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Manipulating The Root Mycobiome To Improve Plant Performance And Reduce Pathogen Pressure In Corn (Zea Mays), Noor F. Saeed Cheema

Electronic Thesis and Dissertation Repository

Crop yield often varies within a field of a single genetically uniform crop plant, with the causes presumed to be a mix of both biotic and abiotic factors. Manipulating crop root mycobiomes could potentially increase yield by reducing pathogen impacts and improving access to soil water and nutrients. This study aimed to identify different fungal inoculation treatments that could increase the growth of corn seedlings sown in low productivity soils to that in high productivity soils and shift the root mycobiome composition. Fungal inoculation treatments did not have significantly different root mycobiome composition than seedlings grown in low yield control …

Developing A Toolbox To Engineer Quantitative Trait Variation In Legume Species Using Crispr/Cas Technologies., Petar Miletic

Electronic Thesis and Dissertation Repository

The impact of intensive agriculture on the environment is immense. This is especially dire with regard to the natural nitrogen (N) cycle, where the human driven interference, primarily associated with industrial fertilization, has reached unsustainable levels. Unlike cereals, legumes, such as soybean, alfalfa and common bean, have the ability to use atmospheric nitrogen, which limits the need for industrial fertilization. A more wide-spread use of legumes could alleviate some of the negative impacts on the biogeochemical cycle while also providing a useful alternative to meat consumption, an important factor in sustainability. To reach this goal, further improvements of legume crops …

Establishment Of A Pesticide Resistance Monitoring Tool For The Two-Spotted Spider Mite, Tetranychus Urticae, Hanna Varonina

Electronic Thesis and Dissertation Repository

Two-spotted spider mite (TSSM), or Tetranychus urticae (Koch), is a major agriculture pest known for its rapid development of resistance to pesticides. The analysis of spider mites’ pesticide resistance demonstrated that resistance patterns and frequencies vary between T. urticae strains collected from different geographic locations and host plants. This research aims at characterization of pesticide resistance patterns in mite populations present in greenhouses in the Southwestern Ontario by identifying genetic and metabolic markers of their pesticide resistance. The establishment of these markers lays the basis for development of a pesticide resistance diagnostic tool that will enable prediction of population resistance …

The Analysis Of Ginsenosides In Ginseng Garden Soil, Karina Marie Kaberi

Electronic Thesis and Dissertation Repository

American ginseng (Panax quinquefolius L.) produces natural products called ginsenosides. The biggest challenge Ontario commercial ginseng farmers face is ginseng replant disease. To understand the function of ginseng root exudates, ginsenoside accumulation and persistence over time were investigated. Currently, no reliable ginsenoside specific extraction method, characterizing the changes in soil chemistry exists. Ginsenoside extraction protocol optimization was required to determine how ginsenoside composition changed over time. Overall, protocol optimization resulted in a 30% increase in yield of ginsenosides compared to previous extraction protocols. In the ginseng gardens, ginsenoside accumulation occurs slowly and did not reach significantly measurable amounts until the …

Dynamics Of The Soil Microbiome In Ginseng Gardens, Megan E. Lambert

Electronic Thesis and Dissertation Repository

Ginseng Replant Disease (GRD) is a syndrome in which ginseng cannot be cultivated in soil previously used to grow ginseng. Since GRD can persist for decades, it severely impacts the Ontario ginseng industry. To better understand the origin of GRD, the impact of ginseng cultivation on the soil microbiome was investigated in bulk soil of three newly planted ginseng gardens in Ontario, from seeding through two years of cultivation. While specific trends in species richness, diversity and composition were unclear, PERMANOVA analyses confirmed that they changed over time. Known GRD-related pathogens, including Ilyonectria mors-panacis and Fusarium oxysporum, were detected in …

Molecular Identification And Characterization Of Viral Pathogens Infecting Sweet Cherry, Aaron J. Simkovich

Electronic Thesis and Dissertation Repository

Stone fruits are a valuable crop grown worldwide, however pathogens such as viruses threaten fruit production by reducing tree health and fruit yield. In an orchard within the Niagara region of Ontario, symptoms typical of viral infection such as chlorosis and leaf deformation were seen on sweet cherry (Prunus avium L.) trees. Next generation sequencing was performed on symptomatic and asymptomatic leaves and four viruses were identified. On the tree displaying the most severe symptoms, Prune dwarf virus (PDV), was the only virus detected. A survey conducted during this work showed 42% of cherry trees on a single …

Bioremediation Of Refinery Desalter Effluent Using Debaryomyces Hansenii And Parachlorella Kessleri, Leila Azimian

Electronic Thesis and Dissertation Repository

Crude oil desalting operations produce an effluent stream which is challenging to treat due to its salt, heavy metal and hydrocarbon content. Consequently, desalter effluent (DE) is usually diluted into other effluent streams and sent to conventional wastewater treatment plants which may lead to upsets the plant operation.

In this study, a novel microbial approach was applied which investigated DE treatment using halotolerant yeast Debaryomyces hansenii (LAF-3 10 U) or the environmentally robust micro-algae Parachlorella kessleri strain CPCC 266. The effect of these two different approaches on both synthetic and actual DE was investigated in both batch and/or continuous mode. …

Cultivating Knowledge: Agrarian Science And Ecological Engagements In Southern Ontario Agriculture, Kelly Linton

Electronic Thesis and Dissertation Repository

Agriculture and climate change are interconnected processes, with agriculture implicated in rising green house gas emissions, deforestation, soil and water pollution, and reductions in biodiversity. Conversely, changes within ecology (including a warming climate), alter growing conditions for farmers. Farmers face changes in both temperature and precipitation, as well as an increase in adverse weather events that significantly threaten productivity and livelihoods.

Based on 40 unstructured interviews as well as informal conversations conducted among farmers in southern Ontario, Canada between the spring of 2014 and the winter of 2017, this dissertation seeks to contribute to a growing body of work that …

Arabidopsis Tt8 Orthologue Of Pinto Bean (Phaseolus Vulgaris L.) Regulates Proanthocyanidin Genes And Seed Coat Darkening, Nishat Shayala Islam

Electronic Thesis and Dissertation Repository

Postharvest darkening of seed coat in pinto bean (Phaseolus vulgaris L.) is an undesirable trait that hinders its market potential. Darkening is more rapid in the cultivars like CDC Pintium than the newly developed slow darkening cultivar 1533-15. A single gene, SLOW DARKENING (Sd), is responsible for the slow darkening in pinto beans and the trait co-segregates with two simple sequence repeat (SSR) markers. The objective of this research is to identify and characterize the Sd gene to understand the slow darkening mechanism in pinto bean seed coat. A search for Sd within the linkage distance from …

Rnai Targeting Of Manduca Sexta Using Chloroplast-Encoded Long Dsrna, William Burke

Electronic Thesis and Dissertation Repository

Pest management strategies are crucial for reducing insect crop damage, and methods with species-specific effect are particularly desirable. Expressing dsRNA with sequence complementarity to a vital gene of feeding pests is a strategy that has shown potential. I modified the chloroplast genome of cultivated tobacco, Nicotiana tabacum, by adding a sequence encoding a 2222 base pair dsRNA complementary to the vacuolar-type H⁺-ATPase subunit A mRNA of Manduca sexta and demonstrated expression of the product in leaves of the plants. Although feeding this same dsRNA that was synthesized in vitro to larvae knocked down expression of the target …

Measurement And Protection Of Lung Health In Poultry Farmers Of Southwestern Ontario, Rose-Marie Doyon Dolinar

Electronic Thesis and Dissertation Repository

Chronic obstructive lung disease (COPD), develops gradually, and tends to be noticed by farmers only after there has been a significant loss of lung function. People with symptoms of COPD often present for health care in later stages of disease, because they accept cough or breathlessness as “normal”.

Measurement of lung health for those at risk of developing COPD provides an opportunity for early detection of lung disease, and deter the progression toward irreversible damage to the lungs. COPD is, however, under-diagnosed.

The importance of bringing primary care providers and farmers together, and acknowledging the high-risk nature of occupational exposures, …

Production Of A Candidate Recombinant Protein Vaccine For Mannheimia Haemolytica In Lettuce And Tobacco Chloroplasts, Coby K. Martin

Electronic Thesis and Dissertation Repository

The cattle industry worldwide is ravaged by bovine respiratory disease (BRD), a bacterial disease caused by Mannheimia haemolytica. Recent efforts to design vaccines against M. haemolytica focus on a virulence factor, leukotoxin, in addition to surface lipoproteins. Plant-based protein production is a safe and inexpensive alternative to traditional methods. Edible vaccines deliver antigens to pharyngeal tissues, which can provide local immunization against M. haemolytica prior to its progression into the lungs. In this project, a chimeric protein containing M. haemolytica antigens was produced in tobacco chloroplasts as a candidate edible vaccine for BRD. Attempts were made to transform lettuce …

Evaluating Plant Root Uptake Of Dsrna For Application In Pest Management, Kaitlyn Ludba

Electronic Thesis and Dissertation Repository

The greenhouse whitefly Trialeurodes vaporariorum is an agricultural pest that damages crops by feeding on plant sap and by vectoring plant viruses. Control of whiteflies has been managed through application of insecticides, but this strategy is not entirely effective and alternative control strategies are needed. In this thesis, I evaluated the efficacy of RNA interference as a means to control whiteflies on greenhouse-grown tomatoes. I found that root uptake of dsRNA synthesized from the v-ATPase subunit A gene caused significant gene knock-down and mortality in feeding whiteflies. This effect was, however, sensitive to the concentration of dsRNA delivery, and concentration …

Gatekeepers Of Nitrogen-Fixing Symbiosis: Cytokinin-Ethylene Crosstalk Regulates Symbiotic Interaction In Lotus Japonicus, Seyedehmandana Miri

Electronic Thesis and Dissertation Repository

Leguminous plants thrive under nitrogen-limited soil conditions because of their ability to symbiotically interact with nitrogen-fixing bacteria, known as rhizobia. In the presence of compatible strains of rhizobia, they develop specialized symbiotic organs, called root nodules, which host the bacteria and provide the appropriate conditions for symbiotic nitrogen fixation to occur. The plant hormone cytokinin is the key endogenous trigger for the inception of root nodule organogenesis. In the model legume Lotus japonicus, analysis of the cytokinin receptor gene Lotus histidine kinase 1 (Lhk1) showed that it is required and also sufficient for the initiation of nodule …

Multi-Gene Resistance To Neonicotinoids In The Colorado Potato Beetle, Leptinotarsa Decemlineata, Emine Kaplanoglu

Electronic Thesis and Dissertation Repository

The Colorado potato beetle, Leptinotarsa decemlineata, is a significant pest of potato, and its impact on agriculture is measured on a global scale. The beetle is mainly controlled by neonicotinoid insecticides, however, resistance development is a growing concern. Resistance to neonicotinoids is thought to involve elevated activity of detoxifying enzymes and xenobiotic transporters that break-down and excrete insecticide molecules. Here, using mRNA sequencing, I identified multiple detoxifying enzyme and xenobiotic transporter genes transcriptionally up-regulated in a neonicotinoid resistant strain of beetles. I then used RNA interference to knock down the transcript levels of the ten most promising genes in …

A Systems Approach To Modelling The Effects Of Climate Change On Agroforestry: A Case Study In Western Tanzania, Elaine M. Samuel

Electronic Thesis and Dissertation Repository

Climate change is anticipated to have significant effects on agricultural production in sub-Saharan Africa as the magnitude of weather events increase in severity. Smallholder farmers in western Tanzania are potentially vulnerable to climate change impact as crops rely on precipitation as the only source of water. It is prudent to evaluate different modes of agricultural adaptations, such as agroforestry, that these farmers can easily adopt to improve their resiliency to the effects of climate change. System dynamics modelling is a cost-effective tool to simulate the long-term behaviour of agroforestry systems under future climate conditions. Water, Nutrient, and Light Capture in …

Planting The Chalcone Reductase Family Tree: Identification And Characterization Of Chalcone Reductase Genes In Soybean, Caroline Julia Sepiol

Electronic Thesis and Dissertation Repository

Soybean (Glycine max [L.] Merr) is an important crop grown in Canada, generating $2.4 billion in sales. Though this number may be promising, soybean farmers lose about $50 million worth of yield annually due to root and stem rot disease caused by Phytophthora sojae. Many strategies have been developed to combat the infection; however, these methods are prohibitively expensive. A ‘cost effective’ approach to this problem is to select a trait naturally found in soybean that can increase resistance. One such trait is the increased production of root glyceollins. One of the key enzymes exclusively involved in glyceollin …

Ginsenosides, Glycosidases And The Ginseng-Pythium Interaction, Dimitre A. Ivanov

Electronic Thesis and Dissertation Repository

Ginsenosides, the triterpenoid saponins produced by American ginseng (Panax quinquefolius L.), have been extensively studied for their medicinal value, however, their function in the rhizosphere remains largely unknown. Like other saponins, ginsenosides possess mild fungitoxic activity toward some common ginseng pathogens. However, numerous oomycete root pathogens of ginseng, most notably Pythium irregulare Buisman, are able to partially deglycosylate the 20 (S)-protopanaxadiol ginsenosides Rb1, Rd and gypenoside XVII via extracellular glycosidases (ginsenosidases), leading to the formation of a common product, ginsenoside F2. In this thesis the potential role(s) of these extracellular ginsenosidases and the ginsenoside products they produce (ie. ginsenoside …

Identification And Characterization Of Cysteine Protease Genes In Tobacco For Use In Recombinant Protein Production, Kishor Duwadi

Electronic Thesis and Dissertation Repository

Plants are an attractive host system for pharmaceutical protein production. Many therapeutic proteins have been produced and scaled up in plants at a low cost compared to the conventional microbial and animal based systems. The main technical challenge during this process is to produce sufficient level of proteins in plants. Low yield is generally caused by proteolytic degradation during expression and downstream processing of recombinant proteins. The yield of a human therapeutic protein interleukin (IL) -10 produced in transgenic tobacco leaves was found to be below the critical level, and is potentially due to degradation by tobacco cysteine proteases (CysPs). …

Two Spotted Spider Mite (Tetranychus Urticae) Selection To Arabidopsis Thaliana, Huzefa Ratlamwala

Electronic Thesis and Dissertation Repository

Spider mite feeding on A. thaliana induces the production of indole glucosinolates (IGs), plant secondary metabolites that negatively affect mite performance. In this study I conducted selection experiments on A. thaliana with varying levels of IGs, to determine if mites could adapt to IGs and other defense compounds. After 12 months, mites reared on host with IGs performed significantly better on A. thaliana than mites maintained on beans. However, an adaptation cost was detected between selected mite lines and their ancestral host. The qRT-PCR data on different mite lines revealed that the detoxification genes previously identified may only be involved …

Heterologous Production And Characterization Of Cell Wall Degrading Enzymes Using Plants As A Bioreactor, Eridan Orlando Rodrigues Pereira

Electronic Thesis and Dissertation Repository

Plants are wonderful living organisms. They are able to store solar energy into carbohydrates by fixing CO₂ through photosynthesis which can be subsequently harvested and used for fuel production. However, one of the major limitations for transforming these carbohydrates into liquid fuels is the recalcitrance of the plant cell wall. Although microorganisms have evolved a series of cell wall degrading enzymes to harvest efficiently this energy and are considered the main source of these biocatalysts, harnessing these microorganisms for the production of enzymes is a costly process and a major factor limiting the commercialization of lignocellulosic biomass-to-ethanol processes. The …

Uncovering The Molecular Link Between Mir156.Spl15 And Carotenoid Accumulation In Arabidopsis, Davood Emami Meybodi

Electronic Thesis and Dissertation Repository

Carotenoid Cleavage Dioxygenases (CCDs) are an enzyme family that cleaves specific double bonds in carotenoids. MicroR156 in Arabidopsis regulates a network of genes by repressing 10 SPL genes, among which, SPL15 was found to regulate shoot branching and carotenoid accumulation. The expression of CCD1, CCD4, CCD7, CCD8, NCED2, NCED3, NCED5, NCED6, NCED9 and SPL15 was evaluated in siliques at 10 days post anthesis and in 10-day-old roots in Arabidopsis wild type, sk156 (miR156 overexpression mutant), RS105 (miR156 overexpression line), spl15 (SPL15 knockout mutant) and two 35S:SPL15 lines. Results showed that most of CCD/NCED genes were …

Modeling Leafhopper Populations And Their Role In Transmitting Plant Diseases., Ji Ruan

Electronic Thesis and Dissertation Repository

This M.Sc. thesis focuses on the interactions between crops and leafhoppers.

Firstly, a general delay differential equations system is proposed, based on the infection age structure, to investigate disease dynamics when disease latencies are considered. To further the understanding on the subject, a specific model is then introduced. The basic reproduction numbers $\cR_0$ and $\cR_1$ are identified and their threshold properties are discussed. When $\cR_0 < 1$, the insect-free equilibrium is globally asymptotically stable. When $\cR_0 > 1$ and $\cR_1 < 1$, the disease-free equilibrium exists and is locally asymptotically stable. When $\cR_1>1$, the disease will persist.

Secondly, we derive another general delay differential equations system to examine how different life stages of leafhoppers affect crops. The basic reproduction numbers $\cR_0$ is determined: when …

Epigenetics Of Stress Adaptation In Arabidopsis: The Case Of Histone Modifications, Behnaz Najafi Majdabadi Farahani

Electronic Thesis and Dissertation Repository

Changes in the epigenetic status of plants, which contribute to changes in gene expression, in response to abiotic stress are well documented. However, their stability and transmission to subsequent generations, and their incorporation into plant stress adaptation are still a matter for debate. Using chromatin immuno-precipitation and Next Generation Sequencing (ChIP-seq), we compared genome wide enrichment of two histone marks, H3K9ac and H3K4me2, in the progeny of salt stressed and control Arabidopsis plants. Data showed less enrichment of the H3K9ac in the chromatin of the progeny of salt stressed plants, but no changes were detected in the enrichment of the …

Cadmium Accumulation And Distribution In Lettuce And Barley, Fardausi Akhter

Electronic Thesis and Dissertation Repository

Cadmium (Cd) is a non-essential trace element and its environmental concentrations are increasing due to human activities. Edible plants can accumulate high concentrations of Cd, which could be toxic to humans. Understanding how and where Cd is stored in plants is important for ensuring lower concentration of Cd in the food. In this thesis, the accumulation and distribution of Cd in three agricultural plants, namely lettuce (Lactuca sativa L.), barley (Hordeum vulgare L.) and radish (Raphanus sativus L.), were investigated with a focus on the potential mechanisms involved in the localization of Cd in the root. The …

Protein Body Formation In Stable Transgenic Plants Of Nicotiana Tabacum Expressing Elastin-Like Polypeptide And Hydrophobin Fusion Proteins, Sonia P. Gutierrez

Electronic Thesis and Dissertation Repository

Plants are recognized as an efficient and inexpensive system to produce valuable recombinant proteins. However, the use of plants still faces two main limitations: the low accumulation levels of some recombinant proteins and the lack of efficient protein purification methods. Two fusion partners, elastin-like polypeptides (ELP) and hydrophobin I (HFBI) were found to increase the accumulation of recombinant proteins and induce the formation of protein bodies (PBs) when targeted to the ER in transient expression assays. In this study I examined the effect of these tags in stable transgenic plants of two Nicotiana tabacum cultivars when fused to green fluorescent …

Impacts Of Agricultural Disturbance On Communities Of Selected Soil Fungi (Agaricomycetes), Jessie R. Wong

Electronic Thesis and Dissertation Repository

The objective of this study was to use phylogeny-based and community-based analyses to compare the community composition of Agaricomycetes among four different agricultural treatments at the Kellogg Biological Station Long Term Ecological Research (KBS LTER) site. A phylogenetic tree that included 591 ribosomal DNA sequences previously obtained from KBS LTER documented the composition of Agaricomycete communities in each treatment. Sequences from KBS LTER were placed into 472 OTUs (putatively species-level operational taxonomic units defined by 99% or greater sequence similarity) and these were dominated by the Agaricales (with 330 OTUs), Cantharellales (39 OTUs), Hymenochaetales (29 OTUs), and Polyporales (23 OTUs). …

Hairy Roots As A Model To Investigate The Role Of Suberin In The Phytophthora Sojae-Soybean Pathosystem, Pooja Sharma

Electronic Thesis and Dissertation Repository

Part of the resistance mechanism of soybean (Glycine max [L.] Merr.) to Phytopthora sojae Kauf. & Gerd. involves pre-formed root suberin. In order to investigate the role of suberin in this host-pathogen interaction, I characterized hairy roots, formed as a result of Agrobacterium rhizogenes (Riker et al.) Conn infection, as a model to be used as a reliable soybean transformation system. I established hairy root cultures and demonstrated that they were a result of A. rhizogenes infection. The anatomy and suberin deposition in soybean hairy roots was examined, and found to be very similar to that of wild-type roots. …