the age of the palm. The composition of the assemblage varied with several taxa being exclusively or more commonly isolated from old tissues (e.g. xylariaceous taxa, Oxydothis sp. nov.) or young tissues (e.g. Stagonospora spp., Phoma multirostrata). Some differences in the composition of the assemblage and in relative frequencies of various species were observed in trees and saplings. Significantly more isolates were recovered from the vein than intervein tissues, independent of leaf age or tree age. Tissue specificity was not exhibited by any taxa isolated from either leaf or petiole tissues, except for xylariaceous taxa in leaf tissues. Some other taxa showed a preference for leaf tissues or petioles, whereas others were equally distributed amongst all tissues. Endophyte assemblages of palms from continuous distributions were similar, but those from disjunct distributions (i.e. outside the natural geographic range of the palm, such as Australia and Switzerland), differed significantly from each other and from assemblages within continuous distributions. The relative importance values of endophyte species at the two sites in China indicates the site-specific nature of the endophyte assemblages. Most previous studies on the endophytes of palm hosts have concentrated on tropical palms. However, this study examines the endophytes of the warm temperate palm Trachycarpus fortunei, and investigates the relative importance of host and climate related processes.

Leaves of a mottled stripe disease-susceptible cultivar (B-4362) and of a mottled stripe disease-resistant cultivar (SP 70–1143) of sugar cane (interspecific hybrids of Saccharum) were inoculated with the diazotrophic endophytes, Herbaspirillum rubrisubalbicans or Herbaspirillum seropedicae, via injection into the apex of the stem. At 7 and 20 d.a.i., H. seropedicae could be isolated only from a small necrotic area around the point of inoculation, where there was considerable degradation of host cells, and was not detected in any other part of the leaves. This suggested a hypersensitive response by the host to this bacterium, and no disease symptoms formed on either cultivar. By contrast, H. rubrisubalbicans could be re-isolated from throughout the infected leaves of both cultivars at both harvests and produced widespread disease symptoms on the leaves of cv. B-4362.

Symptoms consisted of necrotic regions near the point of inoculation, and red stripes and red patches along the vertical axis of the leaves, where the bacteria had spread in the primary and secondary veins. The xylem-conducting elements in diseased regions of leaves were filled with bacteria and, at the edges of disease symptoms, the vessels were filled with a gum which stained blue-green with toluidine blue. This material probably contained phenolic compounds, and was produced as a host defence response. Leaves of cv. SP 70–1143 only developed small red stripes near the point of inoculation. These symptoms did not spread along the leaves, and the infected xylem vessels were never seen to be completely full of bacteria. Instead, the vessels contained encapsulated bacterial colonies attached to secondary wall deposition; these colonies were surrounded by blue-green material that might have been host-defence gums. In cv. B-4362, bacteria were abundant in the intercellular spaces of mesophyll adjacent to infected xylem, and also filled sub-stomatal cavities. Immunogold labelling using polyclonal antisera raised against H. rubrisubalbicans gave a weak signal with the bacteria in cv. SP 70–1143, showing that few binding sites were available to the antibodies. By contrast, bacteria in cv. B-4362 reacted strongly with the antibody, suggesting that they had a denser coating of immunoreactive mucus. In the later stages of infection of cv. B-4362, lysed bacteria were seen within degraded plant cells surrounded by a matrix of plant gums and bacterial mucus. This matrix reacted strongly to the H. rubrisubalbicans antibodies. Immunogold labelling using antibodies against nitrogenase component II showed that nitrogenase was expressed by bacteria in the early stages of infection of cv. B-4362, but not in later stages, or by bacteria infecting cv. SP 70–1143.

The effects of summer drought, dew deposition on leaves and autumn rainfall on plant water relations and diurnal variations of photosynthesis were measured in two evergreen shrubs, rosemary (Rosmarinus officinalis) and lavender (Lavandula stoechas), grown in Mediterranean field conditions. Withholding water for 40 d caused a similar decrease in predawn shoot water potential (ψpd) from c. −0.4 to c. −1.3 MPa in both species, but a 50% decrease in the relative leaf water content in L. stoechas compared with 22% in R. officinalis. A similar decrease in CO2 assimilation rates by c. 75% was observed in water-stressed plants of both species, although L. stoechas showed smaller photosynthesis: stomatal conductance ratio than R. officinalis (35 vs 45 μmol CO2:mol H2O). The relative quantum efficiency of photosystem II photochemistry also decreased by c. 45% at midday in water- stressed plants of both species. Nevertheless, neither L. stoechas nor R. officinalis suffered drought-induced damage to photosystem II, as indicated by the maintenance of the ratio Fv:Fm throughout the experiment, associated with an increase in the carotenoid content per unit of chlorophyll by c. 62% and c. 30%, respectively, in water-stressed plants. Only L. stoechas absorbed dew by leaves. In this species the occurrence of 6 d of dew over a 15-d period improved relative leaf water content by c. 72% and shoot water potential by c. 0.5 MPa throughout the day in water-stressed plants, although the photosynthetic capacity was not recovered until the occurrence of autumn rainfall. The ability of leaves to absorb dew allowed L. stoechas to restore plant water status, which is especially relevant in plants exposed to prolonged drought.

Rice (Oryza sativa) plants were grown with their roots sandwiched between thin layers of phosphorus-deficient soil from which they were separated by fine mesh, and root-induced changes in the soil affecting phosphate solubility were measured. The concentrations of low molecular weight organic anions in the thin layers, particularly citrate, increased in the presence of the plants. Apparent rates of citrate excretion from the roots, calculated from the quantities in the soil and rates of decomposition calculated with a first order rate constant measured independently, varied from 337–155 nmol g−1 root f. wt h−1 over the course of plant growth, equivalent to 2–3% of plant d. wt. Rates of excretion were similar for NH4+ and NO3−-fed plants. The soil pH decreased from its initial value by up to 0.6 units for the NH4+-fed plants and increased by up to 0.4 units for the NO3−-fed ones. The contribution of organic anion excretion to the pH changes was small compared with that of the inorganic cation-anion balance in the plants. The extent to which the observed excretion of citrate and root-induced pH changes could account for the observed phosphate solubilization and uptake was assessed using a mathematical model. Previous work had shown that phosphate solubilization by rice in this soil could not be explained by enhanced phosphatase activity in the rhizosphere, and the roots were not infected with mycorrhizas. The model allows for the diffusion of the solubilizing agent (citrate or H+) away from the roots, its decomposition by soil microbes (citrate only); its reaction with the soil in solubilizing phosphate and diffusion of the solubilized phosphate to the roots. The model contains no arbitrary assumptions and uses only independently measured parameter values. The agreement between the measured time course of phosphorus uptake and that predicted for solubilization by citrate was good. Root-induced acidification by NH4+-fed plants resulted in additional solubilization, the acidification enhancing the solubilizing effect of citrate. However, the final phosphorus uptake by NH4+-fed plants was no greater than that of NO3−-fed plants, presumably because the acidification inhibited plant growth. The mechanism of solubilization by citrate involved formation of soluble metal-citrate chelates rather than displacement of phosphate from adsorption sites.

Summary 196

I. INTRODUCTION 196

1. The neglected dimension 196

2. Basic concepts 197

(a) The heuristic notion of vessel-tiers 197

(b) Ohm's law 199

(c) Conductances, resistances and resistivities 199

(d) Lumen and pit resistances 199

(e) The importance of conduit radius and length in conductance 199

II. EVOLUTIONARY TRENDS IN CONDUIT DIMENSIONS 199

1. Nature and origin of xylem conduits 199

2. Increasing hydraulic conductance with increasing diameter and length 200

(a) Evolutionary trends in tracheid dimensions 200

(b) Origin of the vessel 200

3. Functional limitations to increasing vessel length 201

(a) Safety versus efficiency 201

(b) Containment of cavitation and embolism 202

III. MAXIMUM XYLEM TRANSPORT IN THE PRESENCE OF CAVITATION 203

1. Cavitation is linked to the driving force for transport 203

2. Transport models and extreme assumptions about conduit length 203

(a) Unitary cavitation response (n = 1) 203

(b) Infinitely partitioned response (n = ∞) 204

(c) ΔP and cavitation containment 204

IV. INCLUDING VESSEL LENGTH IN A TRANSPORT MODEL 205

1. Framing questions of optimal conduit length 205

2. A numeric model for flow through n conduit tiers 205

(a) Model structure 205

(b) Model solution 206

3. Optimization when f(P) is linear 206

(a) Isolating the effects of n on cavitation containment 206

(b) Optimal conduit tier-length distributions (OCLDs) 207

(c) Abrupt changes in conduit length 208

(d) Optimal frequency of end walls: incorporating Rpit 208

4. Optimization when f(P) is curvilinear 210

V. CONDUIT LENGTH IN MODERN TAXA: IMPLICATIONS FOR TRANSPORT 210

1. Limitations to the concept of conduit tiers 210

(a) Vessel ends are randomly distributed 210

(b) Dispersion around mean length within each ‘tier’ 210

2. Is the xylem optimally partitioned? 211

(a) Optimal number of end walls 211

(b) Conduit length distribution along the pathway 211

3. Hydraulic segmentation 212

(a) Segmentation in hydraulic resistance 212

(b) Segmentation in cavitation vulnerability 212

VI. CONCLUSIONS 212

1. Anatomy 212

2. Modelling flow 213

VII. APPENDIX: ANALYTICAL SOLUTIONS AND PROOFS 213

1. Analytic solution for Qmaxwith a single tier 213

2. The general case for n tiers 214

3. Analytic solution for Qmaxwith two tiers 214

4. Matric flux and n = ∞ 215

5. Rpit, variable pathway resistance and OCLD 215

6. Proof of Eqn 15 describing limited cavitation containment 215

Acknowledgements 216

References 216

Vascular plants have shown a strong evolutionary trend towards increasing length in xylem conduits. Increasing conduit length affects water transport in two opposing ways, creating a compromise that should ultimately define an optimal conduit length. The most obvious effect of increased length is to decrease the sequential number of separate conduits needed to traverse the entire pathway, and thereby to reduce the number of wall-crossings and the hydraulic resistance to flow within the xylem. This is an essential evolutionary pressure towards the development of the vessel, a conduit of multicellular origin whose length is not restricted by developmental constraints. The vessel has been an essential component in all plant lineages, achieving transport tissues with very high specific conductivity. A countering effect, however, arises from the partitioning of the cavitation response, a process whereby individual xylem conduits drain of water and lose conducting capacity. Flow in the xylem is down a gradient of negative pressure, which is necessarily most negative in the distal regions (i.e. near the foliage). Cavitation can be caused directly by negative pressures, and results in a total loss of the hydraulic conductance of the individual conduits within which it occurs. If cavitation is triggered by low pressure experienced only at the very distal end of a long conduit, the conduit nevertheless loses its conducting capacity along its entire length. Pathways composed of long conduits will therefore suffer greater total conductance loss for equivalent pressure gradients, because the effects of cavitation are not effectively restricted to the tissue regions within which the cavitation events are generated. By contrast, short conduits can restrict cavitation to distal regions, leaving trunk and root tissues less seriously affected. The increased total conductance loss of a system made entirely of very long conduits translates into a lower maximum rate of water transport in the xylem. The loss in hydraulic capacity associated with failure to partition the flow pathway fully, and locally contain the effects of cavitation, theoretically reaches a maximum of 50% for the extreme case in which a single set of conduits traverses the entire pathway. Shorter conduits confine individual cavitation events to smaller regions and permit the pathway as a whole to have a more gradual conductance loss in conjunction with the pressure gradient. A compromise exists between (1) minimizing total conductance loss from cavitation via fine partitioning of the pathway with many tiers of short conduits, and (2) reducing total wall resistance via coarse partitioning with a few tiers of long conduits. An analysis is presented of the optimal number of end walls (i.e. mean conduit length relative to total pathway length) to maximize transport capacity. The principle of optimal containment of cavitation also predicts that conduits should not be of equal length in all portions of the pathway. The frequency of end walls should rather be proportional to the magnitude of the water-potential gradient at each point, and conduits should be longest in the basal portion (roots) and progressively shortened as they move up the stems to the foliage. These concepts have implications for our understanding of the contrasting xylem anatomies of roots and shoots, as well as the limits to evolution for increased hydraulic conductance per xylem cross-sectional area. They also indicate that to model the hydraulic behaviour of plants accurately it is necessary to know the conduit length distribution in the water flux pathway associated with species-specific xylem anatomy.

The objectives of this paper were to review the literature on the responses of root systems to elevated CO2 in intact, native grassland ecosystems, and to present the results from a 2-yr study of root production and mortality in an intact calcareous grassland in Switzerland. Previous work in intact native grassland systems has revealed that significant stimulation of the size of root systems (biomass, length density or root number) is not a universal response to elevated CO2. Of the 12 studies reviewed, seven showed little or no change in root-system size under elevated CO2, while five showed marked increases (average increase 38%). Insufficient data are available on the effects of elevated CO2 on root production, mortality and life span to allow generalization about effects. The diversity of experimental techniques employed in these native grassland studies also makes generalization difficult. In the present study, root production and mortality were monitored in situ in a species-rich calcareous grassland community using minirhizotrons in order to test the hypothesis that an increase in these two measures would help explain the increase in net ecosystem CO2 uptake (net ecosystem exchange) previously observed under elevated CO2 at this site (600 vs 350 μl CO2 l−1; eight 1.2-m2 experimental plots per CO2 level using the screen-aided CO2 control method). However, results from the first 2 yr showed no difference in overall root production or mortality in the top 18 cm of soil, where 80–90% of the roots occur. Elevated CO2 was associated with an upward shift in root length density: under elevated CO2 a greater proportion of roots were found in the upper 0–6-cm soil layer, and a lower proportion of roots in the lower 12–18 cm, than under ambient CO2. Elevated CO2 was also associated with an increase in root survival probability (RSP; e.g. for roots still alive 280 d after they were produced under ambient CO2, RSP = 0.30; elevated CO2, RSP = 0.56) and an increase (48%) in median root life span in the deepest (12–18 cm) soil layer. The factors driving changes in root distribution and longevity with depth under elevated CO2 were not clear, but might have been related to increases in soil moisture under elevated CO2 interacting with vertical patterns in soil temperatures. Thus extra CO2 taken up in this grassland ecosystem during the growing season under elevated CO2 could not be explained by changes in root production and mortality. However, C and nutrient cycling might be shifted closer to the soil surface, which could potentially have a substantial effect on the activities of soil heterotrophic organisms as CO2 levels rise.

Natural abundances of 15N and N concentrations of 34 fruit bodies from 24 species of ectomycorrhizal and saprophytic fungi were measured in a temperate Central European mixed forest stand. The fungi of the two life forms are known to be capable of utilizing different types of N sources (organic N compounds from the humus, inorganic N from the soil and N from litter or wood) differing by their 15N natural abundance values. Based on the two life forms and the three different N sources, four functional groups of fungi were distinguished: (1) ectomycorrhizal fungi capable of utilizing organic N from the humus; (2) ectomycorrhizal fungi known to depend on inorganic N compounds in the soil; (3) saprophytes capable of utilizing organic N from the humus; and (4) saprophytes utilizing N from dead wood or litter. Large differences were found between species in the δ15N values (−3.0 to 3.3‰) and in the N concentrations (0.84 to 6.61 mmol eq N g dw−1) of the fruit bodies. In most cases fungi were more enriched in 15N than their respective bulk N source was. Fungi living in humus, and presumably having access to organic N compounds (groups 1 and 3), were significantly more enriched in 15N than fungi which are known to depend on inorganic N (e.g. Laccaria, group 2), or fungi living on litter or wood (group 4), irrespective of whether they were ectomycorrhizal or saprophytic species. Fungi living in humus had significantly higher N concentrations than fungi living on litter or wood.

Little is known about non-mycorrhizal endophytic fungi in tropical orchids; still less is known about how endophytes vary within and between individual orchid plants. Fungal endophytes were isolated from roots and leaves of epiphytic and lithophytic orchids in the genus Lepanthes; seven species, from rainforests in Puerto Rico, were sampled. The endophytes observed most frequently were Xylaria species and Rhizoctonia-like fungi, found in 29% of roots and 19% of leaves, and 45% of roots and 31% of leaves, respectively. Five deuteromycete genera were also isolated, occurring in 19% of roots and 43% of leaves (combined). At least nine species of Xylaria were found, with several species sometimes occurring in a single plant. Differences between roots and leaves in frequency of Xylaria and Rhizoctonia isolates were not significant, although differences among orchid species in number and types of endophytes were. Heterogeneity of endophytes in single plants and plant organs was greater than differences between species. Many Lepanthes species are very restricted in distribution, and knowledge of their interactions with endophytes might be useful in species management.

Among citrus rootstocks, higher specific root length (SRL, root length/d. wt) has been linked to several specific morphological and physiological traits, including smaller average root diameter, higher root hydraulic conductivity and higher rates of root proliferation. In this study, thickness of the outer tangential exodermal (hypodermal) wall and its suberin layer, number of passage cells, presence of epidermis, and stelar anatomy were examined and related to variation in root diameter of field roots of known maximum age. We also compared root morphology and anatomy of young roots in the field with those of potted rootstock seedlings in the glasshouse. Fibrous roots were measured separately from pioneer (framework) roots. Among the fibrous roots, only the first-order (root links having a root tip) and second-order (root links bearing first-order roots) laterals were examined. Among first-order field roots, larger root diameter was caused by larger rather than more numerous cells in the cortex. Root diameter of first-order roots was positively correlated with both number of passage cells in the exodermis and thickness of the secondary walls of the exodermis in both field and potted plants.

Exodermal walls were about 80% thicker in field- than pot-grown roots. In the field, in more than 50% of the first-order roots examined less than 30% of the root surface was still covered by epidermis, with few differences among rootstocks. In contrast, in roots of 19-wk-old glasshouse plants generally 70–100% of the epidermis was still intact. There was no evidence of secondary xylem development in second-order fibrous roots in the field; in seedling, pot-grown rootsystems, 75–97% of second-order roots had formed secondary xylem despite their small diameter (<0.8 mm).

It is argued that there can be suites of physiological, morphological and anatomical traits in roots that co-vary with specific root length. Investigations of how root morphology and anatomy are linked to root function, moreover, need to recognize trait variability and the potentially important differences between field- and pot- grown (seedling) roots.

A pot experiment was designed to evaluate the interactive effects of multiple microbial inoculation treatments and rock phosphate (RP) application on N and P acquisition by alfalfa plants using 15N and 32P isotopes. The microbial inocula consisted of a wild type (WT) Rhizobium meliloti strain, its genetically modified (GM) derivative, which had an enhanced competitiveness, the arbuscular mycorrhizal (AM) fungus Glomus mosseae (Nicol. and Gerd.) Gerd. and Trappe, and a phosphate-solubilizing rhizobacterium (Enterobacter sp.). Inoculated micro-organisms became established in the root tissues and/or in the rhizosphere soil of alfalfa plants (Medicago sativa L.). The GM Rhizobium strain did not interfere with AM formation. Inoculated phosphate-solubilizing rhizobacteria established in the alfalfa rhizosphere, but the level of establishment was lower where the natural population of phosphate-solubilizing bacteria was stimulated by AM inoculation and RP application. The stimulation of these indigenous bacteria was also greater in the rhizosphere of alfalfa nodulated by the GM Rhizobium. Improvements in N and P accumulation in alfalfa corroborate beneficial effects of the improved GM Rhizobium on AM performance, in RP-amended plants. Inoculation with Enterobacter did not improve the AM effect on N or P accumulation in the RP-added soil, but it did in the non RP-amended controls. Measurements of the 15N[ratio ]14N ratio in plant shoots indicated enhanced N2 fixation rates in Rhizobium-inoculated AM-plants, over that achieved by the same Rhizobium strain in non-mycorrhizal plants. Regardless of the Rhizobium strain and of whether or not RP was added, AM-inoculated plants showed a lower specific activity (32P[ratio ]31P) than did their comparable non-mycorrhizal controls, suggesting that the plant was using otherwise unavailable P sources. The phosphate-solubilizing, AM-associated, microbiota could in fact release phosphate ions, either from the added RP or from the indigenous ‘less-available’ phosphate. Deficiency in Ca concentration in soil solution in the neutral test soil might benefit P solubilization. The proportion of plant P derived either from the labelled soil P (labile P pool) or from RP was similar for AM inoculated and non-mycorrhizal controls (without Enterobacter inoculation) for each Rhizobium strain, but the total P uptake, regardless of the P source, was far higher in AM-plants. Enterobacter inoculation seems to improve the use of RP in the rhizosphere of non-mycorrhizal plants inoculated with the WT Rhizobium.