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Heavy metals are one of the most important environmental pollutants. In recent years, many low cost stretages of bioremediation for contaminated sites by heavy metals, such as fungi, bacteria and plants have been investigated for their biosorption capacity towards heavy metals. The uses of plant species for remediate contaminated sites by heavy metals are so called phytoremediation. The purpose of the first parts of this study are to (1) evaluate bioavailability of Cadmium (Cd) in contaminated soil and phytoremediation potential by three plant species, Vetiveria zizanioides, Pteris multifida, and Alternanthera philoxeroides (Mart.), and (2) realized the influence of photosynthetic bacteria (PSB) on the uptake of Cd in the three species. The results showed that the Alternanthera philoxeroides (Mart.) could accumulate the highest concentration of Cd among the three species, in which the Cd concentration of plant tissue increased with the concentration in soil. The highest concentration of Cd (164.9 mg kg-1) was found in the below-ground parts of Alternanthera philoxeroides (Mart.) at the 8th week of culturing period. However, the species of Vetiveria zizanioides could accumulate the largest total Cd, up to 547.5 £gg/ plant, which thus extracted the greatest amounts of Cd from the soil. Therefore, in the first part of this study the species of Vetiveria zizanioides was concluded to be the best accumulator among the three plant species. In addition, the concentration of Cd in the species of Pteris multifida was found significantly increased after PSB was added into the soil, but the plants died later due to Cd stress. The experimental results also showed that PSB seemed to be not suitable for each species used in this study to accumulate Cd from Cd-contaminated soil. In the second part of this research, both pot and field experiments were conducted to (1) evaluate bioavailability of copper (Cu), lead (Pb) and zinc (Zn) in contaminated soil and phytoremediation potential by domesticated plants, Bidens pilosa and Passiflora foetida inoculated with arbuscular mycorrhizal (AM) fungi, and to (2) compare the results of pot and field experiments. The plant species of Bidens pilosa inoculated with AM fungi had significantly higher Cu concentrations in the shoots and roots than non-inoculated plants. The plant species of Passiflora foetida inoculated with AM fungi also had significantly higher Cu and Pb concentrations in the roots than non-inoculated plants. When we found that the root dry weight of Passiflora foetida inoculated with AM fungi dramatically increased, the concentrations of Cu, Pb and Zn in the root of the plant species increased by 9-14 times, comparing with the plants without inoculation of AM fungi. The AM fungi have potential either to promot plant growth or to increase heavy metal accumulation. The values of element translocation proportion from root to shoot was Zn>Cu>Pb for the plant species in both pot and field experiments. For both experiments, the results of pot test and field test were significantly different. The concentrations of pot tests were found higher than the field tests, and some values of pot tests were even found significantly greater than those in the field tests. In the third part of this study, the field experiments were conducted to test the feasibility of using domesticated vegetations for phytoremediation of the contaminated farmland. The objectives of this study were (1) to acquire information about the ability of five plant species growing wild in the polluted area to accumulate Cu, Pb and Zn, (2) to investigate the season effects on phytoremediaton of five plant species and evaluate the total uptake of heavy metal, and (3) to run both pot tests and a field trial of phytotremediation to compare their differences. The experimental results showed that three maximum toxic elements in a pot were 3020 mg kg-1 Pb, 232 mg kg-1 Cu and 1012 mg kg-1 Zn respectively. The Cu concentrations of the five plant species collected from the polluted plots ranged from 0.7 to 17.43 mg kg-1. The range of variation of Pb in plant tissues was measured varied from 2.29 to 81.65 mg kg−1, while a wide range of Zn concentrations was found from 12.84 to 192.85 mg kg-1 among the plants collected at the contaminated plots. In comparison to winter season, the Zn concentrations in Broussonetia papyrifera, Passiflora foetida and Saccharum sinensis collected in summer season was significantly higher. The higher Cu concentrations were obtained in both plant species of Bidens pilosa and Mimosa diplotricha in summer season. However, Pb concentrations in Saccharum sinensis collected in winter were significantly higher than those in the same plant species collected in summer. Bidens pilosa was also found having the highest total amount of Cu and Zn. The highest total amount of Pb was found in Mimosa diplotricha. For both plant species, both of the pot and field tests were different.

As a potential bioremediation system for contaminated soils, I evaluated the use of an arbuscular mycorrhizal (AM) fungus, Glomus intraradices on roots and shoots uptake of polycyclic aromatic hydrocarbons (PAHs), alkyl PAHs, and toxic metals in Echinacea purpurea, in using a controlled 20-week greenhouse study and a complimentary 2-year field study. E. purpurea seeds were either inoculated with the mycorrhizal fungus (AM) or not inoculated (non-AM) and grown in soil provided by the National Capital Commission (NCC) that have known contamination. In the greenhouse study, AM inoculation increased the uptake of alkyl PAHs in the roots of E. purpurea. The AM inoculation showed no effect on root uptake of PAHs and toxic metals over the 20-week study period. However, when I calculated the uptake rates (k1) for PAHs between both treatments, the AM treated roots ha 10-fold higher k1 values than non-AM treated roots. The soil concentrations of PAHs were found to increase over time with AM inoculation, suggesting, that AM fungi are causing a solvent depletion through root uptake of minerals and carbon, which concentrates the more hydrophobic PAHs in soils. Alkyl PAHs and metals showed no change over time amongst any of the treatments. Assessing the performance of AM fungi on the uptake of contaminants under field conditions, only PAHs showed increased bioaccumulation in the shoots of E. purpurea with AM inoculation. Alkyl PAHs and metals in plant material were unaffected by the AM inoculation, but increased significantly from year 1 to year 2. The uptake rates among treatments were similar, with non-AM roots having slightly greater uptake. Soil concentrations of PAHs and alkyl PAHs were unaffected over the course of the experiment. Our control soil, however, showed significant increases in concentration from year 1 to year 2 with alkyl PAHs. These results quantified the influence of AM hyphae-mediated uptake of organic and inorganic contaminant transfer from soil to plants and the bioaccumulation kinetics for contaminants by E. purpurea that will be useful for environmental models and phytoremediation strategies.

Les HAP sont parmi les substances les plus problématiques parce qu'ils ont un fort pouvoir cancérigène, mutagène et ont, par conséquent, des effets nocifs pour la santé humaine. Parmi les techniques de remédiation des sols contaminés par des HAP, la phytoremédiation a été reconnue comme une méthode prometteuse en raison de ses avantages économiques et écologiques. Toutefois, en raison de la nature récalcitrante des HAP, et de facteurs environnementaux difficile à maîtriser, cette technique est encore limitée en termes d'efficacité, en particulier lorsqu'il s'agit de HAP de poids moléculaires élevés. L'inoculation des plantes avec des champignons mycorhiziens à arbuscules (MA), qui sont omniprésents dans la plupart des sols naturels et anthropiques, est connue pour favoriser l'élimination des HAP. Cependant, des résultats variables ont été rapportés quant à l'effet des plantes et des microorganisms associés notamment les champignons MA, sur la phytoremédiation des HAP et nécessite des études complémentaires. Des expériences été réalisées dans des cultures en pot et en microplaques, pour étudier l'influence de certains paramètres sur la dissipation des HAP dans la rhizosphère: l'espèce végétale, l'espèce fongique, la nutrition minérale, la nature des HAP, leur disponibilité et les interactions entre HAP. Quatre espèces de plantes (luzerne, fétuque élevée, ray-grass et céleri) et deux espèces de champignons MA (Glomus intraradices et Glomus mosseae) ont été testées dans un sol artificiellement contaminé avec trois HAP: le phénanthrène (PHE), le pyrène (PYR) et le dibenzo[a, h] anthracène (DBA). Le poids moléculaire des HAP est un facteur majeur influencant leur élimination. Lorsque le poids moléculaire des HAP était plus élevé, le nombre de bactéries dégradantes cultivables était plus faible, et l'efficacité de la phytoremédiation des HAP limitée. La présence de PHE a diminué la biomasse végétale et la colonisation mycorhizienne, mais il a augmenté la dissipation du DBA par co-métabolisme dans les expériences en pots et en microplaques. En revanche, cet effet n'a pas été observé entre le PYR et le DBA . La dissipation des HAP a varié avec les espèces de plantes et de champignons MA. Bien que les quatre espèces de plantes aient augmenté l'élimination du PHE, seule la luzerne a montré des effets positifs sur la phytoremédiation du DBA. Glomus intraradices a augmenté la biomasse végétale et l'absorption du phosphore par des plantes, et il a également augmenté la dissipation du DBA. Lorsque la co-culture de luzerne et fétuque était colonisée par Glomus mosseae, la biomasse obtenue était plus élevée, et la concentration des gènes de HAP-dioxygénase était significativement plus élevée qu'avec Glomus intraradices. Mais Glomus mosseae n'a montré aucun effet significatif sur la phytoremédiation du DBA. Ainsi cet effet des champignons MA sur la dissipation des HAP n'est pas seulement un effet biomasse. La concentration de phosphore et le régime d'alimentation en eau ont également influencé la colonisation mycorhizienne et la dissipation des HAP. Ainsi la dissipation du DBA en présence de plantes était significativement plus élevée que dans les témoins non plantés lorsque la teneur en eau était élevée et celle en phosphore plus faible, ce qui correspondait à la situation où le taux de mycorhization des plantes était le plus élevé. L'ensemble de ces résultats souligne la complexité des interactions entre plantes, microorganismes et polluants dans les sols. Ils montrent que tous les paramètres considérés affectent significativement la dissipation des HAP dans la rhizosphère des plantes, et méritent d'être pris en compte pour contrôler et améliorer la phytoremédiation / PAHs are among the most problematic substances as they could accumulate in the environment and threaten the development of living organisms because of their acute toxicity, mutagenicity or carcinogenity. Among remediation techniques for PAH contaminated sites, phytoremediation has been recognized as a promising method owing to its economical and ecological benefits. However, due to the recalcitrant nature of PAH, multivariate and changeful environment factors, this technique is still limited in terms of effectiveness, especially when dealing with high molecular weight PAHs. Inoculation of plants with arbuscular mycorrhizal (AM) fungi, which are ubiquitous in natural and most anthropogenically influenced soils, is known to benefit PAH phytoremediation. However, diverging results were reported on PAH dissipation in plant rhizosphere and the parameters affecting the AM fungi assisted PAH phytoremediation needed more investigation. Some of these parameters were considered in the present work: plant species, AM fungi species, phosphorus nutrition and watering regimes, PAH type, availability and interactions between PAHs. Experiments were performed in pot cultures and in microplates, with different plant species (including alfalfa, tall fescue, ryegrass and celery roots), two AM fungi (Glomus intraradices and Glomus mosseae) and three kinds of PAHs (phenanthrene (PHE), pyrene (PYR) and dibenzo[a,h]anthracene (DBA)), spiked to a soil. PAH molecular weight was a major parameter influencing PAH phytoremediation. With the increase of PAH molecular weight, the culturable PAH degraders decreased, so did the efficiency of phytoremediation. PHE decreased the plant biomass and AM fungi colonization, but it increased the DBA dissipation in both pot and microplate experiments. PYR did not increase DBA dissipation, and addition of PYR into PHE+DBA substrate decreased both PHE and DBA dissipation. PAH phytoremediation efficiency varied with the plant and AM fungi species. Although all four species of plants increased the disappearance of PHE, only alfalfa showed a positive effect on high molecular weight (HMW) PAHs. Glomus intraradices increased the plant biomass and phosphorus uptake of plants, and it also increased DBA dissipation in DBA or PHE+PYR+DBA spiked soil. Co-planted alfalfa and tall fescue colonized with Glomus mosseae obtained higher biomass and the concentration of the PAH-ring hydroxylating dioxygenase genes were significant higher, than with Glomus intraradices, but Glomus mosseae showed no or negative effect on DBA phytoremediation. The phosphorus concentration and water regime also influenced the AM fungus colonization and PAH dissipation. The highest AM colonization and a significant positive impact of mycorrhizal plants on the dissipation of DBA was detected in high-water and low-phosphorus treatment. Results indicated that complex interactions between plants, microorganisms and soil control the fate on PAHs. All the studied parameters significantly affected PAH dissipation in plant rhizosphere, and should be considered for controlling and improving phytoremediation efficiency

Plant symbiosis with arbuscular mycorrhizal (AM) fungi provides many benefits for plants, including increased nutrient uptake, drought tolerance, and belowground pathogen resistance. In order to have a better understanding of the genetic architecture of mycorrhizal symbiosis, we conducted a genome-wide association study (GWAS) by phenotyping a diversity panel of cultivated sunflower (Helianthus annuus) for root colonization under inoculation with the AM fungus Rhizophagus intraradices. This mapping panel consists of 261 inbred lines that capture approximately 90% of the genetic diversity present in the cultivated sunflower germplasm. Using a mixed linear model approach with a high-density genetic map, we determined regions of the genome that are likely associated with AM colonization in sunflower. Additionally, we used a ‘core 12' set of twelve diverse lines (representing approximately 50% of the genetic diversity in the cultivated germplasm) to assess the effect that inoculation with AM fungi has on dried shoot biomass and macronutrient uptake. Colonization rate among lines in the mapping panel ranged from 0 to 70% and was not correlated with mycorrhizal growth response, shoot P response, or shoot K response among the core 12 lines. Association mapping yielded three SNPs that were significantly associated with AM colonization rate. These SNPs explained 19.0%, 14.4%, and 27.9% of the variance in three different metrics used to measure the degree of root colonization. Three genes of interest identified from the significant regions that contained these SNPs are potentially related to plant defense. Overall, our data suggests that candidate genes involved in plant defense may affect AM colonization rates within cultivated sunflower, and that these genes have a large effect size.

The broad aim of the work described in this thesis was to use the arbuscular mycorrhizal (AM) defective rmc tomato to explore the development and function of different types of fungus-plant interfaces (phenotypes) and to characterize the cellular modifications preceding colonization of rmc by a range of different AM fungi. Three main patterns of colonization with rmc have been described: 1) Pen- phenotype in which the AM fungus is restricted to the root surface with several attempts to penetrate the epidermal cells without success; 2) Coiphenotype where AM fungi penetrate the epidermis but cannot develop cortical colonization; and 3) Myc+ phenotype (with G. intraradices WFVAM23), where the AM fungus penetrates the cortex and forms a “normal” colonization after a delayed penetration of the epidermal cells (Review of literature). Little is known about cellular interactions, nutrient transfer or the ability of the fungi to complete their life cycles in the different phenotypes. These aspects were the main foci of this work. In addition further fungal isolates were screened to asses their ability to colonize rmc. The first experiments involved compartmented pots to follow the fungal life cycle, production of external mycelium and spores in the different rmc phenotypes (Chapter 3). The results showed that in the Pen- and Coiphenotypes, AM fungi are unable to form spores to complete the life cycle. However, in the Coi-phenotype, the fungus remained alive up to week 18, suggesting that some C transfer occurred. The fungus forming the Myc+ phenotype, G. intraradices WFVAM23, was able to produce spores, although they were significantly smaller than those produced with the wild-type tomato. The results suggested that arbuscules are essential for completion of the fungal life cycle. Labeled 32P was used to determine whether arbuscules are also essential for P transfer (Chapter 4). A compartmented pot system was used in which only fungal hyphae but not roots could obtain 32P. 32P was found in the shoots of rmc inoculated with S. calospora (Coi- phenotype), indicating that interfaces other than arbuscules can be involved in transfer of P. A nurse pot system was used to obtain synchronized colonization to determine how long AM fungi stay alive during the interactions with rmc and to elucidate the cellular modifications preceding colonization of rmc by a range of different AM fungi (Chapter 5). The results showed that rmc did attract the AM fungi, that the plant nucleus moved to the middle of the plant cell only after fungal penetration of plant roots and that callose deposition in rmc was not involved in blocking the AM fungi. Fourteen AM fungi with different taxonomic affiliations and fourteen different G. intraradices isolates were screened to try to relate phylogeny of AM fungi with phenotypes in rmc (Chapter 6). There were a large number of interactions, depending on the inoculated AM fungi, and although there were some similarities in the rmc phenotypes within phylogenetic groups, there was no clear relationship between phylogeny and development of interactions with rmc. This study showed the following. 1) Arbuscules/arbusculate coils are necessary for the completion of the AM fungal cycle. However, intraradical hyphae also participate in transfer of both P and C as demonstrated with the Coi- phenotype. 2) rmc clearly attracted AM fungi and the fungi stay alive and induce plant cellular responses such as nuclear movement only after penetrating rmc roots. 3) Plant defense responses such as callose deposition are not involved in blocking AM fungi in rmc; and 4) there was no relationship between the phenotypes described in rmc and phylogeny of the Glomeromycota. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1292816 / Thesis(Ph.D.)-- School of Earth and Environmental Sciences, 2007.

Striga, a genus of obligate parasitic weeds in the family Orobanchaceae, has been identified as the most important biological factor limiting agricultural productivity in sub-Saharan Africa. Germination of Striga seeds is triggered by strigolactone root exudates from host plants. Strigolactones also induce hyphal branching in arbuscular mycorrhizal (AM) fungi, which are important for plant uptake of phosphorus in low phosphorus soils. Mechanisms of Striga resistance based on reduced strigolactone production may also convey resistance to AM fungi which would require higher inputs of phosphorus fertilizer to attain optimal crop growth. There is evidence for genetic differences in mycorrhizal responsiveness in other grain crops; therefore it is beneficial for breeders to be aware of these differences when developing Striga-resistant sorghum cultivars. This research aims to determine phosphorus and mycorrhizal responsiveness of sorghum genotypes important for or developed by breeders working on Striga resistance. Phosphorus response curves were determined for twelve sorghum genotypes using pasteurized low phosphorus soil amended to achieve four different phosphorus levels. Simple linear regression was performed on root and shoot dry weight data. Results indicate variability in phosphorus responsiveness within Striga resistant and susceptible genotypes. Seven of these genotypes were selected for continued research, which analyzed responsiveness to phosphorous and differences in mycorrhizal responsiveness in relation to reported mechanisms of Striga resistance. Treatments included three levels of phosphorus amendments and the addition of Funneliformis mosseae inoculum. All genotypes were strongly responsive to P amendment when grown without AM fungi and showed a decrease in responsiveness to P when inoculated with F. mosseae. Trends for all genotypes indicate a greater uptake of P, Zn, and Mg by mycorrhizal plants as compared to nonmycorrhizal plants. All seven genotypes were responsive to mycorrhizae, with a significant increase in biomass for all genotypes, especially at the lowest phosphorus level. The responsiveness to the mycorrhizal fungus does not appear to be directly related to the susceptibility of genotypes to the parasitic weed Striga. / Graduation date: 2012

Les prairies indigènes présentent une source importante d'alimentation pour le pâturage du bétail dans les prairies Canadiennes semi-arides. L'addition de légumineuses fixatrices d'azote et de phosphore dans les prairies indigènes peut améliorer la productivité et la valeur nutritive de fourrage. Ces pratiques peuvent induire des modifications de la structure et de la diversité des communautés fongiques du sol, ce qui peut en retour avoir un impact sur la production et le contenu nutritionnel du fourrage. L’objectif de cette étude était de développer un système de pâturage à bas niveau d’intrants, productif, autonome et durable. À court terme, nous voulions 1) déterminer l'effet des légumineuses (Medicago sativa, une légumineuse cultivée ou Dalea purpurea, une légumineuse indigène) et la fertilité en phosphore du sol sur la productivité et la valeur nutritive des graminées indigènes, comparées avec celles de la graminée introduite Bromus biebersteinii en mélange avec le M. sativa, 2) identifier l'effet de ces pratiques sur la diversité et la structure des communautés des champignons mycorhiziens à arbuscules (CMA) et des champignons totaux, 3) identifier l'effet des légumineuses et des CMA sur les interactions compétitives entre les graminées de saison fraîche et les graminées de saison chaude. Les expériences menées au champ ont montré que M. sativa améliorait les teneurs en azote et en phosphore des graminées indigènes au début de l'été, ainsi que la teneur en azote de la graminée de saison chaude Bouteloua gracilis à la fin de l'été de l'année sèche 2009. Par contre, la fertilité en phosphore du sol n'ait pas affecté la productivité des plantes. D'autre part, l'inclusion des légumineuses augmentait la diversité des CMA dans le mélange de graminées indigènes. Cette modification présentait des corrélations positives avec la productivité et la quantité totale d'azote chez le M. sativa et avec la teneur en phosphore des graminées indigènes, au début de l'été. La structure des communautés de champignons totaux était influencée par l'interaction entre le mélange des espèces et la fertilité en phosphore du sol seulement en 2008 (année humide). Cet effet pourrait être lié en partie avec la productivité des plantes et l'humidité du sol. Les expériences menées en chambre de culture ont montré que les CMA peuvent favoriser la productivité des graminées de saison chaude au détriment des graminées de saison fraîche. En effet, Glomus cubense augmentait la productivité de la graminée de saison chaude B. gracilis, en présence de M. sativa. Cet effet pourrait être associé à l’effet négatif du G. cubense sur la fixation de l’azote par le M. sativa et à la diminution de l’efficacité d’utilisation de l'azote de certaines graminées de saison fraîche résultant en une augmentation de la disponibilité de l'azote pour B. gracilis. Par contre, le Glomus sp. augmentait la biomasse de Schizachyrium scoparium, autre graminée de saison chaude, en absence de légumineuse. Ce phénomène pourrait être attribuable à une amélioration de l’efficacité d’utilisation du P de cette graminée. En conclusion, mes travaux de recherche ont montré que la légumineuse cultivée M. sativa peut améliorer la valeur nutritive des graminées indigènes au début de l'été ainsi que celle de la graminée de saison chaude B. gracilis, dans des conditions de sécheresse sévère de la fin de l'été. De plus, l'addition de M. sativa dans le mélange de graminées indigènes peut contribuer à augmenter le nombre des espèces bénéfiques des CMA pour la production et la nutrition du fourrage au début de l'été. / The native grasslands are considered as the main feed source for livestock grazing, in semi-arid regions of the Canadian prairies. The addition of N fixing legumes and phosphorus to semi-arid native grasslands may increase the productivity and nutritive value of forage. However, these practices may also shape the structure and diversity of soil fungal communities which in turn may impact forage production and nutritive value. The global objective of this research was to design productive, self-sustaining, permanent and with low inputs pastures. The specific objectives were 1) to demonstrate the effect of N-fixing legumes (the cultivated legume Medicago sativa or the native legume Dalea purpurea) and soil P fertility on the productivity and nutritive value of native grasses mixes in comparison to the mixture of the introduced grass Bromus biebersteinii and M. sativa, 2) identify the effect of these practices on the diversity and community structure of arbuscular mycorrhizal (AM) fungi and total fungi, and 3) identify the effect of legumes and AM fungi on competitive interactions between native cool-season grasses and native warm-season grasses. The field experiment showed that M. sativa improved the nitrogen and phosphorus concentrations of native grasses mixes early in the summer, as well as the N concentration of the warm-season grass B. gracilis, in late summer of the driest year 2009. In contrast, the soil phosphorus fertility had no effect on plant productivity. On the other hand, the inclusion of legumes to the mix of native grasses generally increased AM fungal diversity. This shift was positively correlated with the productivity and nitrogen uptake by M. sativa and with the phosphorus concentration of native grasses mixes in early summer. The structure of the total fungal community was affected by the interaction between species mixtures and soil P fertility only in the wet year (2008), suggesting that this effect was likely driven in part by plant productivity and soil moisture. The growth chamber experiment showed that the AM fungi may favoured the growth of warm-season grasses under competition with cool-season grasses. However, Glomus cubense increased the productivity of warm-season grass B. gracilis when growing with M. sativa. This effect might be related to a negative impact of G. cubense on the nitrogen-fixing activity of M. sativa and to a lower N-use efficiency of certain cool-season grasses, which resulted in increased soil N availability for B. gracilis. In contrast, Glomus sp. enhanced the growth of S. scoparium, another warm-season grass in the absence of legumes, and this may be related to improved P-use efficiency in this grass. We concluded that the cultivated legume M. sativa can improve the nutritive value of native grasses mixes early in the summer and also of warm season grass under severe drought conditions in late summer. In addition, the inclusion of M. sativa within native grass mixes may contribute to promote beneficial AM fungi taxa that were involved in forage production and nutrition early in the summer.