Influence of fungi on growth and survival of Onychiurus armatus (Collembola) in a metal polluted soil

Summary The influence of food quantity and quality on growth and survival of Onychiurus armatus (Tullb.) in metal polluted environments has been investigated in laboratory experiments. The Collembola was reared on five species of fungi isolated from a metal polluted soil close to a brass mill in SE Sweden.Survival of O. armatus was improved when fungal biomass was continuously added in a polluted mor (1,300 ppm Zn and 200 ppm Cu), and when specimens were fed metal polluted fungi for 1, 3 and 7 days a week, only those that were starved had increased mortality. Allometric growth, on the other hand, was significantly reduced when Collembola was given surplus of metal polluted fungi, whereas growth losses caused by metals were offset by protein rich food. Hence, sufficient food quantities alone could overcome mortality losses but not growth retardation in a metal polluted environment.Feeding preference of O. armatus was not determined by the protein content of the fungi although this was beneficial for growth. Metals changed the relative palatability of fungal species, but one of the metal tolerant species, Paecilomyces farinosus, which was also protein rich, remained reasonably attractive for O. armatus also when it was metal polluted. The mechanisms by which growth and survival of O. armatus were promoted by a combination of protein and Zn/Cu rich fungi seemed to be crucial in understanding the fate of a population of this species in a metal polluted soil.     

Effects of Pisolithus tinctorius and Cenococcum geophilum inoculation on pine in copper-contaminated soil to enhance phytoremediation

We used Pisolithus tinctorius and Cenococcum geophilum to determine the copper (Cu) resistance of ectomycorrhizal (ECM) fungi and their potential for improving phytoremediation of Cu-contaminated soil by Chinese red pine (Pinus tabulaeformis). The results showed that nutrient accumulation in C. geophilum mycelium was significantly lower under higher Cu concentrations in the soil, which was not observed in P. tinctorius. Meanwhile, P. tinctorius exhibited greater Cu tolerance than C. geophilum. Inoculation with ECM fungi significantly improved the growth of pine shoots planted in polluted soil in pot experiments (p < 0.01). The total accumulated Cu in pine seedlings planted in Cu-contaminated soil increased by 72.8% and 113.3% when inoculated with P. tinctorius and C. geophilum, respectively, indicating that ECM fungi may help their host to phytoextract heavy metals. Furthermore, the majority of the total absorbed metals remained in the roots, confirming the ability of ECM fungi to promote heavy metal phytostabilization. There were no differences between the effects of the two fungi in helping the host stabilize and absorb Cu, even though they have different Cu tolerances. Inoculation with ECM fungi can benefit plant establishment in polluted environments and assist plants with phytoremediating heavy-metal-contaminated soils.     

Bioremediation of Heavy Metals in Liquid Media Through Fungi Isolated from Contaminated Sources

Wastewater particularly from electroplating, paint, leather, metal and tanning industries contain enormous amount of heavy metals. Microorganisms including fungi have been reported to exclude heavy metals from wastewater through bioaccumulation and biosorption at low cost and in eco-friendly way. An attempt was, therefore, made to isolate fungi from sites contaminated with heavy metals for higher tolerance and removal of heavy metals from wastewater. Seventy-six fungal isolates tolerant to heavy metals like Pb, Cd, Cr and Ni were isolated from sewage, sludge and industrial effluents containing heavy metals. Four fungi (Phanerochaete chrysosporium, Aspegillus awamori, Aspergillus flavus, Trichoderma viride) also were included in this study. The majority of the fungal isolates were able to tolerate up to 400 ppm concentration of Pb, Cd, Cr and Ni. The most heavy metal tolerant fungi were studied for removal of heavy metals from liquid media at 50 ppm concentration. Results indicated removal of substantial amount of heavy metals by some of the fungi. With respect to Pb, Cd, Cr and Ni, maximum uptake of 59.67, 16.25, 0.55, and 0.55 mg/g was observed by fungi Pb3 (Aspergillus terreus), Trichoderma viride, Cr8 (Trichoderma longibrachiatum), and isolate Ni27 (A. niger) respectively. This indicated the potential of these fungi as biosorbent for removal of heavy metals from wastewater and industrial effluents containing higher concentration of heavy metals.     

Mycorrhizal and Endophytic Fungal Colonization in Arrhenatherum elatius L. Roots According to the Soil Contamination in Heavy Metals

The aim of this work was to jointly study non-mycorrhizal (dark septate fungi) and mycorrhizal (arbuscular mycorrhizae) colonization along a large range of heavy metal pollution in soil in order to determine the effective contribution of each type of endophytes in relation to heavy metal uptake and tolerance. Hence, eight sites were chosen in the mining area of northern France with respect both to a large range of heavy metal contamination (Cd, Pb, Zn) and monospecific colonization by Arrhenatherum elatius. Root colonization with both arbuscular mycorrhizae (AM) and dark septate fungi (DSF) as well as spore density in rhizospheric soil were estimated in relation to soil characteristics. Mycorrhizal infestation (hyphae, arbuscules and vesicles) was adversely affected by soil pollution almost to exclusion. The intensity of colonization with DSF was very low in presence of AM in non-contaminated soils but higher in polluted soils. The effect of the fungal colonization on the heavy metal tolerance of Arrhenatherum elatius is discussed.     

Pollution-related changes in diets of two insectivorous passerines

Insectivorous birds living in polluted areas are not only exposed to pollutants but they may also be affected by changes in their invertebrate food. The populations of many invertebrate species are affected by environmental pollution and such changes may lead to differences in the diet of insectivorous birds. We examined nestling food quality (invertebrate composition and heavy metal levels) and breeding performance of two cavity-nesting passerines, the Great tit, Parus major, and the pied flycatcher, Ficedula hypoleuca, in an area with long-term heavy metal pollution by a copper smelter. There were no differences in feeding frequencies or the amount of food that parents provided to their nestlings between polluted and unpolluted sites, but food quality in a polluted area differed from that of the control area in both bird species. P. major took more beetles and variable “flying insects” and less caterpillars (of smaller size) and moths in the polluted area as compared to the unpolluted one. F. hypoleuca ate more beetles and larvae and less moths and spiders in the polluted area. Breeding success of P. major was better when the nestling diet contained a large proportion of caterpillars and the relationship was especially strong in the polluted area. On the contrary, F. hypoleuca broods succeeded equally well with variable diets. Our data suggest that a more opportunistic forager, F. hypoleuca, is less vulnerable to a changing invertebrate composition caused by human environmental impacts than a caterpillar specialist, P. major. In a heavy metal polluted area, F. hypoleuca seems to be more sensitive to a decreased amount of Ca rich food items (e.g. snails) while P. major suffers especially from the lack of carotenoid rich caterpillars. Our results emphasize the importance of secondary environmental changes, like food quality, in addition to direct impacts of pollutants.     

Heavy metal tolerance potential of Aspergillus strains isolated from mining sites

Increased heavy metal pollution generated through anthropogenic activities into the environment has necessitated the need for eco-friendly remediation strategies such as mycoremediation. With a view to prospecting for fungi with heavy metal remediation potentials, the tolerance of five Aspergillus species isolated from soils of three active gold and gemstone mining sites in southwestern Nigeria to varied heavy metal concentrations was investigated. Isolated Aspergillus strains were identified based on the internal transcribed spacer 1 and 2 (ITS 1 and ITS 2) regions. Growth of Aspergillus strains were challenged with a range of varied concentrations of heavy metals: cadmium (Cd) (0–100), copper (Cu) (0–1000), lead (Pb) (0–400), arsenic (As) (0–500), and iron (Fe) (0–800) concentrations (ppm) incorporated into Malt Extract Agar (MEA) in triplicates. Mycelial radial growths were recorded at intervals of 3 days during a 13-day incubation period. Aspergillus strains were identified as A. tubingensis, A. fumigatus, A. terreus, A. nidulans, and A. nomius. A. tubingensis tolerated Cd, Cu, Pb, As, and Fe at all test concentrations (100–1000 ppm), showing no significant (p > .05) difference compared with the control. Similarly, A. nomius tolerated all concentrations of Cu, Pb, As, and Fe and only 50 ppm Cd concentrations. A. nidulans, A. terreus, and A. fumigatus, on the other hand, tolerated all concentrations of Cu, Pb, and Fe with no statistical significance (p > .05) difference from the controls. Overall, the Aspergillus species showed tolerance to heavy metal concentrations above permissible limits for contaminated soils globally. These heavy metal tolerance traits exhibited by the Aspergillus isolates may suggest that they are potential candidates for bioremediation of heavy metal–polluted environments.     

The parallel expression of metal tolerance in pollen and sporophytes of Silene dioica (L.) Clairv., S. alba (mill.) krause and Mimulus guttatus DC

Summary The purpose of this paper was to determine if heavy metal tolerance was expressed in pollen and if its expression was correlated with the tolerance of the pollen source. Clones of Silene dioica, tolerant to zinc, closely related but nontolerant S. alba and clones of Mimulus guttatus tolerant and sensitive to copper were grown in the greenhouse in either standard potting soil or nutrient culture. Pollen was collected shortly after dehiscence, hydrated, and tested over a broad range of metal concentrations. The tolerance of the pollen source was determined by comparing root growth in solutions with and without heavy metals. In both Silene species and M. guttatus, the tolerance of the parental clone was expressed in its pollen. Pollen from tolerant individuals was able to germinate and grow at concentrations of metals which markedly inhibited pollen from nontolerants.     

Relationships between pasture legumes, rhizobacteria and nodule bacteria in heavy metal polluted mine waste of SW Sardinia

Local populations of the pasture legumes Astragalus hamosus, Lotus edulis, Lotus ornithopodioides, Medicago ciliaris and Scorpiurus muricatus from heavy metal polluted and unpolluted sites in Sardinia were compared for tolerance to Zn, Cd and Pb in hydroponics. Tolerance of plants to heavy metals varied significantly depending on the species, origin of the population and metal. The species L. edulis, L. ornithopodioides and M. ciliaris possessed higher metal tolerance and were used in a pot experiment with Zn, Cd and Pb polluted mine waste. Seeds were inoculated with the metal tolerant plant growth-promoting rhizobacterium Variovorax paradoxus 5C-2 or/and with the corresponding symbiotic nodule bacteria containing the enzyme 1-aminocyclopropane-1-carboxylate deaminase. Co-inoculation with the bacteria had synergistic and additive effects on nodule number, root growth and uptake of elements (N, P, Ca, Mg, Na, Mn, Zn and Pb) in shoots of L. edulis and L. ornithopodioides. Shoot biomass and uptake of K, Fe and Cd was increased by a combined inoculation of L. edulis. The ratio between shoot and root contents of Pb in L. ornithopodioides was above 1, suggesting a characteristic trait of hyperaccumulating species. The results suggest that the development of metal tolerant and efficient plant-bacteria systems might be useful for phytostabilization and revegetation of mine wastes.     

Natural variation of drought response in Brachypodium distachyon

Brachypodium distachyon (Brachypodium) is a temperate wild grass species and is a powerful model system for studying grain, energy, forage and turf grasses. Exploring the natural variation in the drought response of Brachypodium provides an important basis for dissecting the genetic network of drought tolerance. Two experiments were conducted in a greenhouse to assess the drought tolerance of 57 natural populations of Brachypodium. Principle component analysis revealed that reductions in chlorophyll fluorescence (Fv/Fm) and leaf water content (LWC) under drought stress explained most of the phenotypic variation, which was used to classify the tolerant and susceptible accessions. Four groups of accessions differing in drought tolerance were identified, with 3 tolerant, 16 moderately tolerant, 32 susceptible and 6 most susceptible accessions. The tolerant group had little leaf wilting and fewer reductions in Fv/Fm and LWC, while the most susceptible groups showed severe leaf wilting and more reductions in Fv/Fm and LWC. Drought stress increased total water soluble sugar (WSS) concentration, but no differences in the increased WSS were found among different groups of accessions. The large phenotypic variation of Brachypodium in response to drought stress can be used to identify genes and alleles important for the complex trait of drought tolerance.     

Effect of water stress on in vitro mycelium cultures of two mycorrhizal desert truffles

The ability of two species of desert truffle, Terfezia claveryi strain TcS2 and Picoa lefebvrei strain OL2, to tolerate water stress in pure culture has been investigated. Both T. claveryi and P. lefebvrei strains exhibited a mycelium growth pattern characteristic of drought tolerant species. However, they were only tolerant to moderate water stress, below −1.07 MPa, with the P. lefebvrei isolate being slightly more drought tolerant than the T. claveryi isolate. The increased alkaline phosphatase (ALP) activity observed in both fungi at moderate water stress with respect to the control indicated the functional adaptation of these mycelia to these drought conditions. ALP activity can be used as an indicator of the metabolic activity of these fungi. Slight water stress (−0.45 MPa) could improve mycelial inoculum production of these desert truffles. Moreover, P. lefebvrei could be a good candidate for further desert truffle mycorrhizal plant cultivation programmes in semiarid Mediterranean areas.     

Differences in xylem and leaf hydraulic traits explain differences in drought tolerance among mature Amazon rainforest trees

Considerable uncertainty surrounds the impacts of anthropogenic climate change on the composition and structure of Amazon forests. Building upon results from two large‐scale ecosystem drought experiments in the eastern Brazilian Amazon that observed increases in mortality rates among some tree species but not others, in this study we investigate the physiological traits underpinning these differential demographic responses. Xylem pressure at 50% conductivity (xylem‐P₅₀), leaf turgor loss point (TLP), cellular osmotic potential (π_o), and cellular bulk modulus of elasticity (ε), all traits mechanistically linked to drought tolerance, were measured on upper canopy branches and leaves of mature trees from selected species growing at the two drought experiment sites. Each species was placed a priori into one of four plant functional type (PFT) categories: drought‐tolerant versus drought‐intolerant based on observed mortality rates, and subdivided into early‐ versus late‐successional based on wood density. We tested the hypotheses that the measured traits would be significantly different between the four PFTs and that they would be spatially conserved across the two experimental sites. Xylem‐P₅₀, TLP, and π_o, but not ε, occurred at significantly higher water potentials for the drought‐intolerant PFT compared to the drought‐tolerant PFT; however, there were no significant differences between the early‐ and late‐successional PFTs. These results suggest that these three traits are important for determining drought tolerance, and are largely independent of wood density—a trait commonly associated with successional status. Differences in these physiological traits that occurred between the drought‐tolerant and drought‐intolerant PFTs were conserved between the two research sites, even though they had different soil types and dry‐season lengths. This more detailed understanding of how xylem and leaf hydraulic traits vary between co‐occuring drought‐tolerant and drought‐intolerant tropical tree species promises to facilitate a much‐needed improvement in the representation of plant hydraulics within terrestrial ecosystem and biosphere models, which will enhance our ability to make robust predictions of how future changes in climate will affect tropical forests.     

Virulence determines beneficial trade‐offs in the response of virus‐infected plants to drought via induction of salicylic acid

It has been hypothesized that plants can get beneficial trade‐offs from viral infections when grown under drought conditions. However, experimental support for a positive correlation between virus‐induced drought tolerance and increased host fitness is scarce. We investigated whether increased virulence exhibited by the synergistic interaction involving Potato virus X (PVX) and Plum pox virus (PPV) improves tolerance to drought and host fitness in Nicotiana benthamiana and Arabidopsis thaliana. Infection by the pair PPV/PVX and by PPV expressing the virulence protein P25 of PVX conferred an enhanced drought‐tolerant phenotype compared with single infections with either PPV or PVX. Decreased transpiration rates in virus‐infected plants were correlated with drought tolerance in N. benthamiana but not in Arabidopsis. Metabolite and hormonal profiles of Arabidopsis plants infected with the different viruses showed a range of changes that positively correlated with a greater impact on drought tolerance. Virus infection enhanced drought tolerance in both species by increasing salicylic acid accumulation in an abscisic acid‐independent manner. Viable offspring derived from Arabidopsis plants infected with PPV increased relative to non‐infected plants, when exposed to drought. By contrast, the detrimental effect caused by the more virulent viruses overcame potential benefits associated with increased drought tolerance on host fitness.     

AM真菌对重金属污染土壤生物修复的应用与机理

土壤重金属污染威胁人类健康和整个生态系统,而高效、低耗、安全的生物修复技术显示出了极大的应用潜力,特别是利用植物-微生物共生体增强生物修复效应的应用.丛枝菌根(Arbuscular Mycorrhizae,AM)真菌是一类广泛分布于土壤生态系统中的有益微生物,能与90％以上的陆生高等植物形成共生体.研究发现,AM真菌能够增强宿主植物对土壤中重金属胁迫的耐受性.当前,利用AM真菌开展重金属污染土壤的生物修复已经引起环境学家和生态学家的广泛关注.基于此,围绕AM真菌在重金属污染土壤生物修复作用中的最新研究进展,从物理性防御体系的形成、对植物生理代谢的调控、生化拮抗物质的产生、基因表达的调控等角度探究AM真菌在重金属污染土壤生物修复中的作用机理,以期为利用AM真菌开展重金属污染的生物修复提供理论依据,并对本领域未来的发展和应用前景进行了展望.     

Proteomic analysis as a tool for investigating arsenic stress in Pteris vittata roots colonized or not by arbuscular mycorrhizal symbiosis

Pteris vittata can tolerate very high soil arsenic concentration and rapidly accumulates the metalloid in its fronds. However, its tolerance to arsenic has not been completely explored. Arbuscular mycorrhizal (AM) fungi colonize the root of most terrestrial plants, including ferns. Mycorrhizae are known to affect plant responses in many ways: improving plant nutrition, promoting plant tolerance or resistance to pathogens, drought, salinity and heavy metal stresses. It has been observed that plants growing on arsenic polluted soils are usually mycorrhizal and that AM fungi enhance arsenic tolerance in a number of plant species. The aim of the present work was to study the effects of the AM fungus Glomus mosseae on P. vittata plants treated with arsenic using a proteomic approach. Image analysis showed that 37 spots were differently affected (21 identified). Arsenic treatment affected the expression of 14 spots (12 up-regulated and 2 down-regulated), while in presence of G. mosseae modulated 3 spots (1 up-regulated and 2 down-regulated). G. mosseae, in absence of arsenic, modulated 17 spots (13 up-regulated and 4 down-regulated). Arsenic stress was observed even in an arsenic tolerant plant as P. vittata and a protective effect of AM symbiosis toward arsenic stress was observed.     

Laboratory-and Field-Phenotyping for Drought Stress Tolerance and Diversity Study in Lentil (<i>Lens culinaris</i> Medik.)

Drought susceptibility and low genetic variability are the major constraints of lentil (Lens culinaris Medik.) production worldwide. Development of an efficient pre-field drought phenotyping technique and identification of diversified drought tolerant lentil genotype(s) are therefore vital and necessary. Two separate experiments were conducted using thirty diverse lentil genotypes to isolate drought tolerant genotype(s) as well as to assess their diversity. In both of the experiments, significant (p ≤ 0.01) variation in genotype (G), treatment (T) and G X T was observed for most of the studied traits. In experiment I, genotypes were examined for drought tolerance at the seedlings stage under hydroponic conditions by assessing root and shoot traits. Among the 30 genotypes studied, BM-1247, BM-1227 and BM-502 were selected as highly tolerant to drought stress as they showed maximum seedling survivability and minimum reduction in growth parameters under drought stress. In experiment II, the genotypes were assayed for diversity and drought stress tolerance based on morphological traits grown under field condition. Drought stress caused a substantial reduction in yield attributing traits, however, the genotypes BM-1247, BM-981, BM-1227 and BM-502 were categorized as drought tolerant genotypes with less than 20% yield reduction. The field screening result of drought stress tolerance was coincided well with the results of laboratory screening. Genetic divergence study reflected the presence of considerable diversity among the genotypes. Considering laboratory and field screening results, the genotypes, BM-1247, BM-1227, BM-981 and BM- 502 were selected as the best drought tolerant genotypes. This information can be exploited for further breeding in developing drought tolerance in lentil.     

Cadmium removal capacities of filamentous soil fungi isolated from industrially polluted sediments,in La Plata (Argentina)

Aspergillus terreus, Cladosporium cladosporioides, Fusarium oxysporum, Gliocladium roseum, Penicillium spp., Talaromyces helicus and Trichoderma koningii were isolated from heavily polluted streams near an industrial area in La Plata, Argentina. The fungi were obtained from sediments with 0.25–0.50 mg Cd/l and they were isolated in cadmium-basal medium. They were then cultivated to evaluate their Cd detoxification abilities. The biomass developed in static assays represented 5–53% of the yield of stirred cultures, for the different fungal species, although the cadmium absorption were similar in both cases. These soil fungi represented 50% of the total isolates and their mycelial growth was conspicuous in these polluted sediments. Although bacteria have been mentioned as active microorganisms against heavy metals, the mycelial fungi were able to develop a significantly higher mass to sequestrate more metals. Thus, they could be used in remediation biotechnology to improve the Cd detoxification of chronically contaminated habitats.     

Studies of tolerance to heavy metals in the flora of the rivers Ystwyth and Clarach, Wales

A survey of the river Ystwyth demonstrated that it was polluted with metals especially below the areas of old lead mines. Hormidium spp. Kütz in particular Hormidium rivulare Kütz were the most tolerant filamentous green algae present. Scapania undulata (L.) Dum. was a tolerant bryophyte on the Ystwyth and Clarach rivers and was found alone in polluted sites, but was less frequent in cleaner areas. Metal extracts of this bryophyte mirrored the variation in metal concentrations of the environment to some extent. In general, lower levels of Fe, Pb and Mn were found in Scapania as compared with the less tolerant Fontinalis squamosa Hëdw. When the latter was transplanted to polluted sites an increase in its Pb, Cu, Zn and Mn content occurred within 6 weeks and the plant started to die and decay after 18 weeks. Scapania survived when it was transplanted from its natural polluted habitat to a less polluted area, and there was no marked change in its metal content. Zn⁶⁵ tracing experiments, in the laboratory, demonstrated that both plants had similar uptake patterns for the metal, but that the amount taken up was slightly higher in the less tolerant Fontinalis. There was no change in the number of diatoms found upon Scapania when it was transplanted, but a reduction in numbers occurred on the transplanted Fontinalis when it started dying. The composition of these diatom floras was different in polluted sites as compared to that in the cleaner areas.     

Application of in situ cage cultures of phytoplankton for monitoring heavy metal pollution in two Norwegian fjords

A simple apparatus for in situ use of the cage culture technique for growing planktonic algae has been developed and used to follow growth and metal uptake of three diatoms in two Norwegian fjords polluted by heavy metals. Continuous pumping of sea water through a chelating resin and continuous water sampling was used to obtain average values for dissolved and particulate heavy metal content at the various test sites. The metals investigated (zinc, copper, lead, cadmium, and mercury) showed differences in the proportion of dissolved to particulate fractions.The three species of diatoms tested gave systematic growth responses to heavy metal pollution in the moderately polluted fjord; one alga died, one showed reduced growth rate at the more contaminated site compared with those at less polluted sites, while the most tolerant alga was apparently not affected. In the heavily polluted fjord only the most tolerant alga survived, showing decreasing growth rate with increasing pollution.Uptake of heavy metals increased generally with increasing heavy metal content in the sea water. The contents in the algae grown in the most polluted fjord were much higher than those obtained in the less polluted fjord, which were higher than the contents reported for algae from non-contaminated areas.     

Species interactions in three Lemnaceae species growing along a gradient of zinc pollution

Duckweeds (Lemnaceae) are increasingly studied for their potential for phytoremediation of heavy‐metal polluted water bodies. A prerequisite for metal removal, however, is the tolerance of the organism to the pollutant, e.g., the metal zinc (Zn). Duckweeds have been shown to differ in their tolerances to Zn; however, despite them most commonly co‐occurring with other species, there is a lack of research concerning the effect of species interactions on Zn tolerance. Here, we tested whether the presence of a second species influenced the growth rate of the three duckweed species Lemna minor, Lemna gibba, and Lemna turionifera. We used four different Zn concentrations in a replicated microcosm experiment under sterile conditions, either growing the species in isolation or in a two‐species mixture. The response to Zn differed between species, but all three species showed a high tolerance to Zn, with low levels of Zn even increasing the growth rates. The growth rates of the individual species were influenced by the identity of the competing species, but this was independent of the Zn concentration. Our results suggest that species interactions should be considered in future research with duckweeds and that several duckweed species have high tolerance to metal pollution, making them candidates for phytoremediation efforts.