In vitro selection of ecologically adapted ectomycorrhizal fungi through production of fungal biomass and metabolites for use in reclamation of biotite mine tailings.

Mineral weathering plays an important role in poor-nutrient environments such as mine spoils and tailings. Ectomycorrhizal (ECM) fungi are able to enhance mineral weathering through different mechanisms, thereby increasing the availability of minerals and nutrients to plants. Six ECM fungi (Cadophora finlandia, Cenococcum geophilum, Hebeloma crustuliniforme, Lactarius aurantiosordidus, Paxillus involutes, and Tricholoma scalpturatum) were tested here for their tolerance to biotite-quartz-rich mine tailings. Either solid- or liquid-medium methods were used for in vitro selection of ECM fungi for their ability to grow on mine tailings. ECM fungi were selected based on their mycelial radial growth and metabolite production (ergosterol and low-molecular-mass organic acids, LMMOAs). We found a strong correlation between fungal ergosterol content and mycelial radial growth using the solid-medium method. However, the liquid-medium method was more appropriate for ergosterol synthesis and permitted direct measurement of organic acid production. We found that LMMOAs were exuded by ECM fungi, which solubilized mine tailings for their own growth and nutrition. Finally, we concluded that the ECM fungi C. finlandia and T. scalpturatum are the species most tolerant to tailings and could potentially improve the survival rate, growth, and health of white spruce seedlings planted on biotite mine spoils and tailings.

Mycorrhizae and Rhizobacteria on Precambrian Rocky Gold Mine Tailings: I. Mine-Adapted Symbionts Promote White Spruce Health and Growth.

White spruce [Picea glauca (Moench) Voss] is a commercially valuable boreal tree that has been known for its ability to colonize deglaciated rock tailings. Over the last decade, there has been an increasing interest in using this species for the revegetation and successful restoration of abandoned mine spoils. Herein, we conducted a glasshouse experiment to screen mycorrhizal fungi and rhizobacteria capable of improving the health and growth of white spruce seedlings growing directly on waste rocks (WRs) or fine tailings (FTs) from the Sigma-Lamaque gold mine located in the Canadian Abitibi region. After 32 weeks, measurements of health, growth, and mycorrhizal colonization variables of seedlings were performed. Overall, symbionts isolated from roots of healthy white spruce seedlings growing on the mining site, especially Cadophora finlandia Cad. fin. MBN0213 GenBank No. KC840625 and Pseudomonas putida MBN0213 GenBank No. AY391278, were more efficient in enhancing seedling health and growth than allochthonous species and constitute promising microbial symbionts. In general, mycorrhizae promoted plant health and belowground development, while rhizobacteria enhanced aboveground plant biomass. The observed beneficial effects were substrate-, strain-, and/or strains combination-specific. Therefore, preliminary experiments in control conditions such as the one described here can be part of an efficient and integrated strategy to select ecologically well-adapted symbiotic microorganisms, critical for the success of a long-term revegetation program.

Mycorrhizae and Rhizobacteria on Precambrian Rocky Gold Mine Tailings: II. Mine-Adapted Symbionts Alleviate Soil Element Imbalance for a Better Nutritional Status of White Spruce Seedlings.

In the context of a phytorestoration project, the purpose of this study was to assess the respective contribution to the nutritional status of Picea glauca seedlings of ectomycorrhizae and rhizobacteria native or not to the Sigma-Lamaque gold mine wastes in northern Quebec, Canada. In a glasshouse experiment, inoculated plants were grown for 32 weeks on coarse waste rocks or fine tailings obtained from the mining site. The survival, health, growth, and nutritional status of plants were better on coarse waste rocks than on fine tailings. Fe and Ca were especially found at high levels in plant tissues but at much lower concentrations on waste rocks. Interestingly, inoculation of microsymbionts had only minimal effects on N, P, K, and Mg plant status that were indeed close or within the concentration range encountered in healthy seedlings. However, both fungal and bacterial treatments improved Fe and Ca concentrations in plant tissues. Fe concentration in the foliage of plants inoculated with the fungi Tricholoma scalpturatum Tri. scalp. MBN0213 GenBank #KC840613 and Cadophora finlandia Cad. fin. MBN0213 GenBank #KC840625 was reduced by >50%. Both fungi were isolated from the mining site. The rhizobacteria, Azotobacter chroococcum, also improved plant Fe level in some cases. Regarding Ca nutritional status, the native bacterial strain Pseudomonas putida MBN0213 GenBank #AY391278 was the only symbiont that reduced foliar content by up to 23%. Ca concentration was negatively correlated with the fungal mycorrhization rate of seedling roots. This relation was especially strong (r = -0.66, p-value ≤ 0.0001) in the case of C. finlandia. Also, a similar relationship existed with root Fe concentration (r = -0.44, p-value ≤ 0.0001). In fact, results showed that seedling performance was more correlated with elevated Ca and Fe concentration in planta than with nutrient deficiency. Also, native microsymbionts were capable of regulating seedling nutrition in the poor substrate of the Sigma-Lamaque gold mine tailings.

Edaphic Selection Pressures as Drivers of Contrasting White Spruce Ectomycorrhizal Fungal Community Structure and Diversity in the Canadian Boreal Forest of Abitibi-Témiscamingue Region.

Little is known about edaphic selection pressures as drivers of contrasting white spruce ectomycorrhizal fungal community structure and diversity in the Canadian boreal forest. We hypothesized that community composition differs among the four sites sampled-nursery, mining site, forest edge, and natural forest. Ectomycorrhizal (ECM) fungal community structure and diversity was studied at the four locations with soil fertility gradient through morpho-molecular and phylogenetic analyses in relationships with rhizospheric soil chemical properties. 41 different species were identified. Mining site had a significantly different species composition than the surrounding environments. Soil pH and percentage of roots colonized by ECM fungi increased while soil P, N, Fe, C, K, Mg, Al, Ca, and Na contents declined across the soil fertility gradient: nursery → natural forest → forest edge → mining site. Contrary to the preference of acid soils by ECM fungi, a few ecologically adapted to high pH, poor soil chemical fertility, and low organic matter content colonize white spruce roots on the non-acidogenic mining site, allowing natural regeneration of white spruce seedlings. Other ECM fungi are adapted to high fertigation level of commercial nursery. This study clearly shows the contrasting difference in white spruce ectomycorrhizal fungal community structure and diversity driven by edaphic selection pressures.

Performance of ectomycorrhizal alders exposed to specific Canadian oil sands tailing stressors under in vivo bipartite symbiotic conditions.

Canadian oil sands tailings are predominately sodic residues contaminated by hydrocarbons such as naphthenic acids. These conditions are harsh for plant development. In this study, we evaluated the effect of inoculating roots of Alnus viridis ssp. crispa and Alnus incana ssp. rugosa with ectomycorrhizal fungi in the presence of tailings compounds. Seedlings were inoculated with 7 different strains of Paxillus involutus and Alpova diplophloeus and were grown under different treatments of NaCl, Na2SO4, and naphthenic acids in a growth chamber. Afterwards, seedling survival, height, dry biomass, leaf necrosis, and root mycorrhization rate were measured. Paxillus involutus Mai was the most successful strain in enhancing alder survival, health, and growth. Seedlings inoculated with this strain displayed a 25% increase in survival rate, 2-fold greater biomass, and 2-fold less leaf necrosis compared with controls. Contrary to our expectations, A. diplophloeus was not as effective as P. involutus in improving seedling fitness, likely because it did not form ectomycorrhizae on roots of either alder species. High intraspecific variation characterized strains of P. involutus in their ability to stimulate alder height and growth and to minimize leaf necrosis. We conclude that in vivo selection under bipartite symbiotic conditions is essential to select effective strains that will be of use for the revegetation and reclamation of derelict lands.

Integrating GIS components with knowledge discovery technology for environmental health decision support.

This paper presents a new category of decision-support tools that builds on today's Geographic Information Systems (GIS) and On-Line Analytical Processing (OLAP) technologies to facilitate Geographic Knowledge Discovery (GKD). This new category, named Spatial OLAP (SOLAP), has been an R&D topic for about 5 years in a few university labs and is now being implemented by early adopters in different fields, including public health where it provides numerous advantages. In this paper, we present an example of a SOLAP application in the field of environmental health: the ICEM-SE project. After having presented this example, we describe the design of this system and explain how it provides fast and easy access to the detailed and aggregated data that are needed for GKD and decision-making in public health. The SOLAP concept is also described and a comparison is made with traditional GIS applications.