Showing papers on "Mycelium published in 2002"

Extramatrical ectomycorrhizal mycelium contributes one‐third of microbial biomass and produces, together with associated roots, half the dissolved organic carbon in a forest soil

Abstract: Summary • A large-scale tree-girdling experiment enabled estimates in the field of the contribution of extramatrical mycelium of ectomycorrhizal (ECM) fungi to soil microbial biomass and by ECM roots and fungi to production of dissolved organic carbon (DOC). • Tree-girdling was made early (EG) or late (LG) during the summer to terminate the flow of photosynthate to roots and ECM fungi. Determination of microbial C (Cmicr) and microbial N in root-free organic soil was performed by using the fumigation–extraction technique; extractable DOC was determined on unfumigated soil. • Soil Cmicr was 41% lower on LG than on control plots 1 month after LG, whereas at the same time (that is, 3 months after EG), the Cmicr was 23% lower on EG than on control plots. Extractable DOC was 45% lower on girdled plots than control plots. • Our results, which are of particular interest as they were obtained directly in the field, clearly demonstrate the important contribution by extramatrical ECM mycelium to soil microbial biomass and by ECM roots to the production of DOC, a carbon source for other microbes.

In situ13CO2 pulse-labelling of upland grassland demonstrates a rapid pathway of carbon flux from arbuscular mycorrhizal mycelia to the soil

Abstract: Summary • The flux of pulse-derived 13C from upland pasture plants to the external mycelium of their arbuscular mycorrhizal (AM) symbionts was traced and quantified over a 7-d post-labelling period. • Mesh cores, which allowed in-growth of native AM mycelium but were impenetrable to roots, were inserted into unlimed and limed plots and the surrounding vegetation was exposed to 13CO2 at ambient CO2 concentrations. • Release of 13CO2 from cores colonized by AM mycelium peaked 9–14 h after labelling and declined within 24 h after severance of mycelial connections to roots. Between 5 and 8% of carbon lost by plants was respired by AM mycelium over the first 21 h after labelling. Liming increased the amount of carbon fixed by plants and subsequently allocated to fine roots and AM mycelium. • The results demonstrate for the first time under field conditions that AM mycelia provide a rapid and important pathway of carbon flux from plants to the soil and atmosphere.

Characterization of AG-13, a Newly Reported Anastomosis Group of Rhizoctonia solani

Abstract: Rhizoctonia solani anastomosis group (AG)-13 was collected from diseased roots of field grown cotton plants in Georgia in the United States. Isolates of AG-13 did not anastomose with tester isolates of AG-1 through AG-12. Mycelium of all isolates of AG-13 were light brown but darkened as cultures aged. All isolates produced aerial mycelium. Concentric rings were visible after 3 to 4 days of growth but disappeared as cultures aged and darkened. Individual sclerotia were up to 1.5 mm in diameter, similar in color to the mycelium, and generally embedded in the agar. Clumps of sclerotia up to 5 mm in diameter were produced on the agar surface. All attempts to induce basidiospore production were unsuccessful. The 5.8S region of the rDNA from isolates of AG-13 was identical in length and sequence to isolates of all other AGs of R. solani. Length and sequence of the internal transcribed spacer (ITS) regions of rDNA from isolates of AG-13 were unique among AGs of R. solani. Similarity between AG-13 and other AGs of R. solani ranged from 68 to 85% for ITS region 1 and 85 to 95% for ITS region 2. Selected isolates of AG-13 caused minor or no damage to barley, cauliflower, cotton, lettuce, potato, and radish in laboratory or greenhouse studies.

Translocation and Utilization of Fungal Storage Lipid in the Arbuscular Mycorrhizal Symbiosis

Abstract: The arbuscular mycorrhizal (AM) symbiosis is responsible for huge fluxes of photosynthetically fixed carbon from plants to the soil. Carbon is transferred from the plant to the fungus as hexose, but the main form of carbon stored by the mycobiont at all stages of its life cycle is triacylglycerol. Previous isotopic labeling experiments showed that the fungus exports this storage lipid from the intraradical mycelium (IRM) to the extraradical mycelium (ERM). Here, in vivo multiphoton microscopy was used to observe the movement of lipid bodies through the fungal colony and to determine their sizes, distribution, and velocities. The distribution of lipid bodies along fungal hyphae suggests that they are progressively consumed as they move toward growing tips. We report the isolation and measurements of expression of an AM fungal expressed sequence tag that encodes a putative acyl-coenzyme A dehydrogenase; its deduced amino acid sequence suggests that it may function in the anabolic flux of carbon from lipid to carbohydrate. Time-lapse image sequences show lipid bodies moving in both directions along hyphae and nuclear magnetic resonance analysis of labeling patterns after supplying 13C-labeled glycerol to either extraradical hyphae or colonized roots shows that there is indeed significant bidirectional translocation between IRM and ERM. We conclude that large amounts of lipid are translocated within the AM fungal colony and that, whereas net movement is from the IRM to the ERM, there is also substantial recirculation throughout the fungus.

Copper sorption and accumulation by the extraradical mycelium of different Glomus spp. (arbuscular mycorrhizal fungi) isolated from the same polluted soil

Abstract: The form and localisation of Cu accumulation in the extraradical mycelium (ERM) of three arbuscular mycorrhizal fungi (AMF), isolated from the same polluted soil contaminated with the Cu and Arsenate, was studied. There were differences in the capacity of the ERM of the three AMF to sorb and accumulate Cu. Glomus caledonium BEG133 had a significantly lower Cu-sorption capacity than Glomus mosseae BEG132 and Glomus claroideum BEG134 isolated from the polluted soil as well as an isolate of G. mosseae BEG25 from a non-polluted soil. This was directly related to the cation exchange capacity (CEC) of the ERM of these fungi. Transmission Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM) linked to an energy dispersive X-ray spectrometer (EDAX) gave more detailed information, showing that the ERM of AMF from the polluted soil was able to accumulate Cu in the mucilaginous outer hyphal wall zone, cell wall and inside the hyphal cytoplasm. The EDAX spectra showed that the accumulated Cu was mainly associated with Fe in the mucilaginous outer hyphal wall zone and in the cell wall. Cu was associated with traces of arsenate inside the cytoplasm of the ERM of Glomus mosseae BEG132 but this was not visible inside the ERM of Glomus caledonium BEG133 or Glomus claroideum BEG134. This work suggests that the ERM of AMF is able to sorb and accumulate Cu, but different tolerance mechanisms exist between the three AMF isolated from the same polluted soil providing further evidence for functional diversity within populations of AMF in soils.

Antrodia camphorata polysaccharides exhibit anti-hepatitis B virus effects

Abstract: Polysaccharides were extracted from fruiting bodies and cultured mycelia from five Antrodia camphorata strains Polysaccharide profiles of the five strains, as determined by high-performance anion-exchange chromatography, showed varying yields and composition of neutral sugars A camphorata fruiting bodies also had different polysaccharide patterns compared to the cultured mycelium Analysis of 26-day-old mycelia showed that the neutral sugars galactose, glucose, mannose, and galactosamine were predominant All mycelia polysaccharide preparations exhibited anti-hepatitis B virus activity Polysaccharides from strain B86 at a concentration of 50 μg ml−1 showed the highest level of anti-hepatitis B surface antigen effect, which was higher than α-interferon at a dosage of 1000 U ml−1 Only strains B86 and 35398 had substantial anti-hepatitis B e antigen activities None of the polysaccharides exhibited cytotoxic effects

Differential Expression of a Metallothionein Gene during the Presymbiotic versus the Symbiotic Phase of an Arbuscular Mycorrhizal Fungus

Abstract: A full-length cDNA encoding a metallothionein (MT)-like polypeptide, designated GmarMT1, was identified in an expressed sequence tag collection from germinated spores of the arbuscular mycorrhizal fungus Gigaspora margarita (BEG34). The GmarMT1 gene is composed of two exons separated by an 81-bp intron. It codes for a 65-amino acid polypeptide comprising a plant type 1 MT-like N-terminal domain and a C-terminal domain that is most closely related to an as-yet-uncharacterized fungal MT. As revealed by heterologous complementation assays in yeast, GmarMT1 encodes a functional polypeptide capable of conferring increased tolerance against Cd and Cu. The GmarMT1 RNA is expressed in both presymbiotic spores and symbiotic mycelia, even in the absence of metal exposure, but is significantly less abundant in the latter stage. An opposite pattern was observed upon Cu exposure, which up-regulated GmarMT1 expression in symbiotic mycelia but not in germinated spores. Together, these data provide the first evidence, to our knowledge, for the occurrence in an arbuscular mycorrhizal fungus of a structurally novel MT that is modulated in a metal and life cycle stage-dependent manner and may afford protection against heavy metals (and other types of stress) to both partners of the endomycorrhizal symbiosis.

Arbuscular mycorrhizal fungi respond to the substrate pH of their extraradical mycelium by altered growth and root colonization

Abstract: To test the response of arbuscular mycorrhizal (AM) fungi to a difference in soil pH, the extraradical mycelium of Scutellospora calospora or Glomus intraradices, in association with Plantago lanceolata, was exposed to two different pH treatments, while the root substrate pH was left unchanged. Seedlings of P. lanceolata, colonized by one or other of the fungal symbionts, and nonmycorrhizal controls, were grown in mesh bags placed in pots containing pH-buffered sand (pH around 5 or 6). The systems were harvested at approximately 2-wk intervals between 20 and 80 d. Both fungi formed more extraradical mycelium at the higher pH. Glomus intraradices formed almost no detectable extraradical mycelium at lower pH. The extraradical mycelium of S. calospora had higher acid phosphatase activity than that of G. intraradices. Total AM root colonization decreased for both fungi at the higher pH, and high pH also reduced arbuscule and vesicle formation in G. intraradices. In conclusion, soil pH influences AM root colonization as well as the growth and phosphatase activities of extraradical mycelium, although the two fungi responded differently.

Different effectors of dimorphism in Yarrowia lipolytica.

Abstract: Yarrowia lipolytica is an ascomycete with biotechnological potential. In common media, the fungus grows as a mixture of yeast-like and short mycelial cells. The environmental factors that affect dimorphism in the wild-type strain, W29, and its auxotrophic derivative, PO1a, were analyzed. In both strains, pH was the most important factor regulating the dimorphic transition. Mycelium formation was maximal at pH near neutrality and decreased as pH was lowered to become almost null at pH 3. Carbon and nitrogen sources, namely glucose and ammonium, were also important for mycelium formation; and their effect was antagonized by some alternative carbon and nitrogen sources. Citrate was an important positive effector of mycelium growth. Anaerobic stress induced formation of mycelial cells. The importance of the protein kinase A pathway was suggested by the inhibition of mycelium growth by cAMP. We propose that the interplay of these factors regulates the adaptation of the fungus, to better exploit its natural ecological niches.

Suppression of Verticillium Wilt in Eggplant by Some Fungal Root Endophytes

Abstract: One hundred and twenty-three fungal isolates were obtained from 225 root segments of eggplants, melon, tomato, strawberry and Chinese cabbage, grown as bait plants in a mixed soil made up of samples from different fields in Shizuoka, Japan. Isolates belonging to Mycelium radicis atrovirens (MRA), including Phialocephala fortinii, were the most prevalent in all the five bait plants. Eleven of the 123 isolates, after being inoculated onto axenically reared eggplant seedlings, almost completely suppressed the pathogenic effects of a post-inoculated, virulent strain of Verticillium dahliae. Seven of these 11 isolates had come from the roots of eggplant and included Heteroconium chaetospira, P. fortinii, and unidentified species of Fusarium, Penicillium, Trichoderma and MRA. P. fortinii, H. chaetospira, a non-sporulating isolate with white mycelium (SWM) and MRA were easily reisolated from root segments. Hyphae of H. chaetospira, P. fortinii and SWM colonized the root tissues of eggplant without causing apparent pathogenic symptoms. The mechanisms by which these endophytes confer resistance to infection by V. dahliae are unknown but the effectiveness of these fungi in a laboratory setting indicates that they have potential as biocontrol agents and merit further investigation.

Host plant benefit from association with arbuscular mycorrhizal fungi: variation due to differences in size of mycelium

Abstract: Plants differ in the benefit they receive from an association with arbuscular mycorrhizal fungi (AMF) but it is unclear what factors determine host response. We tested whether this difference is due in part to differences among AMF families (Glomaceae, Gigasporaceae, and Acaulosporaceae) in the size of their mycelium. For example, an AMF with a large mycelium might be better able to transfer nutrients to its host. Alternatively, the large mycelium might present a large carbon drain for the host. It might also be the location of AMF mycelium that is important: whether it occurs in the root or in the soil. In a greenhouse experiment, four different host plant species were inoculated with each of 21 AMF isolates representing different mycelial sizes and we measured both host plant biomass and foliar phosphorus concentration over 12 weeks. AMF family significantly affected host biomass with larger internal mycelia conferring greater host benefit. However, both intra-family variation in AMF effect and host identity also influenced host response. For foliar P, AMF family did not affect host response significantly. Our results show that differences in mycelial size among AMF families may contribute to variation in host responsiveness to AMF, but other factors are also important.

Characterization of rhizospheric soil streptomycetes from Moroccan habitats and their antimicrobial activities

Abstract: During an investigation on actinomycetes from rhizospheric soils from Moroccan habitats, 131 streptomycetes were recovered, morphologically characterized and assessed for their antimicrobial activity. Eleven isolates were characterized by the absence of an aerial mycelium. According to the colour of aerial mycelium, the rest were grouped into seven main classes, namely, grey, yellow, cream, white, green, red and polymorphic colours (pink, orange or violet). The grey colour class dominated (40%) and the red one was found only in rhizospheric soil of the Moroccan endemic plant Argania spinosa. About one third of the isolates (34%) produced soluble pigments of various colours and 14% produced melanoid pigments. Most of the isolates (83%) were active against one or more of the organisms tested (one gram-negative bacterium, three gram-positive bacteria, three yeasts and two filamentous fungi). Most antibiotic-producing isolates possess red and white colour. Strong antibiosis was exhibited against Streptomyces scabies, Staphylococcus aureus and Bacillus subtilis (75, 68 and 60% respectively), while only 14 and 8% of isolates displayed an activity against Escherichia coli and Verticillium dahliae respectively.

Transfer of recent photosynthate into mycorrhizal mycelium of an upland grassland: short-term respiratory losses and accumulation of 14C

Abstract: The movement of carbon from plants into their natural communities of arbuscular-mycorrhizal (AM) fungi was investigated. Mesh-bound cores, which allowed in-growth of AM mycelium to be controlled but excluded roots, were inserted into turf monoliths removed from an upland grassland and were exposed to 14 CO 2 . Flux of 14 C-labelled carbon from plants to hyphae of AM fungi for 70 h post-labelling was measured by (a) trapping CO 2 released from soil cores containing AM hyphae linked to the plants compared to cores from which AM hyphal connections to the plant roots had been severed, and (b) quantification of the total amount of 14 C in the cores. Release of 14 CO 2 from the cores colonised by active AM mycelium was highest for 0–28 h from the onset of labelling and declined rapidly thereafter. The amount of 14 C allocated into mycorrhizal mycelium 0–70 h after labelling accounted for 3.4% of the 14 C initially fixed by the plants. The results confirm the rapidity of photosynthate allocation to AM mycelium and demonstrate the importance of the short-term dynamics of C fluxes in undisturbed grasslands.

Phosphorus Effects on Metabolic Processes in Monoxenic Arbuscular Mycorrhiza Cultures

Abstract: The influence of external phosphorus (P) on carbon (C) allocation and metabolism as well as processes related to P metabolism was studied in monoxenic arbuscular mycorrhiza cultures of carrot ( Daucus carota ). Fungal hyphae of Glomus intraradices proliferated from the solid minimal medium containing the colonized roots into C-free liquid minimal medium with different P treatments. The fungus formed around three times higher biomass in P-free liquid medium than in medium with 2.5 mminorganic P (high-P). Mycelium in the second experiment was harvested at an earlier growth stage to study metabolic processes when the mycelium was actively growing. P treatment influenced the root P content and [ 13 C]glucose administered to the roots 7 d before harvest gave a negative correlation between root P content and 13 C enrichment in arbuscular mycorrhiza fungal storage lipids in the extraradical hyphae. Eighteen percent of the enriched 13 C in extraradical hyphae was recovered in the fatty acid 16:1ω5 from neutral lipids. Polyphosphate accumulated in hyphae even in P-free medium. No influence of P treatment on fungal acid phosphatase activity was observed, whereas the proportion of alkaline-phosphatase-active hyphae was highest in high-P medium. We demonstrated the presence of a motile tubular vacuolar system in G. intraradices . This system was rarely seen in hyphae subjected to the highest P treatment. We concluded that the direct responses of the extraradical hyphae to the P concentration in the medium are limited. The effects found in hyphae seemed instead to be related to increased availability of P to the host root.

Antimycotic activities of selected plant flora, growing wild in Lebanon, against phytopathogenic fungi.

Abstract: Petroleum ether (PE) and methanolic extracts of nine wild plant species were tested in vitro for their antimycotic activity against eight phytopathogenic fungi. The efficacy of PE extracts against all pathogens tested was higher than that of methanolic extracts. Wild marjoram (Origanum syriacum) PE extract showed the highest and widest range of activity. It resulted in complete inhibition of mycelial growth of six of eight fungi tested and also gave nearly complete inhibition of spore germination of the six fungi included in the assay, namely, Botrytis cinerea, Alternaria solani, Penicillium sp., Cladosporium sp., Fusarium oxysporum f. sp. melonis, and Verticillium dahlia. The other plant extracts showed differential activities in the spore germination test, but none was highly active against mycelial growth. Inula viscosa and Mentha longifolia were highly effective (>88%) in spore germination tests against five of six fungi tested, whereas Centaurea pallescens, Cichorium intybus, Eryngium creticum, Salvia fruticosa, and Melia azedarach showed >95% inhibition of spore germination in at least two fungi. Foeniculum vulgare showed the least antimycotic activity. Fractionation followed by autobiography on TLC plates using Cladosporium sp. as a test organism showed that O. syriacum PE extracts contained three inhibition zones, and those of Inula viscosa and Cichorium intybus, two, whereas the PE extracts of the remaining plants showed each one inhibition zone. Some of the major compounds present in these inhibition zones were identified by GC-MS. The possibility for using these extracts, or their mixtures, to control plant diseases is discussed.

Interactions Between Ecto-mycorrhizal and Saprotrophic Fungi

Abstract: The microbiota of most forest soils is dominated by ecto-mycorrhizal and saprotrophic decomposer fungi which are the main organisms involved respectively in the supply of nutrients to trees and the decomposition of woody plant litter. In recognition of the pivotal role that these fungi play in plant nutrition and nutrient cycles in plantations and natural forests, the activities of their mycelial systems have been independently investigated in recent years. Such studies have revealed remarkable structural and functional similarities between these organisms, particularly in their active foraging for nitrogen and phosphorus in soils. This has highlighted the need to determine the nature and extent of interactions between the two groups of organisms. The use of radioactive tracers, time-sequence photography and digital image analysis of microcosms containing non-sterile soil in which the two groups of fungi are grown with their natural carbon resources have provided novel insights into their interactions. When mycelia of the fungi meet in soil they often interact strongly, resulting in cessation of growth or even die-back of mycelium. Such inhibition can be accompanied by loss of nutrients to the opposing fungus. This chapter reviews current evidence and presents some new data showing that interactions between the mycelial systems of mycorrhizal and saprotrophic fungi have major effects on the functioning of these organisms and upon nutrient and carbon cycling processes in temperate and boreal forest ecosystems.

Fungal egg-parasites of plant-parasitic nematodes from Spanish soils.

Abstract: We have investigated the presence of fungal egg-parasites in Spanish soils with plant endoparasitic nematodes. Nine out of 68 samples (13%) yielded fungal parasites. The most common (seven strains) was Pochonia chlamydosporia var. chlamydosporia (= Verticillium chlamydosporium var. chlamydosporium), but Lecanicillium lecanii (= Verticillium lecanii) and Paecilomyces lilacinus were also found. Most strains were from cyst nematodes (Heterodera avenae or Heterodera schachtii). Biological factors related with the development and performance of these fungi as biocontrol agents were assessed in laboratory tetsts. Germination for most strains was around 90-100%. Higher biomass values were obtained, for most fungal strains, with complete or yeast extract peptone-glucose liquid media. P. lilacinus and L. lecanii showed the highest sporulation rates (1.0 x 10(9); and 1.5 x 10(10); conidia/g mycelium). All strains had optimum growth at 25 degrees C. High temperature (40 degrees C) was lethal to all fungi but low temperature (5 degrees C) allowed growth of L. lecanii. Most strains showed best growth close to pH 7. Several P. chlamydosporia strains produced diffusible pigments close to pH 3. Lack of moisture (aw = 0.887) in growth medium reduced but never arrested fungus growth. Proteolytic activity was, for all strains, the earliest and highest enzymatic activity. Amylolytic and pectinolytic activities showed the lowest values and the latter was undetectable for most strains. Pathogenicity (70-100percnt; egg infection) and severity (35-40 penetrating hyphae/egg) on Meloidogyne javanica were high for most strains tested. Our results show that agricultural soils in Spain contain fungal parasites susceptible to be biocontrol agents for plant-parasitic nematodes.

Distribution, occurrence and characterization of entomopathogenic fungi in agricultural soil in the Palestinian area.

Abstract: The occurrence of entomopathogenic fungi was investigated in irrigated vegetable fields and citrus orchards soils, over a nine-month period (April-December 1999), using the Galleria bait method (GBM). Entomopathogenic fungi were found to occur in 33.6% of the soil samples studied, with positive samples yielding 70 fungal isolates, belonging to 20 species from 13 genera. Conidiobolus coronatus was the most frequent and abundant entomopathogenic species recovered, comprising 31.4% of the total number of isolates. Soil pH, soil moisture content and the geographical location had minor or no effect on the isolation of entomopathogenic fungi in the fields studied. On the other hand, organic matter content of soil, and vegetation type were found to significantly affect the occurrence of entomopathogenic fungi in soil habitats, with orchard fields yielding larger numbers of isolates than vegetable fields. Using Koch's postulates the pathogenicity of fungal isolates to Galleria larvae was found to range from 16-100% (mortality rate). Isolates of C. coronatus proved to be the most virulent isolates recovered. The effect of media and temperature on mycelial growth rate, conidial production and conidial germination of six entomopathogenic fungal species (C. coronatus, Entomophaga grylli, Erynia castrans, Hirsutella jonesii, Paecilomyces farinosus and Sporodiniella umbellata) was also studied. Mycelial growth rate, spore production and spore germination were significantly affected by media, temperature and isolates. In view of the present results, C. coronatus appears to be a good candidate for pest control in agricultural soils, as it has a wide tolerance to agricultural practices, has frequently been isolated from both vegetable and orchard fields, and is characterized by high mycelial growth rate, conidial production and conidial germination.

Foraging and resource allocation strategies of mycorrhizal fungi in a patchy environment

Abstract: Foraging for nutrients and carbon are essential components of the mycorrhizal symbiosis. Foraging strategies of mycorrhizal fungi have received little attention compared to the interaction with the plant. Proliferation of hyphae, resource allocation (carbon and nutrients) within a mycelium and spatial distribution of resource capturing structures (internal mycelium for carbon and external mycelium for nutrients) can be considered as foraging strategies. The arbuscular mycorrhizal fungi (AMF) form a uniformly distributed mycelium in soil, but hyphal proliferation occurs in response to several types of organic material and near potential host roots. The ecto-mycorrhizal fungi (EMF) normally form denser hyphal fronts than AMF, and they respond to both organic material and inorganic nutrients by increased growth. This is especially evident for the EMF that form extensive mycelia connected by differentiated hyphal strands, so-called rhizomorphs. We hypothesise that the growth strategy of the AM fungal mycelium reflects an evolution towards optimal search for potential new host roots. The growth strategy of EMF instead seems to reflect evolution towards optimised nutrient capture in competition with other mycelia. Foraging behaviour of mycorrhizal fungi is discussed and we suggest two conceptual models for resource allocation in the mycorrhizal mycelium. These models consider both the internal and the external mycelium and the trade-offs between different foraging strategies of mycorrhizal fungi. From the experimental data available, it is clear that mycorrhizal fungi forage. It needs to be investigated whether observed foraging strategies are optimal for the mycelium as one individual.

Studies on the infection process of Fusarium culmorum in wheat spikes: degradation of host cell wall components and localization of trichothecene toxins in infected tissue.

Abstract: After single spikelet inoculation, the infection process of Fusarium culmorum and spread of fungal hyphae in the spike tissues were studied by scanning and transmission electron microscopy. While hyphal growth on outer surfaces of the spike was scanty and no successful penetration was observed, the fungus developed a dense mycelium on the inner surfaces and effectively invaded the lemma, glume, palea and ovary by penetration pegs. During the inter- and intracellular spreading of the fungus, marked alterations in the host tissues were observed, including degeneration of cytoplasm, cell organelles, and depositions of electron dense material between cell wall and plasmalemma. Ultrastructural studies revealed that host cell walls in proximity of the penetration peg and in contact with hyphae were less dense or transparent which suggested that cell wall degrading enzymes were involved in colonisation of host tissues by fungal hyphae. Enzyme- and immunogold-labelling investigations confirmed involvement of extracellular enzymes, that is cellulases, xylanases and pectinases, in degradation of cell wall components. Localization studies of trichothecenes indicated that toxins could be detected in host tissues at an early stage of infection.

Diversity of culturable bacterial populations associated to Tuber borchii ectomycorrhizas and their activity on T. borchii mycelial growth.

Abstract: Isolation and physiological and molecular characterisation of culturable bacterial strains belonging to actinomycetes, pseudomonads and aerobic spore-forming bacteria were carried out on mycorrhizal root tips of Quercus robur var. peduncolata infected by Tuber borchii. Cellular density of the three bacterial groups in ectomycorrhizal root tips was estimated to be 1.3±0.11×106 cfu g−1 dry weight for total heterotrophic bacteria and 1.08±0.6×105 (mean±S.E.), 1.3±0.3×105 and 1.4±0.2×105 cfu g−1 dry weight for pseudomonads, actinomycetes and spore-forming bacteria respectively. Identification of pseudomonads by the Biolog® system indicated, besides the most represented species Pseudomonas fluorescens (biotypes B, F and G), the occurrence of strains belonging to Pseudomonas corrugata. Amplified ribosomal DNA restriction analysis of actinomycetes and spore formers revealed at least three and six different groups of patterns, respectively. Many bacterial isolates were able to induce variations in growth rates of T. borchii mycelium; among these, 101 strains showed antifungal activity, whereas 17 isolates, belonging to spore formers, were able to increase mycelial growth up to 78% when compared to uninoculated mycelial growth. The potential role of these populations in the development and establishment of mycorrhizas is discussed.

Uranium uptake and translocation by the arbuscular mycorrhizal fungus, Glomus intraradices, under root-organ culture conditions

Abstract: Uranium (U) uptake and translocation by the arbuscular mycorrhizal (AM) fungus Glomus intraradices were studied under root-organ culture conditions with Agrobacterium rhizogenes-transformed carrot (Daucus carota) roots as host. Two-compartment Petri plates were used to spatially separate a root compartment (RC) and a hyphal compartment (HC) root growth was restricted to the RC while extraradical hyphae grew in both RC and HC. The HC was labelled with 0.1 muM U-233 at different pH conditions. At the end of the experiment, U was measured in the RC and in the HC. The U absorption by the AM fungus was observed. it included; U uptake by the mycelium developing in the HC, and U translocation from the HC to the RC. The magnitude of this uptake and translocation was highly influenced by the pH of the growth medium, while translocation was highly correlated with the number of hyphae crossing the partition separating the two compartments. These results are the first to show that an AM fungus can take up and translocate U towards roots.

Ectomycorrhizal fungi and exogenous auxins influence root and mycorrhiza formation of Scots pine hypocotyl cuttings in vitro

Abstract: We studied the ability of the ectomycorrhizal (ECM) fungi, Pisolithus tinctorius (Pers.) Coker and Couch and Paxillus involutus (Batsch) Fr. (Strain H), to produce indole-3-acetic acid (IAA) and to affect the formation and growth of roots on Scots pine (Pinus sylvestris L.) hypocotyl cuttings in vitro. Effects of indole-3-butyric acid (IBA) and the auxin transport inhibitor, 2,3,5-triiodobenzoic acid (TIBA), on rooting and the cutting-fungus interaction were also studied. Both fungi produced IAA in the absence of exogenous tryptophan, but the mycelium and culture filtrate of Pisolithus tinctorius contained higher concentrations of free and conjugated IAA than the mycelium and culture filtrate of Paxillus involutus. Inoculation with either fungus or short-term application of culture filtrate of either fungus to the base of hypocotyl cuttings enhanced root formation. Inoculation with either fungus was even more effective in enhancing root formation than treatment of the hypocotyl bases with IBA. Fungal IAA production was not directly correlated with root formation, because rooting was enhanced more by Paxillus involutus than by Pisolithus tinctorius. This suggests that, in addition to IAA, other fungal components play an important role in root formation. Treatment with 5 microM TIBA increased the rooting percentage of non-inoculated cuttings, as well as of cuttings inoculated with Pisolithus tinctorius, perhaps as a result of accumulation of IAA at the cutting base. However, the marked reduction in growth of Pisolithus tinctorius in the presence of TIBA suggests that the effects of TIBA on rooting are complicated and not solely related to IAA metabolism. The high IAA-producer, Pisolithus tinctorius, formed mycorrhizas, and the IBA treatment increased mycorrhizal frequency in this species, whereas TIBA decreased it. Paxillus involutus did not form mycorrhizas, indicating that a low concentration of IAA together with other fungal components were sufficient to stimulate formation and growth of the roots, but not the formation of ECM symbiosis.