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Sorghum Grain Chemistry and Functionality: Effects of Kernel Maturity, Genetic, Environmental and Management Factors SORGHUM GRAIN CHEMISTRY AND FUNCTIONALITY: EFFECTS OF KERNEL MATURITY, GENETIC, ENVIRONMENTAL AND MANAGEMENT FACTORS. by RHETT CHRISTOPHER KAUFMAN B.S., Kansas State University, 2003 M.S., Kansas State University, 2005 AN ABSTRACT OF A DISSERTATION submitted in partial fulfillment of the requirements for the degree DOCTOR OF PHILOSOPHY Department of Grain Science and Industry College of Agriculture KANSAS STATE UNIVERSITY Manhattan, Kansas 2014 Abstract Sorghum (Sorghum bicolor [L.] Moench) is the fifth most important cereal grain grown in the world. Sorghum is an important cereal crop for both animal feed and biofuel production in the United States. The genetic, environmental, and agronomic management influences on sorghum starch and protein chemistry and functionality were evaluated. A method was developed to determine amylose content in cereal starches that achieved the same level of accuracy and precision as traditional methods, but had the capability of analyzing 50 samples per day or approximately a 10-fold increase in throughput. The effect of kernel maturity on sorghum starch properties was conducted by collecting grain from two hybrids at various stages throughout kernel development. The samples ranged from 16.3% amylose in 10 days after anthesis (DAA) to 23.3% amylose in 35 DAA. Starch thermal properties were also altered due to DAA, most notably the ΔH was 16.1 J/g at 14 DAA and 9.45 J/g at 56 DAA. In a separate study using the same developmental samples the protein and starch digestibility was analyzed. The kernel maturity had a notable effect on digestibility with the maximum digestibility occurring at 17 DAA with 82.44% digestible protein. In another study a diverse set of 19 sorghums was grown in three locations in Kansas to evaluate the genetic, location, and genetic x location effect on grain quality attributes. The physical and chemical properties of the sorghums were greatly affected by the genotype, environment, and the GxE interaction. Protein content ranged from 11.09% to 15.17% and digestibility ranged from 45.58% to 62.05% due to genotype. The final study investigates the role of agronomic management on sorghum grain quality. A sorghum hybrid was grown on plots with varying nitrogen fertilization rates and cover cropping systems that are currently used by Kansas producers. Grain attributes such as hardness and size were variable due to the treatments but negative impacts to protein digestibility were not seen due to cropping system. Sorghum grain quality is affected by many variables and a better understanding of the variables will lead to a higher quality product. SORGHUM GRAIN CHEMISTRYAND FUNCTIONALITY: EFFECTS OF KERNEL MATURITY, GENETIC, ENVIRONMENTAL AND MANAGEMENT FACTORS. by RHETT CHRISTOPHER KAUFMAN B.S., Kansas State University, 2003 M.S., Kansas State University, 2005 A DISSERTATION submitted in partial fulfillment of the requirements for the degree DOCTOR OF PHILOSOPHY Department of Grain Science and Industry College of Agriculture KANSAS STATE UNIVERSITY Manhattan, Kansas 2014 Approved by: Co-Major Professor Yong-Cheng Shi Approved by: Co-Major Professor Jeff D. Wilson Copyright RHETT CHRISTOPHER KAUFMAN 2014 Abstract Sorghum (Sorghum bicolor [L.] Moench) is the fifth most important cereal grain grown in the world. Sorghum is an important cereal crop for both animal feed and biofuel production in the United States. The genetic, environmental, and agronomic management influences on sorghum starch and protein chemistry and functionality were evaluated. A method was developed to determine amylose content in cereal starches that achieved the same level of accuracy and precision as traditional methods, but had the capability of analyzing 50 samples per day or approximately a 10-fold increase in throughput. The effect of kernel maturity on sorghum starch properties was conducted by collecting grain from two hybrids at various stages throughout kernel development. The samples ranged from 16.3% amylose in 10 days after anthesis (DAA) to 23.3% amylose in 35 DAA. Starch thermal properties were also altered due to DAA, most notably the ΔH was 16.1 J/g at 14 DAA and 9.45 J/g at 56 DAA. In a separate study using the same developmental samples the protein and starch digestibility was analyzed. The kernel maturity had a notable effect on digestibility with the maximum digestibility occurring at 17 DAA with 82.44% digestible protein. In another study a diverse set of 19 sorghums was grown in three locations in Kansas to evaluate the genetic, location, and genetic x location effect on grain quality attributes. The physical and chemical properties of the sorghums were greatly affected by the genotype, environment, and the GxE interaction. Protein content ranged from 11.09% to 15.17% and digestibility ranged from 45.58% to 62.05% due to genotype. The final study investigates the role of agronomic management on sorghum grain quality. A sorghum hybrid was grown on plots with varying nitrogen fertilization rates and cover cropping systems that are currently used by Kansas producers. Grain attributes such as hardness and size were variable due to the treatments but negative impacts to protein digestibility were not seen due to cropping system. Sorghum grain quality is affected by many variables and a better understanding of the variables will lead to a higher quality product. Table of Contents List of Figures ................................................................................................................................ xi List of Tables ................................................................................................................................ xii Acknowledgements ...................................................................................................................... xiv Preface........................................................................................................................................... xv Chapter 1 - Introduction .................................................................................................................. 1 Enzymatic Pathway ..................................................................................................................... 2 ADP-glucose pyrophosphorylase (AGPase) ........................................................................... 2 Starch Synthases (SSs) ............................................................................................................ 3 Starch Branching Enzymes (SBEs)......................................................................................... 4 Starch Debranching Enzymes (DBEs) .................................................................................... 4 Miscellaneous Enzymes .......................................................................................................... 5 The Starch Granule ..................................................................................................................... 5 Influences on Starch Synthesis and Functionality ...................................................................... 7 Caryopsis Development .......................................................................................................... 7 Environmental Stresses ........................................................................................................... 8 Separation and Detection of Glucose by HPLC ......................................................................... 9 References ..................................................................................................................................... 15 Chapter 2 - Development of a 96-well Plate Iodine Binding Assay for Amylose Content Determination. ....................................................................................................................... 25 Abstract ..................................................................................................................................... 26 Introduction ............................................................................................................................... 27 2. Materials and Methods ......................................................................................................... 28 2.1 Materials ......................................................................................................................... 28 2.2 Conventional Amylose Measurement ............................................................................. 28 2.3 Development of 96-well Plate Method ........................................................................... 29 2.4 Amylose Content Calculation ......................................................................................... 29 2.5 Statistical Analysis .......................................................................................................... 29 3. Results and Discussion ........................................................................................................ 30 vi 3.1 Standard Curve ................................................................................................................ 30 3.2 Measurement of Amylose in Cereal Starches ................................................................. 30 4. Conclusions .......................................................................................................................... 31 References ................................................................................................................................
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