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Identification And Characterization Of Isoflavone Reductase Family Members In Soybean, Negin Azizkhani

Electronic Thesis and Dissertation Repository

Soybean’s yield is threatened by Phytophthora sojae, a pathogen responsible for stem and root rot disease. Glyceollins, unique antimicrobial agents specific to soybeans in partially preventing P. sojae infection, are derived from the isoflavonoid branch of the general phenylpropanoid pathway. One pivotal enzyme exclusively involved in glyceollin synthesis in soybean is the isoflavone reductase (GmIFR), which catalyzes the 2'-hydroxydaidzein conversion to 2'-hydroxy-2,3-dihydrodaidzein as a precursor for glyceollin biosynthesis. To comprehensively identify all members of the GmIFR gene family within the soybean genome, keyword and blast protein searches were conducted, identifying 98 putative GmIFRs. Among these candidates, …

Nitrogen Transfer From Cover Crops To The Subsequent Grain Crop And The Influence Of Variability In Winter Conditions, Spencer Heuchan

Electronic Thesis and Dissertation Repository

Cover crops, which are used to provide ground cover after the harvest of the grain crop, can potentially improve the sustainability of agroecosystems by reducing nutrient losses. However, few data are available to document the extent to which cover crops improve both the retention of soil nitrogen (N) and the transfer of this N to the grain crop. The efficiency of this N transfer may be further influenced by variation in winter soil temperature; for example, reduced snow cover can increase the frequency and intensity of soil freezing, which can affect the survival of cover crops and the timing of …

Cadmium Uptake In Plants As Influenced By Selenium Uptake And Sulphate Availability, Marnie J. Demand

Electronic Thesis and Dissertation Repository

With cadmium uptake by plants posing a risk to plants and consumers alike, strategies to reduce metal uptake are desirable. One strategy may be to apply selenium (as selenate) to the growth medium. I hypothesized selenate would yield greater lignification, with a higher proportion of cadmium bound to root cell walls. Consequently, higher selenium in plants would result in greater tolerance to cadmium. Additionally, since selenate is taken up in place of sulphate, providing the plants with high sulphate would inhibit uptake and translocation of selenium, mitigating selenate’s benefits of reducing cadmium uptake and translocation. Experimental results did not support …

The Biosynthesis Of Non-Protein Sulfur Amino Acid In Developing Seeds Of Common Bean, Zixuan Lu

Electronic Thesis and Dissertation Repository

The protein quality of common bean (Phaseolus vulgaris) is associated with the level of dietary essential sulphur amino acids – methionine and cysteine. Extra sulphur that cannot be stored in the protein pool accumulates as the non-protein amino acid S-methylcysteine (S-methylCys) and its dipeptide γ-glutamyl-S-methylcysteine (γ-Glu-S-methylCys). Previous studies have indicated that S-methylhomoglutathione (S-methylhGSH) is present in the developing seed of common bean. It is hypothesized that S-methylhGSH is the key intermediate in the biosynthetic pathway of the γ-Glu-S-methylCys that leads to …

Long-Term Vs. Short-Term Plant Responses To Warming And Nitrogen Addition In A Temperate Old Field, Breanna Craig

Electronic Thesis and Dissertation Repository

Climate warming and atmospheric nitrogen (N) deposition are expected to have strong effects on plant productivity in temperate ecosystems over the next century. However, short-term experiments may not adequately address their potential effects because of lags in changes in species composition. I added new plots to a pre-existing field experiment to compare the short-term (1-2 year; new plots) vs. long-term (14-15 year; old plots) effects of warming and N addition on plant productivity, relative species abundances, plant tissue N content, and litter decomposition. In 2020, N addition increased aboveground plant productivity most in old plots and only increased belowground biomass …

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 …

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 …

Floral Traits And Carbon Dynamics Of Cucumber In Response To Climate Change, Sarah Josina Mcdonald

Electronic Thesis and Dissertation Repository

Pollination by animals is an important ecosystem service that contributes to the reproduction of many angiosperms. Climate change may alter this mutualism by affecting floral traits that are important to pollinators. Using Cucumis sativus, I tested the effects of experimentally elevated temperature and CO₂ concentration on flowering onset, flower number, flower size, and floral rewards. Additionally, to better understand plant carbon balance and investment in reproduction, I measured biomass partitioning and leaf carbon fluxes of plants under their growth conditions. Carbon dynamics were similar across treatments, and plants grown under high [CO₂] and temperature showed similar …

Investigating The Role Of Carbon Stress In The Mortality Of Tamarack Seedlings Under A Warming Environment, Bridget K. Murphy

Electronic Thesis and Dissertation Repository

Climate warming is increasing the frequency of climate-induced tree mortality events. While drought combined with heat is considered the primary cause of this tree mortality, little is known about whether high temperatures alone can induce mortality, or whether rising CO₂ will increase survival. I grew tamarack in two experiments combining warming (0-8 ˚C above ambient) and CO₂ (400-750 ppm) to investigate whether high growth temperatures led to carbon limitations and mortality. Using glasshouses, +8 ˚C warming with ambient CO₂ (8TAC) led to 40% mortality despite thermal acclimation of respiration. Dying 8TAC seedlings had lower needle carbon concentrations …

Soybean Root Exudates Increase The Physiological Diversity Of Bacteria In Cadmium-Treated Soil, Leily Kazemi Movahed

Electronic Thesis and Dissertation Repository

Three soybean cultivars with contrasting retention of cadmium (Cd) in the root were grown in Cd-spiked nutrient solution and used to determine that symplastic compartmentalization of Cd in roots is probably responsible for retention of Cd in roots. Roots of the low Cd-accumulator AC Hime treated with 30 mM Cd exuded up to 10-fold higher concentrations of citric, succinic, fumaric and malic acids into the hydroponic solution when compared to control; concentrations of the same organic acids from the high Cd-accumulator Westag 97 increased by up to 3-fold. The same cultivars were grown in Cd-spiked soil and the physiological profiles …

Microrna156: A Short Rna With A Major Role In Abiotic Stress Tolerance In Alfalfa, Biruk Ayenew Feyissa

Electronic Thesis and Dissertation Repository

The highly conserved plant microRNA156, miR156, affects various aspects of plant development and stress response by silencing SQUAMOSA-PROMOTER BINDING PROTEIN-LIKE (SPL) transcription factors. Our understanding of the role of miR156 and its mode of action in alfalfa’s (Medicago sativa L.) response to drought and flooding is still elusive, and thus this study was aimed at filling this gap in knowledge. Physiological parameters, metabolite and transcriptional analyses showed an interplay between miR156/SPL13 and WD40-1/DFR to mitigate drought stress. Low to moderate levels of miR156 overexpression suppressed SPL13 and increased WD40-1 to fine-tune the DIHYDROFLAVONOL-4-REDUCTASE (DFR) level for …

Effects Of Elevated Temperature And Elevated Co2 On Leaf Carbon Fluxes In Boreal Conifers: Lab And Field Studies, Mirindi Eric Dusenge

Electronic Thesis and Dissertation Repository

Rising CO₂ may warm northern latitudes up to 10 °C by the end of the century. However, responses of plant physiological processes (such as photosynthesis and respiration) and growth to climate change remain uncertain. Seedlings and mature trees of tamarack (a deciduous species) and black spruce (an evergreen species), North America dominant conifers, were exposed to combined warming (up to +9 ˚C) and elevated CO₂ (up to +300 ppm). In seedlings, stomatal conductance (g_s) tended to increase with warming in tamarack seedlings, while g_sdeclined with warming in spruce. In both species, CO₂ had …

Genome-Wide Occupancy Of Polycomb Group Proteins And Chromatin Remodeler Splayed And Their Interplay In Arabidopsis, Jie Shu

Electronic Thesis and Dissertation Repository

The Polycomb group (PcG) proteins form two protein complexes, Polycomb repressive complex 1 (PRC1) and PRC2, which are key epigenetic regulators in eukaryotes. PRC2 represses gene expression by catalyzing trimethylation of histone H3 lysine 27 (H3K27me3). In Arabidopsis thaliana, CURLY LEAF (CLF) and SWINGER (SWN) are two major H3K27 methyltransferases, playing essential roles in plant growth and development. Despite their importance, genome-wide occupancy profiles of CLF and SWN have not yet been determined and compared. In this thesis, I generated transgenic lines expressing GFP-tagged CLF/SWN and determined the genome-wide distributions of CLF and SWN in Arabidopsis seedlings. I also …

Carbon And Nitrogen Dynamics In Plants Grown At Low Co2 Conditions Of The Past, Andre Galvao Duarte

Electronic Thesis and Dissertation Repository

For most of the recent evolutionary history of plants, atmospheric CO₂ concentrations have been far below modern values. However, little is known about how plants grown in a low CO₂ environment balance their carbon demand for growth while meeting their requirement for nitrogen acquisition, the establishment of mycorrhizal fungal associations, and the production of defense compounds. Here, I investigated how low CO₂ affects Elymus canadensis and Picea mariana by comparing their growth at low and current CO₂ concentrations. I found that reduced N availability exacerbated low CO₂ effects on growth, and reduced stomatal index and …

Identifying Brachypodium Distachyon Proteins Interacting With Histone Deacetylase Bdhd1, Alberto Giovanni Torrez

Electronic Thesis and Dissertation Repository

Current evidence has revealed the involvement of epigenetic mechanisms, including histone deacetylases (HDACs), in plant stress responses. In Arabidopsis thaliana, HDA19, belonging to the RPD3/HDA1 class, interacts with transcription factors to form repressor complexes. HDAC research mainly exists for dicotyledons, whereas research on monocotyledons is limited. Brachypodium distachyon is used as a model plant to investigate questions unique to monocot crops. BdHD1 is the closest homologous gene to HDA19 in B. distachyon. This study investigated potential protein-protein interactions between BdHD1 and each of BdMYB22, BdWRKY24, and BdHOS15. Protein interactions were investigated using yeast two-hybrid (Y2H) and bimolecular fluorescence …

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Partial Characterization Of Putative Cyp86a Genes From Soybean, Trish Tully

Electronic Thesis and Dissertation Repository

Soybean (Glycine max [L.] Merr.) is a globally cultivated crop that is important to the sustainability of many industries. However, like all plants, optimal cultivation of soybean is threatened by detrimental environmental factors. For example, high yield of soybean is threatened by soil-borne pathogens like Phytophthora sojae. Resistance against P. sojae was previously positively correlated with aliphatic suberin deposition in soybean. As such, a deeper understanding of the biosynthesis of suberin may assist in engineering a resistant form of soybean, based on enhanced suberin content. In soybean, the ω-OH fatty acid monomers are predominant and most strongly correlated …

Quantitative Proteome Analysis Of Alfalfa In Drought Stress Under The Influence Of Mir156, Alpa Puri

Electronic Thesis and Dissertation Repository

Medicago sativa is one of the most produced perennial forage crops in Canada. Drought stress is a major form of abiotic stress affecting the productivity and annual yield of alfalfa. A small noncoding RNA, miR156 has a role in abiotic stress tolerance, while regulating downstream SPL genes. In this study, whole leaf proteome and amino acid concentrations of miR156OE (A8) genotype of alfalfa under drought stress were quantified. In results, 3,000 proteins groups were identified using MaxQuant when searched against Medicago truncatula protein sequence database. Gene ontology analysis showed enriched proteins were involved in biological and molecular processes such as …

Suberin Biosynthesis And Deposition In The Wound-Healing Potato (Solanum Tuberosum L.) Tuber Model, Kathlyn Natalie Woolfson

Electronic Thesis and Dissertation Repository

Suberin is a heteropolymer comprising a cell wall-bound poly(phenolic) domain (SPPD) covalently linked to a poly(aliphatic) domain (SPAD) that is deposited between the cell wall and plasma membrane. Potato tuber skin contains suberin to protect against water loss and microbial infection. Wounding triggers suberin biosynthesis in usually non-suberized tuber parenchyma, providing a model system to study suberin production. Spatial and temporal coordination of SPPD and SPAD-related metabolism are required for suberization, as the former is produced soon after wounding, and the latter is synthesized later into wound-healing. Many steps involved in suberin biosynthesis remain uncharacterized, and the mechanism(s) that regulate …

Investigating The Role Of Brachypodium Distachyon Cellulose Synthase 8 In Gluconacetobacter Diazotrophicus Colonization, Xuan Yang

Electronic Thesis and Dissertation Repository

Nitrogen is an essential nutrient for plant growth. Significant amount of nitrogen fertilizer is applied to crop field to maintain high yield. Alternatives to chemical nitrogen fertilizer are needed to reduce the costs of crop production and offset environmental damage. Gluconacetobacter diazotrophicus is a nitrogen fixing bacterium that was originally isolated from sugarcane and has been proposed as a possible biofertilizer for monocot crop production. However, the colonization of G. diazotrophicus in most monocot crops is limited and deep understanding of the response of the host plants to G. diazotrophicus colonization is still lacking. In this study, research was conducted …

Defective Aba-Mediated Sugar Signalling Pathway In An Established Arabidopsis Thaliana Cell Suspension Culture Explains Its Stay-Green Phenotype At High Sugar Concentrations, Avery Mccarthy

Electronic Thesis and Dissertation Repository

An unusual sugar insensitive phenotype was identified in an established cell suspension culture of Arabidopsis thaliana. We characterized the physiology, biochemistry and genetics of the sugar insensitive cell culture, in order to identify factors contributing to the phenotype. Chlorophyll levels of the cell suspension culture were insensitive to high sucrose (6-15% w/v) and maintained a green phenotype. Immunoblotting indicated that levels of key photosynthetic proteins (PsaA, Lhcb2 and Rubisco) increased as a function of external sucrose concentration. The green cell culture was photosynthetically competent based on light-dependent, CO₂-saturated rates of O₂ evolution as well as Fv/Fm …

Characterization Of Acaricide Resistance, Plant-Mediated Rnai Against Two-Spotted Spider Mites (Tetranychus Urticae Koch), And Assessing Off- And Non-Target Effects, Hooman Hosseinzadeh Namin

Electronic Thesis and Dissertation Repository

The two-spotted spider mite (TSSM), Tetranychus urticae (Koch), is one of the most damaging agricultural pests in the world. It feeds on over 150 crops, causing considerable yield losses in greenhouses and agricultural fields. Currently, using synthetic acaricides is the main method to control TSSM. However, it can develop resistance to acaricides with repeated exposure, and typically resistance can occur within two to four years. To understand the underlying mechanisms of spider mite adaptation to acaricides is an essential part of resistance management strategy. The resistance ratio of the pyridaben-selected strain compared with the pre-selection strain was estimated at greater …

Deciphering Sulfur Amino Acid Metabolism In Developing Seeds Of Common Bean, Jaya Joshi

Electronic Thesis and Dissertation Repository

With increasing food insecurity in the populated world, the number of people affected by chronic undernourishment is also increasing. Alone, protein energy malnutrition is linked to 6 million deaths annually. Despite being a good source of protein and dietary fibre, the quality of bean protein is limited because of sub optimal levels of essential sulfur amino acids: methionine and cysteine. Levels of cysteine and methionine in developing seeds have an inverse relationship with the non-protein sulfur amino acid S-methyl-cysteine (S-methylCys) and dipeptide g-glutamyl-S-methyl-cysteine (g-Glu-S-methylCys).

One of the strategies to improve protein quality in …

Genetic Analysis Of Nitrate Transporter 2 Family In Brachypodium Distachyon, With An Essential Role Of Bdnrt2.1 To Nitrogen Use Efficiency, Jiang Wang

Electronic Thesis and Dissertation Repository

Plant agricultural productivity relies heavily on nitrogenous fertilizers. Excess N fertilizer application can often lead to lower nitrogen use efficiency (NUE) along with energy waste and environmental problems. Therefore, understanding and improving NUE in plants are of key importance. The small monocot plant Brachypodium distachyon (Brachypodium) is rapidly emerging as a powerful model system to study questions unique to monocot crops (wheat, maize, rice, etc.).

Here, through an intensive BLAST search, six putative orthologues of the Arabidopsis NRT2 gene family were identified in the fully sequenced Brachypodium genome (Phytozome v11.0), among which I isolated a T-DNA mutant ( …

Developing A Plant Virus-Based Expression System For The Expression Of Vaccines Against Porcine Reproductive And Respiratory Syndrome Virus, Hong Hanh Tran

Electronic Thesis and Dissertation Repository

Virus-based expression systems have been widely exploited for the production of recombinant proteins in plants during the last three decades. Advances in technology have boosted scale-up manufacturing of plant-made pharmaceuticals to high levels, via the complementation of transient expression and viral vectors. This combination allows proteins of interest to be produced in plants within a matter of days and thus, is well suited for the development of plant-made vaccines or therapeutics against emerging infectious diseases and potential bioterrorism agents. Several plant-based products are currently in varying stages of clinical development. To investigate the viability of virus-based expression systems for plant-made …

Gmmyb176 Interactome And Regulation Of Isoflavonoid Biosynthesis In Soybean, Arun Kumaran Anguraj Vadivel

Electronic Thesis and Dissertation Repository

MYB transcription factors are one of the largest transcription factor families characterized in plants. They are classified into four types: R1 MYB, R2R3 MYB, R3 MYB and R4 MYB. GmMYB176 is an R1MYB transcription factor that regulates Chalcone synthase (CHS8) gene expression and isoflavonoid biosynthesis in soybean. Silencing of GmMYB176 suppressed the expression of the GmCHS8 gene and reduced the accumulation of isoflavonoids in soybean hairy roots. However, overexpression of GmMYB176 does not alter either GmCHS8 gene expression or isoflavonoid levels suggesting that GmMYB176 alone is not sufficient for GmCHS8 gene regulation. I hypothesized that GmMYB176 acts cooperatively with another …

The Influence Of Adjacent Forest And Agriculture On Restored Grassland Diversity And Composition, Rachel A. Chambers

Electronic Thesis and Dissertation Repository

Little native grassland remains in North America due to land-use changes. Conversion to agriculture is a common means of loss. This fragmentation creates edges in the landscape and associated edge effects. Grassland plant communities are susceptible to edge effects, directly via dispersal and indirectly through environment. This study took place in Norfolk County, Ontario, Canada, in a landscape of agriculture, forest, rural development, and patches of grassland restored by Nature Conservancy Canada. I examined restored grassland edges bordering forest and crops. An intensive study at a single site identified spatial and soil environmental influences on plant diversity and composition. I …

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 …

Cytosolic Acetyl-Coa Promotes Histone H3 Lysine 27 Acetylation In Arabidopsis, Chen Chen

Electronic Thesis and Dissertation Repository

Acetyl-coenzyme A (acetyl-CoA) serves as a central metabolite in energy metabolism and biosynthesis. High level of acetyl-CoA can fuel the tricarboxylic acid (TCA) cycle to generate energy and store excess energy in fatty acids. Meanwhile, it also provides acetyl groups for protein acetylation, which normally occurs at the lysine or arginine residues. Acetylation regulates protein functions largely due to the change of total charges. Acetylation of histones, for example, can lead to loss of the interaction between histone and DNA, thus relaxing chromatin structure and potentially promoting gene expression. However, whether and how acetyl-CoA regulates plant chromatin remains unexplored. Here, …

Identification And Characterization Of The Isoflavonoid-Specific Prenyltransferase Gene Family To Prevent Stem And Root Rot In Soybean, Arjun Sukumaran

Electronic Thesis and Dissertation Repository

Soybean is one of the most predominantly grown legumes worldwide, however, one deterrent to maximizing its yield is the pathogen, Phytophthora sojae, which causes stem and root rot disease. Many strategies have been implemented to combat this pathogen such as use of pesticides and certain agricultural practices. However, these have been largely ineffective in completely preventing P. sojae infection. An alternative strategy would be to improve the innate resistance of soybean by promoting increased glyceollin production. Glyceollins are soybean-specific antimicrobial agents which are derived from the isoflavonoid branch of the general phenylpropanoid pathway. Soybeans produce 3 forms of glyceollin: …

Evapotranspiration From Extensive Green Roofs: Influence Of Climatological Conditions, Vegetation Type, And Substrate Depth, Maria Eloisa Sia

Electronic Thesis and Dissertation Repository

Green roofs are gaining popularity worldwide as a low impact development tool to mitigate increasing stormwater runoff within dense urban areas. Evapotranspiration (ET) is the key hydrologic process governing the capacity of a green roof to retain rainfall as it regenerates available water storage space in the green roof substrate (soil) between rainfall events. To date, there are limited data on how the interaction between different climatological conditions and design parameters (e.g., vegetation type, substrate depth) affect ET rates. This currently limits the ability to optimize green roof design for stormwater management. In this field study, the impact of climatological …