Heavy metal uptake by arbuscular mycorrhizas of Elsholtzia splendens and the potential for phytoremediation of contaminated soil

A pot culture experiment and a field experiment were carried out separately to study heavy metal (HM) uptake from soil contaminated with Cu, Zn, Pb and Cd by Elsholtzia splendens Nakai ex F. Maekawa inoculated with arbuscular mycorrhizal (AM) fungi and the potential for phytoremediation. The HM-contaminated soil in the pot experiment was collected from the field experiment site. Two AM fungal inocula, MI containing only one AM fungal strain, Glomus caledonium 90036, and M II consisting of Gigaspora margarita ZJ37, Gigaspora decipens ZJ38, Scutellospora gilmori ZJ39, Acaulospora spp. andGlomus spp., were applied to the soil under unsterilized conditions. In the pot experiment, the plants were harvested after 24 weeks of growth. Mycorrhizal colonization rate, plant dry weight (DW) and P, Cu, Zn, Pb, Cd concentrations were determined. MI-treated plants had higher mycorrhizal colonization rates than MII-treated plants. Both MI and MII increased shoot and root DW, and MII was more effective than MI. In shoots, the highest P, Cu, Zn and Pb concentrations were all observed in the plants treated with MII, while MI decreased Zn and Pb concentrations and increased P but did not alter Cu, and Cd concentrations were not affected by either of two inocula. In roots, MII increased P, Zn, Pb concentrations but did not alter Cu and Cd, and MI did not affect P, Cu, Zn, Pb, Cd concentrations. Cu, Zn, Pb, Cd uptake into shoots and roots all increased in MII-treated plants, while in MI-treated plants, Cu and Zn uptake into shoots and Cu, Zn, Pb, Cd into roots increased but Pb and Cd uptake into shoots decreased. In general, MII was more effective than MI in promoting plant growth and HM uptake. The field experiment following the pot experiment was carried out to investigate the effects of MII under field conditions. The 45-day-old nonmycorrhizal and MII-colonized seedlings of E. splendens were transplanted to HM-contaminated plots and harvested after 5 months. MII-inoculation increased shoot DW and shoot P, Cu, Zn, Pb concentrations significantly but did not alter shoot Cd concentrations, which led to higher uptake of Cu, Zn, Pb, Cd by E. splendens shoots. These results indicate that the AM fungal consortium represented by MII can benefit phytoextraction of HMs and therefore play a role in phytoremediation of HM-contaminated soils.     

Arbuscular mycorrhizal contribution to heavy metal uptake by maize (Zea mays L.) in pot culture with contaminated soil

In two pot-culture experiments with maize in a silty loam (P2 soil) contaminated by atmospheric deposition from a metal smelter, root colonization with indigenous or introduced arbuscular mycorrhizal (AM) fungi and their influence on plant metal uptake (Cd, Zn, Cu, Pb, Mn) were investigated. Soil was -irradiated for the nonmycorrhizal control. In experiment 1, nonirradiated soil provided the mycorrhizal treatment, whereas in experiment 2 the irradiated soil was inoculated with spores of a fungal culture from P2 soil or a laboratory reference culture, Glomus mosseae. Light intensity was considerably higher in experiment 2 and resulted in a fourfold higher shoot and tenfold higher root biomass. Under the conditions of experiment 1, biomass was significantly higher and Cd, Cu, Zn and Mn concentrations significantly lower in the mycorrhizal plants than in the nonmycorrhizal plants, suggesting a protection against metal toxicity. In contrast, in experiment 2, biomass did not differ between treatments and only Cu root concentration was decreased with G. mosseae-inoculated plants, whereas Cu shoot concentration was significantly increased with the indigenous P2 fungal culture. The latter achieved a significantly higher root colonization than G. mosseae (31.7 and 19.1%, respectively) suggesting its higher metal tolerance. Zn shoot concentration was higher in both mycorrhizal treatments and Pb concentrations, particularly in the roots, also tended to increase with mycorrhizal colonization. Cd concentrations were not altered between treatments. Cu and Zn, but not Pb and Cd root-shoot translocation increased with mycorrhizal colonization. The results show that the influence of AM on plant metal uptake depends on plant growth conditions, on the fungal partner and on the metal, and cannot be generalized. It is suggested that metal-tolerant mycorrhizal inoculants might be considered for soil reclamation, since under adverse conditions AM may be more important for plant metal resistance. Under the optimized conditions of normal agricultural practice, however, AM colonization even may increase plant metal absorption from polluted soils.     

Morphological and molecular analyses in Scleroderma species associated with some Caesalpinioid legumes, Dipterocarpaceae and Phyllanthaceae trees in southern Burkina Faso

A combination of morphotypes, polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP) analyses and internal transcribed spacer (ITS) sequencing was used to investigate Scleroderma species that were collected from woodlands in Burkina Faso. We harvested 52 specimens from 20 sites during rainy seasons between 1997 and 2000. According to their morphological features, these specimens were initially characterised, and we then identified six species of Scleroderma. Two of the species were clearly determined as Scleroderma dictyosporum Pat. and S. verrucosum Pers. The four remaining species were characteristically described as (1) displaying big spores with spines up to 2 μm (Scleroderma sp1), (2) producing spores without ornamentation (Scleroderma sp2), (3) spores with very small spines (Scleroderma sp3) and (4) with yellow sporocarps and sub-spherical spores (Scleroderma sp4). The specimens were then analysed using PCR/RFLP of the intergenic regions of rDNA, ITS and IGS1 and ITS sequencing. The restriction fragments obtained with two endonucleases, HinfI and MboI on ITS and IGS1 regions, showed that some isolates of S. dictyosporum had the same patterns as isolates and basidiocarps of Scleroderma sp4 (IR265, IR408, SP4-2903). Isolates of Scleroderma sp3 (IR252) had common restriction fragments as isolates of S. verrucosum (IR500, IR600). Intraspecific differences were observed in the two previously determined species, as well as in Scleroderma sp2. The ITS sequencing and phylogenetic analyses showed that the ribotypes identified by PCR/RFLP within these species might be phylogenetic species. Combining these molecular results allowed regrouping the six morphological species in three sets of cryptic species: a first set with two species including S. dictyosporum Pat., a second set with four species, including both S. verrucosum Pers. and Scleroderma sp1 and a third set with two species, including Scleroderma sp2. These investigations and the combined morphological and molecular analyses used to sort out species paved the way for identifying larger populations of Scleroderma species in Burkina Faso and other tropical zones.     

<Emphasis Type="Italic">Pterostylis nutans</Emphasis> (Orchidaceae) has a specific association with two <Emphasis Type="Italic">Ceratobasidium</Emphasis> root-associated fungi across its range in eastern Australia

In this study, we have identified the root-associated fungi of a common species of terrestrial orchid across its range in eastern Australia. We have amplified and cloned fungal ITS DNA extracted from roots of 15 Pterostylis nutans R. Br. plants from six separate geographic localities. Sequencing and GenBank comparison demonstrated two species of Ceratobasidium fungi as the main fungal partners of the orchid. Uncommon fungal associates included homobasidiomycete species such as a Gymnomyces sp. and a Tricholoma sp., Leptodontidium orchidicola, and an unidentified soil fungus. These results demonstrate that specificity for fungal partners occurs in P. nutans and reinforces the idea that conservation measures for endangered Australian orchids must include ex situ perpetuation of fungal symbionts as well as plant material.     

Relationship between copper- and zinc-induced oxidative stress and proline accumulation in<Emphasis Type="Italic"> Scenedesmus</Emphasis> sp.

A 4-h exposure of Scenedesmus sp. to Cu or Zn enhanced intracellular levels of both test metals and proline. The level of intracellular proline increased markedly up to 10 µM Cu, but higher concentrations were inhibitory. However, intracellular proline consistently increased with increasing concentration of Zn in the medium. Cu and Zn induced oxidative stress in the test alga by increasing lipid peroxidation and membrane permeability, and by reducing SH content. Pretreatment of the test alga with 1 mM proline for 30 min completely alleviated Cu-induced lipid peroxidation, minimized K⁺ efflux and also reduced depletion of the SH pool. But proline pretreatment could only slightly reduce Zn-induced oxidative stress. Interestingly, proline pretreatment increased the level of Cu (25–54%) and Zn (19–49%) inside the cells. It did not affect the activities of superoxide dismutase, ascorbate peroxidase or catalase, but improved glutathione reductase activity under Cu and Zn stress. A comparison of the effects of proline pretreatment on lipid peroxidation by Cu, Zn, methyl viologen and ultraviolet-B radiation suggests that proline protects cells from metal-induced oxidative stress by scavenging reactive oxygen species rather than by chelating metal ions. Pretreatment of cells with a known antioxidant (ascorbate) and a hydroxyl radical scavenger (sodium benzoate) considerably reduced metal-induced lipid peroxidation and proline accumulation. However, sodium benzoate had a very mild effect on Zn-induced lipid peroxidation and proline accumulation. The present study demonstrates that proline possibly acts by detoxifying reactive oxygen species, mainly hydroxyl radicals, rather than by improving the antioxidant defense system under metal stress.Abbreviations APOX Ascorbate peroxidase - CAT Catalase - GR Glutathione reductase - MDA Malondialdehyde - MV Methyl viologen - ROS Reactive oxygen species - SH Sulphydryl - SOD Superoxide dismutase - UV-B Ultraviolet-B radiation     

Trace elements found to be variable in two coral reef species,Heliofungia actiniformis andGalaxea fascicularis, collected from the ryukyu islands

Biominerals and metals of intertidal corals of two species (Heliofungia actiniformis, Quoy and Gaimard;Galaxea fascicularis, Linnaeus), collected from the Iriomote Island of Ryukyu, were examined with an inductively coupled plasma atomic emission spectrometer (ICP-AES). Twelve elements were detectable in the coralline skeletons dissected radially along the growth axis. The relative content (RC) of Hg periodically fluctuated and was minimum at the hollow sites of the coralline slab ofHeliofungia sp., corresponding to the cyclic growth. There were two types of elements: constant elements and variable elements along the growth axis. RCs of Ca, Mg, A1, Si, and P were nearly constant. RCs of Fe, Mn, Cu, and Ba were variable, but not as regularly changed as Hg. There were positive mass correlations of Hg to Mn, Cu and Zn, but not to Ba and Fe. In contrast, these relationships were not prominent and were likely degraded by aging in the skeleton ofGalaxea sp., suggesting a different mode from that of theHeliofungia sp.     

Lignosulfonate promotes the interaction between Scots pine and an ectomycorrhizal fungus Pisolithus tinctorius in vitro

Lignosulfonate (LS) is a lignin-based polymer obtained as a by-product from paper industry, which may have potential as an amendment with macronutrients. We studied effects of LS on the interaction between Scots pine (Pinus sylvestris L.) seedlings and hypocotyl cuttings and the ectomycorrhizal (ECM) fungusPisolithus tinctorius (Pers.) Coker and Couch. The experiments were performed in vitroon the MMN agar medium containing Fe–LS chelate at the concentrations of 0, 5, 10 and 25 mg/L. Inoculation with P. tinctoriusincreased root growth of the seedlings. Fe–LS enhanced P. tinctorius induced formation of lateral roots and had a dose-dependent positive effect on the establishment of mycorrhizas on the seedlings. The growth of the fungal mycelium was improved by Fe–LS, which might cause faster and more intensive contact with the roots and, thus, better root growth and mycorrhiza formation. P.tinctorius enhanced also adventitious root formation and subsequent root growth of the hypocotyl cuttings but without any synergistic effect with Fe–LS. Our study with P. tinctorius and Scots pine in vitro indicates that a low-cost by-product Fe–LS, obtained from paper industry, may be a potential tool to improve the efficiency of fungal inoculations, thus, facilitating the early interaction between an ECM fungus and host seedling.     

In vitro weathering of phlogopite by ectomycorrhizal fungi

The ways in which ectomycorrhizal fungi benefit tree growth and nutrition have not been fully elucidated. Whilst it is most probably due to improved soil colonization, it is also likely that ectomycorrhizal fungi could be directly involved in nutrient cycling of soil reserves. This study assessed access by two species of ectomycorrhizal fungi to soil nonexchangeable K⁺ reserves. The incubation of ectomycorrhizal fungi in bi-compartment Petri dishes with phlogopite led to cation exchange reactions and to crystal lattice weathering. Paxillus involutus COU led to irreversible phlogopite transformations, while Pisolithus tinctorius 441 led to reversible ones. Simultaneous depletion in K⁺ and Mg²⁺ led to an enhanced weathering of phlogopite by P. tinctorius 441. The observation of phlogopite evolution shows that some specific Al³⁺ immobilization occurred under P. tinctorius 441. The data suggest that these bio-weathering mechanisms could be related to the release of fungal organic acids or other complex forming molecules.     

Effect of time of inoculation on in vitro ectomycorrhizal colonization and nodule initiation in Acacia holosericea seedlings

The complex interactions that occur in systems with more than one type of symbiosis were studied using one isolate of Bradyrhizobium sp. and the ectomycorrhizal fungus Pisolithus tinctorius (Pers.) Coker and Couch inoculated on to the roots of Acacia holosericea A. Cunn. ex G. Don in vitro. After a single inoculation with Bradyrhizobium sp., bacteria typically entered the roots by forming infection threads in the root hair cells via the curling point of the root hair and/ or after intercellular penetration. Sheath formation and intercellular penetration were observed on Acacia roots after a single inoculation with Pisolithus tinctorius but no radial elongation of epidermal cells. Simultaneous inoculation with both microorganisms resulted in nodules and ectomycorrhiza on the root system, occasionally on the same lateral root. On lateral roots bearing nodules and ectomycorrhiza, the nodulation site was characterized by the presence of a nodule meristem and the absence of an infection thread; sheath formation and Hartig net development occurred regularly in the region of the roots adjacent to nodules. Prior inoculation with Bradyrhizobium sp. did not inhibit ectomycorrhizal colonization in root segments adjacent to nodules in which nodule meristems and infection threads were clearly present. Conversely, in ectomycorrhizae inoculated by bacteria, the nodule meristem and the infection thread were typically absent. These results show that simultaneous inoculation with both microorganisms inhibits infection thread development, thus conferring an advantage on fungal hyphae in the competition for infection sites. This suggests that fungal hyphae can modify directly and/or indirectly the recognition factors leading to nodule meristem initiation and infection thread development.     

Stress responses and metal tolerance of <Emphasis Type="Italic">Chlamydomonas acidophila</Emphasis> in metal-enriched lake water and artificial medium

Chlamydomonas acidophila faces high heavy-metal concentrations in acidic mining lakes, where it is a dominant phytoplankton species. To investigate the importance of metals to C. acidophila in these lakes, we examined the response of growth, photosynthesis, cell structure, heat-shock protein (Hsp) accumulation, and metal adsorption after incubation in metal-rich lake water and artificial growth medium enriched with metals (Fe, Zn). Incubation in both metal-rich lake water and medium caused large decreases in photosystem II function (though no differences among lakes), but no decrease in growth rate (except for medium + Fe). Concentrations of small Hsps were higher in algae incubated in metal-rich lake-water than in metal-enriched medium, whereas Hsp60 and Hsp70A were either less or equally expressed. Cellular Zn and Fe contents were lower, and metals adsorbed to the cell surface were higher, in lake-water-incubated algae than in medium-grown cells. The results indicate that high Zn or Fe levels are likely not the main or only contributor to the low primary production in mining lakes, and multiple adaptations of C. acidophila (e.g., high Hsp levels, decreased metal accumulation) increase its tolerance to metals and permit survival under such adverse environmental conditions. Supposedly, the main stress factor present in the lake water is an interaction between low P and high Fe concentrations.     

Detecting the heavy metal tolerance level in ectomycorrhizal fungi <Emphasis Type="Italic">in vitro</Emphasis>

Summary Eight isolates of ectomycorrhizal fungi namely, Laccaria fraterna (EM-1083),Laccaria laccata (EM-1191), Pisolithus tinctorius (EM-1081),Pisolithus tinctorius (EM-1293), Scleroderma cepa (EM-1233),Scleroderma flavidum (EM-1235),Scleroderma verucosum, (EM-1283) and Hysterangium incarceratum (EM-1185) were grown on specially designed cocktail media prepared by adding various concentrations of different heavy metals namely Al, As, Cd, Cr, Ni and Pb. The heavy metals were selected keeping in view their relative abundance in coal ash and potential toxicity. The fungal isolates were grown on such designed cocktail media. The colony diameter was used for the measurement of the fungal growth. Total heavy metal accumulated in the mycelia was assayed by atomic absorption spectrophotometry.In relation to metal tolerance ability in general,Hysterangium incarceratum (EM-1185) showed maximum tolerance with respect to growth, Laccaria fraterna (EM-1083) and Pisolithus tinctorius (EM-1293) also showed considerable tolerance to the heavy metals tested. In relation to metal uptake in particular, Pisolithus tinctorius (EM-1293), has reported maximum uptake of Al (34642.58 ppm), Cd (302.12 ppm) and Pb (3501.96 ppm). In Laccaria fraterna (EM-1083), As (130.57 ppm) and Cr (402.38 ppm) uptake was recorded maximum; and Hysterangium incarceratum (EM-1185) has recorded maximum Ni (2648.59 ppm) uptake among the three suitable isolates documented here.     

Fungal Infection of Mantis Shrimp (<Emphasis Type="Italic">Oratosquilla oratoria</Emphasis>) Caused by Two Anamorphic Fungi Found in Japan

Two fungal pathogens of the mantis shrimp (Oratosquilla oratoria) in Yamaguchi and Aichi Prefectures, Japan are described as the new species Plectosporium oratosquillae and Acremonium sp. (a member of the Emericellopsis marine clade). Both fungi infect the gills of the mantis shrimp, which become brown or black due to melanization. The former species is characterized by its slow growth on artificial seawater yeast extract peptone glucose (PYGS) agar, pale yellow to pale orange or grayish yellow colonies, short cylindrical solitary phialides with a wavy tip, and one-celled ellipsoidal conidia. Although lacking the two-celled conidia demonstrated by the type species Plectosporium tabacinum, the taxonomic placement of the new species was confirmed by DNA sequence analysis of the internal transcribed spacer (ITS) region of ribosomal DNA (ITS1, 5.8S rDNA and ITS2). Acremonium sp., the other causal pathogen, differs from P. oratosquillae by its fast growth on PYGS agar, pale orange to salmon-colored colonies, long, slender conidiophores consisting of solitary phialides with tips lacking an undulate outline, and typically cylindrical conidia. Analysis of ITS and β-tubulin gene sequences placed this fungus within the phylogenetically distinct Emericellopsis (anam. Acremonium) marine clade. Various physiological characteristics of both pathogens were also investigated. This is the first report of a fungal infection found on the mantis shrimp in Japan.     

Differential effects of AM fungal isolates on Medicago truncatula growth and metal uptake in a multimetallic (Cd, Zn, Pb) contaminated agricultural soil

Toxic metal accumulation in soils of agricultural interest is a serious problem needing more attention, and investigations on soil–plant metal transfer must be pursued to better understand the processes involved in metal uptake. Arbuscular mycorrhizal (AM) fungi are known to influence metal transfer in plants by increasing plant biomass and reducing metal toxicity to plants even if diverging results were reported. The effects of five AM fungi isolated from metal contaminated or non-contaminated soils on metal (Cd, Zn) uptake by plant and transfer to leachates was assessed with Medicago truncatula grown in a multimetallic contaminated agricultural soil. Fungi isolated from metal-contaminated soils were more effective to reduce shoot Cd concentration. Metal uptake capacity differed between AM fungi and depended on the origin of the isolate. Not only fungal tolerance and ability to reduce metal concentrations in plant but also interactions with rhizobacteria affected heavy metal transfer and plant growth. Indeed, thanks to association with nodulating rhizobacteria, one Glomus intraradices inoculum increased particularly plant biomass which allowed exporting twofold more Cd and Zn in shoots as compared to non-mycorrhizal treatment. Cd concentrations in leachates were variable among fungal treatments, but can be significantly influenced by AM inoculation. The differential strategies of AM fungal colonisation in metal stress conditions are also discussed.     

In vitro weathering of phlogopite by ectomycorrhizal fungi

Oxalate accumulation in external medium under hyphal mats of two ectomycorrhizal species is strongly stimulated (1.7 to 35 fold) by a simultaneous depletion of available K⁺ and Mg²⁺. Pisolithus tinctorius strain 441 accumulates oxalate both on NH₄–N and on NO₃–N whereas Paxillus involutus strain COU only accumulates oxalate on NO₃–N. On NO₃–N, under a simultaneous K⁺ and Mg²⁺ deficiency, P. involutus COU is a very active oxalate producer compared to P. tinctorius 441. The present results could explain the various mineralogical evolutions of a phlogopite mica previously recorded under P. involutus COU or P. tinctorius 441 and suggest a key role for fungal oxalic acid during mineral weathering in response to nutrient deficiency.     

Aluminum-mycorrhizal interactions in the physiology of pitch pine seedlings

Aluminum (Al) in the rhizosphere adversely affects plant nutrition and growth. Although many conifer species, and pitch pine (Pinus rigida) in particular, have evolved on acidic soils where soluble Al is often high, controlled environment studies often indicate that Al interferes with seedling growth and nutrient relations. Under normal field conditions, conifer roots grow in a symbiotic relationship with ectomycorrhizal fungi, and this association may modulate the effects of Al on root physiology. To investigate the influence of mycorrhizal infection on Al toxicity, pitch pine seedlings were grown with or without the ectomycorrhizal symbiont Pisolithus tinctorius and were exposed to low levels of Al in sand culture. Aluminum at 50 μM reduced nonmycorrhizal seedling growth and increased foliar Al concentrations, but did not alter photosynthetic gas exchange or other aspects of seedling nutrition. Nonmycorrhizal seedlings exposed to 200 μM Al exhibited decreased growth, increased transpiration rates, decreased water use efficiency, increased foliar Al and Na levels, and reduced foliar P concentrations. Seedlings inoculated with P. tinctorius exhibited unaltered growth, physiological function, and ionic relations when exposed to Al. The fungal symbiont evidently modulated ionic relations in the rhizosphere, reducing Al-P precipitation reactions, Al uptake, and subsequent root and shoot tissue Al exposure.     

Variation in soft tissue chemistry among scyphozoan and cubozoan jellyfishes from the Great Barrier Reef,Australia

Bioaccumulation of trace elements in jellyfish has so far received little attention, despite their being prey for many animals from multiple trophic levels and targeted by commercial jellyfish fisheries. Scyphozoan and cubozoan jellyfish were collected over a three year period from across-shelf and along the northern and central Great Barrier Reef, Australia. To test the hypotheses that jellyfishes were able to accumulate elements above ambient background levels, and if there were spatial or temporal variations among species, soft tissue concentrations of 14 trace elements were compared with ambient seawater concentrations. Most elements, including aluminium, arsenic, barium, cadmium, chromium, copper, iron, manganese and zinc were measured at concentrations above ambient seawater levels indicating bioaccumulative capacity. Results showed some regulation of lithium in Cassiopea sp., Cyanea sp. and Mastigias sp., while calcium, magnesium and strontium reflected ambient conditions for all species. Accumulation varied significantly among species and sampling locations. For Mastigias sp. and Netrostoma sp., tissue concentrations of Al, As, Cu, Fe and Zn decreased with distance from the mainland. The hypothesis that jellyfishes are capable of accumulating trace elements was accepted, and their use as biomonitors should be investigated further.     

Differential pulse polarography: a method for the direct study of biosorption of metal ions by live bacteria from mixed metal solutions

The technique of differential pulse polarography is shown here to be applicable to the monitoring directly the biosorption of metal ions from solution by live bacteria from mixed metal solutions. Biosorption of Cd(II), Zn(II) and Ni(II) by P. cepacia was followed using data obtained at the potential which is characteristic of the metal ion in the absence and presence of cells. Hepes buffer (pH 7.4, 50 mM) was used as a supporting electrolyte in the polarographic chamber and metal ion peaks in the presence of cells of lower amplitude were obtained due to metal-binding by the cells. Well defined polarographic peaks were obtained in experiments involving mixtures of metal ions of Cd(II)-Zn(II), Cu(II)-Zn(II), Cu(II)-Cd(II) and Cd(II)-Ni(II). Biosorption of Cd(II), Zn(II) increased with solution pH. The method was also tested as a rapid technique for assessing removal of metal ions by live bacteria and the ability of the polarographic technique in measuring biosorption of metal ions from mixed metal solutions is demonstrated. Cu(II) was preferentially bound and removal of metals was in the order Cu(II) > Ni(II) > Zn(II), Cd(II) by intact cells of P. cepacia. This revised version was published online in August 2006 with corrections to the Cover Date.     

Influence of aluminum on in vitro formation of Pinus caribaea mycorrhizae

The ability of Pinus caribaea var. hondurensis to form mycorrhizae was determined in vitro with seven species of ectomycorrhizal fungi in the presence of six levels of Al (added as AlCl₃) in a nutrient solution. The time required for mycorrhizal formation, the number of mycorrhizal root tips and the percent mycorrhizae were measured after 15, 30, 60, 90 and 120 days. Cenococcum graniforme was susceptible to Al toxicity at all Al concentrations. Pisolithus tinctorius and Suillus sp. were depressed at lower but stimulated at higher Al concentrations. The inverse was shown for Rhizopogon reaii and Hebeloma cylindrosporum. Tolerance to Al was verified for R. nigrescens and H. crustuliniforme. Pisolithus tinctorius had the largest mycorrhizal capacity, defined as the sum of the values for time, percent and number of mycorrhizae. Ectomycorrhizal fungi appeared to ameliorate Al damage to plant roots even in treatments where no mycorrhizae formed. Inoculation of pine seedlings with Al-tolerant mycorrhizal fungi is likely to improve reforestation efforts in highly-weathered tropical soils.     

Comparative structural study ofQuercus serrata andQ. acutissima formed byPisolithus tinctorius andHebeloma cylindrosporum

Ectomycorrhizas were synthesized in pots and growth pouches betweenQuercus serrata, Q. acutissima, and two ectomycorrhizal fungi,Pisolithus tinctorius andHebeloma cylindrosporum. Root morphology and the structure of the mantle and Hartig net were compared using light, fluorescence, scanning and transmission electron microscopy.P. tinctorius initially colonized root cap cells, and eventually produced a highly branched lateral root system with a complete mantle, whereasH. cylindrosporum promoted root elongation with few hyphae on the root apex surface indicating that interaction between roots differs with fungal species. Hartig net structure and hyphal inclusions varied between all the combinations tested. There were structural differences between mycorrhizas ofH. cylindrosporum/Q. acutissima grown in soil and growth pouches, which indicate that the growth pouch environment can induce artefacts in roots. Fruit bodies ofH. cylindrosporum developed in pots withQ. acutissima. AlthoughP. tinctorius has been used to inoculate oak seedlings in the nursery, results of this study indicate thatH. cylindrosporum may also be an effective ectomycorrhizal fungus forQ. serrata andQ. acutissima.     

Arbuscular mycorrhizas and ectomycorrhizas of <Emphasis Type="Italic">Uapaca bojeri</Emphasis> L. (Euphorbiaceae): sporophore diversity,patterns of root colonization,and effects on seedling growth and soil microbial catabolic diversity

The main objectives of this study were (1) to describe the diversity of mycorrhizal fungal communities associated with Uapaca bojeri, an endemic Euphorbiaceae of Madagascar, and (2) to determine the potential benefits of inoculation with mycorrhizal fungi [ectomycorrhizal and/or arbuscular mycorrhizal (AM) fungi] on the growth of this tree species and on the functional diversity of soil microflora. Ninety-four sporophores were collected from three survey sites. They were identified as belonging to the ectomycorrhizal genera Afroboletus, Amanita, Boletus, Cantharellus, Lactarius, Leccinum, Rubinoboletus, Scleroderma, Tricholoma, and Xerocomus. Russula was the most frequent ectomycorrhizal genus recorded under U. bojeri. AM structures (vesicles and hyphae) were detected from the roots in all surveyed sites. In addition, this study showed that this tree species is highly dependent on both types of mycorrhiza, and controlled ectomycorrhization of this Uapaca species strongly influences soil microbial catabolic diversity. These results showed that the complex symbiotic status of U. bojeri could be managed to optimize its development in degraded areas. The use of selected mycorrhizal fungi such the Scleroderma Sc1 isolate in nursery conditions could be of great interest as (1) this fungal strain is very competitive against native symbiotic microflora, and (2) the fungal inoculation improves the catabolic potentialities of the soil microflora.