In order to predict future range shifts for invasive species it is important to explore their ability to acclimate to the new environment and understand physiological and reproductive constraints controlling their distribution. My dissertation studied mechanisms by which temperature may affect the distribution of two of the most aggressive plant invaders in North America, Bromus tectorum and Bromus rubens. While Bromus tectorum is dominant in the “cold desert” steppes of the Intermountain region of western North America, B. rubens is one of the severe grass invaders in the “hot deserts” of southwestern North America. I first evaluated whether winter freezing tolerance is the mechanism responsible for the distinct northern range limits of Bromus species. Bromus rubens has a slower rate of freezing acclimation that leads to intolerance of sudden, late-autumn reductions in temperature below -12°C, Bromus tectorum, by contrast, cold hardens rapidly and is not impacted by the sudden severe late-autumn cold. Photosynthetic response to temperature does not explain their current range separation. Bromus species differ little in their photosynthetic temperature responses and the acclimation pattern of photosynthesis. Both species acclimated to a broad range of temperature through the amelioration of Pi regeneration limitation at sub-optimal temperatures and improved carboxylation capacity above the thermal optimum which probably resulted from increased thermostability of Rubisco activase. The effect of elevated temperatures during flowering on the seed yield of Bromus species demonstrates that neither species produces seed at 36°C and above. These thresholds are close to temperatures encountered during flowering in their natural environment. In summary, climatic changes will cause northward range expansion of Bromus species due to less severe autumn and winter, while reproductive failure could cause range contraction at their southern margins.

Mon projet de maîtrise avait pour but d’évaluer la faisabilité d’une solution de phytoremédiation par phyto-extraction d’un sol modérément contaminé en zinc et en arsenic. Cette évaluation s’est basée sur les résultats d’une expérience en pots réalisée dans un tunnel de croissance de l’Institut de recherche en biologie végétale (IRBV) durant l’été 2015. Le sol utilisé provenait directement du site contaminé. Les plantes choisies pour cette expérience étaient deux cultivars de saule, Salix purpurea ‘Fish Creek’ et Salix miyabeana ‘SX67’ ainsi que trois herbacées Medicago sativa, Festuca arundinacea et un mélange de plantes utilisé pour la stabilisation de sol. L’impact d’un agent chélateur (acide nitrilotriacétique, NTA) sur la phytoremédiation et le développement des espèces a également été étudié. Les saules ont extrait deux à trois fois plus de zinc que les herbacées. La tendance était inversée pour l’arsenic avec une extraction deux à trois fois plus grande chez les herbacées. Le chélateur NTA a eu un effet positif sur la biomasse aérienne des plantes et sur la phyto-extraction du zinc, mais pas sur la phyto-extraction de l’arsenic. Une diminution importante de la quantité de zinc et d’arsenic a été observée dans les sols. Cette diminution provient des effets combinés de la phyto-extraction et d’autres facteurs, comme la variabilité intrinsèque du sol ainsi que son lessivage. La phyto-extraction de l’arsenic a donné des résultats médiocres, quels que soient les traitements et les plantes utilisés. Les résultats obtenus démontrent l’efficacité des saules et le potentiel du chélateur NTA pour la phyto-extraction du zinc. / My master project aimed to evaluate if phytoremediation by phytoextraction was a viable solution in cleaning up a moderately contaminated soil in zinc and arsenic. This evaluation was based on the results of a potted experiment done within Montreal Botanical Garden, in a growth tunnel set up by Institut de recherche en biologie végétale (IRBV). The soil was collected from a contaminated site. We chose the following plants species: two willow cultivars, Salix purpurea ‘Fish Creek’ and Salix miyabeana ‘SX67’, two grasses Medicago sativa L., Festuca arundinacea and a mix of grasses used for soil stabilization. The impact of a chelating agent (nitrilotriacetic acid, NTA) on phytoextraction and plant growth was also assessed. Salix extracted two to three time more zinc than the grasses. For arsenic, the opposite trend was observed, with two to three times more arsenic extracted by grasses. Chelator NTA had a positive effect on above ground biomass and overall phytoextraction of zinc, but not for arsenic phytoextraction. A significant diminution of zinc and arsenic was observed in soil after the experiment. This diminution comes from the combined effects of phytoextraction and other factors, such as intrinsic soil variability and its leaching. Arsenic phytoextraction was overall mediocre regardless of the species and treatments used. These results show the effectiveness of willows and potential from chelator NTA for zinc phytoextraction.

Master of Science / Department of Biology / Jyoti Shah / Plant defense responses are constitutively activated in the Arabidopsis thaliana ssi2 mutant plant. In addition, the ssi2 mutant allele confers a dwarf phenotype. The SSI2 gene encodes a stearoyl-ACP-desaturase, which converts stearic acid (18:0) to oleic acid (18:1), suggesting a role for lipids in plant defense. Microarray analysis identified several genes which encode putative acyl hydrolases/lipases that are expressed at elevated levels in the leaves of ssi2, in comparison to the wild type plant. One gene in particular, At5g14180, was expressed at 60-fold greater level in ssi2 than in the wild type plant. To study the involvement of At5g14180 in plant defense and lipid metabolism, two transgenic lines containing T-DNA insertions within the At5g14180 gene were identified. These two T-DNA insertional alleles of the At5g14180 gene attenuate the ssi2-conferred heightened resistance to a virulent strain of Pseudomonas syringae pv. maculicola in the ssi2 At5g14180 double mutant plant. Furthermore, pathogen growth was enhanced in the At5g14180 single mutant plants, as compared to the wild type plant. Profiling of lipid composition in leaf tissue identified changes in the lipid composition between the At5g14180 mutant and wild type plants, suggesting that the At5g14180 encoded protein may impact lipid metabolism in Arabidopsis leaves.

Master of Science / Department of Agronomy / William T. Schapaugh Jr / Crops research moving forward faces many challenges to improve crop performance. In breeding programs, phenotyping has time and economic constraints requiring new phenotyping techniques to be developed to improve selection efficiency and increase germplasm entering the pipeline. The objectives of these studies were to examine the changes in spectral reflectance with soybean breeding from 1923 to 2010, evaluate band regions most significantly contributing to yield estimation, evaluate spectral reflectance data for yield estimation modeling across environments and growth stages and to evaluate the usefulness of spectral data as an optimized phenotyping technique in breeding programs. Twenty maturity group III (MGIII) and twenty maturity group IV (MGIV) soybeans, arranged in a randomized complete block design, were grown in Manhattan, KS in 2011 and 2012. Spectral reflectance data were collected over the growing season in a total of six irrigated and water- stressed environments. Partial least squares and multiple linear regression were used for spectral variable selection and yield estimation model building. Significant differences were found between genotypes for yield and spectral reflectance data, with the visible (VI) having greater differences between genotypes than the near-infrared (NIR). This study found significant correlations with year of release (YOR) in the VI and NIR portions of the spectra, with newer released cultivars tending to have lower reflectance in the VI and high reflectance in the NIR. Spectral reflectance data accounted for a large portion of variability for seed yield between genotypes using the red edge and NIR portions of the spectra. Irrigated environments tended to explain a larger portion of seed yield variability than water-stressed environments. Growth stages most useful for yield estimation was highly dependent upon the environment as well as maturity group. This study found that spectral reflectance data is a good candidate for exploration into optimized phenotyping techniques and with further research and validation datasets, may be a suitable indirect selection technique for breeding programs.

Doctor of Philosophy / Department of Agronomy / Scott A. Staggenborg / Grain sorghum yield has notably increased from the beginning of hybrid production and commercialization in the late 1950s. The yield increases were the result of improved agronomic practices and hybrid advancement. The objectives of my research were: (1) to determine the magnitude of yield change in the hybrid era in irrigated and rain fed sorghum production, (2) to determine the contribution of agronomic and hybrid changes for yield in the hybrid era, (3) to investigate changes in sorghum morphology, physiology, and water use that contributed to yield increases, (4) to investigate changes that accompanied yield increase with hybrid improvement, and (5) to understand sorghum water and nutrient use and variations between hybrids in these regards. Fifty-two years of grain sorghum hybrid performance trial data (1957-2008), were analyzed and greenhouse and field studies were conducted on five selected hybrids to meet our objectives. The greenhouse and field studies were conducted from the summer of 2007 to the fall of 2009 on five selected hybrids, each representing a decade from the past fifty years. Results indicated that there was an increase in hybrid yield of nearly 50 kg ha-1 yr-1 in dryland sites over the 52 yrs (1957-2008) analyzed. Irrigated grain sorghum yields, however, remained unchanged over the same period. Agronomic practices such as planting date, phosphorus fertilizer use, and planting density changed over these years but were not found to contribute to increased dryland sorghum yields. There was no difference found between old and new hybrids tolerance to different densities. Hybrid advancement and increased nitrogen fertilizer application were responsible for changes in dryland yields. Total water use changed with hybrid advancement. New hybrids used the greatest total water and also had greater root-to-total biomass ratio than the old hybrids. Leaf biomass was also greater for the newest hybrid. There was a difference in amount of total nutrients extracted by hybrids, and there were differences among hybrids in allocation of nutrients to different tissues. In general the yield focus of sorghum hybrid development was effective in dryland sorghum production, likely because of intentional or inadvertent selection of hybrids with better drought tolerance. Results indicated that breeding programs created hybrids with improved morphological characteristics that might have resulted in better resource use (water and nutrient) and ultimately increased yield.

Doctor of Philosophy / Department of Biochemistry and Molecular Biophysics / Lawrence C. Davis / Synthetic dyes are capable of producing the whole color spectrum on account of their structural diversity but this diversity poses challenges in the degradation of dyeing wastes. Laccases and peroxidases from bacterial or fungal sources and parts of plants in the presence of hydrogen peroxide (H₂O₂) plus a mediator have been exploited in the bioremediation of synthetic dyes. However, intact plants have not found much favor despite their phytoremediation potential. The goal of this research was to further clarify ways by which whole plants bring about decolorization of different types of synthetic dyes. Hydroponically cultivated plants from two dicot families namely Arabidopsis thaliana and sunflowers (Helianthus annuus) were exposed to representative dyes from several classes: monoazo (Methyl Red and Methyl Orange), disazo (Trypan Blue, Evans Blue and Chicago Blue 6B), and arylmethane (Brilliant Blue G, Bromocresol Green, Malachite Green and Phenol Red). Tests were done in presence or absence of externally added H₂O₂, with or without a free radical mediator, 1-hydroxybenzotriazole, using UV-Visible spectrophotometry. The initial rate of decolorization and the overall percentage decolorization was calculated for each dye in the different treatments. Decolorization of the dyes from different classes varied between plant species and depending on the treatment. Except for Methyl Red, all dyes required added H₂O₂ as well as mediator to achieve rapid decolorization. Added H₂O₂ was found to be the limiting factor since it was degraded by plants within a few hours. Both species were able to slowly decolorize dyes upon daily addition of fresh dye even in the absence of added H₂O₂ and mediator, provided that nutrients were supplied to the plants with the dye. A. thaliana was found to be more effective in dye decolorization per gram tissue than sunflower when treated under similar conditions. Analysis of the residual dye solution by ESI/MS did not reveal any potential by-products following the decolorization treatment with plants, suggesting that the plant roots might be trapping the by-products of dye decolorization and preventing their release into the solution. All these findings support the potential application of whole plants for larger scale remediation.

ABA insensitive mutants (abi1 - abi5) have been isolated in germination screens that use high concentrations of exogenous ABA, and this method is believed to be saturated. To overcome this problem, a sensitized screen that used much lower concentration of exogenous ABA was performed to isolate new ABA insensitive mutants. Some of the isolated mutants had defects in light or retrograde signalling. One particular mutant (18-11) developed long hypocotyls under normal light condition. Based on its response to blue, red and far-red light conditions, this mutant is likely a novel hy mutant. Genetic analysis revealed that while ABA insensitivity in this mutant is recessive, the long hypocotyl phenotype is dominant. Positional cloning is currently being carried out to identify the gene. Findings from this study supports that ABA signalling interacts with light signalling networks.

ABA insensitive mutants (abi1 - abi5) have been isolated in germination screens that use high concentrations of exogenous ABA, and this method is believed to be saturated. To overcome this problem, a sensitized screen that used much lower concentration of exogenous ABA was performed to isolate new ABA insensitive mutants. Some of the isolated mutants had defects in light or retrograde signalling. One particular mutant (18-11) developed long hypocotyls under normal light condition. Based on its response to blue, red and far-red light conditions, this mutant is likely a novel hy mutant. Genetic analysis revealed that while ABA insensitivity in this mutant is recessive, the long hypocotyl phenotype is dominant. Positional cloning is currently being carried out to identify the gene. Findings from this study supports that ABA signalling interacts with light signalling networks.

The Arabidopsis cyclic nucleotide-gated ion channels (AtCNGCs) form a large family consisting of 20 members. It has been suggested that CNGCs contribute to a wide array of biological functions such as pollen tube growth and pathogen defence signalling. However, the precise mechanisms by which AtCNGCs act, and the extent of their biological roles, have yet to be fully elucidated. AtCNGC11/12, the chimeric CNGC that resulted from the fusion of AtCNGC11 and 12, induces a number of pathogen defence related phenotypes in the Arabidopsis mutant cpr22. Spontaneous lesion formation is one such phenotype. Interestingly, when AtCNGC11/12 is transiently expressed in N. benthamiana it causes cell death which was characterized in this study. Also, AtCNGC11/12 was used to investigate the structural features responsible for the proper function and regulation of AtCNGCs. Electron microscopic analysis of the AtCNGC11/12-induced cell death showed similar characteristics to programmed cell death (PCD), such as plasma membrane shrinkage and vesicle formation. Interestingly caspase-1 inhibitors and the silencing of vacuolar processing enzyme, a plant enzyme with caspase-1 activity, suppressed the induction of cell death. Additionally, pharmacological analyses indicated that the AtCNGC11/12-indiced cell death was also dependent on Ca2+. Furthermore, 3 amino acid residues, R190, A225, and G287, were demonstrated to be essential for AtCNGC11/12-induce cell death. Taken together, these results indicate that the cell death that develops in the cpr22 mutant is indeed PCD and that AtCNGC11/12, is at the point of, or up-stream of, the Ca2+ signal necessary for the development of HR. Furthermore, the functionality of AtCNGC11/12 as a model for AtCNGC structure-function analyses was demonstrated by the identification of several amino acids necessary for cell death development. Yoshioka et al. (2006) demonstrated that the loss of AtCNGC11 or 12 results in decreased resistance to avirulent isolates of the oomycete pathogen, H. arabidopsidis. Thus, the present biological role suggested for AtCNGC11 and 12 is in pathogen defence, specifically within effector triggered immunity (ETI). Like AtCNGC11 and 12, AtCNGC2 has been demonstrated to contribute to pathogen defence signalling but has also been implicated in other physiological responses such as ion stress and senescence. To better understand the roles of AtCNGC11 and 12 in both pathogen defence and other Ca2+ dependent signalling processes, I have investigated promoter:GUS reporter lines, as well as, AtCNGC11 and 12 KO and RNAi silenced lines subjected to various treatments. From this work, I have demonstrated that AtCNGC11 and 12 have similar expression patterns during pathogen defence, development, and dark-induced senescence. Additionally, the findings presented here further characterize AtCNGC11 and 12 as contributors to ETI rather than PAMP triggered immunity. Furthermore, I demonstrated that AtCNGC11 and 12 are likely involved in the endogenous movement of Ca2+, contributing to a range of Ca2+ associated signalling pathways including gravitropism and senescence. Taken together, these results have greatly improved the characterization of AtCNGC11 and 12; significantly contributing to the understanding of a large and increasingly important channel family.

The Arabidopsis cyclic nucleotide-gated ion channels (AtCNGCs) form a large family consisting of 20 members. It has been suggested that CNGCs contribute to a wide array of biological functions such as pollen tube growth and pathogen defence signalling. However, the precise mechanisms by which AtCNGCs act, and the extent of their biological roles, have yet to be fully elucidated. AtCNGC11/12, the chimeric CNGC that resulted from the fusion of AtCNGC11 and 12, induces a number of pathogen defence related phenotypes in the Arabidopsis mutant cpr22. Spontaneous lesion formation is one such phenotype. Interestingly, when AtCNGC11/12 is transiently expressed in N. benthamiana it causes cell death which was characterized in this study. Also, AtCNGC11/12 was used to investigate the structural features responsible for the proper function and regulation of AtCNGCs. Electron microscopic analysis of the AtCNGC11/12-induced cell death showed similar characteristics to programmed cell death (PCD), such as plasma membrane shrinkage and vesicle formation. Interestingly caspase-1 inhibitors and the silencing of vacuolar processing enzyme, a plant enzyme with caspase-1 activity, suppressed the induction of cell death. Additionally, pharmacological analyses indicated that the AtCNGC11/12-indiced cell death was also dependent on Ca2+. Furthermore, 3 amino acid residues, R190, A225, and G287, were demonstrated to be essential for AtCNGC11/12-induce cell death. Taken together, these results indicate that the cell death that develops in the cpr22 mutant is indeed PCD and that AtCNGC11/12, is at the point of, or up-stream of, the Ca2+ signal necessary for the development of HR. Furthermore, the functionality of AtCNGC11/12 as a model for AtCNGC structure-function analyses was demonstrated by the identification of several amino acids necessary for cell death development. Yoshioka et al. (2006) demonstrated that the loss of AtCNGC11 or 12 results in decreased resistance to avirulent isolates of the oomycete pathogen, H. arabidopsidis. Thus, the present biological role suggested for AtCNGC11 and 12 is in pathogen defence, specifically within effector triggered immunity (ETI). Like AtCNGC11 and 12, AtCNGC2 has been demonstrated to contribute to pathogen defence signalling but has also been implicated in other physiological responses such as ion stress and senescence. To better understand the roles of AtCNGC11 and 12 in both pathogen defence and other Ca2+ dependent signalling processes, I have investigated promoter:GUS reporter lines, as well as, AtCNGC11 and 12 KO and RNAi silenced lines subjected to various treatments. From this work, I have demonstrated that AtCNGC11 and 12 have similar expression patterns during pathogen defence, development, and dark-induced senescence. Additionally, the findings presented here further characterize AtCNGC11 and 12 as contributors to ETI rather than PAMP triggered immunity. Furthermore, I demonstrated that AtCNGC11 and 12 are likely involved in the endogenous movement of Ca2+, contributing to a range of Ca2+ associated signalling pathways including gravitropism and senescence. Taken together, these results have greatly improved the characterization of AtCNGC11 and 12; significantly contributing to the understanding of a large and increasingly important channel family.