The photosynthate costs of processes (amino acid and protein synthesis and turnover, and pH regulation) associated with the utilization of nitrate (NO3−), ammonium (NH4+) or glutamine (Gln) for plant growth were estimated. Based on these estimates, the effects of these forms of nitrogen (N) on the carbon balance of plants and on shoot–root biomass allocation were evaluated. The results indicated that NO3− as an N source for plant growth is not substantially more expensive to utilize than either NH4+ or Gln, particularly in the long term when costs due to protein turnover dominate the total costs of N utilization. It is also suggested that the photosynthate use in processes associated with N assimilation has little impact on the carbon balance of plants, and hence on shoot–root biomass allocation.

To develop further the methods for estimation of NOx absorption by plants supplied with 15N-labelled fertilizer, we proposed a new calculation method, total N fixed method (TNF), and compared with the 15N dilution method and the classical mass balance method (MB).

Hydroponically grown soybean plants were supplied with 15N-labelled nitrate and exposed to 200–250 nl l−1 NO2 for 7 d. The proportions of the N derived from NO2 to total N in exposed plants were estimated by the three methods.

The reported rates of NO2 absorption by several plant species, estimated by the 15N dilution method, were recalculated using the TNF method. The results of the two methods were compared and showed that: (1) The 15N dilution method overestimated the content of NO2-N in exposed plants compared with the MB method whilst the TNF method produced estimations of NO2-N closer to those by the MB method when the plants were supplied with 5 mM nitrate. (2) The differences in estimations between the MB method and either the 15N dilution method or the TNF method increased with decreasing supply of 15N-labelled nitrate to roots.

Mean δ15N of whole-plants of lettuce and barley varied by [les ]3‰ when given a chemically and isotopically uniform N source. This variation was related to the presence, absence and species of arbuscular mycorrhizal (AM) fungi and to external N concentration. A highly AM-susceptible plant (lettuce) responded to treatments differently than a less susceptible one (barley). The largest change in whole plant δ15N was related to the experimental combination most likely to be found in field conditions: species of fungus interacting with varying external concentrations of N.

The mechanisms underlying observed variations of plant δ15N are not understood, nor can they be approached directly using δ15N. However, descriptive data, such as presented here, are important to the development of a mechanistic model, because they suggest relationships for future research, using inter alia15N-enriched tracers. They also confirm that plant N sources, cannot be identified using plant δ15N, even when the type of mycorrhizal association (endo vs. ecto) is known.

Two cvs of alfalfa (Medicago sativa L.), Gilboa and Moapa 69, were inoculated in glasshouse pots with three arbuscular mycorrhizal (AM) fungi to investigate the efficacy of mycorrhizas with respect to the extent of colonization and sporulation. Paspalum notatum Flugge also was inoculated to describe fungal parameters on a routine pot culture host. Percentage root length of P. notatum colonized by Glomus mosseae (Nicol. & Gerd.) Gerdemann & Trappe, Glomus intraradices Schenck & Smith, and Gigaspora margarita Becker & Hall increased from 10 to 21 wk, and all fungi sporulated during that period. In alfalfa, only colonization by G. intraradices increased over that time period, and it was the only fungus to sporulate in association with alfalfa at 10 wk. Glomus mosseae did not sporulate after 16–21 wk despite having colonized 30–35% of the root length of both alfalfa cvs. In vitro experiments in which Ri T-DNA-transformed roots of alfalfa were inoculated with AM fungi showed normal mycorrhizal formation by G. intraradices and a hypersensitivity-like response to Gi. margarita. Colonized cells became necrotic, and HPLC analysis indicated increased concentrations of phenolics and isoflavonoids in these root segments. These data strongly support the existence of a degree of specificity between AM fungi and host that might rely on specific biochemical regulatory processes initiated in the host as a result of the attempts at colonization by the fungus.

Using field plots, three species (Mercurialis perennis L., Rubus fruticosus L., and Trientalis europaea L.) were tested for their potential to emit gaseous ammonia to the atmosphere. Canopies were misted with 5 mM methionine sulphoximine (MSO) to inhibit glutamine synthetase (GS), the enzyme of ammonium assimilation. Leaf tissue NH4+ concentration of control plants was 0·03–0·1 μmol g−1 f. wt. Although NH4+ accumulated in the leaf tissue of MSO-treated plants of all three species to similar concentrations (6-10 μmol g−1 f. wt after 4 d), emissions were only detected from the leaves of M. perennis, with potential rates of 2·5 nmol m−2 leaf s−1. Experiments carried out in a controlled environment confirmed this rate of emission over 9 d, during which time leaf tissue ammonium increased to 66 μmol g−1 f. wt. Comparisons with Hordeum vulgare grown under the same conditions showed that tissue NH4+ concentration reached a plateau of about 40 μmol g −1f. wt after 2 d. Emissions of NH3 during the 5 d of treatment reached a maximum rate of 10 nmol m−2 s−1 by the third day.

Apoplastic pH of the plants was determined, and it is suggested that this is an important factor explaining the differences in NH3 emission between species. The higher the apoplastic pH, the greater the likelihood of loss of NH3 from sub-stomatal spaces to the atmosphere. T. europaea (non-emitter) had an apoplastic pH of 5, R. fruticosus (non-emitter) a pH of c. 5·6, whereas that of M. perennis (emitter) was c. pH 6·3. The apoplastic pH is thought to be dictated in part by the N nutrition of a species, nitrophilous species tending to have high pH. Without NO3− fertilization, H. vulgare had an apoplastic pH of 6·8 but this increased to 7·3, 3 d after feeding with NO3−.

Short-term fumigation (2 h) of shoots of H. vulgare with 60 μg (≈32 mg NH3 m−3) of labelled gaseous 15N-NH3 (in the absence of MSO) showed that a substantial proportion (60%) of the applied label was found in the leaves, as well as in stems and roots (3%). There was also a change in amino acid pools, with an increase in shoot amino acids and a decrease in those in the root, while tissue NH4+ was very low in both shoots and roots. This provided indirect evidence that some of the applied label had been incorporated into an organic form. Following the fumigation treatment, emissions of NH3 were collected for 3 h, then c. 6·5 μg of N was recovered, of which c. 17% was 15N-labelled. Some of this label could have resulted from desorption of NH3 from leaf surfaces, but it was more likely that the remaining 14N isotope was from sub-stomatal emissions of NH3.

It is argued that non-nitrophilous plants tend to rely on mixed sources of N (NO3−, NH4+ or organic-N) and are more likely to favour root rather than shoot assimilation. Under these circumstances, their apoplastic pH is relatively low (compared with that of nitrophiles, which tend to assimilate NO3− mainly in their shoots), and at atmospheric concentrations most wild species are likely to be net assimilators, rather than emitters, of atmospheric ammonia.

TIP1 is a gene defined by an X-ray induced allele tip1–2 and a previously described EMS-induced allele tip1−1. TIP1 is involved in plant cell growth. tip1–2 plants display growth defects throughout the plant and exhibit defects in both root-hair and pollen-tube growth. tip1–2 plants are partly male sterile resulting from a combination of pollen germination and pollen-tube defects; their root-hairs are short, exhibit a tendency to branch and 2–4 hairs can initiate from each hair cell. They are also slightly dwarf in stature as a result of a general decrease in cell growth indicating that TIP1 activity is required for general cell growth. We propose a role for TIP in both the initiation and maintenance of growth in tip-growing cells. In addition TIP1 activity is required for normal cell expansion (non-tip cell growth) indicating that TIP1 is not exclusively involved in tip-growth.

To study the response of non-mycorrhizal and mycorrhizal maize plants to drought, the changes in the pools of non-structural carbohydrates and amino acids were analysed in leaves and roots of two maize cvs. Plants well colonized by the arbuscular mycorrhizal fungus Glomus mosseae (Nicol. & Gerd.) (60% of root length infected) and comparable non-mycorrhizal plants were subjected to moderate drought stress by reducing the water supply. This stress induced a conspicuous increase in the trehalose pool in the mycorrhizal roots, probably because it was accumulated by the fungal symbiont. Furthermore, glucose and fructose were accumulated in leaves and roots of non-mycorrhizal plants but not in the mycorrhizal ones. Starch disappeared completely from the leaves of both mycorrhizal and non-mycorrhizal plants in response to drought. Activities of soluble acid invertase and trehalase were also measured. Acid invertase activity increased during drought in the leaves of both non-mycorrhizal and mycorrhizal plants whilst in the roots it was unaffected in non-mycorrhizal plants and decreased in the mycorrhizal ones. Without drought stress, trehalase activity was considerably higher in the leaves and roots of mycorrhizal plants than in those of non-mycorrhizal plants. It increased conspicuously during drought, primarily in the leaves of non-mycorrhizal plants. A drought-induced accumulation of amino acids as well as imino acids was found in roots and leaves of both mycorrhizal and non-mycorrhizal plants; leaves of mycorrhizal plants accumulated more imino acids than those of non-mycorrhizal ones. Our results show that drought stress and the presence of a mycorrhizal fungus have a considerable effect on carbon partitioning, imino acid and amino acid accumulation in maize plants.

Maize (Zea mays L.) plants parasitized by the root hemi-parasitic angiosperm, Striga hermonthica (Del.) Benth., consistently display a range of symptoms similar to those found in droughted plants. The mechanisms by which these changes occur are largely unknown. However, S. hermonthica has unusually high rates of transpiration, and stomata which are relatively insensitive to water deficit. Consequently, it has often been suggested that the parasite might cause a severe depletion of the available water in the host's rooting zone. To determine whether the lower stomatal conductance and retarded growth of infected plants could be a result of parasite-induced water deficit, we have monitored the matric potential of the growth medium, water use, growth and stomatal conductance of infected vs. uninfected maize plants.

Host plant height and stomatal conductance of parasitized plants were significantly lower than those of control plants from 31 or 37 d after planting (d.a.p.) respectively. However, there was no indication of an increase in the rate of water depletion in the rooting zone of infected plants until approx. 63 d into the parasitic association. In fact, from 39 until 59 d.a.p. infected plants used less water than uninfected control plants, probably the result of the plants having fewer expanded leaves during part of this period, combined with the lower stomatal conductance exhibited by the infected plants from day 37 onwards. Leaf RWC of infected plants was unchanged in comparison with that of uninfected plants, therefore the change in stomatal conductance was not a response to dehydration of the leaf tissue. Our results indicate that parasitism by S. hermonthica does not cause an increase in water uptake/use in the host until well after most of the symptoms of infection have become fully established. It is highly unlikely, therefore, that the observed effects on the host are primarily due to soil water deficit.

Developmental changes in the root apex and accompanying changes in lateral root growth and root hydraulic conductivity were examined for Opuntia ficus-indica (L.) Miller during rapid drying, as occurs for roots near the soil surface, and more gradual drying, as occurs in deeper soil layers. During 7 d of rapid drying (in containers with a 3-cm depth of vermiculite), the rate of root growth decreased sharply and most root apices died; such a determinate pattern of root growth was not due to meristem exhaustion but rather to meristem mortality after 3 d of drying. The length of the meristem, the duration of the cell division cycle, and the length of the elongation zone were unchanged during rapid drying. During 14 d of gradual drying (in containers with a 6-cm depth of vermiculite), root mortality was relatively low; the length of the elongation zone decreased by 70%, the number of meristematic cells decreased 30%, and the duration of the cell cycle increased by 36%. Root hydraulic conductivity (LP) decreased to one half during both drying treatments; LP was restored by 2 d of rewetting owing to the emergence of lateral roots following rapid drying and to renewed apical elongation following gradual drying. Thus, in response to drought, the apical meristems of roots of O. ficus-indica near the surface die, whereas deeper in the substrate cell division and elongation in root apices continue. Water uptake in response to rainfall in the field can be enhanced by lateral root proliferation near the soil surface and additionally by resumption of apical growth for deeper roots.

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

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

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

The leaf surfaces of beech, oak and ginkgo have been investigated with respect to contamination with particles during one growing season. Based on the observation that particles are removed from water-repellent leaves by rain (Lotus effect) the three species were selected because they differ in leaf surface micromorphology and wettability. Leaves of beech are smooth, lacked wax crystals and were ±wettable. Those of ginkgo were rough because their cells were convex and were densely covered by wax crystals, resulting in permanent water repellency. Leaves of oak were covered by waxes and were water repellent when young, but, a few weeks after leaf expansion had ceased the waxes were rapidly eroded. These differences in wettability resulted in different amounts of contamination. Ginkgo collected a very small number of particles during the whole vegetation period. In beech the contamination was significantly higher, but fairly constant, whereas oak leaves accumulated particles with age.

For the first time the well known drought stress hormone abscisic acid, which is involved in regulating processes increasing desiccation tolerance in many plant systems, was analysed in lichens. ABA was detected in all 26 species investigated. In contrast to higher plants and liverworts, the ABA content increased after hydration of air dry lichen thalli and decreased in desiccating lichen material. Experiments with Baeomyces rufus (Huds.) Rebent indicated that the mycobiont might be the major site of ABA biosynthesis. After incubation of hydrated lichen thalli with radioactive ABA for up to 72 h no metabolism to phaseic acid and dihydrophaseic acid could be detected. Fluctuations of internal ABA might be a result of ABA release to the external medium.

A study was carried out to determine whether expansion of marram-grass stands (Ammophila arenaria (L.) Link) on acidic inner Dutch coastal dunes was caused by suppressiveness of soils from these stands against three potential pathogenic fungi of marram grass, namely Fusarium culmorum (W. G. Sm.) Sacc., Phoma exigua Desm. and a Ulocladium sp. The suppressiveness of the acidic inner dune soils was compared with that of lime-rich dune soils from vigorous and declining marram-grass stands. Suppressiveness of the dune soils against the saprotrophic fungi Chaetomium globosum Kunze: Fr, Mucor hiemalis Wehmer and Trichoderma harzianum Rifai was also determined. All fungi had been isolated from marram-grass roots. Suppressiveness was determined by comparing the formation of hyphae from potato–dextrose agar discs into (layer method) or on top of (surface method) dune soils with that of controls consisting of sterile, acid-washed beach sand. The growth of the three root-infecting fungi was strongly inhibited in all soils regardless of the method used. Hence, there were no indications that the potential pathogenic fungi were selectively suppressed by the acidic dune soils and, consequently, the results did not give any indication for the involvement of a fungal component in the decline of marram grass. Growth of the saprotrophs C. globosum and M. hiemalis was much less inhibited than that of the root-infecting fungi. Growth of T. harzianum was strongly inhibited in alkaline soils but not in the acid ones. The suppression of fungal growth could be partly or completely eliminated by a microwave treatment, indicating that biological components of the soil were essential to suppressiveness. The suppression of the fungi by colonies of dune soil micro-organisms on water-agar differed considerably from soil alone. Yet, all methods indicated the occurrence of general suppressiveness against fungi by dune soils, irrespective of the origin of the soil samples. This suppressiveness was probably not due to direct competition with other soil micro-organisms for nutrients but to inhibiting compounds produced by the soil micro-organisms.

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.

Roots of bluebell (Hyacinthoides non-scripta (L.) Chouard ex Rothm.) are colonized by a range of fungal symbionts from several genera of the order Glomales. Using the identification scheme described in Merryweather & Fitter (1998), arbuscular mycorrhizal (AM) fungi in bluebell roots were quantified throughout the single growing season of 1994–5 and compared with populations of spores found in the soil around the roots.

In the early part of the growing season, when its activity is entirely subterranean (autumn and winter), bluebell habitually associates with a Scutellospora morphotype which is almost certainly S. dipurpurescens Morton & Koske (emend. Walker, 1993) whose spores occur in the root zone. This is the time of maximum phosphorus inflow and bulb-stored carbohydrate utilization by this mycorrhiza. A diverse flora of other AM fungal morphotypes (Acaulospora and Glomus), which might also form mycorrhizas with other plants in the vicinity of bluebells, invade the roots later in the season (spring), when P inflow is reduced and carbohydrate is available as fresh photosynthate. Their contribution to the mycorrhiza might be less than that of Scutellospora, particularly in terms of P assimilation.

Both AM fungi in roots and glomalean spores recovered from soil around bluebell roots showed a significant degree of correlation with the vegetation within which the test plants grew. In the case of AM fungi in roots, Scutellospora showed no special preference for either, but Glomus correlated with a canopy of sycamore (Acer pseudoplatanus L.) and Acaulospora with oak (Quercus petraea (Mattuschka) Liebl.). Spores which most closely resembled S. dipurpurescens and Acaulospora gerdemannii Schenck & Nicolson were significantly more numerous under sycamore, but a spore like Acaulospora koskei Blaskowski, the most numerous and frequently encountered glomalean spore in the system, showed no preference for areas dominated by either tree. There was no significant relationship between AM fungal populations in bluebell roots and glomalean spores recovered from associated soil.

The two spore taxa most frequently found in the vicinity of bluebell roots (A. koskei and S. dipurpurescens) were also found in lower numbers in soil from a region of the field site in which bluebell was absent, indicating that the main bluebell AM fungi do not exclusively associate with that host.

We have studied the chitin and ergosterol contents of ectomycorrhizal roots in three sets of experiments to evaluate them as indicators of fungal biomass. The first set of experiments showed that ageing had a marked effect on ergosterol concentrations. The ergosterol content of 7-month-old, brown, shrunken Pinus sylvestris L.–Paxillus involutus (Fr.) Fr. mycorrhizas was found to be only 10% of that found in white, turgid, 1- or 4-month-old specimens. This supports the hypothesis that the compound is a good indicator of living fungal biomass. Ageing had a lesser effect on chitin concentrations since the chitin levels found in 7-month-old mycorrhizas were still 60% of the levels found in 1- and 4-month-old specimens.

Consequently, the chitin[ratio ]ergosterol ratio increased from about 14 to 19 in 1- and 4-month-old mycorrhizas respectively to about 110 in 7-month-old mycorrhizas. In the second set of experiments, we found that variation in plant growth had no effect on the chitin[ratio ]ergosterol ratio in whole root systems of either Alnus incana (L.) Moench or Pinus sylvestris mycorrhizal with Paxillus involutus. In the third set of experiments, we found a constant relationship between the two marker concentrations in 10-month-old root systems of Pinus sylvestris, regardless of fungal species involved, using Paxillus involutus, Piloderma croceum Erikss. & Hjorts and Suillus variegatus (Fr.) O. Kuntze as test organisms. Taken together, the results of this study suggest that both chitin and ergosterol give reliable, but different, relative measures of fungal biomass in mycorrhizal roots. Furthermore, we demonstrate that, in combination, the two chemical markers can be used to estimate both total and living fungal biomass (derived from the chitin[ratio ]ergosterol ratio).

Sixty-two isolates of Pisolithus were obtained from a carpophore population collected from a 90-km2 area of New South Wales, Australia. Random amplified polymorphic DNA (RAPD) analysis was used initially to screen isolates for variation, and restriction fragment length polymorphism (RFLP) analysis of the internal transcribed spacer (ITS) region and ITS sequence analysis were subsequently performed on selected isolates. ITS–RFLP and ITS sequence analysis indicated that two isolates (LJ07 and WM01) from different sites were considerably different from all other isolates, a strict consensus tree generated by bootstrap analysis of sequence data strongly suggesting that isolates LJ07 and WM01 represent a separate Pisolithus species. Although the general morphology of carpophores and basidiospores of the majority of collected isolates was consistent with previous descriptions of P. tinctorius (Pers.) Coker and Couch, basidiospore characteristics suggest that isolates LJO7 and WMO1 might represent a Pisolithus species distinct from P. tinctorius and the previously described Australian species, P. microcarpus (Cooke & Massee) G. Cunn.

A recent re-analysis of the data of Salisbury (1974) claims his data do not support the hypothesis that seeds of species from shaded habitats are heavier than those from unshaded habitats, partly because the original analysis was inappropriate and partly because of bias in the dataset. We show first that the re-analysis itself contains errors, and second that the charge of bias is based largely on a misunderstanding. We also show that analysis of a larger dataset, drawn from Salisbury (1942) and from Salisbury (1974), provides convincing support for the hypothesis and suggests that the relationship is independent of life history.

Studies on whether or not species which need more light for establishment have smaller seeds should be made between plants of similar mature height living in the same soil and macroclimate. Salisbury's classic comparisons confused differences in light-requirement during establishment with differences in mature height and in the supply of soil resources.