Thioredoxins are ubiquitous small-molecular-weight proteins (typically 100–120 amino-acid residues) containing an extremely reactive disulphide bridge with a highly conserved sequence -Cys-Gly(Ala/Pro)-Pro-Cys-. In bacteria and animal cells, thioredoxins participate in multiple reactions which require reduction of disulphide bonds on selected target proteins/enzymes. There is now ample biochemical evidence that thioredoxins exert very specific functions in plants, the best documented being the redox regulation of chloroplast enzymes. Another area in which thioredoxins are believed to play a prominent role is in reserve protein mobilization during the process of germination. It has been discovered that thioredoxins constitute a large multigene family in plants with different subcellular localizations, a unique feature in living cells so far. Evolutionary studies based on these molecules will be discussed, as well as the available biochemical and genetic evidence related to their functions in plant cells. Eukaryotic photosynthetic plant cells are also unique in that they possess two different reducing systems, one extrachloroplastic dependent on NADPH as an electron donor, and the other one chloroplastic, dependent on photoreduced ferredoxin. This review will examine in detail the latest progresses in the area of thioredoxin structural biology in plants, this protein being an excellent model for this purpose. The structural features of the reducing enzymes ferredoxin thioredoxin reductase and NADPH thioredoxin reductase will also be described. The properties of the target enzymes known so far in plants will be detailed with special emphasis on the structural features which make them redox regulatory. Based on sequence analysis, evidence will be presented that redox regulation of enzymes of the biosynthetic pathways first appeared in cyanobacteria possibly as a way to cope with the oxidants produced by oxygenic photosynthesis. It became more elaborate in the chloroplasts of higher plants where a co-ordinated functioning of the chloroplastic and extra chloroplastic metabolisms is required.

Sequential harvests of cotton seedlings grown in soil cores enabled the quantification of the density of arbuscular mycorrhizal fungi to detect the effects of time, cultivation and periodic wetting of the soil. Cotton seedlings grown in soil cores from three locations formed arbuscular mycorrhizas at similar rates when cores were stored dry for up to 18 months. Disturbance of dry cores followed by dry storage for 18 months did not reduce the rate of establishment of mycorrhizas. Periodic wetting and drying of the cores, especially if the cores had first been disturbed, significantly reduced the rate of establishment of mycorrhizas. We suggest that long fallow disorder is possibly caused by falls of rain in clay soils of eastern Australia used to grow cotton. The proportion of the root with mycorrhizas at 3 wk was strongly correlated with the infection at 8 wk. We also suggest that it might be possible to predict maximum levels of infection and early uptake of phosphate of seedlings by determining the proportion of roots that are mycorrhizal 3 wk after emergence of cotton seedlings.

Safflower (Carthamus tinctorius L. cv. S555) and spring wheat (Triticum aestivum L. cv. Anza) were grown with or without the arbuscular mycorrhizal fungus Glomus etunicatum Becker & Gerd., under environmentally controlled conditions. Soil phosphate concentrations were adjusted before planting to produce mycorrhizal (M) and non-mycorrhizal (NM) plants that had similar leaf areas and root length densities at the same stage of development before initiating drought stress treatments. Drought did not affect the amount of mycorrhizal infection in safflower or wheat. Interactions between water stress treatments and mycorrhizal infection on plant growth and phosphorus uptake were limited and only occurred in wheat. NM wheat plants had 28% greater shoot d. wt, slightly greater root length densities, and 39% greater P acquisition than M plants when grown under well watered conditions, but under droughted conditions plant size and tissue P contents of M and NM wheat plants were similar. Mycorrhizas did not affect stomatal behaviour during drought stress in either safflower or wheat, i.e., transpiration and stomatal conductance declined independently of infection as soil water was depleted and leaf water potentials declined. Therefore, mycorrhizal infection did not alter the intrinsic hydraulic properties of the plant/soil system. Whilst wheat maintained turgor of recently expanded leaves during severe drought and safflower did not, mycorrhizal infection had no effect on leaf turgor during drought in either plant species.

The objective of this study was to determine if infection by arbuscular mycorrhizal fungi alters water uptake by roots under well watered to severely droughted conditions. Safflower and wheat plants were grown with and without the mycorrhizal fungi, Glomus etunicatum or G. intraradices in nutrient-amended soil under environmentally controlled conditions to yield mycorrhizal and non-mycorrhizal plants with similar leaf areas, root length densities, d. wt, and adequate tissue phosphorus and nitrogen. Specific water uptake rates (cm3 of water cm−1 root length d−1) were estimated non-destructively at various depths in the soil from changes in the soil water content measured using a gamma attenuation method. When soil water was severely depleted, changes in soil water potentials were also measured with soil psychrometers. Roots from both plant species extracted water at the fastest rate from the upper soil layers when the soil water content was high, and later, extracted water primarily from deeper depths as water in the upper soil layers was depleted. Mycorrhizal infection did not affect the rates at which roots extracted water from soil whether soil moisture conditions were at their wettest condition, at container capacity, or at the driest extreme when soil water potentials ranged from −1·5 to −2·0 MPa and the plants were completely wilted. Plant water relations were also largely unaffected by infection. Mycorrhizal infection did not alter the ability of plants to extract water from soil even during extreme drought.

The resurrection plant Chamaegigas intrepidus Dinter (Scrophulariaceae) grows as a typical hydrophyte in shallow rock pools on granitic outcrops in arid areas of Namibia. During the rainy season, the rock pools are temporarily filled with water. When the pools dry up, C. intrepidus desiccates and survives in an air-dry condition for at least 8 months. After rewatering, the plants regain their metabolic activity in under 2 h. The desiccation of the vegetative organs is accompanied by a dramatic accumulation of abscisic acid (ABA). Beyond this, desiccation of roots is accompanied by the occurrence of specific dehydration-related proteins, whereas the leaves of C. intrepidus show high levels of dehydrins in the dehydrated as well as in the hydrated state.

Investigations in Namibia showed drastic diurnal fluctuations in the pH of the rock pools. The pH value increased from slightly acidic or neutral conditions during the morning to alkaline conditions (up to pH 12) during late afternoon. Since compartmental ABA distribution depends strongly on pH gradients across membranes, the external pH would be expected to affect the ABA relations in the plant. According to the anion trap concept, an alkaline pH in the surrounding medium should cause a release of ABA from the roots, although C. intrepidus appeared to release less ABA than the terrestrial rosettes of Valerianella locusta.

The effects of prolonged simulated acid rain on percentage cover of ground vegetation, and on growth and reproduction of two dominating dwarf shrubs (Empetrum nigrum and Vaccinium vitis-idaea) were examined in a field experiment in the Finnish Subarctic, in an area with low ambient levels of sulphur and nitrogen deposition. Acid rain treatments included moderate (pH 3·8) and high (pH 2·9) concentrations of either H2SO4, or HNO3, or a mixture of them, and were compared with irrigated (pH 6) and dry control plots.

Long-term application of acid rain caused significant alteration in the cover and composition of ground vegetation. Effects of acid rain depended on the accompanying anion and on pH. Sub-plots under different canopy tree species differed in responses indicating that spatial heterogeneity is important in predicting the effect of acidifying pollution on this plant community.

In the bottom layer, acid rain caused significant reduction in cover of the cyanobacterial lichens Nephroma arcticum and Peltigera spp. Decrease in cover of fruticose lichens, mainly composed of Cladina spp., more likely resulted from additional watering.

In the field layer, acid rain containing moderate concentrations of NO3− caused an increase in cover of graminoid species. There were only slight alterations in growth and cover of the two dominant evergreen dwarf shrubs, E. nigrum and V. vitis-idaea, indicating that these species are tolerant to acid rain of as low as pH 3. Even some positive responses of dwarf shrubs were observed, depending on canopy tree. Application of acid rain of pH 3 to plots under pine trees caused an increase in cover of V. vitis-idaea and, when the nitric acid only was applied, a short-term increase in the number of new shoots of E. nigrum.

In contrast to vegetative growth, reproduction of the dwarf shrubs was more strongly affected by acid rain, but this also depended on local conditions and anion composition of acid rain. On ‘pine’ plots, rain of pH 3 reduced the number of berries and flower buds on terminal current shoot of E. nigrum, however, this was partially compensated by an increase in berry production at the ramet level. Simulated acid rain had mainly negative effects on berry production by V. vitis-idaea.

Exposure to ozone (O3) has been shown to decrease the allocation of carbon to tree roots. Decreased allocation of carbon to roots might disrupt root metabolism and rhizosphere organisms. The effects of soil type and shoot O3 exposure on below-ground respiration and soil microbial populations were investigated using container-grown ponderosa pine (Pinus ponderosa Laws.) growing in a low-nutrient soil, or a fertilizer-amended organic potting media, and exposed to one of three levels of O3 for two growing seasons in open-top exposure chambers. A closed system, designed to measure below-ground respiratory activity (CO2 production, O2 consumption and RQ-Respiration Quotient; (CO2[ratio ]O2) of plants growing in pots, was used monthly to monitor below-ground respiration of 3-yr-old ponderosa pine.

Although seasonal differences were detected, CO2 production (μmol h−1 g−1 total root d. wt), O2 consumption (μmol h−1 g−1 total root d. wt) and RQ (CO2[ratio ]O2) increased with increasing O3 exposure level. Seasonal patterns showed increased respiration rates during periods of rapid root growth in spring and early fall. Respiration quotient tended to decrease during known periods of active root growth in control seedlings, but a similar response was not observed in O3-treated seedlings. Responses to O3 were greatest in the soil-grown plants, which had a lower fertility level than media-grown plants. Although root d. wt was decreased, root[ratio ]shoot ratios did not change in response to O3. Soil-grown plants had higher root-shoot ratios than media-grown plants, reflecting the lower fertility of the soil.

Plant exposure to O3 was found to affect both active and total populations of soil organisms. In both organic potting media and in soil, biomass of active soil fungi, and the ratio of active-fungal to active-bacterial biomass increased with increasing plant exposure to O3. The effect of O3 on total fungal and bacterial biomass was not linear: at low O3 levels, total fungal and bacterial biomass increased; at the high O3 level, total fungal and bacterial biomass decreased compared with those of controls.

Our results show that O3 exposure to shoots significantly disrupts CO2 production and O2 consumption of soil and roots of ponderosa pine seedlings. Below-ground respiratory differences were thought to be a result of changes in respiratory substrates, carbon refixation within the plant and soil microbial activity. Ozone also changes below-ground RQ, suggesting that O3 substantially disrupts root metabolism and interactions with rhizosphere organisms. Ozone exposure of ponderosa pine grown in different soil types can disrupt below-ground respiration and influence populations of soil organisms without alteration of biomass partitioning between above- and below-ground plant components. Collectively, the effect of O3 on the below-ground system is of concern since it is likely that these changes are accompanied by a change in the ability of root systems to acquire nutrient and water resources and possibly to synthesize amino acids and proteins necessary for normal plant function.

Spring wheat (Triticum aestivum cv. Nandu) cultivated under glasshouse conditions was exposed to ozone in large fumigation chambers for 2 wk. Different exposure regimes were applied as constant concentrations as well as with ozone peaks, partly under equal dose-conditions, in times of high solar radiation during different stages of development (seedling, late tillering, anthesis). Chlorophyll fluorescence was monitored and amounts of carbohydrates (hexoses, sucrose, starch) and chlorophyll were measured in young leaves (seedling) and flag leaves (late tillering, anthesis) during and after ozone exposure. Although seedlings showed no significant response in photosynthesis, strong effects on photosynthesis and carbohydrate accumulation were measured when plants were fumigated during anthesis, especially after a heat stress period preceding ozone treatments. Under equal dose conditions chlorophyll fluorescence parameters (Fv[ratio ]Fm) and electron transport rate decreased and sucrose content of flag leaves increased significantly if ozone at a concentration of 220 μg m−3 was supplied for 4 h, indicating that peak concentrations show stronger effects than constant concentrations. The reaction of wheat plants is dependent on environmental conditions such as preceding heat stress and on the developmental stage during exposure. The results favour the hypothesis that photoinhibition and disturbance of photosynthesis are only secondary effects as a consequence of retarded sucrose export from the leaf, because of damage at the plasma membrane.

Development of mycelial cord systems of Phanerochaete velutina (DC.: Pers.) Parmasto from 4-cm3 inocula on a nutrient-depleted non-sterile soil was studied in laboratory microcosms using image analysis techniques. Cord systems were ‘baited’ after 13 d growth with either fresh, non-sterile 4-cm3 wood baits or control Perspex® blocks of the same contact area placed behind the foraging mycelial front. After 26 d growth, mycelial ‘patches’ arose by dedifferentiation of consolidated mycelial cords in both wood- and Perspex-baited cord systems. ‘Patches’ comprised fine, highly branched separate hyphae extending radially from points of aggregated hyphae in cords. ‘Patches’ and cords could be readily distinguished by image analysis and the areas covered by patches and cords could be measured and compared. Whilst the total hyphal cover of Perspex- and wood-baited systems did not differ significantly (P>0·05), patch cover in wood-baited systems was up to 10 times greater than in Perspex-baited systems. Patches were temporary structures, regressing more rapidly with age than mycelial cords. Patch development ceased after application of a nutrient solution which replenished phosphate levels in the soil. Wood-baited mycelial systems displayed significant developmental polarity (P[les ]0·05) of both total hyphal cover (patches plus cords) and hyphae in patches towards the ‘baited’ sector of cord systems after 42 d, which corresponded with peak patch development. However, significant (P[les ]0·05) developmental polarity of the mycelial systems along the bait-inoculum line could be detected 8 d before patch formation when assessed by fractal geometry. Radiotracer studies showed that mycelial patches were not sinks for supplied 32P, but that they were sites of increased nutrient uptake capacity compared with that of mycelial cords. We discuss the need for mycelial cord systems to balance allocation of mycelial biomass between the two essential processes of colonizing wood resource units, and the acquisition of soluble inorganic nutrients from soil.

Arbuscular mycorrhizal (AM) fungi, Rhizobium bacteria and plant-growth-promoting rhizobacteria (PGPR) were isolated from a representative area of a desertified semi-arid ecosystem in the south-east of Spain. Microbial isolates were characterized and screened for effectiveness by a single-inoculation trial in soil microcosms. Anthyllis cytisoides L., a mycotrophic pioneer legume, dominant in the target mediterranean ecosystem, was the test plant. Several microbial cultures from existing collections were also included in the screening process. Two AM fungi (Glomus coronatum, native, and Glomus intraradices, exotic), two Rhizobium bacteria (NR4 and NR9, both native) and two PGPR (A2, native, and E, exotic) were selected. A further screening for the appropriate double and triple combinations of microbial inoculants was then performed. The parameters evaluated were biomass accumulation and allocation, N and P uptake, N2-fixation (15N) and specific root length. Overall, G. coronatum, native in the field site was more effective than the exotic G. intraradices in co-inoculation treatments. In general, our results support the importance of physiological and genetic adaptation of microbes to the whole environment, thus local isolates must be involved. Many microbial combinations were effective in improving either plant development, nutrient uptake, N2-fixation or root system quality. Selective and specific functional compatibility relationships in plant response between the microbial inoculants, were observed. Despite the difficulty of selecting a multifunctional microbial inoculum, appropriate microbial combinations can be recommended for a given biotechnological input related to improvement of plant performance. This could be exploited in nursery production of target plant species endowed with optimized rhizosphere/mycorrhizosphere systems that can be tailored to help plants to establish and survive in nutrient-deficient, degraded habitats. The relevance of this microbial-based approach in the context of a reclamation strategy addressed to environmental sustainability purposes is discussed.

Mineral nutrient concentrations were determined in leaves of 83 mostly herbaceous species collected from central England. Most samples were analysed for N, P, K, Ca, Mg, Na, Fe, Al, Mn, Cu and Zn. Concentrations of K, N and P showed similar levels of interspecific variability, with the highest concentrations being 6–9 times the lowest. Mg and (especially) Ca were much more variable, with the highest concentrations being 24 and 49 times the lowest respectively. Only in the case of P concentration was the majority of the variance in the data found at or below the species level. Most of the variance in Ca and Mg concentrations was between monocots and dicots. Concentrations of N and P were strongly positively correlated with each other. Only Ca and Mn were consistently associated with soil pH, positively and negatively respectively. Dicots tended to accumulate more Ca and Mn from high soil concentrations than did monocots. Concentration of P was significantly positively correlated with maximum potential relative growth rate. Plants of woodland and arable habitats contained high concentrations of P, and those of pasture and skeletal habitats contained low concentrations of P. The P[ratio ]N ratio was higher in plants of arable habitats. Species with P-rich leaves tended to be currently increasing in abundance. The results suggest that plants with nutrient-rich foliage grow quickly, dominate nutrient-rich ecosystems and are generally increasing as a result of the eutrophication and disturbance arising from human exploitation.

Measurements of the chitin content of the rooting horizons of a typical mor-humus heathland soil, indicate that chitin can contain in excess of 20% of the total nitrogen in the litter (L) horizon and 30% in the fermentation (F) horizon. Much of this chitin-nitrogen is thought to be contained in the mycelial walls of soil fungi.

Experiments were therefore designed to test the hypothesis that such sources of N could be rendered accessible to the ericaceous plants by their fungal endophytes. Mycelium of the ericoid endophyte Hymenoscyphus ericae (Read) Korf & Kernan and of the ectomycorrhizal fungus Suillus bovinus (Fr.) O. Krantze were grown in liquid culture before being killed and added either in the intact condition, or after fractionation, as sole sources of N to sterile media upon which were grown H. ericae in pure culture, or mycorrhizal and non-mycorrhizal plants of Vaccinium macrocarpon Ait. and Calluna vulgaris L. The abilities of the test organisms to utilize the nitrogen contained in the intact mycelial necromass, or in its fractions, were assessed by determining their yields and nitrogen concentrations of their tissues.

It was revealed that H. ericae was able to produce significantly higher yield when grown on intact fungal necromass than when provided with equivalent concentrations of N in the form of ammonium. Its yields on mycelial fractions were lower, but still significantly greater than those obtained in the controls lacking N. Significantly greater yields and N contents were also found in the ericaceous plants grown with these nitrogenous substrates in the mycorrhizal condition. Without H. ericae they had no access to the substrates. The possible ecological implications of these results are discussed.

Hydrogen sulphide emission in lichens as a response to low concentration SO2 fumigation was investigated. In an open flow-through system several lichen species were fumigated with 36 ppb SO2. Two species were also fumigated with higher concentrations (72, 119, 122 and 198 ppb SO2). Hydrogen sulphide emission was monitored concurrently by cryogenic trapping and analysis with gas chromatography. All tested species increased H2S emission significantly in response to fumigation with 36 ppb SO2. Parmelina tiliacea (L.) Hale and Cladina rangiferina (L.) Wigg. released significantly more H2S (0·098±0·015 and 0·073±0·013 pmol H2S g−1 d. wt s−1, respectively) than Parmelina quercina (Ach.) Hale, Ramalina menziesii Tayl. and Parmelia sulcata Tayl. (0·028±0·01, 0·025±0·014 and 0·023±0·013 pmol H2S g−1 d. wt s−1, respectively). Release of H2S in Hypogymnia physodes was enhanced by increasing SO2 concentrations up to 72 ppb SO2. No significant difference in H2S emission in the dark vs. in the light was found. Generally, no correlation was found between photosynthetic activity and H2S emission for the tested species. Uptake of SO2 was similar for all species, at 24·7±5·6 pmol SO2 g−1 d. wt s−1 in 36 ppb SO2 and increasing at greater SO2 concentrations. Therefore, H2S-S release represents only 0·11–0·74% of SO2-S uptake.

The 15N natural abundance and N concentrations of fruit bodies from 70 species (23 genera) of ectomycorrhizal fungi found in boreal forests are presented. Large intraspecific and intrageneric differences were found, e.g. 8·3‰ 15N in the species Dermocybe crocea and 12·6‰ in the genus Cortinarius. In addition, significant differences in both δ15N and %N were found between different parts of fruit bodies, with cap material giving consistently higher values. Proteins and amino acids were enriched by 9·7±0·4‰ (mean±1 SE) relative to chitin, irrespective of the part of the fruit body examined. Chitin had δ15N values similar to that of plant hosts. The higher δ15N and %N values of the caps than of the stipes probably reflect a higher portion of proteins and amino acids in the caps. The δ15N of mycorrhizal fungi can be a function of the N species used (organic N, NH4+, NO3−), the depth of soil at which the mycelium occurs, and metabolic fractionations. The metabolic fractionations, e.g. potential transaminations during the flux of N from the soil through the fungus to the plant, make it difficult, at present, to make inferences about sources of N based on δ15N values alone. No effect of sample drying temperature on δ15N values of fungal material was detected.