Binucleate Rhizoctonia endophytes from the roots of nursery-grown Pinus sylvestris (Scots pine) seedlings and the orchid Goodyera repens from Scots pine forests were characterized on the basis of morphological characters, anastomosis group membership and PCR-assisted ribosomal DNA fingerprinting. Common hyphal and colony morphological traits displayed by the Finnish binucleate Rhizoctonia isolates and a range of Canadian orchid root endophytes enabled them to be placed in the anamorphic genus Ceratorhiza. Five main anastomosis groups were identified and included groups that contained different combinations of Scots pine, G. repens and Canadian Ceratorhiza spp. isolates. Two Scots pine root endophytes anastomosed with a phytopathogenic Japanese tester isolate, confirming their membership of anastomosis group I, which is known to include the teleomorphic species Ceratobasidium cornigerum. Hierarchical cluster analysis of RFLPs in the internal transcribed spacer of ribosomal DNA enabled the division of isolates into one of five RFLP groups. The RFLP and anastomosis groupings were closely correlated; isolates within each of four RFLP groups, which shared 100% RFLP identity, anastomosed in the cross-pairing anastomosis group tests. However, all represented different vegetatively compatible populations (clones) because the diagnostic killing reaction, a cellular vegetative incompatibility response, was identified at hyphal fusion junctions. These findings indicate a high degree of intraspecific variation within Ceratobasidium cornigerum, which includes isolates able to enter into either mutualistic or pathogenic root association with susceptible host plants. The common anastomosis group/RFLP groupings identified also strongly support the hypothesis that conifer tree roots can act as large inoculum reservoirs for these orchid endophytes, allowing the development of inter-plant connections, via commonly shared hyphal linkages, in boreal forest ecosystems.

A walk through the forest is a good way to become aquainted with the variety of terpenes emitted by plants. The most fragrant of these are the monoterpenes, which lend a distinctive smell to trees (e.g. α- and β-pinene from pines), mints (e.g. menthol from peppermint), fruits (e.g. limonene from citrus) and flowers (e.g. geraniol from roses). In addition to adding fragrance, phytogenic hydrocarbons play an important role in atmospheric chemistry. This was recognized as early as 1960 by Fritz Went who noted that blue hazes often formed above forested areas (Went, 1960). This idea was picked up by US President Ronald Reagan who accused trees of polluting the atmosphere, although in truth phytogenic hydrocarbons only contribute to pollution in air that is already contaminated by industry or automobile exhaust (Feshenfeld et al., 1992).

Terpenes serve a variety of functions in plants including deterring herbivores and attracting pollinators. Until recently, the function of terpene emission from some groups of plants, which do not store the terpenes in their tissues, has been a mystery. Following the discovery that isoprene, a hemiterpene, increases the thermotolerance of photosynthesis in some species (Sharkey & Singsaas, 1995; Singsaas et al., 1997), Loreto et al. (1998) discovered that thermotolerance of the monoterpene-emitting oak, Quercus ilex, increases when leaves are fumigated with monoterpenes. Now, a report by Delfine et al. in this issue (pp. 27–36) provides further support for the hypothesis that terpenes play a role in thermotolerance of leaves and suggests, intriguingly, that the mechanism by which this occurs is not limited to plants that make these compounds.

The effects of elevated concentrations of atmospheric carbon dioxide and ozone on diurnal patterns of photosynthesis have been investigated in field-grown spring wheat (Triticum aestivum). Plants cultivated under realistic agronomic conditions, in open-top chambers, were exposed from emergence to harvest to reciprocal combinations of two carbon dioxide and two ozone treatments: [CO2] at ambient (380 μmol mol−1, seasonal mean) or elevated (692 μmol mol−1) levels, [O3] at ambient (27 nmol mol−1, 7 hr seasonal mean) or elevated (61 nmol mol−1) levels. After anthesis, diurnal measurements were made of flag-leaf gas-exchange and in vitro Rubisco activity and content. Elevated [CO2] resulted in an increase in photoassimilation rate and a loss of excess Rubisco activity. Elevated [O3] caused a loss of Rubisco and a decline in photoassimilation rate late in flag-leaf development. Elevated [CO2] ameliorated O3 damage. The mechanisms of amelioration included a protective stomatal restriction of O3 flux to the mesophyll, and a compensatory effect of increased substrate on photoassimilation and photosynthetic control. However, the degree of protection and compensation appeared to be affected by the natural seasonal and diurnal variations in light, temperature and water status.

The contribution of the extramatrical mycelium to N and P nutrition of mycorrhizal Norway spruce (Picea abies (L.) Karst.) was investigated. Seedlings either inoculated with Paxillus involutus (Batsch) Fr. or non-mycorrhizal were grown in a two compartment sand culture system where hyphae were separated from roots by a 45 μm nylon net. Nutrient solution of the hyphal compartment contained either 1.8 mm NH4+ and 0.18 mm H2PO4− or no N and P. Aluminium added to the hyphal compartment as a tracer of mass flow was not detected in the plant compartment, indicating that measurements of N and P transfer by the mycelium were not biased by solute movement across the nylon net.

The addition of N and P to the hyphal compartment markedly increased dry weight, N and P concentration and N and P content of mycorrhizal plants. Calculating uptake from the difference in input and output of nutrient in solution confirmed a hyphal contribution of 73% and 76% to total N and P uptake, respectively. Hyphal growth was increased at the site of nutrient solution input.

An experiment was carried out in open-top chambers located in eastern Spain. One-yr-old Pinus halepensis Mill. seedlings were exposed during three consecutive summers to the following ozone (O3) treatments: charcoal-filtered air (CFA), non-filtered air (NFA) or non-filtered air plus 40 nl l−1 O3, 9 h d−1, 5 d wk−1 (NFA+40). Seasonal variations in Aleppo pine performance were observed since reductions in chlorophyll and cellular peroxidase levels associated with increases in superoxide dismutase activity, were recorded during the summer. Similarly, a reduction in epoxidation state was found at midday during the summer, derived from an activation of the xanthophyll cycle associated to an increment in radiation and temperature levels.

The first O3-induced effects were recorded in previous-year needles (1991) during the first summer exposure as an increase in extracellular and total peroxidase activities and in zeaxanthin levels in the NFA+40 treatment along with a trend to a higher SOD activity in this treatment. A carry-over effect was detected since a lower winter recovery of chlorophyll levels was found in the NFA+40 seedlings along with a reduction of xanthophyll levels. A reduction in chlorophyll levels was observed in the previous-year needles (1992) from the NFA+40 treatment at the end of the second fumigation period. Realistic ozone exposures induced alterations in plant antioxidative systems and plant pigments as shown in this paper. These observations together with the reductions in stomatal conductance and net photosynthesis recorded in the same experiment, indicate that Aleppo pine is a species sensitive to ozone.

The effects of aluminium (Al) on dividing root-tip cells of Triticum turgidum were investigated with tubulin immunolabelling and electron microscopy. Aluminium affects the mechanisms controlling the organization of microtubule (MT) cytoskeleton, as well as tubulin polymerization, and induces the following aberrations in mitotic cells. (1) It delays the MT disassembly during mitosis, resulting in the persistence of preprophase MT bands in the late prophase cells, the presence of prophase spindles in prometaphase cells, and a disturbance in the shortening of kinetochore MT bundles in anaphase cells. (2) It interferes with the self-organization process of MTs into bipolar systems, inhibiting the formation of prophase and metaphase spindles. (3) Aluminium induces the formation of atypical MT arrays, which in the immunofluorescent specimens appear as ring-like tubulin aggregations in the cortical cytoplasm of the preprophase/prophase cells and as endoplasmic tubulin bundles in prophase and metaphase/anaphase cells; abnormal preprophase MT bands are assembled, consisting of atypical cortical and endoplasmic MT bundles, the latter clearly lining the nuclear envelope on the preprophase MT band plane. (4) It disorders the chromosome movements carried out by the mitotic spindle. In addition, after prolonged Al treatments chromatin condensation is inhibited. The outcome is greatly disturbed organization and function of the mitotic apparatus, as well as inhibition of cells from entering mitosis. This study shows that the MT cytoskeleton is a target site of Al toxicity in mitotic root-tip cells of T. turgidum. The possible mechanisms by which Al exerts its toxicity on MT organization and function are discussed.

Two-yr-old Norway (Picea abies (L.) Karst.), red (P. rubens Sarg.), black (P. mariana (Mill.) B.S.P.) and white (P. glauca (Moench.) Voss) spruce seedlings were exposed from bud break for 35 or 63 d (4·5 h irradiance d−1) to a gradient of biologically effective u.v.-B radiation (λ=280–315 nm) ranging from 0·61 kJ m−2 d−1 to 5·99 kJ m−2 d−1. No visible symptoms of u.v.-B injury were observed. Epicuticular wax production was not affected by needle exposure to increasing u.v.-B irradiance. Seven constituent classes were resolved by GC and confirmed by GC–MS in wax recovered from needles of the four species. Wax composition of Norway, black and red spruce was altered following needle exposure to increasing u.v.-B dose. White spruce wax composition was unaffected. Direction and magnitude of wax composition response was species-dependent. The proportion of nonacosane diols on Norway spruce needles increased (P<0·05) whereas that of alkyl esters decreased with increasing u.v.-B dose. The proportion of fatty acids in black spruce needle wax increased (P<0·05), and that of estolides (GC-identified) in red spruce needle wax increased (P<0·05) with increasing u.v.-B dose. Changes in wax chemical composition reported were induced following to daily, 4-h duration exposures of needles to u.v.-B centred on 1200 hours. Affected variables exhibited a continuum of response. The highest dose applied was within the range of measured or predicted increases in mid-northern latitudes. Such changes in conifer needle epicuticular wax chemical composition might result in increased seedling sensitivity to the changing atmospheric environment, especially from co-exposure to tropospheric ozone in mid-northern latitudes where much of Canada's productive forest is located.

Seeds of cherry (Prunus avium) were germinated and grown for two growing seasons in ambient (∼350 μmol mol−1) or elevated (ambient+∼350 μmol mol−1) CO2 mole fractions in six open-top chambers. The seedlings were fertilized once a week, following Ingestad principles in order to supply mineral nutrients at free-access rates. In the first growing season gradual drought was imposed on rapidly growing cherry seedlings by withholding water for a 6-wk drying cycle. In the second growing season, the rapid onset of drought was imposed at the height of the growing season on the seedlings which had already experienced drought in the first growing season. Elevated [CO2] significantly increased total dry-mass production in both water regimes, but did not ameliorate the growth response to drought of the cherry seedlings subjected to two sequential drying cycles. Water loss did not differ in either well watered or droughted seedlings between elevated and ambient [CO2]; consequently whole-plant water- use efficiency (the ratio of total dry mass produced to total water consumption) was significantly increased. Similar patterns of carbon allocation between shoot and root were found in elevated and ambient [CO2] when the seedlings were the same size. Thus, elevated [CO2] did not improve drought tolerance, but it accelerated ontogenetic development irrespective of water status.

Sorghum (Sorghum bicolor) synthesizes a complex mixture of 3-deoxyanthocyanidin phytoalexins in response to inoculation with the non-pathogenic fungus Bipolaris maydis. The anthocyanin cyanidin 3-dimalonyl glucoside, is also synthesized naturally in response to light. To determine the order and time of appearance of these compounds, etiolated sorghum mesocotyls were inoculated with B. maydis and tissue extracts were analysed by photodiode array-HPLC every 2 h for the first 24 h and at 48 h post inoculation (hpi). Uninoculated and inoculated etiolated mesocotyls were also allowed to incorporate L-[U-14C] phenylalanine. Apigeninidin appeared at 10 hpi, followed by luteolinidin and apigeninidin 5-O-arabinoside at 14 hpi. Luteolinidin 5-methylether was not detected until 18 hpi and apigeninidin 7-methylether not until 20 hpi. The concentrations of the primary phytoalexins, apigeninidin, luteolinidin and apigeninidin 5-O-arabinoside, rose steadily between 12 and 24 hpi, and the levels of apigeninidin and luteolinidin were approximately equivalent by 24 hpi. However, between 24 and 48 hpi luteolinidin and luteolinidin 5-methylether accumulated rapidly so that by 48 hpi the amounts of luteolinidin and luteolinidin 5-methylether had increased approximately twofold. Radiolabelling also showed that 14C was incorporated into the 3-deoxyanthocyanidins and cyanidin 3-dimalonyl glucoside. Several other unidentified phenolic compounds also accumulated radioactivity.

An homologous series of hydrocarbons ranging from C21 to C37 (was detected in foliar cuticular waxes of Austrocedrus chilensis (D. Don) Pic.-Ser. & Bizz., by gas chromatography. Qualitatively, the chromatograms were similar for all individuals sampled from 29 natural populations in Chile and Argentina, showing C33 and C35 alkanes to be dominant. Quantitatively, the chromatograms varied significantly among populations. Multivariate analyses showed that arid zone populations from mediterranean Chile and the Patagonian steppe were different from one another and were distinct from mesic populations close to the Valdivian rainforest vegetation zone. Mediterranean Chilean populations were characterized by higher concentrations of the longer carbon-chain alkanes, but retained equal amounts of shorter-chain homologues as the mesic populations. A greater variation in chain lengths and higher concentrations of longer carbon chains in the mediterranean populations is consistent with a model for the adaptation to reduced cuticular permeability. Partial Mantel matrix tests revealed significant climatic and inter-population distance effects with taxonomic distances based on multivariate and univariate hydrocarbon data. Annual rainfall was overall the most significant factor, particularly in regressions with the shorter-chain hydrocarbons. Annual mean temperature was most significant for the longer-chain hydrocarbons. This suggests, on the one hand, ecogenic adaptation to both temperature and precipitation of cuticular hydrocarbon composition, and, on the other hand, a weaker, but important effect of gene flow in determining hydrocarbon composition in this species. The northernmost populations at San Felipe and San Gabriel were the most distinctive. This could result from the effects of random changes in allele frequencies and/or to founder effects in isolated and small populations. These populations show some East-Andean affinities presumably due to historic migration patterns.

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.

Bulbous rush (Juncus bulbosus) initiates plant colonization in extremely acid lakes resulting from coal mining operations. Various analytical techniques (methylene blue/agar method, Ti3+-citrate solution) X-ray diffraction (XRD), scanning electron microscopy (SEM), and Energy-dispersive X-ray (EDX) were used to assess the mechanisms and strategies employed by J. bulbosus to overcome the extreme conditions. The plant releases oxygen into the rhizosphere in turn increasing the redox potential and inducing iron oxide plaque formation. XRD showed that the iron oxide of the plaque is mainly goethite that has been developed in the presence of CO2; SEM showed that there is a micro-space between the roots and sand grains which is inhabited by microorganisms. Furthermore, SEM-EDX studies on internal iron distribution demonstrate that iron toxicity is delayed by the physiological and biochemical structure of the plant. It is suggested that J. bulbosus uses a variety of mechanisms and strategies (morphological, physiological and biochemical adaptation) which are mainly complementary and which interact with each other to help J. bulbosus to manage its growth and survival in an extreme environment.

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 range of processes regulated by gibberellins (GAs) covers all aspects of the life history of the plant from seed germination to vegetative growth and flowering. In seeds there has been an intensive search, using the techniques of both biochemistry and cell biology, for the regulatory molecules linking GA perception to gene regulation and the events of germination. Although a GA receptor has yet to be identified, the site of perception has been localized to the plasma membrane. Calmodulin, Ca2+ and cGMP have also been identified as elements of the GA signal transduction pathway. These regulators parallel many of the signalling elements identified in the transduction of other signals such as phytochrome and ABA. Studies of GA-regulated gene expression, principally of the α-amylases of cereal aleurone, have identified core GA-responsive promoter elements, such as the gibberellin response element (GARE), box-1 and pyrimidine boxes, as well as elements that may lend specificity to GA-regulated expression, such as the Opaque-2-similar element (O2S), and TRE and CRE motifs. One of the most striking features of all of these studies of the molecular basis of GA action is the interaction of GA-dependent regulatory elements with those of other factors such as ABA. GA-response elements also appear to be conserved between disparate GA-response systems. For example, Myb transcription factors appear to regulate a multitude of GA-induced genes in cereal aleurone as well as to alter GA responses when expressed in Arabidopsis. Thus the study of GA signal transduction and response systems is highlighting the conservation of regulatory elements used by plants. These common factors, used by distinct signal transduction systems, provide a molecular basis for the integration of the GA signal with other growth regulators that is the hallmark of plant growth and development.

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.

Genotypic variability was studied in two Mediterranean grass species, Bromus erectus and Dactylis glomerata, with regard to the response to CO2 of leaf total non-structural carbohydrate concentration ([TNC]lf), specific leaf area (SLA), and leaf carbon and nitrogen concentrations ([C]lf and [N]lf, respectively). Fourteen genotypes of each species were grown together on intact soil monoliths at ambient and elevated CO2 concentrations (350 and 700 μmol mol−1, respectively). In both species, the most consistent effect of elevated CO2 was an increase in [TNC]lf and a decrease in leaf nitrogen concentration when expressed either as total dry mass [Nm]lf, structural dry mass [Nmst]lf or leaf area [Na]lf. The SLA decreased only in D. glomerata, due to an accumulation of total non-structural carbohydrates and to an increase in leaf density. No genotypic variability was found for any variable in B. erectus, suggesting that genotypes responded in a similar way to elevated CO2. In D. glomerata, a genotypic variability was found only for [Cst], [Nm]lf, [Nmst]lf and [Na]lf. Since [Nm]lf is related to plant growth and is a strong determinant of plant–herbivore interactions, our results suggest evolutionary consequences of elevated CO2 through competitive interactions or herbivory.

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.

A fructan polymerase activity was partially purified and concentrated by sequential acid and salt precipitation from extracts of excised, illuminated leaves of timothy grass (Phleum pratense). The polymerase catalysed the de novo synthesis of oligo- and polyfructan from sucrose as sole substrate at near-physiological rates (0.5–0.9 mg g−1 fresh mass h−1; 0.9–1.5 nkat g−1). Rates of in vitro polymerisation were high, at up to 4.1 mg cm−3 h−1 (7.1 nkat cm−3) of total products of degree of polymerization greater than 2 (DP>2). The trisaccharides 1-kestose and 6-kestose together with oligosaccharides of up to DP[les ]c. 10 were synthesized in under 2 h at 30°C. In longer incubations, ethanol-precipitable polymers of DP=c. 10–35 (1.6–5.7 kDa) were detected by anion-exchange chromatography and pulsed amperometry. When this polymeric product was used as a primer and re-incubated with fresh enzyme and sucrose, abundant polymers of up to DP=50 (8.1 kDa) were formed. The structure of the polymeric enzyme product was compared with native fructan from timothy leaves and with standard inulin, using glycosyl-linkage analysis followed by identification of partially methylated alditol acetate derivatives by GC–MS. The deduced structure was a linear (unbranched) 2,6-linked fructose chain terminated with glucose and fructose. The linkage structures of the native and enzyme-generated polymers were identical, increasing confidence in the physiological relevance of the activity. After ultracentrifugation of tissue homogenates at 265 000 gav, the polymerase remained in the supernatant, demonstrating no tight association with particulate components. The polymerizing reaction was dependent on enzyme concentration, requiring at least 3 g fresh mass equivalent cm−3 (c. 2.7 nkat cm−3) for the efficient in vitro generation of fructans of DP>3. In common with other trisaccharide-synthesizing and oligofructan-glycosylating enzymes from grasses, the polymerase reaction exhibited both a maximal velocity at pH 5.0–5.5 and a low affinity for sucrose. The polymerization reaction did not saturate fully even at 1.5 M sucrose, and the concentration causing half maximal velocity (apparent Km) was c. 560 mM. The preparation contained substantial invertase activity (1.8 mg sucrose g−1 fresh mass h−1=1.5 nkat g−1 fresh mass) with a Km for sucrose hydrolysis of 5 mM. A single peak of polymerase activity with an Mr of 51 kDa was recovered from size-exclusion chromatography (SEC). Invertases of Mr 51 and 110 kDa were identified in the preparation. The 110-kDa invertase isoform exhibited no polymerase activity, but synthesized trisaccharide (mainly 1-kestose) from sucrose. The 51-kDa isoform co-eluted with the polymerase. The trisaccharide fraction produced by this isoform contained abundant 1- and 6-kestose. After SEC, the purification of the polymerase was 41-fold relative to the original tissue homogenate. The properties of enzymatic polymerization of fructan are discussed with respect to the physiology of accumulation in grass leaves and other systems.

The developmental regulation and organ-specific expression of shikimate dehydrogenase was studied in seedlings of Capsicum annuum L. during their early development. The results obtained suggest that there is both a developmental regulation and an organ-specific expression of shikimate dehydrogenase in seedlings of C. annuum, seen mainly in the cotyledons. For example, shikimate dehydrogenase isoenzyme 3 was differentially expressed in cotyledons, but not at all in roots or hypocotyls. When the amounts of shikimate dehydrogenase were compared with the end-products of the shikimate pathway, chlorogenic acid and lignins, it was observed that the fall in shikimate dehydrogenase activity in cotyledons coincided with a decrease in the concentrations of phenolics during primary leaf development. However, this did not seem to be the case with the hypocotyls, where the fall in phenolics was not related to the amount of shikimate dehydrogenase but rather to an increase in lignin deposition, supporting the view that chlorogenic acid might act as a precursor of the aromatic moiety of cinnamoyl alcohols during cell wall lignification.

During their symbiotic phase, white truffles are barely distinguishable morphologically, and molecular probes are needed for their identification. Here we report the design of species-specific primers for two white truffles (Tuber magnatum and T. borchii) on the basis of their ITS sequence. Their efficiency has been successfully tested on fruit bodies of the same species, many related and unrelated fungal species, as well as mycorrhizal roots in direct, nested and multiplex PCR experiments. They have allowed us to identify T. magnatum mycorrhizas for the first time.