Differences between individuals collected from sun and shade populations could result from either somatic or genetic differences, particularly in populations of perennial plants. Our objective in this study was to separate somatic from genetic differences. We collected Anthoxanthum odoratum and Plantago lanceolata from sun and shade populations and made measurements on both vegetatively propagated clones and seed progeny from each clone. The parental populations differed in a wide range of physiological and morphological traits. However, only photosynthetic capacity was significantly different between both the original sun and shade populations and their seed progeny. In both species, plants from the sun population had higher photosynthetic capacities than those from the shade population when grown in a common environment. This demonstrates that there was genetic differentiation between the sun and shade populations. Photosynthetic capacity of parents and offspring also differed, suggesting a somatic effect. Since many of the original clones were virus-infected, but all but one of the offspring were virus-free, this might have been a result of virus infection. However, in spite of the fact that the parents and offspring clones were propagated vegetatively so that the plants were at the same developmental stage at the time of measurement, we cannot rule out the possibility that differences in age of cell lines could also have been a factor.

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

The authoritative talk by Professor Fowler (Fowler et al., 1998), emphasized the huge increase in the rate of NOx (NO and NO2) emissions into the atmosphere due to fossil fuel combustion, from 1 Tg N y−1 to over 20 Tg N y−1 during the 100 yr between 1880 and 1980. He went on to predict that this rate of emission from anthropogenic sources would increase to 46 Tg N y−1 by the year 2025. In addition, NO can also be released from the soil following microbial action, a process that is very dependent upon soil temperature, nitrogen availability and water content. Later in the meeting, Professor Raven (Raven & Yin, 1998) pointed out that terrestrial plants, though not necessarily each individual species, have over the past 450 million yr coped with large changes in nitrogenous compounds in the environment. Nevertheless, this is no basis for complacency about the current situation because the rates of change caused by man's activities are probably unprecedented. Furthermore, the fact that terrestrial plant life in some form can continue, despite massive changes in environmental chemistry, does not necessarily indicate that the systems on which we ourselves are dependent will be conserved.

The morphology and anatomy of ectomycorrhizas of Rhizopogon parksii, Rhizopogon vinicolor and Rhizopogon subcaerulescens, and a hypogeous relative, Truncocolumella citrina, synthesized on Douglas fir in glasshouse conditions using spore slurries as inoculum, are described and compared. Mycorrhizas formed with R. parksii and R. vinicolor did not exhibit their characteristic subtuberculate morphology in these tests, but rather had a pinnate form. All species had diagnostic features of ectomycorrhizas: a well-developed Hartig net and a fungal mantle. In addition, several species exhibited crystal inclusions in the outer mantle, usually at the interface between the mantle and soil. Truncocolumella citrina had crystal-like inclusions within the mantle but external to fungal hyphae, a feature rarely described in ectomycorrhizas.

Fructans (polyfructosylsucrose) are synthesized by a number of plants and micro-organisms. Plant fructans are localized in the vacuole and have a low degree of polymerization (DP), whereas the fructans synthesized by micro-organisms are usually much bigger. There is an increasing interest in fructans for both food and non-food applications. In order to accumulate fructans of high DP in the plant vacuole, the levansucrase protein of Bacillus subtilis was fused to the vacuolar targeting sequence of sporamin and expressed in plants. Transgenic tobacco plants in which this fusion gene is expressed accumulate fructans to levels up to 21% of the d. wt. They showed a reduced translocation of carbohydrates, bleaching of the leaves, stunted growth and increased levels of hexoses and starch. The levansucrase protein was not translocated to the plant vacuole, but retained in the endomembrane system, even though the same targeting signal was able to translocate the E. coli GUS protein to the plant vacuole.

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.

The non-reducing disaccharide trehalose is a major storage compound in most fungi. For a better understanding of carbon metabolism in the ectomycorrhizal-forming basidiomycete Amanita muscaria, trehalase activity was analysed from mycelium growing in liquid culture or on agar plates, and from mycorrhizas. Trehalase activity was found in both the culture medium and in mycelial extracts. The excreted trehalase was purified to apparent homogeneity by anion-exchange chromatography, gel filtration and preparative gel electrophoresis. The identified enzyme belongs to the group of acid trehalases. The apparent molecular mass of the native enzyme was estimated to be 165 kDa by gel filtration and 135 kDa by SDS–PAGE. Isoelectrofocusing indicated an isoelectric point (pI) of approx. 3.7, which is typical for acid trehalases. The enzyme is highly specific for its substrate trehalose, with an apparent Km of 0.38 mM. Validamycin and sucrose act as competitive inhibitors with Ki values of 45 nM and 15 mM, respectively. The activity of acid trehalase excreted into the growth medium is independent of the carbon source (glucose or trehalose), revealing that the enzyme is not regulated by its substrate trehalose. Nevertheless, fungal hyphae grown in the absence of an external carbon source showed enhanced enzyme excretion. The biochemical characteristics of the trehalase activity measured in the mycelium are in the same range as those determined for the excreted enzyme. The enzyme is localized in the cell wall and its activity is strongly decreased in ectomycorrhizas.

The biomass of Mycelis muralis (L.) Dumort plants increased with increases in growth irradiance and nitrate supply. Comparisons of the data from this study with published information indicate that biomass production and growth rate of M. muralis are comparable to that of a number of slow-growing species from resource-poor environments. Biomass allocation varied with growth irradiance and nitrate supply. At high irradiances and low nitrate supplies there was a significantly greater allocation of plant mass to roots, resulting in a lower leaf area ratio (LAR) and leaf mass ratio (LMR). In contrast to the findings of a number of other studies, light level and nitrate supply had interactive effects on leaf mass per unit area (LMA). Changes in LAR due to variations in irradiance were mainly a consequence of changes in LMR. Given the ability of M. muralis to grow in deep shade, this suggests that large changes to within-leaf allocation (LMA) are not an essential requirement for growth at low irradiances. Conversely, because M. muralis can also grow in fully exposed situations, these traits might not exclude some shade species from high irradiance habitats. However, a low growth rate, or traits which result in a low growth rate, might be advantageous in a number of resource-poor environments.

Dr Willem Asman concluded that the major global sources of atmospheric NH3 are excreta from domestic animals and fertilizers. A question raised was: how reliable are the emission estimates and extrapolations? The answer was that emission estimates are surrounded by uncertainty, which is a major handicap to sound modelling of NH3 dry deposition and, consequently, to obtaining good estimates of critical load exceedences.

Major uncertainties in emission estimates seem to be related to the use of simple emission factors, many of which are highly empirical or have been derived from measurements carried out under conditions which deviate considerably from those following modern practices of handling and applying manure and fertilizers. An example is provided by the commonly used emission factors for synthetic fertilizers (see e.g. Bouwman et al. (1997)), which are much higher than recent micrometeorological assessments seem to suggest. Thus, emission from urea, the most widespread fertilizer used in the world (currently around 55% of world N consumption) can be completely avoided if the fertilizer is incorporated into the upper soil layers. Similarly, a growing crop can reduce losses to well below 10% of the applied amount of urea-N, i.e. to less than half of the generally used emission factors of 15% for Europe and 25% for the tropics. The emission factor for NPK-fertilizer is set at 4%, whereas that for pure calcium-ammonium-nitrate, the same N compound as is present in NPK-fertilizers, is assumed to be only 2%.

The role of external Ca2+ in the homeostasis of intracellular pH (pHi) of Anabaena sp. strain PCC7120 in response to a decrease in the external pH (pHex) has been studied in cell suspensions. Increase in cytoplasmic pH after acid shock is dependent on the presence of Ca2+ in the medium. The observed Ca2+-mediated alkalization of the cytoplasm depends on the extent of the shift in external pH. Acid pH shifts resulted in an increased permeability of the cytoplasmic membrane to protons, which could be reversed by increasing the concentration of Ca2+ in the medium. Thus, the ability of Ca2+ to increase cytoplasmic pH might be correlated with an inhibition of net proton uptake by increasing concentrations of external Ca2+ under these conditions. This combined response resulted in the generation and maintenance of a larger pH gradient (ΔpH) at acid external pH values. All Ca2+ channel blockers tested, such as verapamil and LaCl3, inhibited the observed Ca2+-mediated response. On the other hand, the Ca ionophore calcimycin (compound A23187) was agonistic, and stimulated both cytoplasmic alkalization and inhibition of net proton uptake. The protonophorous uncoupler carbonylcyanide m-chlorophenyl hydrazone, inhibited this Ca2+-mediated response, whereas monensin, an inhibitor of the Na+/H+ antiporter, had no significant effect. The results of the present study suggest that an influx of Ca2+ from the extracellular space is required for the regulation of cytoplasmic pH in Anabaena sp. strain PCC7120 exposed to low external pH values.

Under ecologically realistic environmental conditions, the water-use efficiency (WUE) of Peperomia scandens, a CAM plant, was higher than that of the C3 congener P. obtusifolia. This difference has been attributed to differences in stomatal activity between C3 and CAM plants, coupled with differences in the evaporative demand of the atmosphere during which the stomata are open. This explanation has apparently not, however, been experimentally tested. Thus, WUEs were compared in these species in two experiments in which the atmospheric evaporative demand was identical (or nearly so) during the period of stomatal opening (i.e. during the night for the CAM plant and during the day for the C3 species). In both experiments, the WUE of the CAM species was higher than that of the C3 species. These results suggest that factors other than differences in atmospheric environmental conditions must also be responsible for the observed differences in WUE. Because CO2 uptake rates of the CAM species were substantially lower than those of the C3 species, the lower WUE in the CAM species resulted primarily from lower transpiration rates. Lower rates of water loss in P. scandens, relative to rates in P. obtusifolia, were ascribed, in part, to lower stomatal densities. Thus, leaf morphological differences, in addition to differences in atmospheric evaporative demand, help to explain the high WUE typically measured in CAM plants.

Four pot experiments are reported in which Norway spruce (Picea abies (L.) Karst) seedlings, of different nutrient status, were treated with acid mist for one growing season in open-top chambers (OTCs). Combinations of H+, SO42−, NH4+ and NO3− were applied at different frequencies of application and supplying different doses of S and N kg ha−1. Plant growth, visible injury, frost hardiness and nutrient status were observed. These experiments were undertaken to improve our understanding of the interaction of environmental factors such as nutrition and mist-exposure frequency on seedling response to N and S deposition.

Both acidity ([les ]pH 2·7) and SO42− ions were necessary to induce visible injury. Mist containing SO42−, H+ and to a lesser extent NH4+ significantly reduced winter frost hardiness. Increasing the misting frequency, and to a lesser extent the overall dose, increased the likelihood of acid mist causing visible injury and reducing frost hardiness. Post-planting stress, low N status and needle juvenility increased the likelihood of acid mist causing visible injury. Increased plant vitality, adequate N status and growth rate reduced the likelihood of acid-mist-induced reductions in frost hardiness.

Principles underlying the responses of spruce seedlings treated in controlled conditions to acid mist are discussed.

Fructans are linear or branched polymers containing a single sucrose and repeating fructose residues. An early model for fructan biosynthesis in higher plants suggested that partial synthesis of the polymer occurred in the cell cytosol. The current model suggests that synthesis requires the interaction of two separate fructosyltransferases located in the vacuole. Tobacco lines containing a chemically induced promoter, directing expression of the Bacillus amyloliquefaciens SacB gene in the present study, provided an opportunity to regulate and target fructan synthesis to the cytosol of transgenic plants. Induced expression of the gene led to rapid destruction of leaf tissue. Amino acid substitution at a highly conserved site (Arg331) in the SacB gene reduced the fructosyltransferase efficiency without reducing the invertase activity of the enzyme. Expression of the mutant gene in transgenic tobacco also resulted in leaf damage. However, the appearance of necrotic tissue was greatly delayed. The results suggest that the phenotype is due to accumulation of fructan in the cytosol. Fructan metabolism in the cytosol of potato tubers was also detrimental to development. Tuber size and starch synthesis was significantly reduced in lines containing the untargeted gene. Transgenic tobacco and potato containing the SacB gene offer an opportunity to study the metabolism of fructan and the effect of accumulation on plant cell development.

Companion cells in the secondary phloem of 125 Indian dicotyledonous species belonging to 43 families were examined by light microscopy. Four types of companion cell were identified: E-, S- and L-types which were equal in length, shorter, or longer, respectively, than the associated sieve tube element; and R-type, in which two or more companion cells in a vertical row were associated with the sieve tube element. The commonest was the E-type and the rarest was the L-type. E-type companion cells were most frequently found associated with short sieve tube elements (50–250 μm) which had a high frequency of simple sieve plates, considered phylogenetically advanced. R-type companion cells were most frequently associated with long sieve tube elements (>400 μm) with a high frequency of compound sieve plates, considered phylogenetically the least advanced. A strong positive correlation was found between the average number of companion cells associated with a sieve tube element and the lengths of the sieve tube elements. There was also a strong negative correlation between the average number of companion cells associated with a sieve tube element and companion cell lengths.

Oxygen evolution was measured from mesophyll tissues in spinach leaves using a photoacoustic technique. The photosynthetic capacity of individual cell layers was measured by directing microscopic beams of light, 40 μm wide, to cells exposed within a leaf cross section. The resulting profile for oxygen-evolution potential was relatively flat, indicating a uniform capacity for photosynthesis in leaf mesophyll tissues. Two experimental approaches were used to estimate the photosynthetic performance of individual mesophyll cell layers when white light was applied to the adaxial leaf surface. These experiments indicated that oxygen was produced relatively uniformly across the mesophyll and that oxygen evolution increased with irradiance of the white light applied to the leaf surface. The measured profiles for oxygen evolution and capacity are flatter than previous measurements of profiles of fixed carbon and estimates of profiles for absorbed light within spinach leaves.

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

This study investigated the mechanisms which control the partitioning between roots and shoots in plants subjected to changes in environment. Two types of analyses were used: firstly, an examination of the cost and revenue associated with investment in different plant parts, and secondly, a test of the principle of functional equilibrium between roots and shoots, i.e. whether root dry matter × root specific activity balances shoot dry matter × shoot specific activity. Measurements were made on individual plants of Lolium perenne in sunlit controlled environments, grown from germination to canopy closure under optimal nitrogen supply. At the final harvest, increased air temperature (+4 °C above ambient) reduced whole-plant dry matter by 12% relative to the control, whereas elevated CO2 mole fraction (700 μmol mol−1) led to a 38% gain. The combined treatment yielded an intermediate result (+19%). Plants grown at +4 °C maintained balanced activity between roots and shoots throughout the experimental period, irrespective of CO2 concentration. This required enhanced allocation to roots in young plants to compensate for a strong negative effect of higher temperature on root specific activity, which suggests that plants conserve balanced activity by adjusting dry matter partitioning. The extra cost involved with the adjustment at +4 °C significantly enhanced the cost[ratio ]revenue ratio of plant investment. In ambient temperature, the balance between roots and shoots departed from equilibrium, slightly at ambient but substantially at elevated CO2: the plants accumulated excess carbon relative to nitrogen, and this imbalance increased with plant age. At elevated CO2, the cost[ratio ]revenue ratio increased in young plants but this was later reversed owing to loss of root specific activity, which explains the gradually declining CO2 stimulation with time. The strategies in equilibrating root and shoot functioning observed in the different treatments are discussed in the light of whole plant performance.

In contrast to that of Anabaena sp. PCC 7120, the fdxN gene in Anabaena variabilis ATCC 29413 is not interrupted by a 55-kb DNA element, making this strain more suitable for genetic analysis of fdxN independent of the developmentally regulated excision during heterocyst formation. As a basis for mutational analysis, the fdxN gene of A. variabilis was cloned and sequenced. The deduced FdxN protein sequence was highly homologous to the Anabaena 7120 fdxN gene product including eight cysteine residues that are known to be conserved among ferredoxins containing two [4Fe-4S] clusters. The fdxN gene of A. variabilis was disrupted by insertion of an interposon within the fdxN coding region resulting in mutant strain KG29. Diazotrophic growth and in vivo nitrogenase activity of KG29 were similar to those of the wild-type, indicating that FdxN was not essential for N2 fixation in A. variabilis.

The revised local curvature distribution model, which provides accurate computer simulations of the gravitropic response of mushroom stems, was found to produce accurate simulations of the gravitropic reaction of wheat (Triticum aestivum) coleoptiles. The key feature of the mathematical model that enables it to approach universality of application is the assumption that the stem has an autonomic straightening reaction (curvature compensation or ‘autotropism’). In the model, the local bending rate for any segment of the organ is determined by the difference between the ‘bending signal’ (generated by the gravitropic signal perception system) and a ‘straightening signal’ (which is proportional to the local curvature of the segment). The model reveals three major differences between the gravitropic reactions of wheat coleoptiles and Coprinus mushroom stems. First, in Coprinus, the capacity for autonomic straightening is much more concentrated in the apical region of the stem. Second, local perception of the gravitropic signal, which is necessary for exact simulation in Coprinus, is not needed in wheat coleoptiles (the corresponding constant in the model can be set to zero). Third, the transmission rate of the gravitropic signal is about seven times faster in wheat coleoptiles than in the mushroom stem. Thus, we demonstrate that a single model, depending on the values given to its parameters, is able to simulate the spatial organization of the gravitropic reaction of wheat coleoptiles and Coprinus mushroom stems. The model promises to be a valuable predictive tool in guiding future research into the gravitropic reaction of axial organs of all types.