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<p>Lipids are important cellular biomolecules that perform essential functional and biological roles. For instance, lipids in the cell are the compartmentalizer for the cytoplasm and an energy storage unit. The knowledge surrounding lipids is abundant, yet there is still so much to uncover. There are many categories of lipids and within each category the structural composition is extremely diverse. In turn, the dramatic structural complexity of lipids demands analytical methods capable of providing in-depth structural characterization of individual molecular structures. However, lipid structural elucidation has remained challenging, namely due to the presence of isomeric and isobaric species with a complex mixture. In particular, isomeric/isobaric lipid structures arise from variations in class, headgroup, fatty acyl chain, <i>sn</i>-position, and/or carbon-carbon double bond (C=C) position(s). Recently, recent research suggests C=C composition impacts lipid physical properties, metabolic fate, and intermolecular interactions. Thus, analytical strategies capable of localizing sites of unsaturation are of interest in the lipidomics community.</p> <p>Mass spectrometry (MS) is a leading tool for lipid analysis. Electrospray ionization (ESI), a soft ionization method, is the most commonly used method for lipid ionization as a means of taking the ions from liquid-phase to gas-phase without extensive decomposition of the species. Utilizing ESI-MS, lipids can be identified at a sum compositional level via accurate mass measurements. . With tandem mass spectrometers, lipid ions can be further probed, utilizing tandem-MS (MS/MS) to generate structurally informative product ion spectra that facilitate the assignment of lipid molecular structure. More so, gas-phase ion/ion reactions represent a unique MS-based technique that has improved the analysis of lipids structures. Gas-phase ion/ion reactions allow for lipid species to be charge inverted from one polarity to the opposite polarity. This reaction enables lipids to be ionized in a polarity that is optimal for class identification and further investigated in the opposite polarity where more structural information is obtained. All the information provided is captured without the requirement of multiple solution conditions which is necessary when analyzing in both polarities. In the case of charge inverted lipids from positive ion mode to negative ion mode, fatty acyl composition can be obtained; however, C=C information is lacking.</p> <p>MS can also be paired with other analytically technologies to assist with lipid analysis. One of those technologies is liquid chromatography (LC), which allows for the separation of lipids based on different characteristic depending on the column type being used. Reverse-phase LC (RPLC) allows for the separation of lipid molecular species based on structural composition. RPLC-MS/MS benefits from the ability to separate lipids and determine their fatty acyl chain composition but it is difficult to specify C=C location with the use of a synthetic standard that is identical to each molecular species being analyzed.</p> <p>Commonality between the gas-phase ion/ion reactions for charge inversion of lipids and RPLC-MS/MS approaches is the inability to provide C=C coverage. In-solution and unique ion activation techniques have been developed for seeking such information. The Paternò–Büchi reaction is a UV-initiated [2 + 2]-cycloaddition of an excited carbonyl containing compound onto an olefin group. This reaction can be initiated onto the alkene group within an unsaturated lipid aliphatic chain to form an oxetane ring modification. There are two product ions that can be formed upon each unsaturation site due to a lack of regioselectivity the reagent can attach at either side of the C=C. The modified lipids can be taken into gas-phase and collisionally activated via low-energy collision induced dissociation, generating product ions indictive of C=C position(s). The work herein shows the incorporation of the PB reaction into the gas-phases ion/ion reaction and RPLC-MS/MS apparatuses for C=C localization. The methods have been applied to the lipid extracts of bovine liver and human plasma for confident molecule species determination.</p>

Although cytoplasmic lipid droplets (LDs) are the major reserves for energy-dense neutral lipids in plants, the cellular mechanisms for packaging neutral lipids into LDs remain poorly understood. To gain insights into the cellular processes of neutral lipid accumulation and compartmentalization, a necessary step forward would be to characterize functional roles of lipogenic proteins that participate in the compartmentalization of neutral lipids in plant cells. In this study, the lipogenic proteins, Arabidopsis thaliana SEIPIN homologues and mouse (Mus Musculus) fat storage-inducing transmembrane protein 2 (FIT2), were characterized for their functional roles in the biogenesis of cytoplasmic LDs in various plant tissues. Both Arabidopsis SEIPINs and mouse FIT2 supported the accumulation of neutral lipids and cytoplasmic LDs in plants. The three Arabidopsis SEIPIN isoforms play distinct roles in compartmentalizing neutral lipids by enhancing the numbers and sizes of LDs in various plant tissues and developmental stages. Further, the potential applications of Arabidopsis SEIPINs and mouse FIT2 in engineering neutral lipids and terpenes in plant vegetative tissues were evaluated by co-expressing these and other lipogenic proteins in Nicotiana benthamiana leaves. Arabidopsis SEIPINs and mouse FIT2 represent effective tools that may complement ongoing strategies to enhance the accumulation of desired neutral lipids and terpenes in plant vegetative tissues. Collectively, our findings in this study expand our knowledge of the broader cellular mechanisms of LD biogenesis that are partially conserved in eukaryotes and distinct in plants and suggest novel targets that can be introduced into plants to collaborate with other factors in lipid metabolism and elevate oil content in plant tissues.

Groat oil content and composition are important quality traits in oats (Avena sativa L). These traits are controlled by many genes with additive effects. The chromosomal regions containing these genes, known as quantitative trait loci (QTL), can be discovered through their close association with markers. This study investigated total oil content and fatty acid components in an oat breeding population derived from a cross between high oil ('Dal') and low oil ('Exeter') parents. A genetic map consisting of 475 DArT (Diversity Array Technology) markers spanning 1271.8 cM across 40 linkage groups was constructed. QTL analysis for groat oil content and composition was conducted using grain samples grown at Aberdeen, ID in 1997. QTL analysis for multiple agronomic traits was also conducted using data collected from hill plots and field plots in Ottawa, ON in 2010. QTLs for oil content, palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), linoleic acid (18:2) and linolenic acid (18:3) were identified. Two of the QTLs associated with oil content were also associated with all of the fatty acids examined in this study, and most oil-related QTL showed similar patterns of effect on the fatty acid profile. These results suggest the presence of pleiotropic effects on oil-related traits through influences at specific nodes of the oil synthesis pathway. In addition, 12 QTL-associated markers (likely representing nine unique regions) were associated with plant height, heading date, lodging, and protein content. The results of this study will provide information for molecular breeding as well as insight into the genetic mechanisms controlling oil biosynthesis in oat.

Issues de ressources renouvelables, les huiles végétales (mélanges de triacylglycérols) sont des matières premières aux multiples applications dans divers secteurs, tels que l’industrie alimentaire, les lubrifiants, les solvants, les cosmétiques, la pharmacie, parmi autres. Pour faciliter le développement de produits utilisant de telles matières premières, le travail de thèse s’intéresse à la modélisation de l'équilibre de phases pour mélanges de triacylglycérols et la conception d’un logiciel prédictif de leurs propriétés. Les modèles employés sont totalement prédictifs et tiennent compte des polymorphes. La phase liquide et le polymorphe a sont supposés idéaux à l’inverse des polymorphes b et b’ dont les coefficients d’activités sont calculés à l’aide du modèle de Margules où les coefficients d’interactions binaires sont prédits à partir de la similarité des chaines d’acide gras sur le squelette glycérol. L'optimisation directe de l'énergie libre de Gibbs permet de calculer la quantité de solide dans les graisses ; propriété fondamentale corrélée à de nombreuses fonctionnalités de produits basés sur des huiles végétales. Le calcul de l'équilibre solide - liquide à plusieurs températures permet de simuler des courbes de fusion et des courbes de calorimétrie différentielle à balayage. L’influence sur ces courbes de modifications dans la structure moléculaire et de la composition des mélanges est étudier pour plusieurs systèmes et comparés avec des données expérimentales: triacylglycérols purs, mélanges binaires et ternaires de triacylglycérols, huiles végétales naturelles et mélanges d'huiles végétales impliquant des centaines de triacylglycérols. Des réactions d'interestérification chimique, largement utilisées pour modifier la composition et propriétés d'huiles et graisses, sont simulées et comparés avec des données expérimentales. Le bon accord avec les données expérimentales permet de valider l'outil informatique et son caractère prédictif permet d’envisager son utilisation pour des mélanges, températures et compositions encore non évaluées de façon à orienter les efforts expérimentaux vers les mélanges les plus prometteurs. / The search for sustainable development is an incentive for renewable resources usage in chemical industry. Vegetable oils (triacylglycerol-based mixtures) are raw-materials that match this criterion and they are currently and potentially used in a wide range of sectors, such as: food industry, lubricants, solvents, cosmetics, pharmaceutical, among others. Aiming to aid product design using such raw-materials, this work deals with phase equilibrium modeling and the development of a computational tool for properties prediction. The models used are totally predictive and they deal with the existence of solid polymorphism. The liquid state and the crystals are treated as ideal phases. The activity coefficients of phases ’ and (notideal) are estimated using Margules model, for which the binary molecular interaction parameters are calculated in a predictive manner by means of correlations with the similarity degree between fatty acids chains in the glycerol. By direct optimization of Gibbs Free Energy, the Solid Fat Content is computed, a fundamental property for vegetable oils–based products functionality. The results of phase equilibrium in different temperatures allow simulating melting curves and Differential Scanning Calorimetry curves, allowing a computational evaluation of how molecular structure and composition changes affect desired properties. The calculated results were applied and compared with experimental data in a wide range of systems: pure systems, binary and ternary triacylglycerol mixtures, natural vegetable oils and mixture of vegetable oils (hundreds of components). Chemical interesterification reaction, widely applied to change fats and oils composition and properties, were also simulated and the results compared with experimental data in different compositions and temperatures. The good agreement between computational results and experimental data enables the computational tool validation and its predictive nature makes it suitable for the study of mixtures in temperatures and compositions not yet evaluated. Consequently, the experimental efforts (cost and time) can be concentrated on the most promised molecules/mixtures. / A busca pelo desenvolvimento sustentável é um incentivo para o uso de recursos renováveis na indústria química. Óleos vegetais (misturas de triacilgliceróis) são matérias-primas que atendem a este critério e apresentam aplicações atuais e potenciais em diversos setores, tais como indústria de alimentos, lubrificantes, solventes, cosméticos, farmacêutica, dentre outros. Com o objetivo de auxiliar no desenvolvimento de produtos utilizando tais matérias-primas, o presente trabalho trata da modelagem do equilíbrio de fases em misturas de triacilgliceróis e do desenvolvimento de uma ferramenta computacional preditiva de propriedades. Os modelos usados são totalmente preditivos e consideram a existência de polimorfismo na fase sólida. O estado líquido e o polimorfo são considerados ideais. Os coeficientes de atividade das fases e (não-ideais) são estimados através do modelo de Margules, no qual os coeficientes de interação molecular binária são calculados de forma preditiva através de correlações com o grau de similaridade entre as cadeias de ácidos graxos no glicerol. Através da otimização direta da Energia Livre de Gibbs, é calculado o Conteúdo de Gordura Sólida, uma propriedade fundamental para funcionalidade de produtos baseados em óleos vegetais. Através do cálculo do equilíbrio de fases em diversas temperaturas são simuladas curvas de fusão e curvas de Calorimetria Exploratória Diferencial, permitindo uma avaliação computacional do efeito que alterações na estrutura molecular e composição apresentam nas propriedades de interesse. Os resultados calculados foram aplicados e comparados com dados experimentais em diversos sistemas: triacilgliceróis puros, misturas binárias e ternárias de triacilgliceróis, óleos vegetais naturais e misturas de diversos óleos vegetais (centenas de triacilgliceróis). Reações de interesterificação química, amplamente utilizadas para alterar a composição e propriedades de óleos e gorduras, foram igualmente simuladas e os resultados comparados com dados experimentais em diversas composições e temperaturas. A boa concordância entre os dados simulados e experimentais permite validar a ferramenta computacional e seu caráter preditivo a torna útil no estudo de misturas em temperaturas e composições ainda não avaliadas, de forma que os esforços experimentais (custo e tempo) possam ser direcionados às misturas mais promissoras.

<p>The glycaemic responses to various carbohydrate-rich foods are partly dependent on the rate at which the carbohydrate is digested and absorbed. The glycaemic index (GI) is a way of ranking foods according to their glycaemic response and is recommended as a useful tool in identifying starch-rich foods that give the most favourable glycaemic response. This investigation was undertaken to determine whether carbohydrate-rich foods with a low GI and a high content of dietary fibre (DF) could have beneficial metabolic effects in the insulin resistance syndrome. This question was addressed both in single-meal studies and in randomised controlled clinical trials. Starch-rich foods with low GI values incorporated into composite meals resulted in lower postprandial responses of both glucose and insulin than foods with a high GI in meals with an identical macronutrient and DF composition, in subjects with type 2 diabetes. After three weeks on a diet including low GI starchy foods metabolic profile was improved in subjects with type 2 diabetes, compared with a corresponding high GI diet. The glucose and insulin responses throughout the day were lower, the total and low density lipoprotein cholesterol was decreased, and the fibrinolytic activity was normalised. In subjects with impaired insulin sensitivity and diabetes low GI foods rich in soluble DF for breakfast gave a more favourable metabolic profile, with smaller glucose fluctuations from baseline during the day, than a breakfast with high GI foods low in DF. A low GI breakfast high in DF also resulted in lower responses of insulin and C-peptide after breakfast and a lower triacylglycerol response after a standardised lunch. However, none of the tested breakfasts improved the glucose and insulin responses after lunch. Similar results were obtained in obese subjects after including a breakfast with a low GI high in soluble DF for a period of four weeks in comparison with a breakfast with a high GI and low content of DF.</p><p>These results support the therapeutic potential of a diet with a low GI in the treatment of diabetes and also in the treatment of several of the metabolic disturbances related to the insulin resistance syndrome.</p>

The glycaemic responses to various carbohydrate-rich foods are partly dependent on the rate at which the carbohydrate is digested and absorbed. The glycaemic index (GI) is a way of ranking foods according to their glycaemic response and is recommended as a useful tool in identifying starch-rich foods that give the most favourable glycaemic response. This investigation was undertaken to determine whether carbohydrate-rich foods with a low GI and a high content of dietary fibre (DF) could have beneficial metabolic effects in the insulin resistance syndrome. This question was addressed both in single-meal studies and in randomised controlled clinical trials. Starch-rich foods with low GI values incorporated into composite meals resulted in lower postprandial responses of both glucose and insulin than foods with a high GI in meals with an identical macronutrient and DF composition, in subjects with type 2 diabetes. After three weeks on a diet including low GI starchy foods metabolic profile was improved in subjects with type 2 diabetes, compared with a corresponding high GI diet. The glucose and insulin responses throughout the day were lower, the total and low density lipoprotein cholesterol was decreased, and the fibrinolytic activity was normalised. In subjects with impaired insulin sensitivity and diabetes low GI foods rich in soluble DF for breakfast gave a more favourable metabolic profile, with smaller glucose fluctuations from baseline during the day, than a breakfast with high GI foods low in DF. A low GI breakfast high in DF also resulted in lower responses of insulin and C-peptide after breakfast and a lower triacylglycerol response after a standardised lunch. However, none of the tested breakfasts improved the glucose and insulin responses after lunch. Similar results were obtained in obese subjects after including a breakfast with a low GI high in soluble DF for a period of four weeks in comparison with a breakfast with a high GI and low content of DF. These results support the therapeutic potential of a diet with a low GI in the treatment of diabetes and also in the treatment of several of the metabolic disturbances related to the insulin resistance syndrome.

Groat oil content and composition are important quality traits in oats (Avena sativa L). These traits are controlled by many genes with additive effects. The chromosomal regions containing these genes, known as quantitative trait loci (QTL), can be discovered through their close association with markers. This study investigated total oil content and fatty acid components in an oat breeding population derived from a cross between high oil ('Dal') and low oil ('Exeter') parents. A genetic map consisting of 475 DArT (Diversity Array Technology) markers spanning 1271.8 cM across 40 linkage groups was constructed. QTL analysis for groat oil content and composition was conducted using grain samples grown at Aberdeen, ID in 1997. QTL analysis for multiple agronomic traits was also conducted using data collected from hill plots and field plots in Ottawa, ON in 2010. QTLs for oil content, palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), linoleic acid (18:2) and linolenic acid (18:3) were identified. Two of the QTLs associated with oil content were also associated with all of the fatty acids examined in this study, and most oil-related QTL showed similar patterns of effect on the fatty acid profile. These results suggest the presence of pleiotropic effects on oil-related traits through influences at specific nodes of the oil synthesis pathway. In addition, 12 QTL-associated markers (likely representing nine unique regions) were associated with plant height, heading date, lodging, and protein content. The results of this study will provide information for molecular breeding as well as insight into the genetic mechanisms controlling oil biosynthesis in oat.

Motivation: The prevalence of non-communicable diseases such as diabetes mellitus (DM) and cardiovascular disease (CVD) is rapidly increasing in industrialized societies. Experts believe that lifestyle, and in particular its nutritional aspects, plays a decisive role in increasing the burden of these chronic conditions. Dietary habits would, therefore, be modified to exert a positive impact on the prevention and treatment of chronic diseases of lifestyle. It is believed that the state of hyperglycaemia that is observed following food intake under certain dietary regimes contributes to the development of various metabolic conditions. This is not only true for individuals with poor glycaemic control such as some diabetics, but could also be true for healthy individuals. It would, therefore, be helpful to be able to reduce the amplitude and duration of postprandial hyperglycaemia. Selecting the correct type of carbohydrate (CHO) foods may produce less postprandial hyperglycaemia, representing a possible strategy in the prevention and treatment of chronic metabolic diseases. At the same time, a key focus of sport nutrition is the optimal amount of CHO that an athlete should consume and the optimal timing of consumption. The most important nutritional goals of the athlete are to prepare body CHO stores pre-exercise, provide energy during prolonged exercise and restore glycogen stores during the recovery period. The ultimate aim of these strategies is to maintain CHO availability to the muscle and central nervous system during prolonged moderate to high intensity exercise, since these are important factors in exercise capacity and performance. However, the type of CHO has been studied less often and with less attention to practical concerns than the amount of CHO. The glycaemic index (GI) refers to the blood glucose raising potential of CHO foods and, therefore, influences secretion of insulin. In several metabolic disorders, secretion of insulin is inadequate or impossible, leading to poor glycaemic control. It has been suggested that low GI diets could potentially contribute to a significant improvement of the conditions associated with poor glycaemic control. Insulin secretion is also important to athletes since the rate of glycogen synthesis depends on insulin due to it stimulatory effect on the activity of glycogen synthase. Objectives: Three main objectives were identified for this study. The first was to conduct a meta-analysis of the effects of the GI on markers for CHO and lipid metabolism with the emphasis on randomised controlled trials (RCT's). Secondly, a systematic review was performed to determine the strength of the body of scientific evidence from epidemiological studies combined with RCT's to encourage dieticians to incorporate the GI concept in meal planning. Finally, a systematic review of the effect of the GI in sport performance was conducted on all available literature up to date to investigate whether the application of the GI in an athlete's diet can enhance physical performance. Methodology: For the meta-analysis, the search was for randomised controlled trials with a cross-over or parallel design published in English between 1981 and 2003, investigating the effect of low GI vs high GI diets on markers of carbohydrate and lipid metabolism. The main outcomes were serum fructosamine, glycosylated haemoglobin (HbA1c), high-density lipoprotein cholesterol (HDL-c), low-density lipoprotein cholesterol (LDL-c), total cholesterol (TC) and triacylglycerols (TG). For the systematic review, epidemiological studies as well as RCT's investigating the effect of LGI vs HGI diets on markers for carbohydrate and lipid metabolism were used. For the systematic review on the effect of the GI on sport performance, RCT's with either a cross-over or parallel design that were published in English between January 1981 and September 2004 were used. All relevant manuscripts for the systematic reviews as well as meta-analysis were obtained through a literature search on relevant databases such as the Cochrane Central Register of Controlled Trials, MEDLINE (1981 to present), EMBASE, LILACS, SPORTDiscus, ScienceDirect and PubMed. This thesis is presented in the article format. Results and conclusions of the individual manuscripts: For the meta-analysis, literature searches identified 16 studies that met the strict inclusion criteria. Low GI diets significantly reduced fructosamine (p<0.05), HbA1c, (p<0.03), TC(p<0.0001) and tended to reduce LDL-c (p=0.06) compared to high GI diets. No changes were observed in HDL-c and TG concentrations. Results from this meta analysis, therefore, support the use of the GI concept in choosing CHO-containing foods to reduce TC and improve blood glucose control in diabetics. The systematic review combined the results of the preceding meta-analysis and results from epidemiological studies. Prospective epidemiological studies showed improvements in HDL-c concentrations over longer time periods with low GI diets vs. high GI diets, while the RCT's failed to show an improvement in HDL-c over the short-term. This could be attributed to the short intervention period during which the RCT's were conducted. Furthermore, epidemiological studies failed to show positive relationships between LDL-c and TC and low GI diets, while RCT's reported positive results on both these lipids with low GI diets. However, the epidemiological studies, as well as the RCT's showed positive results with low GI diets on markers of CHO metabolism. Taken together, convincing evidence from RCT's as well as epidemiological studies exists to recommend the use of low GI diets to improve markers of CHO as well as of lipid metabolism. 3 From the systematic review regarding the GI and sport performance it does not seem that low GI pre-exercise meals provide any advantages over high GI pre-exercise meals. Although low GI pre-exercise meals may better maintain CHO availability during exercise, low GI pre-exercise meals offer no added advantage over high GI meals regarding performance. Furthermore, the exaggerated metabolic responses from high GI compared to low GI CHO seems not be detrimental to exercise performance. However, athletes who experience hypoglycaemia when consuming CHO-rich feedings in the hour prior to exercise are advised to rather consume low GI pre-exercise meals. No studies have been reported on the GI during exercise. Current evidence suggests a combination of CHO with differing Gl's such as glucose (high GI), sucrose (moderate GI) and fructose (low GI) will deliver the best results in terms of exogenous CHO oxidation due to different transport mechanisms. Although no studies are conducted on the effect of the GI on short-term recovery it is speculated that high GI CHO is most effective when the recovery period is between 0-8 hours, however, evidence suggests that when the recovery period is longer (20-24 hours), the total amount of CHO is more important than the type of CHO. Conclusion: There is an important body of evidence in support of a therapeutic and preventative potential of low GI diets to improve markers for CHO and lipid metabolism. By substituting high GI CHO-rich with low GI CHO-rich foods improved overall metabolic control. In addition, these diets reduced TC, tended to improve LDL-c and might have a positive effect over the long term on HDL-c. This confirms the place for low GI diets in disease prevention and management, particularly in populations characterised by already high incidences of insulin resistance, glucose intolerance and abnormal lipid levels. For athletes it seems that low GI pre-exercise meals do not provide any advantage regarding performance over high GI pre-exercise meals. However, low GI meals can be recommended to athletes who are prone to develop hypoglycaemia after a CHO-rich meal in the hour prior to exercise. No studies have been reported on the effect of the GI during exercise. However, it has been speculated that a combination of CHO with varying Gl's deliver the best results in terms of exogenous CHO oxidation. No studies exist investigating the effect of the GI on short-term recovery, however, it is speculated that high GI CHO-rich foods are suitable when the recovery period is short (0-8 h), while the total amount rather than the type of CHO is important when the recovery period is longer (20-24 h). Therefore, the GI is a scientifically based tool to enable the selection of CHO-containing foods to improve markers for CHO and lipid metabolism as well as to help athletes to prepare optimally for competitions. Recommendations: Although a step nearer has been taken to confirm a place for the GI in human health, additional randomised, controlled, medium and long-term studies as well as more epidemiological studies are needed to investigate further the effect of low GI diets on LDL-c. HDL-c and TG. These studies are essential to investigate the effect of low GI diets on endpoints such as CVD and DM. This will also show whether low GI diets can reduce the risk of diabetic complications such as neuropathy and nephropathy. Furthermore, the public at large must be educated about the usefulness and application of the GI in meal planning. For sport nutrition, randomised controlled trials should be performed to investigate the role of the GI during exercise as well as in sports of longer duration such as cricket and tennis. More studies are needed to elucidate the short-term effect of the GI post-exercise as well as to determine the mechanism of lower glycogen storage with LGI meals post-exercise. / Thesis (Ph.D. (Dietetics))--North-West University, Potchefstroom Campus, 2005.

Soybean (Glycine max (L.) Merr) is the largest oil and protein crop in the world and it is grown for both oil and protein. To address the needs of both the edible oil market and industrial applications of soybean oil, fatty acid modification has been a focus of soybean breeding programs. Natural variation, mutagenesis and genetic engineering have been used to alter the fatty acid profile. Several genes, mostly desaturases, have been associated with altered fatty acid profiles but enzymes in the Kennedy Pathway have yet to be studied as another source of genetic variation for altering the fatty acid profiles. The Kennedy Pathway is also known as the oil producing pathway and consists of four enzymes: glycerol-3-phosphate acyltransferase (G3PAT); lysophosphatidic acid acyltransferase (LPAAT); phosphatidic acid phosphatase (PAP); and diacylglycerol acyltransferase 1 (DGAT1). The starting material for this pathway is glycerol-3-phosphate, which is produced from glycerol by glycerol kinase (GK), and the product of this pathway is triacylglycerol (TAG). The overall objective of this study was to elucidate the role that the Kennedy Pathway plays in determining the fatty acid profile in two ways: (1) sequencing the transcribed region of the genomic genes encoding the enzymes of GK, G3PAT, LPAAT, and DGAT1 in soybean genotypes with altered fatty acid profiles; and (2) studying their expression over seed development, across three growing temperatures. The genetic material for the study consisted of four soybean genotypes with altered fatty acid profile: RG2, RG7, RG10, and SV64-53. Results from sequencing showed that the mutations identified in G3PAT, LPAAT, and DGAT1 in the four soybean genotypes did not explain the differences in the fatty acid profiles. The expression of G3PAT, LPAAT, and DGAT1 over seed development showed that G3PAT had the lowest levels, followed by LPAAT, then DGAT1, across the growing temperatures. The differences in expression among genotypes corresponded to differences in fatty acid accumulation, suggesting that expression rather than genetic mutations in the transcribed region of the genes influenced the fatty acid profile of the genotypes in this study. In conclusion, the enzymes of the Kennedy Pathway appear to contribute to the altered fatty acid profiles observed in the soybean mutant genotypes. / Ontario Ministry of Economic Development and Innovation (formerly Ontario Ministry of Research and Innovation), BioCar Initiative, Grain Farmers of Ontario, SeCan