In this paper, the response to temperature of three physiological processes has been examined in detail in seedlings of sorghum and pearl millet. These have been compared with their field performance under high temperature conditions with the aim of developing laboratory-based screening techniques for seedling thermotolerance. Membrane thermostability, as assessed by electrolyte leakage, seedling re-growth and protein synthetic ability were measured in seedlings exposed to a range of temperatures using a thermal gradient bar. The effect of genotype, seedling age and pre-treatment was examined. Differences between genotypes were only apparent after seedlings had been given a brief high temperature pre-treatment (2 h at 43°C). Similar results were found with all three methods used and their suitability for use in the screening of large numbers of entries is discussed. Significant correlation was found between the ability of membrane thermostability to acclimate and seedling survival in the field.

Many hypotheses have been developed to explain the adaptive nature of insect galls. One of these, the nutrition hypothesis, states that gall formers have advantages over other insects because gall tissue provides a better (higher quality) food source than unmodified tissue. However, this has rarely been experimentally tested. In a test of this hypothesis, we grew plants of Cirsium arvense (L.) Scop. in a factorial design with two main treatments: the addition of nitrogen (to enhance foliar N levels) and of fungicide (to reduce colonization of roots by arbuscular mycorrhizal fungi). Mycorrhizal fungi have been shown previously to reduce the N concentration of host plants. Plants were exposed to adult gall flies, Urophora cardui L., and maintained through one season to allow maturation of galls.

Reduction of the percentage mycorrhizal colonization by fungicide resulted in an elevation of total stem N comparable to that achieved by N addition, but gall N concentration remained unchanged in all treatments. Nitrogen application elevated stem N levels when mycorrhizal fungi were present, but application of both compounds together did not result in any increase over either single treatment. Fungicide application resulted in larger galls, which contained more larval chambers, with more live, and heavier, larvae. However, the main effects of N were not significant, as N addition only increased fly performance on plants where mycorrhizas were not reduced.

It is suggested that U. cardui gall inhabitants can manipulate N at an optimal level and thus might conform to a modified version of the nutrition hypothesis. Mycorrhizal colonization might reduce gall fly performance by delaying the appearance, or impairing the quality, of secondary nutritive tissue in the gall. Future tests of the nutrition hypothesis should include a consideration of the plant's mycorrhizal status.

Copper and zinc tolerances of 10 micropropagated birch (Betula pendula and B. pubescens) clones were studied in hydroponic culture. Tolerance indices were determined, based on the mean growth rate of the longest root in 1 wk. A seed-derived clone (142A), from a lead/Zn-contaminated site showed more tolerance to Cu and Zn than bud-derived clones (HA02 and HA18) from a Cu/nickel-contaminated site or an ozone-tolerant clone (KL-2-M) from an uncontaminated area. For Cu, the EC50 values were 30, 14, 8 and 11 μm in clones 142A, HA02, HA18 and KL-2-M, respectively. For Zn, the EC50s were 4000 and 350 μm in clones 142A and KL-2-M, respectively. The relative Cu and Zn tolerances of the other clones were estimated by growing the plants in 30 μm CuSO4, and in 2000 or 350 μm ZnSO4, respectively. It is of interest that the Zn-tolerant clone 142A was tolerant to Cu, although this metal was present at a very low concentration in the soil where the parent tree grows. Another clone (142B), from another seed of the same parent tree, was tolerant neither to Zn nor Cu. Compared with their own EC50s for root growth for Cu, 142A took up more Cu than KL-2-M, suggesting that the higher tolerance of the former clone is not explained by reduced uptake of Cu. The Zn uptake in clones 142A and KL-2-M was studied at 4000 μm and 800 μm Zn, respectively. Interestingly, the roots of both clones contained the same amount of Zn, even though clone 142A was exposed to a fivefold concentration of Zn.

This paper reviews the results of several years' study by a multidisciplinary team comprising geneticists, physiologists, ecologists and statisticians of abiotic stress tolerance in barley. The strategy involves genetic fingerprinting of wild barley lines (Hordeum spontaneum C. Koch) from a range of habitats and testing these for responses to abiotic stresses under controlled conditions. Multiple regression analysis is used to identify genetic markers associated with experimentally determined stress responses. These data are related to site-of-origin ecogeographic data and used to identify areas of useful wild variation. Markers associated with traits of interest can be mapped in genetic mapping populations, revealing areas of the barley genome carrying genes controlling stress responses. This paper highlights our work on associations of amplied fragment length polymorphisms with salt tolerance and describes some initial results of the use of SSRs in studying drought tolerance in barley.

The ability of ryegrass (Lolium perenne L.) to take up and utilize aspartic acid (Asp) and serine (Ser), and the effect of colonization of the roots by the arbuscular mycorrhizal (AM) fungus Glomus fasciculatum (Thax. sensu Gerd.) were studied. The seedlings were grown under controlled conditions in a series of micro-lysimeters. All plants were fed with a nutrient solution containing either nitrate, Asp or Ser as the sole N source. After 49 d, they were supplied with 15N labelled nitrate, Asp or Ser for 1 h and harvested.

AM colonization increased the growth and total N content of the plants in all cases. Similarly, the amount of Asp or Ser taken up was higher in AM than in control plants. There were no differences in biomass production between the nitrate and Ser-fed plants. However uptake rates were lower for Ser than for nitrate. Growth of the Asp-fed plants was significantly less than the other two treatments, and uptake of 15N-Asp was lower than uptake of 15N-Ser. Analysis of 15N incorporation into the amino acids extracted from the roots suggests the hydrolysis of Ser followed by re-assimilation of the resulting ammonia via the GS-GOGAT cycle. There were no differences in the patterns of accumulation of amino acids in the root-zone of control and AM-ryegrass. The implication of these results for the pathway of nitrogen transfer between plants is discussed.

A number of ectomycorrhizal (ECM) fungi, from sites uncontaminated by toxic metals, were investigated to determine their sensitivity to Cd2+, Pb2+, Zn2+ and Sb3−, measured as an inhibition of fungal biomass production. Isolates were grown in liquid media amended with the metals, individually (over a range of concentrations) and in combination (at single concentrations) to determine any significant interactions between the metals. Significant interspecific variation in sensitivity to Cd2+ and Zn2+ was recorded, while Pb2+ and Sb3− individually had little effect. The presence of Pb2+ and Sb3− in the media did however, ameliorate Cd2+ and Zn2+ toxicity in some circumstances. Interactions between Cd2+ and Zn2+ were investigated further over a range of concentrations. Zn2+ was found to significantly ameliorate the toxicity of Cd2+ to three of the four isolates tested. The influence of Zn2+ varied between ECM species and with the concentrations of metals tested.

Amplified fragment length polymorphisms (AFLPs) offer a reproducible, multiplex DNA assay by which to genotype mapping populations. We have evaluated physiological traits in barley seedlings grown in an hydroponic system and given a salt treatment. Multiple regression was used to show associations between AFLPs and quantitative traits. Effects at different loci were detected in stress treatments in comparison to the control implying that either novel gene action was induced by salt stress or that normal activity was reduced to a low level where alternate gene action is revealed. The QTLs occurred on all chromosomes but there appeared to be clusters of loci on chromosomes 1 (7H), 4 (4H), 5 (1H) and 6 (6H). The significance of our results is discussed in the context of studies to explore the barley genome and the application of the results of these genetical analyses to barley breeding.

The amounts of cyclic AMP (cAMP), fructose-2,6-bisphosphate (F26BP), trehalose and glycogen were determined in cell suspension cultures of the ectomycorrhiza-forming fungus Amanita muscaria (L. ex Fr.) Hooker. For the assay of cAMP a protocol was developed that enabled the detection of as little as 50 fmol of this secondary messenger by an enzyme-linked immuno assay (EIA). Values varied from <1 and up to 5 pmol cAMP mg−1 d. wt according to the age of the fungal culture. Typically, a transient increase in cAMP occurred after c. 4 d of culture of the fungus on glucose-containing medium. This increase (up to 100%) was followed by the start of the logarithmic growth phase, and by a more persistent increase in F26BP. In parallel, glucose in the medium started to decrease, whilst the amounts of fungal carbohydrates, especially the disaccharide trehalose, increased. From these data we assume that a high initial rate of glucose uptake caused an increase in the fungal pools of storage carbohydrates and, via activation of an adenylate cyclase, of cAMP. According to data reported for yeast cells this should enhance the formation of F26BP by phosphorylation of relevant enzymes. In animal and yeast cells an increase in the concentration of F26BP stimulates glycolysis by activation of the ATP-dependent phosphofructokinase (PFK). A. muscaria also possesses an F26BP activated PFK and, under conditions of symbiosis, host-derived carbohydrates are supplied mainly in the form of glucose. The implications of these findings to the regulation of carbohydrate metabolism of symbiotic plant root/fungus structures (ectomycorrhiza) are discussed.

The identity of black alder (Alnus glutinosa (L.) Gaertn.) ectomycorrhizas was investigated using PCR/RFLP analysis of the ITS region from 16 morphotypes sampled at a 60-yr-old black alder stand. A comparison was made with restriction patterns from sporocarps of 28 mycobionts, of which 16 originated from the same stand, the remaining 12 came from two geographically distant alder stands. Eight of the mycorrhizal types could thus be identified, whereas eight mycorrhizal types remained unidentified. The identified mycorrhizas belonged to the genera Russula, Lactarius, Naucoria and Cortinarius. Four of the identified ectomycorrhizal types had identical PCR/RFLP profiles to corresponding fruit bodies from all investigated stands with no detectable intraspecific variation, despite the geographical distance of c. 300 km between the sampling locations. By contrast, intraspecific variation between sporocarps from the different locations was detected in Paxillus rubicundulus, mycorrhizas of which were not found. The diversity of fruiting alder mycobionts at the main experimental plot only partly matched the diversity observed from mycorrhizas when comparing their PCR/RFLP profiles. The results are discussed regarding sampling techniques, PCR/RFLP analyses and ecological aspects.

This paper describes a developing approach for correlating molecular markers with phenotypic expression in barley, and research examples are given. This approach is intended to elucidate the connection between defined parts of the barley genome and their physiological expressions. To illustrate our approach (up to the point of marker correlation), the materials, methods and results of an experiment are presented. Briefly, the natural abundance levels of carbon and nitrogen isotopes (δ13C and δ15N) serve as the primary screening tools for assessing barley genotypes for their responses to imposed abiotic stresses. The isotopic responses are then correlated with molecular markers as the first step in identifying the mechanisms relating genetic variation with phenotypic expression.

A well-described mechanistic model exists for interpreting plant δ13C, and this enables partitioning of the physiological processes contributing to the whole plant δ13C. Although plant δ15N appears to be equally well correlated with genotype and environment, there is no mechanistic model explaining this isotopic signature. Hence, δ15N is used empirically and in parallel with attempts to understand its underlying mechanism(s).

This is a review of genetic mapping with molecular markers aimed at the non-specialist who wishes to use, or at least grasp the concepts behind, this powerful analytical tool. Restriction fragment length polymorphisms (RFLPs) are defined and used to illustrate the different aspects of mapping. The principles of segregation, recombination and linkage are considered and related to the idea of a molecular marker map. A description of a typical mapping population and how it is analysed follows. Traits to be mapped are divided into those controlled by ‘major’ genes and those governed by quantitative trait loci (QTLs). Exploitation of the map for marker-assisted selection, gene cloning and synteny comparisons is discussed, as are some of the limitations to the usefulness of molecular marker maps. Finally other marker systems are introduced, namely minisatellites or variable number tandem repeats (VNTRs); randomly amplified polymorphic DNA (RAPDs); microsatellites or simple sequence repeats (SSRs); and amplified fragment length polymorphisms (AFLPs).