Arbuscular mycorrhizal fungi (AMF) are important components of agro-ecosystems and are especially significant for productive low-input agriculture. Traditional spore morphology-based identification of AMF in biodiversity studies is subjective and requires expertise and time. Researchers have used molecular techniques to investigate community composition of AMF in uncultivated, disturbed, or contaminated soils, but this approach to community analysis of AMF in agricultural soils has not been reported. In this study, a polymerase chain reaction and denaturing gradient gel electrophoresis (PCR-DGGE) procedure for the detection of fungal 18S ribosomal RNA gene was developed with reference cultures. Five AMF species were procured from the International Culture Collection of Arbuscular and Vesicular-Arbuscular Mycorrhizal Fungi (INVAM). These reference cultures were chosen because isolates of their species were putatively identified in a previous survey of farm field soils in Saskatchewan, Canada. A reference PCR-DGGE profile was generated using DNA extracted and amplified from the spores of these INVAM cultures. The methods technical limitations were investigated. The optimized procedures effectiveness was tested by its application to soil samples from 38 farms. Bands from the PCR-DGGE profiles of these samples were excised for sequence analysis. The total number of species recovered was low in comparison to other AMF community surveys of temperate climate locations. The majority of the sequences recovered were Glomus species. Scutellospora calospora, a previously undetected AM fungus in Saskatchewan was found. A trend in AMF distribution in Saskatchewan was observed and it was relatable to their phylogenetic taxonomy. Though not without its drawbacks, this approach to community composition analysis of AMF was faster than conventional trap cultivation methods.

Arbuscular mycorrhizal fungi (AMF) are important components of agro-ecosystems and are especially significant for productive low-input agriculture. Traditional spore morphology-based identification of AMF in biodiversity studies is subjective and requires expertise and time. Researchers have used molecular techniques to investigate community composition of AMF in uncultivated, disturbed, or contaminated soils, but this approach to community analysis of AMF in agricultural soils has not been reported. In this study, a polymerase chain reaction and denaturing gradient gel electrophoresis (PCR-DGGE) procedure for the detection of fungal 18S ribosomal RNA gene was developed with reference cultures. Five AMF species were procured from the International Culture Collection of Arbuscular and Vesicular-Arbuscular Mycorrhizal Fungi (INVAM). These reference cultures were chosen because isolates of their species were putatively identified in a previous survey of farm field soils in Saskatchewan, Canada. A reference PCR-DGGE profile was generated using DNA extracted and amplified from the spores of these INVAM cultures. The methods technical limitations were investigated. The optimized procedures effectiveness was tested by its application to soil samples from 38 farms. Bands from the PCR-DGGE profiles of these samples were excised for sequence analysis. The total number of species recovered was low in comparison to other AMF community surveys of temperate climate locations. The majority of the sequences recovered were Glomus species. Scutellospora calospora, a previously undetected AM fungus in Saskatchewan was found. A trend in AMF distribution in Saskatchewan was observed and it was relatable to their phylogenetic taxonomy. Though not without its drawbacks, this approach to community composition analysis of AMF was faster than conventional trap cultivation methods.

This document is a project document based on finding a more economical way to use buttermilk at Synlait Milk Ltd. Buttermilk is a by-product from the Anhydrous Milk Fat (AMF), which is the concentration of cream. It is a problem for many dairy companies in New Zealand, including Synlait. the problems arise due to the opportunity cost of using it. It is a low value product, but made in substantial quantities at a ratio of 55% buttermilk to 45% AMF. This feasibility study contains an analysis on buttermilk at Synlait, including opportunity costs and benefits with processing buttermilk into buttermilk powder. It provides insight into the feasibility of implementing a ‘washed cream’ process at Synlait which would provide an alternative method for using buttermilk. The feasibility of separating buttermilk components for separate use is also examined. Overall, this project provides a more economical solution for buttermilk use at Synlait Milk Ltd.

Upstream oil and gas production has caused soil salinity problems across western Canada. In this work we investigated the use of ACC (1-aminocyclopropane-1-carboxylate) deaminase-producing plant growth-promoting rhizobacteria (PGPR) and the arbuscular mycorrhizal fungus (AMF) Glomus intraradices to enhance the efficiency and feasibility of phytoremediation of saline soils. This work involved laboratory and field research for three sites in south east Saskatchewan, Canada. The three research sites were Cannington Manor South (CMS), Cannington Manor North (CMN) and Alameda (AL). CMS and AL were highly saline, while the CMN site had moderate salinity. Indigenous PGPR were isolated from these sites and tested in greenhouse experiments using authentic salt-contaminated soils taken from the research sites. Increased plant biomass by PGPR and/or AMF was observed. This growth promotion effect varied with plant species, soil salinity and soil fertility. The combination treatment of two previously isolated PGPR Pseudomonas putida UW3 and UW4 (noted as UW3+4) from farm soil in Ontario consistently promoted shoot growth of both barley and oats grown in saline soils by approximately 100%. The indigenous PGPR Pseudomonas corrugata (CMH3) and Acinetobacter haemolyticus (CMH2) also promoted plant growth on par with UW3+4. In addition, in one experiment where alfalfa was tested, UW3+4, CMH2 and CMH3 treatments not only enhanced shoot biomass but also increased root nodulation. For AMF effects, G. intraradices enhanced biomass of oats and barley. Furthermore, the AMF+CMH3 was effective in promoting growth of Topgun ryegrass, while AMF+CMH2 was beneficial for Inferno tall fescue growth in salt impacted soils. The concentration of NaCl in the plants grown in salt-impacted soils ranged from 24 – 83 g/kg. There was no evidence of an increase in NaCl concentrations of plant tissue by PGPR and/or AMF treatments. In addition, to determine the importance of nutrient addition to research sites, liquid fertilizer was applied to 2-week old plants. Results demonstrated that fertilizer effectively increased biomass, and more importantly the biomass of PGPR treated plants supplied with fertilizer was approximately 20% higher than that of plants treated with fertilizer alone. Therefore, research sites were then amended with compost before planting of the 2007 field trial. Plant growth promotion by UW3+4 and CMH3 was tested in the summer of 2007 in the field. Prior to planting, soils were sampled from each site for soil salinity analysis. Barley, oats, tall fescue and ryegrass treated with and without PGPR were sown in plots. The plant coverage condition, NaCl concentrations and biomass of plant shoots were assessed to evaluate the PGPR effect. The results showed that PGPR promoted shoot dry weight by 30% - 175%. The NaCl concentrations of barley, oats and tall fescue averaged 53 g/kg, 66 g/kg and 35 g/kg, respectively. There was no evidence of an increase in NaCl concentrations of plant tissue by PGPR in the field. The salt removal of the CMN site was the highest among three sites due to the large amount of shoot biomass produced. The amount of salt accumulated in the shoots on the CMN site is estimated to be 1580 kg per hectare per year when both barley and ryegrass are planted together as a mix and treated with PGPR. Based on the field data, the estimated time required to remove 50% salt in the top 50 cm soil is seven years with PGPR treatments, while it takes fifteen years to do so without PGPR. In conclusion, PGPR-promoted phytoremediation was proven to be a feasible and effective remediation technique for soils with moderate salinity.

Upstream oil and gas production has caused soil salinity problems across western Canada. In this work we investigated the use of ACC (1-aminocyclopropane-1-carboxylate) deaminase-producing plant growth-promoting rhizobacteria (PGPR) and the arbuscular mycorrhizal fungus (AMF) Glomus intraradices to enhance the efficiency and feasibility of phytoremediation of saline soils. This work involved laboratory and field research for three sites in south east Saskatchewan, Canada. The three research sites were Cannington Manor South (CMS), Cannington Manor North (CMN) and Alameda (AL). CMS and AL were highly saline, while the CMN site had moderate salinity. Indigenous PGPR were isolated from these sites and tested in greenhouse experiments using authentic salt-contaminated soils taken from the research sites. Increased plant biomass by PGPR and/or AMF was observed. This growth promotion effect varied with plant species, soil salinity and soil fertility. The combination treatment of two previously isolated PGPR Pseudomonas putida UW3 and UW4 (noted as UW3+4) from farm soil in Ontario consistently promoted shoot growth of both barley and oats grown in saline soils by approximately 100%. The indigenous PGPR Pseudomonas corrugata (CMH3) and Acinetobacter haemolyticus (CMH2) also promoted plant growth on par with UW3+4. In addition, in one experiment where alfalfa was tested, UW3+4, CMH2 and CMH3 treatments not only enhanced shoot biomass but also increased root nodulation. For AMF effects, G. intraradices enhanced biomass of oats and barley. Furthermore, the AMF+CMH3 was effective in promoting growth of Topgun ryegrass, while AMF+CMH2 was beneficial for Inferno tall fescue growth in salt impacted soils. The concentration of NaCl in the plants grown in salt-impacted soils ranged from 24 – 83 g/kg. There was no evidence of an increase in NaCl concentrations of plant tissue by PGPR and/or AMF treatments. In addition, to determine the importance of nutrient addition to research sites, liquid fertilizer was applied to 2-week old plants. Results demonstrated that fertilizer effectively increased biomass, and more importantly the biomass of PGPR treated plants supplied with fertilizer was approximately 20% higher than that of plants treated with fertilizer alone. Therefore, research sites were then amended with compost before planting of the 2007 field trial. Plant growth promotion by UW3+4 and CMH3 was tested in the summer of 2007 in the field. Prior to planting, soils were sampled from each site for soil salinity analysis. Barley, oats, tall fescue and ryegrass treated with and without PGPR were sown in plots. The plant coverage condition, NaCl concentrations and biomass of plant shoots were assessed to evaluate the PGPR effect. The results showed that PGPR promoted shoot dry weight by 30% - 175%. The NaCl concentrations of barley, oats and tall fescue averaged 53 g/kg, 66 g/kg and 35 g/kg, respectively. There was no evidence of an increase in NaCl concentrations of plant tissue by PGPR in the field. The salt removal of the CMN site was the highest among three sites due to the large amount of shoot biomass produced. The amount of salt accumulated in the shoots on the CMN site is estimated to be 1580 kg per hectare per year when both barley and ryegrass are planted together as a mix and treated with PGPR. Based on the field data, the estimated time required to remove 50% salt in the top 50 cm soil is seven years with PGPR treatments, while it takes fifteen years to do so without PGPR. In conclusion, PGPR-promoted phytoremediation was proven to be a feasible and effective remediation technique for soils with moderate salinity.

Master of Science / Department of Agronomy / Charles W. Rice / Maize (Zea mays L.) is a major staple crop whose wild ancestor was domesticated about 9,000 years ago (Beadle, 1939). Long-term breeding for more desirable traits ultimately resulted in the maize we see today. This long-term breeding likely impacted the processes within the rhizosphere of maize, however, to what extent is not well understood. This study examined the microbial communities between an inbred maize line (B73), a hybrid of two isogenic lines (B73xMo17), and two genetically modified maize hybrids (DKC63-55RIB and DKC64-69RIB) to determine if the plant’s ability to attract beneficial microbes changed with breeding. The hypothesis was that the isogenic cultivar forms better relationships with bacteria and fungi compared to the newer cultivars, especially in low P soil. It was also expected that the greater the difference between the cultivars the more distinct their soil microbiome. To test these hypotheses, experiments were conducted under greenhouse and field conditions. Analyses consisted of root staining to test symbiotic relationships, phospholipid fatty acid analysis (PLFA) for microbial communities, total plant and root biomass, and nutrient content to understand plant responses. Based on the field results, there was no impact on root and shoot biomass and nutrient content by differences in cultivar. Differences in cultivar did impact arbuscular mycorrhizal fungi (AMF) colonization, which decreased over time and depth for all. Soil AMF also saw a significant effect by cultivar. Other microbial groups were not impacted by cultivar, were greatest in the control, and decreased over time. Greenhouse results showed a cultivar by time interaction for root and shoot biomass. Soil P also impacted shoot biomass, but not root biomass. Shoot nutrient content was greater in high P soil, while roots only saw an impact for root P. No cultivar effect was found for soil microbial groups except for fungi, while all microbial groups were reduced in the control soil. Most soil microbial groups were also impacted by soil P as indicated by reduced concentrations in low P soil. AMF was the only microbial group that was not negatively impacted by limited soil P. In addition, all soil microbial groups increased over time, although fungi saw a decrease at R1. No significances were observed for percent AMF colonization.

No description available.

Arbuscular mycorrhizal fungi (AMF) are obligate plant symbionts that colonize the roots of the majority of vascular land plants. These fungi have a unique nuclear organization, in which thousands of nuclei co-exist among an unsegmented fungal body. In individual strains these nuclei can all be genetically similar (homokaryotic) or be derived from two distinct parents (dikaryotic). In other fungal groups the presence of two distinct nuclei in one cell (fungal dikaryons) can change their fitness, function, and symbiotic relationship; begging the question, what impact does the presence of two parental genotypes have on the arbuscular mycorrhizal symbiosis? I am investigating this by measuring the mycorrhizal response (MR) of potato cultivars with different degrees of domestication using representative AMF homokaryons (4) and AMF dikaryons (4). I found that the genetic organization (dikaryotic vs homokaryotic) and domestication status of the host (modern vs old) are both significant factors in the mycorrhizal response of host plants. Specifically, biomass is significantly greater when inoculated with homokaryotic AMF compared to dikaryotic AMF. Dikaryotic strains have low arbuscule colonization in modern cultivars and higher in old, although there are not significant differences in other fungal responses between homokaryotic and dikaryotic AMF. Furthermore, nutrient uptake (N and P) is greater in old cultivars than modern cultivars, although the root:shoot ratio is lower in old cultivars. Analyses of single spores using digital droplet PCR (ddPCR) confirm that nucleotype ratio of dikaryotic spores shifts depending on the host identity. This research provides novel insights into the role of AMF genetic organization in the mycorrhizal symbiosis in greenhouse conditions. In particular, this work shows that the presence of two distinct nucleotypes results in the fungi being more readily adaptable to the host leading to a more stable MR and a potentially selfish strategy, when in symbiosis with potato cultivars.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Arbuscular mycorrhizal fungi (AMF) are widespread soil dwelling microorganisms that associate with plant hosts. AMF receive carbon from the host as a result of the mutualism, while the plant’s ability to acquire nutrients is enhanced by AMF. Additionally, AMF benefit their host in the form of pathogen protection. While it is known that increased AMF species richness positively correlates with aboveground plant productivity, the relationship between AMF diversity and pathogen protection is not well understood. In a growth chamber study, the plant host Leucanthemum vulgare, a non-native plant species in North America, was introduced to all combinations of three AMF species either in the presence or absence of the plant root pathogen Rhizoctonia solani. In the presence of the pathogen, the plant host increased its dependence on the AMF symbiosis. However, the richest AMF species assemblage did not provide the greatest pathogen protection. Understanding how diverse groups of AMF protect plants from pathogen attack provides insight into how plant communities are formed and structured. / NSERC