Effects of arbuscular mycorrhizal inoculation on uranium and arsenic accumulation by Chinese brake fern (Pteris vittata L.) from a uranium mining-impacted soil

A glasshouse experiment was conducted to investigate U and As accumulation by Chinese brake fern, Pteris vittata L., in association with different arbuscular mycorrhizal fungi (AMF) from a U and As contaminated soil. The soil used contains 111 mg U kg(-1) and 106 mg As kg(-1). P. vittata L. was inoculated with each of three AMF, Glomus mosseae, Glomus caledonium and Glomus intraradices. Two harvests were made during plant growth (two and three months after transplanting). Mycorrhizal colonization depressed plant growth particularly at the early stages. TF (transfer factor) values for As from soil to fronds were higher than 1.0, while those for roots were much lower. Despite the growth depressions, AM colonization had no effect on tissue As concentrations. Conversely, TF values for U were much higher for roots than for fronds, indicating that only very small fraction of U was translocated to fronds (less than 2%), regardless of mycorrhizal colonization. Mycorrhizal colonization significantly increased root U concentrations at both harvests. Root colonization with G. mosseae or G. intraradices led to an increase in TF values for U from 7 (non-inoculation control) to 14 at the first harvest. The highest U concentration of 1574 mg kg(-1) was recorded in roots colonized by G. mosseae at the second harvest. The results suggested that P. vittata in combination with appropriate AMF would play very important roles in bioremediation of contaminated environments characterized by a multi-pollution.     

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.     

Pteris umbrosa R. Br. as an arsenic hyperaccumulator: accumulation, partitioning and comparison with the established As hyperaccumulator Pteris vittata

The capacity of the Australian native fern Pteris umbrosa to function as an arsenic (As) hyperaccumulator (shoot:soil As concentration >1) was examined by growing plants under glasshouse conditions in an inert medium supplemented with As. Arsenic preferentially accumulated in the fronds, a trait of a hyperaccumulator. The As concentration of fronds decreased with age, being particularly high in the croziers and low in the senesced fronds. Below ground, rhizomes accumulated more As than adventitious roots. Uptake from a range of solution concentrations followed Michaelis Menten kinetics up to a soil solution As concentration of 400mgl(-1). The K(m) for As uptake by roots suggested the operation of a low-affinity carrier. The predicted Nernst membrane potential indicated that uptake was against the electrochemical gradient of As. At 600mgl(-1), the rate of As uptake increased and phytotoxic effects were indicated by a significant decline in biomass. Arsenic uptake and translocation in P. umbrosa and Pteris vittata were similar at low exposure to As. At higher exposure, As uptake and translocation by P. vittata increased more than in P. umbrosa. The growth rate of both ferns was similar, whereas the biomass distribution was not, with P. vittata having a much larger root mass. This suggests that As uptake by P. umbrosa roots was very efficient and may be improved by stimulating root growth to enhance its potential.     

Effects of arsenic on concentration and distribution of nutrients in the fronds of the arsenic hyperaccumulator Pteris vittata L

Pteris vittata was the first terrestrial plant known to hyperaccumulate arsenic (As). However, it is unclear how As hyperaccumulation influences nutrient uptake by this plant. P. vittata fern was grown in soil spiked with 0-500 mg As kg(-1) in the greenhouse for 24 weeks. The concentrations of essential macro- (P, K, Ca, and Mg) and micro- (Fe, Mn, Cu, Zn, B and Mo) elements in the fronds of different age were examined. Both macro- and micronutrients in the fronds were found to be within the normal concentration ranges for non-hyperaccumulators. However, As hyperaccumulation did influence the elemental distribution among fronds of different age of P. vittata. Arsenic-induced P and K enhancements in the fronds contributed to the As-induced growth stimulation at low As levels. The frond P/As molar ratios of 1.0 can be used as the threshold value for normal growth of P. vittata. Potassium may function as a counter-cation for As in the fronds as shown by the As-induced K increases in the fronds. The present findings not only demonstrate that P. vittata has the ability to maintain adequate concentrations of essential nutrients while hyperaccumulating As from the soil, but also have implications for soil management (fertilization in particular) of P. vittata in As phytoextraction practice.     

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.     

Uptake and accumulation of arsenic by 11 Pteris taxa from southern China

A field survey was conducted at a deserted arsenic (As) mine in Guangxi Province, China to explore new potential As hyperaccumulators. In addition, young plants of 11 Pteris taxa were grown in glasshouse conditions for 12 weeks on As-amended soils with 0, 50 and 200 mg As kg(-1). Results of the field survey showed that the fern Pteris fauriei accumulated over 1000 mg As kg(-1) in its fronds. Of the 11 Pteris taxa, Pteris aspericaulis, Pteris cretica var. nervosa, P. fauriei, Pteris multifida, P. multifida f. serrulata, and Pteris oshimensis were all found to hyperaccumulate As in addition to P. cretica 'Albo-Lineata' and Pteris vittata (already reported as As hyperaccumulators). However, Pteris ensiformis, Pteris semipinnata and Pteris setuloso-costulata showed no evidence of As hyperaccumulation. Results also revealed a constitutive property of As hyperaccumulation in different populations of P. cretica var. nervosa, P. multifida, P. oshimensis and P. vittata.     

Effects of the arbuscular mycorrhizal fungus Glomus mosseae on growth and metal uptake by four plant species in copper mine tailings

A greenhouse experiment was conducted to evaluate the potential role of arbuscular mycorrhizal fungi (AMF) in encouraging revegetation of copper (Cu) mine tailings. Two native plant species, Coreopsis drummondii and Pteris vittata, together with a turf grass, Lolium perenne and a leguminous plant Trifolium repens associated with and without AMF Glomus mosseae were grown in Cu mine tailings to assess mycorrhizal effects on plant growth, mineral nutrition and metal uptake. Results indicated that symbiotic associations were successfully established between G. mosseae and all plants tested, and mycorrhizal colonization markedly increased plant dry matter yield except for L. perenne. The beneficial impacts of mycorrhizal colonization on plant growth could be largely explained by both improved P nutrition and decreased shoot Cu, As and Cd concentrations. The experiment provided evidence for the potential use of local plant species in combination with AMF for ecological restoration of metalliferous mine tailings.     

Arsenic speciation, and arsenic and phosphate distribution in arsenic hyperaccumulator Pteris vittata L. and non-hyperaccumulator Pteris ensiformis L

This study examined the roles of arsenic translocation and reduction, and P distribution in arsenic detoxification of Pteris vittata L. (Chinese Brake fern), an arsenic hyperaccumulator and Pteris ensiformis L. (Slender Brake fern), a non-arsenic hyperaccumulator. After growing in 20% Hoagland solution containing 0, 133 or 267 microM of sodium arsenate for 1, 5 or 10 d, the plants were separated into fronds, rhizomes, and roots. They were analyzed for biomass, and concentrations of arsenate (AsV), arsenite (AsIII) and phosphorus. Arsenic in the fronds of P. vittata was up to 20 times greater than that of P. ensiformis, yet showing no toxicity symptoms as did in P. ensiformis. While arsenic was concentrated primarily in the fronds of P. vittata as arsenite it was mainly concentrated in the roots of P. ensiformis as arsenate. Arsenic reduction in the plants took longer than 1-d. P. vittata maintained greater P in the roots while P. ensiformis in the fronds. The high arsenic tolerance of the hyperaccumulator P. vittata may be attributed to its ability to effectively reduce arsenate to arsenite in the fronds, translocate arsenic from the roots to fronds, and maintain a greater ratio of P/As in the roots.     

Arsenic hyperaccumulation by Pteris vittata from arsenic contaminated soils and the effect of liming and phosphate fertilisation

Pot experiments were carried out to investigate the potential of phytoremediation with the arsenic hyperaccumulator Pteris vittata in a range of soils contaminated with As and other heavy metals, and the influence of phosphate and lime additions on As hyperaccumulation by P. vittata. The fern was grown in 5 soils collected from Cornwall (England) containing 67-4550 mg As kg(-1) and different levels of metals. All soils showed a similar distribution pattern of As in different fractions in a sequential extraction, with more than 60% of the total As being associated with the fraction thought to represent amorphous and poorly-crystalline hydrous oxides of Fe and Al. The concentration of As in the fronds ranged from 84 to 3600 mg kg(-1), with 0.9-3.1% of the total soil As being taken up by P. vittata. In one soil which contained 5500 mg Cu kg(-1) and 1242 mg Zn kg(-1), P. vittata suffered from phytotoxicity and accumulated little As (0.002% of total). In a separate experiment, neither phosphate addition (50mg P kg(-1) soil) nor liming (4.6 g CaCO3 kg(-1) soil) was found to affect the As concentration in the fronds of P. vittata, even though phosphate addition increased the As concentration in the soil pore water. Between 4 and 7% of the total soil As was taken up by P. vittata in 4 cuttings in this experiment. The results indicate that P. vittata can hyperaccumulate As from naturally contaminated soils, but may be suitable for phytoremediation only in the moderately contaminated soils.     

Antioxidative responses to arsenic in the arsenic-hyperaccumulator Chinese brake fern (Pteris vittata L.)

This study measured antioxidative responses of Chinese brake fern (Pteris vittata L.) upon exposure to arsenic (As) of different concentrations. Chinese brake fern was grown in an artificially-contaminated soil containing 0 to 200 mg As kg(-1) (Na2HAsO4) for 12 weeks in a greenhouse. Soil As concentrations at < or =20 mg kg(-1) enhanced plant growth, with 12-71% biomass increase compared to the control. Such beneficial effects were not observed at >20 mg As kg(-1). Plant As concentrations increased with soil As concentrations, with more As being accumulated in the fronds (aboveground biomass) than in the roots and with maximum frond As concentration being 4675 mg kg(-1). Arsenic uptake by Chinese brake enhanced uptake of nutrient elements K, P, Fe, Mn, and Zn except Ca and Mg, whose concentrations mostly decreased. The contents of non-enzymatic antioxidants (glutathione, acid-soluble thiol) followed similar trends as plant As concentrations, increasing with soil As concentrations, with greater contents in the fronds than in the roots especially when exposed to high As concentrations (>50 mg kg(-1)). The activities of enzymatic antioxidants (superoxide dismutase, catalase, ascorbate peroxidase, guaiacol peroxidase) in Chinese brake followed the same trends as plant biomass, increasing with soil As up to 20 mg kg(-1) and then decreased. The results indicated though both enzymatic and non-enzymatic antioxidants played significant roles in As detoxification and hyperaccumulation in Chinese brake, the former is more important at low As exposure (< or =20 mg kg(-1)), whereas the latter is more critical at high As exposure (50-200 mg kg(-1)).     

Factors influencing arsenic accumulation by Pteris vittata: a comparative field study at two sites

This study compared the factors influencing arsenic (As) accumulation by Pteris vittata at two sites, one containing As along with Au mineralization and the other containing Hg/Tl mineralization. The soils above these two sites contained high As concentrations (26.8-2955 mg kg(-1)). Although the As concentration, pH, soil cation exchange capacity and plant biomass differed significantly between the two sites, no differences were observed in the As concentrations in the fronds and roots, or the translocation factors, of P. vittata, suggesting that this species has consistent As hyperaccumulation properties in the field. The As concentration in the fronds was positively related to phosphorus (P) and potassium (K), but negatively related to calcium (Ca), at one site. This suggested that P, K and Ca influenced As accumulation by P. vittata in the field.     

Phytoremediation of an arsenic-contaminated site using Pteris vittata L. and Pityrogramma calomelanos var. austroamericana: a long-term study

This field study investigated the phytoremediation potential of two arsenic (As) hyperaccumulating fern species, Pityrogramma calomelanos var. austroamericana and Pteris vittata over 27-month duration at a disused As-contaminated cattle-dip site located at Wollongbar, NSW, Australia. Ferns planted in January 2009 were harvested following 10, 22 and 27 months of growth. A detailed soil sampling was undertaken in June 2009 (initial, n?=?42 per plot) and limited sampling in April 2011 (after 27 months, n?=?15 per plot) to measure total and phosphate-extractable As concentrations in soil at 0?-?20-, 20?-?40- and 40?-?60-cm depths. The choice of the limited number of samples was considered sufficient to estimate the changes in soil As concentration following phytoremediation based on a geostatistical model. The average frond dry biomass, As concentration and As uptake were significantly (P??0.05), respectively, by P. vittata. Our results show that phytoremediation time based on observed changes in soil As based on limited sampling is not reliable; hence, it is recommended that the frond As uptake should be considered in order to evaluate the phytoremediation efficiency of the two fern species at the experimental site. Using As uptake of the two fern species, we estimate that with P. calomelanos var. austroamericana it would take 55?-?125 years to decrease mean total As content below the ecological investigation level (20 mg kg(-1)) in the surface and subsurface soils, whereas with P. vittata 143?-?412 years would be required to achieve this target.     

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.     

A modified glass bead compartment cultivation system for studies on nutrient and trace metal uptake by arbuscular mycorrhiza

A modified glass bead compartment cultivation system is described in which glass beads continue to be used in the hyphal compartment but are replaced by coarse river sand in the compartments for host plant roots and mycorrhizal hyphae. Arbuscular mycorrhizal (AM) associations were established using two host plant species, maize (Zea mays L.) and red clover (Trifolium pratense L.) and two AM fungi, Glomus mosseae and G. versiforme. When the standard and modified cultivation systems were compared, the new method yielded much more fungal tissue in the hyphal compartment. Using G. versiforme as the fungal symbiont, up to 30 mg of fungal dry matter (DM) was recovered from the hyphal compartment of mycorrhizal maize and about 6 mg from red clover. Multi-element analysis was conducted on samples of host plant roots and shoots and on harvested fungal biomass. Concentrations of P, Cu and Zn were much higher in the fungal biomass than in the roots or shoots of the host plants but fungal concentrations of K, Ca, Mg, Fe and Mn were similar to or lower than those in the plants. There were also significant differences in nutrient concentrations between the two AM fungi and these may be related to differences in their proportions of extraradical mycelium to spores. The high affinity of the fungal mycelium for Zn was very striking and is discussed in relation to the potential use of arbuscular mycorrhiza in the phytoremediation of Zn-polluted soils.     

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.     

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.     

Enhanced phytoremediation of arsenic contaminated land

In an attempt to clean up arsenic (As) contaminated soil, the effects of phosphorus (P) fertilizer and rhizosphere microbes on arsenic accumulation by the silverback fern, Pityrogramma calomelanos, were investigated in both greenhouse and field experiments. Field experiments were conducted in Ron Phibun District, an As-contaminated area in Thailand. Soil (136-269 microg As g(-1)) was collected there and used in the greenhouse experiment. Rhizosphere microbes (bacteria and fungi) were isolated from roots of P. calomelanos growing in Ron Phibun District. The results showed that P-fertilizer significantly increased plant biomass and As accumulation of the experimental P. calomelanos. Rhizobacteria increased significantly the biomass and As content of the test plants. Thus, P-fertilizer and rhizosphere bacteria enhanced As-phytoextraction. In contrast, rhizofungi reduced significantly As concentration in plants but increased plant biomass. Therefore, rhizosphere fungi exerted their effects on phytostabilization.     

Arbuscular mycorrhizal fungi differentially affect the response to high zinc concentrations of two registered poplar clones

The effects of a high concentration of zinc on two registered clones of poplar (Populus alba Villafranca and Populus nigra Jean Pourtet), inoculated or not with two arbuscular mycorrhizal fungi (Glomus mosseae or Glomus intraradices) before transplanting them into polluted soil, were investigated, with special regard to the extent of root colonization by the fungi, plant growth, metal accumulation in the different plant organs, and leaf polyamine concentration. Zinc accumulation was lower in Jean Pourtet than in Villafranca poplars, and it was mainly translocated to the leaves; the metal inhibited mycorrhizal colonization, compromised plant growth, and, in Villafranca, altered the putrescine profile in the leaves. Most of these effects were reversed or reduced in plants pre-inoculated with G. mosseae. Results indicate that poplars are suitable for phytoremediation purposes, confirming that mycorrhizal fungi can be useful for phytoremediation, and underscore the importance of appropriate combinations of plant genotypes and fungal symbionts.     

Saprobe fungi decreased the sensitivity to the toxic effect of dry olive mill residue on arbuscular mycorrhizal plants

We studied the influence of olive mill dry residue (DOR) treated with saprobe fungi on growth of tomato and alfalfa colonized by Glomus deserticola. The application of 25g kg(-1) of dry DOR to soil decreased the shoot and root dry weight of tomato and alfalfa. Plants were more sensitive to the toxicity of DOR when colonized with the arbuscular mycorrhizal (AM) fungi. The sensitivity of both plants to the toxicity of DOR differed according to whether they were colonized by G. deserticola or by indigenous AM fungi. The phytotoxicity of DOR towards tomato and alfalfa was decreased by incubation the residue before planting with saprobe fungi for 20wk. The beneficial effects of AM fungi on plant growth added with DOR incubated with saprobe fungi depend of the type of the plant and AM fungi. The contribution of AM fungi to the beneficial effect of DOR incubated with saprobe fungi varied according to the type of the plant and AM fungi. G. deserticola increased the shoot and root dry weight of plants when they were grown in the presence of DOR incubated with saprobe fungi for 20wk. The beneficial effect of saprobe fungi on the dry weight and the level of AM mycorrhization of plants seem to be related to the decrease caused by these fungi in the phenol concentration in DOR. However, the toxicity of DOR due to substances other than phenols can not be ignored. The use of certain saprobe and AM fungi allows the possibility of using DOR as an organic fertilizer.     

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.