Doctor of Philosophy / In this thesis, a set of wheat varieties (Triticum aestivum L.) produced by the Value Added Wheat Cooperative Research Centre with lower swelling power as compared to commercial Australian wheat varieties were studied to enhance our understanding of the role of amylose in starch functionality. These starches originated from a heterogeneous genetic background and had a narrow range of elevated amylose content (35 to 43%) linked with diverse functional properties. Small-angle X-ray scattering together with complementary techniques of differential scanning calorimetry and X-ray diffraction have been employed to investigate the features of starch granular structure at the nanometer scale. Starch chemical structure was characterized in terms of amylose content and amylopectin chain length distribution. Starch functionality was studied by a series of swelling, pasting and enzymic digestion methods. This study showed that swelling power of flour is a simple test that reflects a number of industrially relevant characteristics of starch, and therefore can be used as an indicator of amylose content and pasting properties of starch. In contrast to waxy starches and starches with normal amylose content, wheat starches with increased amylose content displayed characteristic pasting properties that featured decreasing peak, breakdown and final viscosities with increasing amylose contents. Existence of a threshold value in amylose content, above which final viscosity of starch paste does not further increase with increasing amylose content, was proposed. Variability in amylopectin chain length distribution was shown to have an additional effect on the swelling and pasting properties of the starches. On the molecular level, increased amylose content was correlated with increased repeat spacing of the lamellae present in the semicrystalline growth rings. In agreement with current understanding of starch synthesis, amylose was shown to accumulate in both crystalline and amorphous parts of the lamella. Using waxy starch as a distinctive comparison with the other samples confirmed general trend of increasing amylose content being linked with the accumulation of defects within crystalline lamellae. Amylose content was shown to directly influence the architecture of semicrystalline lamellae, whereas thermodynamic and functional properties were proposed to be brought about by the interplay of amylose content and amylopectin architecture. Subjecting starch granules with varying amylose content to pancreatic α-amylase showed differences in their digestion patterns. Pancreatic α-amylase preferentially attacked amorphous regions of waxy starch granules, whereas these regions for initial preferential hydrolysis gradually diminished with increasing amylose content. Observed variations in the extent of enzymic digestion were concluded to be primarily determined by the level of swelling of amorphous growth rings, which can also explain observed morphologies of partly digested granules with varying amylose content. It was confirmed that access to the granular components is not a function of the extent of crystallinity but rather the spatial positioning of the crystalline regions within the granule. Digestion kinetics is governed by factors intrinsic to starch granules, whereas influence of enzyme type was shown to be critical in determining the absolute rate of hydrolysis. Wheat starches with increased amylose content offer the potential to be used as slow digestible starch, mostly in their granular form or when complexed with lipids. Differences among varieties largely diminished when starches were gelatinized or allowed to retrograde demonstrating the importance of granular structure on starch hydrolysis. Wheat varieties used in this study displayed widely differing pasting properties in a Rapid Visco Analyser (RVA) and textural characteristics of the respective retrograded starch gels. Varietal differences in starch chemical composition among wheat varieties were shown to have significant effect on the extent of the response of starch viscoelastic characteristics to the addition of monopalmitin. Amylose content was positively correlated with the increase in final viscosity, which was attributed to the presence of more amylose in non-aggregated state contributing to higher apparent viscosity of the starch paste. Comparison of stored gels obtained from amylose-rich starches with gel prepared from waxy wheat varieties confirmed the critical role of amylose on the formation of starch network and thus providing the strength of the gel. Lack of correlation between textural properties of stored gels with amylose content or rheological characteristics measured by the RVA indicated that subtle differences in starch structure may have far-reaching consequences in relation to the strength of the gels, although these differences may have only limited effect on pasting properties in the RVA Viscoelastic properties of starch paste prepared from commercial wheat starch were significantly altered depending on the chain length and saturation of the fatty acid of the monoglyceride added during repeated heating and cooling in the Rapid Visco Analyser. Varying effects of different monoglycerides on the paste viscosity were attributed to different complexation abilities of these lipids with starch. It was proposed that stability and structure of the starch-lipid complexes formed affect the viscosity trace of the paste subjected to multiple heating and cooling. Our study indicated that differing monoglycerides in combination with the number of heat-cool cycles can be used to induce form I or form II starch-lipid complexes and thus manipulate paste rheology, gel structure and resistant starch content.

Thesis (Ph. D.)--Oregon Graduate Institute of Science and Technology, 1990.

Thesis (Ph.D.)--University of Western Australia, 2004.

Thesis (M.S.)--Ohio State University, 2008. / Title from first page of PDF file. Includes bibliographical references (p. 57-62).

Thesis (Ph. D.)--Drexel University, 2004. / Includes abstract and vita. Includes bibliographical references (leaves 135-137).

Thesis (M.Sc.)--Memorial University of Newfoundland, 2004. / Bibliography: leaves 79-90.

Thesis (Ph. D.)--University of Virginia, 2000. / Spine title: Phospholipase A₂ binding. Includes bibliographical references (p. 245-258). Also available online through Digital Dissertations.

Thesis (Ph. D.)--Drexel University, 2007. / Includes abstract and vita. Includes bibliographical references (leaves 101-108).

Thesis (Ph. D.)--University of Washington, 2007. / Vita. Includes bibliographical references (leaves 112-125).

Doctor of Philosophy / Biochemistry and Molecular Biophysics Interdepartmental Program / Timothy Durrett / In plants including the members of Brassicaceae family, such as Arabidopsis thaliana and Brassica juncea, seed storage reserves, which include lipids and proteins, accumulate in seeds during development. Triacylglycerols (TAG) are the major storage lipids found in the developing seeds, petals, pollen grains, and fruits of plants. In Arabidopsis seeds, acyl-CoA: diacylglycerol acyltransferase 1 (DGAT1) is the major enzyme contributing to TAG biosynthesis. In Arabidopsis, dgat1-1 mutants retain 60-80% seed TAG content due to the involvement of phospholipid: diacylglycerol acyltransferase (PDAT1) in acyl-CoA independent TAG biosynthesis. My study focuses on the elucidation and functional characterization of novel genes involved in the regulation of the TAG biosynthesis pathway. In developing seeds of the dgat1-1 mutant, altered fatty acid composition was observed with reduced TAG content and increased polar lipid content as compared to wild type. RNA-Seq of developing Arabidopsis seeds was employed to detect differentially expressed genes in dgat1-1. An empirical analysis for differential gene expression revealed a significant number of differentially expressed genes among all developmental stages in dgat1-1. Significant changes in gene expression profile were detected in lipid-related genes such as lipases and desaturases. RT-PCR was used to confirm the differential expression of major lipid-related genes including DGAT1, PDAT, and FAD2. Lipid profiling of T-DNA insertion mutants for differentially expressed genes revealed significant changes in lipid content and composition. Mutations in a member of the α, β-hydrolase family, encoded by gene named PLIP1, resulted in smaller seed and an altered seed oil phenotype. Also, combining the dgat1-1 and plip1-2 mutations resulted in a lethal phenotype, demonstrating the important role of this enzyme in embryo development and TAG biosynthesis. To identify key components in the regulation of storage lipid biosynthesis, correlation analysis using differential transcript abundance and lipid profile during different stages of seed development from dgat1-1 and wild type lines of Arabidopsis was performed. Using clustering analysis with Pearson correlation coefficient and single linkage identified one cluster of genes which included PLIP1, FAD2, FAD3, and PDCT . Similar analysis using combined data from the neutral and polar fractions resulted in clustering of lipids containing polyunsaturated fatty acids. To investigate the reduced seed germination phenotype for mature seeds of dgat1-1 and non-germinating green seed phenotype of dgat1-1 plip1-2 lines, differential expression (DE) analysis for genes involved in hormone metabolism was performed. Upregulation of expression was observed for genes involved in promoting abscisic acid (ABA) response, which led us to specuate the role of altered hormone metabolism in delayed germination of dgat1-1 seeds. Development of allopolyploid Brassica species from its diploid progenitors involves duplication, loss, and reshuffling of genes leading to massive genetic redundancy. It leads to selective expression or newly acquired role for duplicated homeologs. Differential expression (DE) analysis for homoeologous genes from A and B subgenomes of allopolyploid B. juncea implicated in FA synthesis, acyl editing, and TAG biosynthesis and metabolism was performed. Differential expression (DE) analysis identified the transcriptional dominance of A subgenome homoeologs. Identification of these homoeologs will enable their use in breeding programs directed towards improvement of lipid content and composition in seeds.