The role of arbuscular mycorrhiza in zinc uptake by red clover growing in a calcareous soil spiked with various quantities of zinc

Three pot experiments were conducted to investigate the role of arbuscular mycorrhiza (AM) in Zn uptake by red clover. Plants inoculated with Glomus mosseae and uninoculated controls were grown in a sterile calcareous soil in 'Plexiglas' (Acrylic) containers with two nylon net partitions (30 microm mesh) to separate the central root zone from the two outer hyphal zones. The effects of mycorrhiza on plant growth and Zn uptake changed dramatically with increasing Zn addition level (range 0-1200 mg kg(-1)) in the root zone. With Zn addition levels <300 mg kg(-1), added Zn did not affect plant yield and above the critical level plant yield gradually decreased but was always higher for mycorrhizal than for controls. Below the critical Zn application rate (50 mg kg(-1)), Zn uptake was enhanced while above this level Zn translocation to the shoots decreased. At all Zn addition levels, mycorrhizal colonization increased Zn absorption and accumulation in the roots, and this may help to explain the alleviation of Zn toxicity at high Zn application rates. As expected, AM colonization enhanced P nutrition and hence yield at all added Zn levels studied. Efforts were made to obtain direct evidence for hyphal contribution to Zn uptake by applying both Zn to the hyphal growth zone and additional P to the root zone to avoid the 'growth dilution effect'. The data demonstrate that mycorrhizal hyphae could absorb Zn directly from the soil and then transfer it to the plant roots. The hyphal contribution to Zn uptake by the host plant reached its maximum value at the Zn addition level of 50 mg kg(-1), in which Zn uptake via the extramatrical hyphae comprised 22% of total uptake, thus confirming the critical Zn application level found previously.     

The effect of mycorrhiza on the growth and elemental composition of Ni-hyperaccumulating plant Berkheya coddii Roessler

The effect of arbuscular mycorrhizal fungi (AMF) on growth and element uptake by Ni-hyperaccumulating plant, Berkheya coddii, was studied. Plants were grown under laboratory conditions on ultramafic soil without or with the AM fungi of different origin. The AM colonization, especially with the indigenous strain, significantly enhanced plants growth and their survival. AMF affected also the elemental concentrations that were studied with Particle-induced X-ray emission (PIXE). AMF (i) increased K and Fe in shoots, Zn and Mn in roots, P and Ca both, in roots and shoots; (ii) decreased Mn in shoots, Co and Ni both, in shoots and roots. Due to higher biomass of mycorrhizal plants, total Ni content was up to 20 times higher in mycorrhizal plants compared to the non-mycorrhizal ones. The AMF enhancement of Ni uptake may therefore provide an improvement of a presently used technique of nickel phytomining.     

Uptake of Zn by arbuscular mycorrhizal white clover from Zn-contaminated soil

A randomised block glasshouse pot experiment compared the growth and Zn uptake of mycorrhizal and nonmycorrhizal white clover plants grown in a sterile soil/sand mixture containing 25 mg Zn kg(-1) to which five application rates of Zn (as ZnSO4) from 0 to 400 mg kg(-1) were made. Two mycorrhizal inocula infected roots from the field and from clover trap cultures, were compared. Mycorrhizal infection (ranging from 33% to 46% of total root length) and Zn application had little effect on plant growth. Increasing Zn application rate led to increased uptake of Zn in roots and shoots (especially roots), but the increases were significantly greater in non-mycorrhizal controls than in mycorrhizal treatments. In contrast, P uptake was higher in mycorrhizal than in non-mycorrhizal plants. Plants that received trap culture inoculum had significantly lower Zn uptake than those that received field inoculum. The results indicate that mycorrhizal infection may have exerted some protective effect against plant Zn accumulation at the range of soil Zn concentrations studied and may have immobilised Zn in or near the roots to some extent. However, this mycorrhizal effect cannot be explained simply by tissue dilution, hyphal sequestration or root immobilisation of Zn.     

Arbuscular mycorrhizal fungi can decrease the uptake of uranium by subterranean clover grown at high levels of uranium in soil

Subterranean clover inoculated or not with the arbuscular mycorrhizal (AM) fungus Glomus intraradices was grown on soil containing six levels of 238U in the range 0-87 mg kg(-1). Increasing U concentration in soil enhanced the U concentration in roots and shoots of both mycorrhizal and nonmycorrhizal plants but had no significant effects on plant dry matter production or root AM colonization. Mycorrhizas increased the shoot dry matter and P concentration in roots and shoots, while in most cases, it decreased the Ca, Mg and K concentrations in plants. The AM fungus influenced U concentration in plants only in the treatment receiving 87 mg U kg(-1) soil. In this case, U concentration in shoots of nonmycorrhizal plants was 1.7 times that of shoots of mycorrhizal plants. These results suggested that mycorrhizal fungi can limit U accumulation by plants exposed to high levels of U in soil.     

Growth and nutrient uptake of arbuscular mycorrhizal maize in different depths of soil overlying coal fly ash

Application of topsoil over phytotoxic mine wastes is often practised to establish perennial plant communities on minespoil areas. In China, population pressure encourages attempts to remediate such areas by growing arable crop plants, but efforts to establish agricultural crops often fail. We report an outdoor pot experiment that compared the effects of two arbuscular mycorrhizal (AM) fungi, Glomus mosseae (Nicol. and Gerd.) Gerdemann and Trappe and G. versiforme (Karsten) Berch, on the growth and nutrient uptake of maize (Zea mays L.) grown in different depths of soil layer overlying coal fly ash. Colonization by both AM fungi increased plant growth compared with non-mycorrhizal controls, with G. mosseae giving higher yields of maize than G. versiforme at the same depths of soil. Increasing soil depth led to increased plant yields. Mycorrhizal plants absorbed more nutrients than non-mycorrhizal controls, and translocated less Na to the shoots, perhaps protecting the plants from excessive Na accumulation. These preliminary results indicate that arbuscular mycorrhizas may make a substantial contribution to successful crop establishment in soils overlying areas of coal fly ash.     

Arbuscular mycorrhizal fungi mediated uptake of lanthanum in Chinese milk vetch (Astragalus sinicus L.)

A glasshouse experiment was carried out to study the effect of mycorrhizal formation by Gigaspora margarita, Glomus intraradices or Acaulospora laevis on plant growth and lanthanum (La) uptake of Astragalus sinicus L. in soils spiked with La at five levels (0, 1, 5, 10 and 20 mg kg(-1)). La application decreased mycorrhizal infection frequency and activity of fungal succinate dehydrogenase and alkaline phosphatase. Increasing La concentrations in soil led to increased La accumulation in tissues of both mycorrhizal and non-mycorrhizal plants, but inoculation with Gig. margarita or G. intraradices reduced La concentrations in shoots and roots at higher concentrations of La in the soil. Plant biomass and P concentrations in shoots and roots were reduced by La application, but increased by inoculation with Gig. margarita or G. intraradices. The results showed that appropriate AM fungi colonization could be effective in alleviating La toxicity in A. sinicus.     

Survival strategies of plants associated with arbuscular mycorrhizal fungi on toxic mine tailings

A field survey of metal concentrations and arbuscular mycorrhizal (AM) components of plants growing on five mining sites was conducted in Chenzhou City, Hunan Province, Southern China and a control site in Hong Kong. Significant differences were observed in the average concentrations of total heavy metals (Pb, Zn, Cu, Cd) and one metalloid (As) in contaminated soils compared with the control site. Gramineae and Compositae were the dominant plant families growing on mine tailings, with Chrysanthemum moritolium (common chrysanthemum), Cynodon dactylon (Bermuda grass), Miscanthus florodulus (Sword grass) and Pteris vittata (Ladder brake fern) commonly found at all sites. AM fungal colonization was detected in most of the plants. Comparing the four common plant species, three components of mycorrhizal colonization (arbuscules, vesicles and coiled hyphae) were found in the roots of C. dactylon and P. vittata growing at Do Shun Long (DSL) mine site. Concentrations of As in fronds were 24-fold higher than in roots of P. vittata with the highest mycorrhizal colonization rate (73%) among all sampling sites. Extensive mycorrhizal colonization (85%) was also recorded in the roots of C. dactylon with As accumulation 57 times higher than in shoots. The four common plants found in metal contaminated sites had developed different strategies for survival in the contaminated sites with the aid of indigenous AM fungi.     

Changes in soil solution Zn and pH and uptake of Zn by arbuscular mycorrhizal red clover in Zn-contaminated soil

Red clover plants inoculated with Glomus mosseae were grown in a sterile pasture soil containing 50 mg Zn kg(-1) in 'Plexiglas' (acrylic) containers with nylon net partitions (30 microm mesh) designed to separate the soil into a central root zone and two outer zones for hyphal growth with no root penetration. Two porous plastic soil moisture samplers were installed in each pot, one in the root compartment and the other in one of the hyphal compartments. The soil in the outer compartments was amended with one of the four application rates of Zn (as ZnSO4) ranging from 0 to 1000 mg kg(-1). Non-mycorrhizal controls were included, and there were five replicates of each treatment in a randomised block in a glasshouse. Uninoculated plants received supplementary P to avoid yield limitation due to low soil P status. Plants grew in the central compartment for nine weeks. Soil moisture samples were collected 4, 24 and 62 days after sowing to monitor changes in the Zn concentration and pH of the soil solution. At harvest, the mean mycorrhizal infection rate of inoculated plants ranged from 29% to 34% of total root length and was little affected by Zn application. Root and shoot yields were not affected by mycorrhizal infection. Plant Zn concentration and uptake were lower in mycorrhizal plants than non-mycorrhizal controls, and this effect was more pronounced with increasing Zn application rate to the soil. Soil solution Zn concentrations were lower and pH values were higher in mycorrhizal treatments than non-mycorrhizal controls and the mycorrhiza effect was more pronounced at higher Zn application rates. The protective effect of mycorrhiza against plant Zn uptake may have been associated with changes in Zn solubility mediated by changes in the soil solution pH, or by immobilisation of Zn in the extraradical mycelium.     

Arbuscular mycorrhiza enhanced arsenic resistance of both white clover (Trifolium repens Linn.) and ryegrass (Lolium perenne L.) plants in an arsenic-contaminated soil

In a compartmented cultivation system, white clover (Trifolium repens Linn.) and ryegrass (Lolium perenne L.), with their roots freely intermingled, or separated by 37 microm nylon mesh or plastic board, were grown together in an arsenic (As) contaminated soil. The influence of AM inoculation on plant growth, As uptake, phosphorus (P) nutrition, and plant competitions were investigated. Results showed that both plant species highly depended on mycorrhizas for surviving the As contamination. Mycorrhizal inoculation substantially improved plant P nutrition, and in contrast markedly decreased root to shoot As translocation and shoot As concentrations. It also showed that mycorrhizas affected the competition between the two co-existing plant species, preferentially benefiting the clover plants in term of nutrient acquisition and biomass production. Based on the present study, the role of AM fungi in plant adaptation to As contamination, and their potential use for ecological restoration of As contaminated soils are discussed.     

Influence of early stages of arbuscular mycorrhiza on uptake of zinc and phosphorus by red clover from a low-phosphorus soil amended with zinc and phosphorus

In a pot experiment, red clover (Trifolium pratense) was grown in sterilized Zn-amended low available P soil (0, 50 or 400 mg Zn kg(-1)) with or without 100 mg kg(-1) added P and with or without inoculation with the arbuscular mycorrhizal (AM) fungus G. mosseae. When the plants were harvested after 40 days, AM colonization of the roots was still at an early stage, with only 14-38% of total root length colonized on average. AM colonization was highest in low-P soil, and was lowest in soil amended with 400 mg Zn kg(-1). Shoot yields were highest in AM plants with added P, but root yields were unaffected by AM inoculation. Shoot and root yields were higher with 100 mg added P kg(-1) soil, but lower with 400 mg Zn kg(-1) than 50 mg Zn kg(-1) or controls unamended with Zn. Shoot and root P concentrations were seldom higher in AM plants, but shoot P offtakes were higher in AM plants with added P. Concentrations of Zn and Cu were much higher in the roots than in the shoots. Shoot and root Zn and shoot Cu were lower, but root Cu was higher, in AM plants. Soil residual pH after plant growth was higher in AM treatments, and residual total Zn was also higher, indicating lower Zn uptake by AM plants. Soil solution pH was higher in AM treatments, and soil solution Zn was lower in the presence of mycorrhiza. The results are discussed in terms of AM protection of the plants against excessive shoot Zn uptake.     

Effects of mycorrhizae and other soil microbes on revegetation of heavy metal contaminated mine spoil

The effects of mycorrhizal fungi and other soil microorganisms on growth of two grasses, Andropogon gerardii Vitm. and Festuca arundinacea Schreb., in heavy metal-contaminated soil and mine tailings were investigated. A. gerardii is highly dependent on mycorrhizal fungi in native prairie, while F. arundinacea is a facultative mycotroph and relies on mycorrhizal symbiosis only in extremely infertile soils. Regardless of microbial amendments, neither plant species was able to establish and grow in the mine tailings. Both plant species grew in the moderately contaminated or non-contaminated soils, although A. gerardii grew in these soils only when mycorrhizal. Other soil microbes significantly improved growth of A. gerardii only in uncontaminated soil, but to a lesser extent than mycorrhizae. Although F. arundinacea was more highly colonized by mycorrhizal fungi than A. gerardii, neither microbial amendment affected growth of fescue in any soil. In several treatments mycorrhizal fungi adapted to uncontaminated soil stimulated plant growth more than mycorrhizae adapted to the moderately contaminated soil. However, mycorrhizal fungi adapted to contaminated soil did not increase the productivity of plant growth in contaminated soil more than fungi adapted to uncontaminated soil. A. gerardii plants inoculated with mycorrhizal fungi retained more Zn in roots than in shoots, confirming earlier reports that mycorrhizal fungi alter the translocation pattern of heavy metals in host plants. In contrast, mycorrhizae did not affect translocation patterns in F. arundinaceae, suggesting that the mycorrhizal dependence of a plant species is correlated with the retention of metals in roots. The correlation between mycorrhizal dependence of a plant species and mycorrhizal alteration of translocation pattern may also explain the inconsistent reports of mycorrhizal effects on translocation of heavy metals in plants. Plant response to mycorrhizal symbiosis may therefore provide a useful criterion for the selection of the plant species to be used in revegetation of contaminated sites.     

Plant accumulation of potentially toxic elements in sewage sludge as affected by soil organic matter level and mycorrhizal fungi.

Leek (Allium ameloprasum) was grown in pot trials in two clay loams of contrasting organic contents, with and without indigenous mycorrhizal propagules. Sewage sludges containing varying levels of Cd, Cu and Zn were added. Extractable soil metals, plant growth, major nutrient content and accumulation of metals, and soil microbial indices were investigated. The aim was to establish whether soil organic content and mycorrhizal status affected plant and microbial exposure to these metals. Extractable metals were higher and responses to inputs more pronounced in the arable, lower organic matter soil, although only Cd showed a soil difference in the CaCl2 fraction. There were no metal toxic effects on plants and some evidence to suggest that they promoted growth. Uptake of each metal was higher in the larger plants of the grassland, higher organic matter soil. Inoculation with arbuscular mycorrhizal fungi increased root Cd and Zn concentrations. With the exception of Cd (roots) and Zn (shoots), higher inputs of sludge metals did not increase plant metals. Zn and Cu, but not Cd, concentrations were higher in roots than in shoots.     

The arbuscular mycorrhizal fungus Glomus mosseae can enhance arsenic tolerance in Medicago truncatula by increasing plant phosphorus status and restricting arsenate uptake

A pot experiment examined the biomass and As uptake of Medicago truncatula colonized by the arbuscular mycorrhizal (AM) fungus Glomus mosseae in low-P soil experimentally contaminated with different levels of arsenate. The biomass of G. mosseae external mycelium was unaffected by the highest addition level of As studied (200 mg kg⁻¹) but shoot and root biomass declined in both mycorrhizal and non-mycorrhizal plants, indicating that the AM fungus was more tolerant than M. truncatula to arsenate. Mycorrhizal inoculation increased shoot and root dry weights by enhancing host plant P nutrition and lowering shoot and root As concentrations compared with uninoculated plants. The AM fungus may have been highly tolerant to As and conferred enhanced tolerance to arsenate on the host plant by enhancing P nutrition and restricting root As uptake.     

Effects of mycorrhizae and fertilizer amendments on zinc tolerance of plants

In soils containing elevated levels of zinc, plant growth may be impaired because of Zn interference with P uptake by plants and because of detrimental effects of Zn toxicity itself. Because mycorrhizal fungi are known to improve uptake of plant P, the beneficial effects of mycorrhizal symbiosis on Zn tolerance of Andropogon gerardii Vitm. were assessed in soil amended with various levels of Zn and P. In the absence of P amendment, mycorrhizal fungi stimulated plant growth, but the degree of benefit depended on the inoculum source and the soil Zn level. Mycorrhizal fungi from a Zn contaminated site were more effective in increasing plant biomass at higher levels of Zn in the soil, whereas plant growth at lower levels of soil Zn was greater with mycorrhizal fungi from a non-contaminated site. Mycorrhizal fungus inoculation had no effect on shoot Zn concentration; however, inoculation significantly improved the plant P nutrition and therefore resulted in a high shoot P/Zn concentration ratio at all the soil Zn levels. To a certain extent, addition of P to the soil alleviated the Zn toxicity that had inhibited plant growth, but plant biomass tended to decrease with increasing soil Zn levels. Although P amendment improved P uptake, it also resulted in increased shoot Zn uptake.     

Effects of arbuscular mycorrhizal fungi on the root uptake and translocation of radiocaesium

Because mycorrhizal fungi are intimately associated with plant roots, their importance in radionuclide (RN) recycling and subsequent dispersion into the biosphere has received an increasing interest. Recently, the capacity of arbuscular mycorrhizal fungi to take up and translocate radiocaesium to their host was demonstrated. However, the relative contribution of these processes in comparison to the ones of roots remains unknown. Here, the respective contributions of the hyphae of a Glomus species and the transformed carrot (Daucus carota L.) roots on radiocaesium uptake and translocation were compared and quantified. We observed that radiocaesium uptake by hyphae was significantly lower as compared to that of the roots, while the opposite was noted for radiocaesium translocation/uptake ratio. We also observed that the intraradical fungal structures might induce a local accumulation of radiocaesium and concurrently reduce its translocation within mycorrhizal roots. We believe that intraradical fungal structures might induce the down-regulation of radiocaesium channels involved in the transport processes of radiocaesium towards the xylem.     

Arbuscular mycorrhizae enhance metal lead uptake and growth of host plants under a sand culture experiment

A sand culture experiment was conducted to investigate whether mycorrhizal colonization and mycorrhizal fungal vesicular numbers were influenced by metal lead, and whether mycorrhizae enhance host plants tolerance to metal lead. Metal lead was applied as Pb(NO3)2 in solution at three levels (0, 300 and 600 mg kg(-1) sand). Five mycorrhizal host plant species, Kummerowia striata (Thunb.) Schindl, Ixeris denticulate L., Lolium perenne L., Trifolium repens L. and Echinochloa crusgalli var. mitis were used to examine Pb-mycorrhizal interactions. The arbuscular mycorrhizal inoculum consisted of mixed spores of mycorrhizal fungal species directly isolated from orchard soil. Compared to the untreated control, both Pb concentrations reduced mycorrhizal colonization by 3.8-70.4%. Numbers of AM fungal vesicles increased by 13.2-51.5% in 300 mg Pb kg(-1) sand but decreased by 9.4-50.9% in 600 mg Pb kg(-1) sand. Mycorrhizae significantly enhanced Pb accumulation both in shoot by 10.2-85.5% and in root by 9.3-118.4%. Mycorrhizae also enhanced shoot biomass and shoot P concentration under both Pb concentrations. Root/shoot ratios of Pb concentration were higher in highly mycorrhizal plant species (K.striata, I. denticulate, and E. crusgalli var. mitis) than that in poorly mycorrhizal ones (L. perenne and T. repens,). Mycorrhizal inoculation increased the root/shoot ratio of Pb concentration of highly mycorrhizal plant species by 7.6-57.2% but did not affect the poorly mycorrhizal ones. In the treatments with 300 Pb mg kg(-1) sand, plant species with higher vesicular numbers tended to show higher root/shoot ratios of the Pb concentration. We suggest that under an elevated Pb condition, mycorrhizae could promote plant growth by increasing P uptake and mitigate Pb toxicity by sequestrating more Pb in roots.     

Potential contribution of arbuscular mycorrhiza to cadmium immobilisation in soil

The contribution of arbuscular mycorrhiza (AM) to immobilisation of Cd in substrate was studied in two experiments. In the first experiment, substrates prepared by cultivating tobacco, either non-mycorrhizal or inoculated with the AM fungus Glomus intraradices were enriched with a range of Cd concentrations, and Cd toxicity in the substrates was assessed using root growth tests with lettuce as a test plant. The tests revealed lower Cd toxicity in the mycorrhizal than in the non-mycorrhizal substrate, and the difference increased with increasing total Cd concentration in the substrates. In the second experiment, extraradical mycelium (ERM) of G. intraradices exposed in vivo to Cd was collected and analysed on Cd concentration. The ERM accumulated 10–20 times more Cd per unit of biomass than tobacco roots. While Cd concentrations were lower in the biomass of mycorrhizal plants than of non-mycorrhizal plants, Cd immobilisation by ERM did not affect the total Cd content in mycorrhizal tobacco.

It is concluded that mycorrhiza may decrease Cd toxicity to plants by enhancing Cd immobilisation in soil. The results therefore suggest a potential role of AM symbiosis in the phytostabilisation of contaminated soils, where high Cd availability inhibits plant growth.     

Influence of arbuscular mycorrhizal fungi on the growth and nutrient status of bermudagrass grown in alkaline bauxite processing residue

A nursery experiment was conducted to evaluate the potential role of arbuscular mycorrhizal (AM) fungi in encouraging the vegetation cover on bauxite residue (red mud) sites. An alkali tolerant bermudagrass (Cynodon dactylon) adapted to local conditions were grown in red mud with different amendments with and without AM fungi to assess mycorrhizal effects on plant growth, mineral nutrition, metal uptake and neutralization of bauxite residue. Inoculation of AM fungi significantly increased the plant growth, nutrient uptake and reduced Fe, Al accumulation in plant tissue and also improved the soil physico-chemical and biochemical properties. Gypsum and sludge amended treatments inoculated with AM fungi had maximum biomass, nutrient uptake and reduced accumulation of metals. The neutralization of red mud was significant in presence of AM fungi than control. The experiment provided evidence for the potential use of bermudagrass in combination with AM fungi for ecological restoration of bauxite residue sites.     

Role of microbial inoculation and chitosan in phytoextraction of Cu, Zn, Pb and Cd by Elsholtzia splendens--a field case

A field experiment was carried out to study the effect of microbial inoculation on heavy metal phytoextraction by Elsholtzia splendens and whether chitosan could have a synergistic effect with the microbial inocula. The microbial inocula consisted of a consortium of arbuscular mycorrhizal fungi and two Penicillium fungi. Three treatments were included: the control, inoculation with microbial inocula, and the inoculation combined with chitosan. Microbial inoculation increased plant biomass especially shoot dry weight, enhanced shoot Cu, Zn and Pb concentrations but did not affect Cd, leading to higher shoot Cu, Zn, Pb and Cd uptake. Compared with microbial inoculation alone, chitosan application did not affect plant growth but increased shoot Zn, Pb and Cd concentrations except Cu, which led to higher phytoextraction efficiencies and partitioning to shoots of Zn, Pb and Cd. These results indicated synergistic effects between microbial inocula and chitosan on Zn, Pb and Cd phytoextraction.     

Testing nickel tolerance of Sorghastrum nutans and its associated soil microbial community from serpentine and prairie soils

Ecotypes of Sorghastrum nutans from a naturally metalliferous serpentine grassland and the tallgrass prairie were assessed for Ni tolerance and their utility in remediation of Ni-polluted soils. Plants were inoculated with serpentine arbuscular mycorrhizal (AM) root inoculum or whole soil microbial communities, originating from either prairie or serpentine, to test their effects on plant performance in the presence of Ni. Serpentine plants had marginally higher Ni tolerance as indicated by higher survival. Ni reduced plant biomass and AM root colonization for both ecotypes. The serpentine AM fungi and whole microbial community treatments decreased plant biomass relative to uninoculated plants, while the prairie microbial community had no effect. Differences in how the soil communities affect plant performance were not reflected in patterns of root colonization by AM fungi. Thus, serpentine plants may be suited for reclamation of Ni-polluted soils, but AM fungi that occur on serpentine do not improve Ni tolerance.