Life Sciences Commons^™

Molecular Analysis Of Epigenetic Memory Of Stress Establishment And Long-Term Maintenance In A Perennial Woody Plant, Jia Wen Tan

Theses and Dissertations--Plant and Soil Sciences

Plants adapt to extreme environmental conditions through physiological adaptations, which are usually transient. Recent research has suggested that environmental conditions can activate a memory of stress that can result in a primed response to subsequent stress events. While the effect of priming has been observed in many plants, the underlying mechanisms are puzzling and seldom studied. A large body of research has been developed in the last decade linking response to stress, stress priming, and memory of stress with epigenetic mechanisms. This understanding of plant epigenetics has opened the door to the application of epigenetics to crop improvement, such as …

The Impact Of Plant Secondary Metabolites On Auxin And Cytokinin Signaling, Timothy E. Shull

Theses and Dissertations--Plant and Soil Sciences

Secondary metabolites are a broad class of specialized compounds that mediate plant-environment interactions and mitigate stress. It is increasingly clear that many phenylalanine-derived secondary metabolites are nearly indispensable for plant survival and that plants adjust their growth according to their secondary metabolic outputs. Consequently, many phenylalanine-derived secondary metabolites have influence over hormone activity. For instance, multiple phenylpropanoid intermediates and catecholamines alter the sensitivity of plants to the central hormone auxin, which in concert with cytokinin directs most aspects of plant growth and development. This dissertation reviews previous research on the influence of phenylpropanoid intermediates and catecholamines on plants, with a …

Regulatory Mechanisms In Arabidopsis Embryogenesis, Sanjay Joshi

Theses and Dissertations--Plant and Soil Sciences

Plants have amazing regeneration properties with single somatic cells, or groups of cells able to give rise to fully formed plants. One means of regeneration is Somatic Embryogenesis (SE), by which an embryonic structure is formed that “converts” into a plantlet. Somatic embryogenesis has been used as a model for zygotic processes that are buried within layers of maternal tissues. Understanding mechanisms of somatic embryo induction and development are important as a more accessible model for seed development. We rely on seed development not only for most of our caloric intake, but also as a delivery system for engineered crops …

Efficacy And Mechanism Of Acs811 In Tomato Immune Responses, Layne Ellen Harris

Theses and Dissertations--Plant and Soil Sciences

Biologicals are increasingly becoming an important part of sustainable integrated pest management in agricultural crop production and encompass a wide variety of products with varying degrees of efficacy and available research data. As biologicals become more integrated into commercial production systems, it is critical to rigorously evaluate biological product efficacy and mode(s) of action in crops. The better that biologicals are understood in terms of effective application, integration into commercial programs, and their specific mechanisms in crop growth and protection, then the greater the opportunity for increasing yields and food security.

ACS811 is a microbial fermentation-based biological product from Alltech …

The Balancing Act Of Cytokinin In Environmental Stress Tolerance, Sumudu Sandeepani Karunadasa

Theses and Dissertations--Plant and Soil Sciences

Cytokinin, long known as a phytohormone that regulates plant growth and development, has been recently recognized as an important regulator of stress responses. However, our current knowledge about the mechanisms by which cytokinin regulates stress responses is fragmentary, as many of the studies in this field yielded conflicting results. Most of the work described here has focused on analyses of the molecular mechanisms of cytokinin-dependent regulation of growth and development under stress conditions, with an emphasis on the role of cytokinin-dependent regulation of protein synthesis in development and stress tolerance.

One of the important contributions of this study is the …

Bacterial Inoculants, Endophytic Bacteria And Their Influence On Nicotiana Physiology, Development And Microbiome, Andrea Marisa Sanchez Barrios

Theses and Dissertations--Plant and Soil Sciences

Soil and root microbial communities have been studied for decades, and the incorporation of high-throughput techniques and analysis has allowed the identification of endophytic/non-culturable organisms. This has helped characterize and establish the core microbiome of many model plant species which include underground and aboveground organs. Unfortunately, the information obtained from some of these model plants is not always transferable to other agronomic species. In this project, we decided to study the microbiome of the Nicotiana genus because of its importance in plant physiological and plant-microbe interactions studies. The data obtained was used as baseline information that allowed us to better …

Transcriptional And Post-Translational Regulation Of Terpenoid Indole Alkaloid Biosynthesis In Catharanthus Roseus, Priyanka Paul

Theses and Dissertations--Plant and Soil Sciences

Catharanthus roseus (Madagascar periwinkle) is the exclusive source of an array of terpenoid indole alkaloids (TIAs) that are used in the treatments of hypertension and certain types of cancer. TIA biosynthesis is under stringent spatiotemporal control and is induced by jasmonate (JA) and fungal elicitors. Tryptamine, derived from the indole branch, and secologanin from the iridoid branch are condensed to form the first TIA, strictosidine. Biosynthesis of TIA is regulated at the transcriptional level and several transcription factors (TFs) regulating the expression of genes encoding key enzymes in the pathway have been isolated and characterized. The JA-responsive APETALA2/ETHYLENE RESPONSE FACTOR …

Increasing Renewable Oil Content And Utility, William Richard Serson

Theses and Dissertations--Plant and Soil Sciences

Since the dawn of agriculture man has been genetically modifying crop plants to increase yield, quality and utility. In addition to selective breeding and hybridization we can utilize mutant populations and biotechnology to have greater control over crop plant modification than ever before. Increasing the production of plant oils such as soybean oil as a renewable resource for food and fuel is valuable. Successful breeding for higher oil levels in soybean, however, usually results in reduced protein, a second valuable seed component. We show that by manipulating a highly active acyl-CoA: diacylglycerol acyltransferase (DGAT) the hydrocarbon flux to oil in …

Insights Into Triterpene Metabolism In Model Monocotyledonous And Oilseed Plants Genetically Engineered With Genes From Botryococcus Braunii, Chase F. Kempinski

Theses and Dissertations--Plant and Soil Sciences

Isoprenoids are one of the most diverse classes of natural products and are all derived from universal five carbon, prenyl precursors. Squalene and botryococcene are linear, hydrocarbon triterpenes (thirty carbon compounds with six prenyl units) that have industrial and medicinal values. Squalene is produced by all eukaryotes as it is the first committed precursor to sterols, while botryococcene is uniquely produced by the green algae, Botryococcus braunii (race B). Natural sources for these compounds exist, but there is a desire for more renewable production platforms. The model plant Arabidopsis thaliana was engineered to accumulate botryococcene and squalene in its oil …

Investigation Into The Cell Wall And Cellulose Biosynthesis In Model Species And In The C4 Model Plant Setaria Viridis, Mizuki Tateno

Theses and Dissertations--Plant and Soil Sciences

A uniform feature of all plant cells is the presence of a cell wall. The cell wall functions in facilitating directional expansion and is therefore important for cell shape and morphogenesis. All plant cell walls contain cellulose microfibrils embedded in a network of polysaccharides, lignin and protein. Cellulose is evolutionarily conserved and is made by all plants as well as other members of various taxonomic kingdoms. From a human perspective, the field of renewable energy has had an ever increasing interest in using the cell wall for production of renewable platform chemicals and fuels. However, the biosynthesis of these components …

Probing The Plant Cell Wall With Herbicides: A Chemical Genetics Approach, Chad B. Brabham

Theses and Dissertations--Plant and Soil Sciences

The primary cell wall is a highly organized multi-layered matrix of polysaccharides (cellulose, hemi-cellulose, and pectin). The ability of the rigid cell wall to sufficiently loosen to allow growth is a complex process that differs considerably between grasses monocots and dicots. Cellulose is the major structural component required for anisotropic cell expansion and is synthesized by CELLULOSE SYNTHASE A (CesA) proteins. Here, our objectives were two-fold: 1) dissect cell walls and cellulose biosynthesis in dicots and grasses using chemical biology and reverse genetic approaches 2) characterize and classify the inhibitory mechanisms of cellulose biosynthesis inhibitors (CBIs). A reverse genetics TILLING …

Transcriptional Regulation Of Specialized Metabolites In Arabidopsis Thaliana And Catharanthus Roseus, Craig M. Schluttenhofer

Theses and Dissertations--Plant and Soil Sciences

For millennia humans have utilized plant specialized metabolites for health benefits, fragrances, poisons, spices, and medicine. Valued metabolites are often produced in small quantities and may command high prices. Understanding when and how the plant synthesizes these compounds is important for improving their production. Phytohormone signaling cascades, such as jasmonate (JA) activate or repress transcription factors (TF) controlling expression of metabolite biosynthetic genes. TFs regulating specialized metabolite biosynthetic genes can be manipulated to engineer plants with increased metabolite production.

WRKY transcription factor are known components of both JA signaling cascades and regulation of specialized metabolism. The presence of WRKY binding …

Unveiling Novel Aspects Of D-Amino Acid Metabolism In The Model Bacterium Pseudomonas Putida Kt2440, Atanas D. Radkov

Theses and Dissertations--Plant and Soil Sciences

D-amino acids (D-AAs) are the α-carbon enantiomers of L-amino acids (L- AAs), the building blocks of proteins in known organisms. It was largely believed that D-AAs are unnatural and must be toxic to most organisms, as they would compete with the L-counterparts for protein synthesis. Recently, new methods have been developed that allow scientists to chromatographically separate the two AA stereoisomers. Since that time, it has been discovered that D-AAs are vital molecules and they have been detected in many organisms. The work of this dissertation focuses on their place in bacterial metabolism. This specific area was selected due to …

Engineering Triterpene Metabolism In Tobacco, Zuodong Jiang

Theses and Dissertations--Plant and Soil Sciences

Terpenes comprise a large diverse class of natural products and many of them attract interest because of their physiological function, therapeutic and industrial values. Triterpene oils including squalene (C30), botrycococcene (C30) and their methylated derivatives (C31-C37) generated by the green algae Botryococcus braunii race B, which have recently received significant attention because of their utility for advanced biofuels. However, the slow growth habit of B. braunii makes it impractical as a robust biofuel production system. In this thesis, we firstly evaluated the potential of generating high levels of triterpene (C30) production in tobacco plants by diverting carbon flux from cytosolic …

Understanding The Role Of Membrane Localized Ugt80b1 Encoding For Udp-Glucose: Sterol Glucosyltransferase In Plant Development, Meera Nair

Theses and Dissertations--Plant and Soil Sciences

Sterols have been identified as major components of membrane lipids that are part of specialized membrane domains necessary for organizing events such as polar protein targeting and signal transduction in plants, fungi and animals. However a common modification of sterols is the addition of sugar moieties via glycosylation abundantly found in plants. An exact physiological role for such diversification of sterols in plants is still unknown. Using reverse genetics and transcriptomics we show that UDP-glucose: sterol glucosyltransferase encoded by UGT80B1 is necessary for correct epidermal patterning in Arabidopsis root. Patterning of hair cells (trichoblasts) and non-hair cells (atrichoblasts) in the …

Boron Nutrition Of Burley And Dark Tobacco, Laura Ann Frakes Mitchell

Theses and Dissertations--Plant and Soil Sciences

The incidences of suspected Boron (B) deficiency have increased recently in Kentucky tobacco fields, potentially due to recent changes in management practices. The symptoms observed in the field include; hollow stalk, stunted growth, deformed or no bud formation, small slits on the lower leaf midrib and uncontrollable breaking of the midrib approximately two inches from the stalk. B is a micronutrient tobacco needs in minute amounts, however excessive additions of B could cause toxicity. The objectives of this work were to 1) establish critical points for B sufficiency, 2) describe and define B deficiency and toxicity symptoms and 3) develop …

Dna-Binding Site Recognition By Bhlh And Mads-Domain Transcription Factors, Joshua R. Werkman

Theses and Dissertations--Plant and Soil Sciences

Herewithin, two transcription factor (TF) regulatory complexes were investigated. A bHLH–MYB–WDR (BMW) DNA-binding complex from maize was the first complex to be studied. R, a maize bHLH involved in the activation of genes in the anthocyanin pathway, had been characterized to indirectly bind DNA despite the presence of a functional DNA-binding domain. Findings presented here reveal that this is only partially correct. Direct DNA-binding by R was found to be dependent upon two distinct dimerization domains that function as a switch. This switch-like mechanism allows R to be repurposed for the activation of promoters of differing cis-element structure.

The …

Ecophysiological Responses Of Tall Fescue Genotypes To Endophyte Infection And Climate Change, Marie Bourguignon

Theses and Dissertations--Plant and Soil Sciences

Tall fescue is a widely used forage grass in the eastern USA and can form a symbiosis with a fungal endophyte, which can be beneficial for the plant but can cause livestock health issues. Little is known regarding the symbiotic response to predicted climate change. To address this knowledge gap, I analyzed tall fescue variety trial data collected throughout the U.S., exploring relationships between climate variables and yield for two different fescue cultivars that were either endophyte-free or infected. This study showed no endophyte or cultivar effect on fescue yield, but identified temperature, precipitation and location as significant predictors of …

The Role Of Polyadenylation In Seed Germination, Liuyin Ma

Theses and Dissertations--Plant and Soil Sciences

Seed germination has many impacts on the uses of seeds, and is an important subject for study. Seed germination is regulated at both transcriptional and post-transcriptional levels. Therefore, it is important to study how polyadenylation regulates gene expression during seed germination. To this end, a modified Illumina GAIIx sequencing protocol (described in Chapter Two) was developed that allows deep coverage of poly(A) site position and distribution.

Alternative polyadenylation (APA) regulates gene expression by choosing one potential poly(A) site on a precursor RNA consequentially shortening/lengthening the mRNA relative to other possible sites. To further explore this phenomenon, genes affected by APA …

The Role Of Auxin Resistant 1 (Axr1) In Arabidopsis Cytokinin Signaling, Yan Li

Theses and Dissertations--Plant and Soil Sciences

The plant hormone cytokinin plays essential roles in many aspects of growth and development. The cytokinin signal is transmitted by a multistep phosphorelay to the members of two functionally antagonistic classes of Arabidopsis response regulators (ARRs): the type-B ARRs (response activators) and type-A ARRs (negative-feedback regulators). Previous studies have shown that mutations in AXR1, encoding a subunit of the E1 enzyme in the related to ubiquitin (RUB) modification pathway, leads to decreased cytokinin sensitivity. This research shows that the cytokinin resistance of axr1 seedlings is suppressed by loss-of-function of type-A ARRs and that the cytokinin resistance caused by …

Elucidating The Biochemical Wizardry Of Triterpene Metabolism In Botroycoccus Braunii, Thomas Daniel Niehaus

Theses and Dissertations--Plant and Soil Sciences

B. braunii is a green alga that has attracted attention as a potential renewable fuel source due to its high oil content and the archeological record of its unique contribution to oil and coal shales. Three extant chemotypes of B. braunii have been described, namely race A, race B, and race L, which accumulate alkadienes and alkatrienes, botryococcene and squalene and their methylated derivatives, and lycopadiene, respectively. The methylated triterpenes, particularly botryococcenes, produced by race B can be efficiently converted to high quality combustible fuels and other petrochemicals; however, botryococcene biosynthesis has remained enigmatic.

It has been suggested that botryococcene …

Molecular And Chemical Dissection Of Cellulose Biosynthesis In Plants, Darby M. Harris

Theses and Dissertations--Plant and Soil Sciences

Plant cell walls are complex structures that must not only constrain cellular turgor pressure but also allow for structural modification during the dynamic processes of cell division and anisotropic expansion. Cell walls are composed of highly glycosylated proteins and polysaccharides, including pectin, hemicellulose and cellulose. The primary cell wall polysaccharide is cellulose, a polymer composed of high molecular weight !- 1,4-glucan chains. Although cellulose is the most abundant biopolymer on Earth, there is still a lot to learn about its biosynthesis and regulation. This research began by applying a variety of analytical techniques in an attempt to understand differences in …