Jeff D. Wilson

Effect of temperature on expression of genes encoding enzymes for starch biosynthesis in developing wheat endosperm

The effect of high temperature on starch accumulation, starch granule populations, and expression of genes encoding key enzymes for starch biosynthesis was examined during grain development in wheat (Triticum aestivum L. cv. Butte 86). High temperature applied from anthesis to maturity reduced the duration of starch accumulation. Starch accumulation ceased approximately 6 days earlier for grain produced under a 37/17 8C (day/night) regimen and 21 days earlier under a 37/28 8C (day/night) regimen than for grain produced under a 24/17 8C (day/night) regimen. Compared to the 24/17 8C regimen, starch content was approximately 19% less for mature grain produced under the 37/17 8C regimen and 58% less under the 37/28 8C regimen. Based on relative volume, the smaller type B starch granules were the predominant class in mature grain produced under the 24/17 and 37/17 8C regimens, whereas the larger type A granules were predominant in grain produced under the 37/28 8C regimen. Under the 24/17 8C regimen, steady state transcript levels for ADP-glucose pyrophosphorylase, starch synthases I, II, and III, granule-bound starch synthase, and starch branching enzymes I and II were highest from 12/16 days post-anthesis (dpa). Under the 37/17 8C regimen, steady state levels of these transcripts followed the same temporal pattern, but were substantially lower. Under the 37/28 8C regimen, transcript levels peaked earlier, at 7 dpa. The high temperature regimens reduced the relativ el evels of transcripts for starch synthase more than the other starch biosynthetic enzymes. Published by Elsevier Science Ireland Ltd.

Amyloplast Formation and Starch Granule Development in Hard Red Winter Wheat

ABSTRACT Plastids in the coenocytic endosperm of young wheat caryopses were mostly in the form of pleomorphic proplastids with a few of the plastids containing small starch granules. Following cellularization of the coenocytic cytoplasm, the outer one or two endosperm cell layers became meristematic and continued to divide until about 14 days after flowering (DAF). During the first week of endosperm development, newly divided cells had plastids that were pleomorphic in shape, while subaleurone cells interior to the meristematic region contained amyloplasts that contained a single-size class of starch granules (incipient A-type starch granules). The pleomorphic plastids exhibited tubular protrusions that extended a considerable distance through the cytoplasm. Amyloplasts in cells interior to the meristematic region did not exhibit protrusions. Both subaleurone and central endosperm cells had amyloplasts that exhibited protrusions at 10–12 DAF, and some of the protrusions contained small starch granules (in...

Associations of Starch Gel Hardness, Granule Size, Waxy Allelic Expression, Thermal Pasting, Milling Quality, and Kernel Texture of 12 Soft Wheat Cultivars

ABSTRACT Starches were isolated from 12 soft wheat (Triticum aestivum L.) cultivars and were characterized for waxy (Wx) allelic expression, thermal pasting characteristics, and starch granule size. Gels were produced from the thermally degraded starches and were evaluated using large deformation rheological measurements. Data were compared with cultivar kernel texture, milling characteristics, starch chemical analyses, and flour pasting characteristics. Larger flour yields were produced from cultivars that had larger starch granules. Flour yield also was correlated with lower amylose content and greater starch content. Harder starch gels were correlated with higher levels of amylose content and softer kernel texture. The cultivar Fillmore, which had a partial waxy mutation at the B locus, produced the highest peak pasting viscosity and the lowest gel hardness. Softer textured wheats had greater lipid-complexed amylose and starch phosphorus contents and had less total starch content. Among these wheats of...

Comparison of Waxy vs. Nonwaxy Wheats in Fuel Ethanol Fermentation

ABSTRACT Fermentation performance of eight waxy, seven nonwaxy soft, and 15 nonwaxy hard wheat cultivars was compared in a laboratory dry-grind procedure. With nitrogen supplements in the mash, the range of ethanol yields was 368–447 L/ton. Nonwaxy soft wheat had an average ethanol yield of 433 L/ton, higher than nonwaxy hard and waxy wheat. Conversion efficiencies were 91.3–96.2%. Despite having higher levels of free sugars in grain, waxy wheat had higher conversion efficiency than nonwaxy wheat. Although there was huge variation in the protein content between nonwaxy hard and soft wheat, no difference in conversion efficiency was observed. Waxy cultivars had extremely low peak viscosity during liquefaction. Novel mashing properties of waxy cultivars were related to unique pasting properties of starch granules. With nitrogen supplementation, waxy wheat had a faster fermentation rate than nonwaxy wheat. Fermentation rates for waxy cultivars without nitrogen supplementation and nonwaxy cultivars with nitro...

Development of a 96-well plate iodine binding assay for amylose content determination

Cereal starch amylose/amylopectin (AM/AP) is critical in functional properties for food and industrial applications. Conventional methods of AM/AP are time consuming and labor intensive making it difficult to screen the large sample sets necessary for evaluating breeding samples and investigating environmental impact on starch development. The objective was to adapt and optimize the iodine binding assay in a 96-well plate format for measurement at both λ 620 nm and λ 510 nm. The standard curve for amylose content was scaled to a 96-well plate format and demonstrated R(2) values of 0.999 and 0.993 for single and dual wavelengths, respectively. The plate methods were applicable over large ranges of amylose contents: high amylose maize starch at 61.7±2.3%, normal wheat starch at 29.0±0.74%, and a waxy maize starch at 1.2±0.9%. The method exhibited slightly greater amylose content values than the Concanavalin A method for normal type starches; but is consistent with cuvette scale iodine binding assays.

Position of modifying groups on starch chains of octenylsuccinic anhydride-modified waxy maize starch

Octenylsuccinic anhydride (OSA)-modified starches with a low (0.018) and high (0.092) degree of substitution (DS) were prepared from granular native waxy maize starch in aqueous slurry. The position of OS substituents along the starch chains was investigated by enzyme hydrolysis followed by chromatographic analysis. Native starch and two OS starches with a low and high DS had β-limit values of 55.9%, 52.8%, and 34.4%, respectively. The weight-average molecular weight of the β-limit dextrin from the OS starch with a low DS was close to that of the β-limit dextrin from native starch but lower than that of the β-limit dextrin from the OS starch with a high DS. Debranching of OS starches was incomplete compared with native starch. OS groups in the OS starch with a low DS were located on the repeat units near the branching points, whereas the OS substituents in the OS starch with a high DS occurred both near the branching points and the non-reducing ends.

Variability in a Starch Isolation Method and Automated Digital Image Analysis System Used for the Study of Starch Size Distributions in Wheat Flour

ABSTRACT A starch isolation method and digital image analysis system were developed to accurately measure size distributions of the starch populations in wheat. The image analysis system was coupled directly to a light microscope equipped with computer controlled step stage and automatic focus. Automation of data acquisition and processing eliminated some of the labor intensive steps previously required for analyzing starch granule size distributions. This system was used to standardize starch isolation methods and compare variation and precision of the system. Operational variations were determined and statistically assessed. The number of fields of view required for low standard errors and acceptable speed of analysis was determined to be fifty. A major advantage of the system has been the increased resolution. The use of higher magnifications and stage automation allowed the analysis of starch granules as small as 0.84 μm in diameter while analyzing thousands of starch granules per sample.

Variability in tannin content, chemistry and activity in a diverse group of tannin containing sorghum cultivars.

BACKGROUND Tannins are large polyphenolic polymers and are known to bind proteins, limiting their digestibility, but are also excellent antioxidants. Numerous studies investigating the functional properties of sorghum tannin have been conducted by comparing grain samples from different sorghum lines without considering the other intrinsic characteristics of the grain. The purpose of this study was to remove the confounding intrinsic factors present in the endosperm so the effect of the tannins could be evaluated utilizing a unique decortication/reconstitution procedure. RESULTS The tannin content of the 14 cultivars tested ranged from 2.3 to 67.2 catechin equivalents. The bran fractions were studied for their impact on protein binding and antioxidant capacity. Protein digestibility by pepsin ranged from 8% to 58% at the highest tannin level addition. Protein binding ranged from 3.11 to 16.33 g blue bovine serum albumin kg⁻¹ bran. Antioxidant capacity ranged from 81.33 to 1122.54 µmol Trolox equivalents g⁻¹ bran. High-performance size-exclusion chromatography detailed molecular size distributions of the tannin polymers and relationship to tannin functionality. CONCLUSION The tannin content and composition play a significant role in determining tannin functionality. These differences will allow for selections of high-tannin sorghums with consideration of the biological activities of the tannins.

Ultrastructure of Developing Hard and Soft Red Winter Wheats After Air- and Freeze-Drying and Its Relationship to Endosperm Texture

ABSTRACT A transmission electron microscopic study was conducted on air- and freeze-dried developing wheat to determine the effects of drying on the structure of the starchy endosperm. Field-grown hard red winter wheat (Karl) and soft red winter wheat (Clark) were harvested at 15, 18, 21, 23, 25, 28, and 35 days after flowering (DAF). Wheat was dried by either air-drying in the spike at 28°C or freeze-drying following freezing in liquid nitrogen. Dried wheat was prepared for microscopy. Fresh samples of Karl and Clark were also harvested on the same days and prepared immediately for microscopy. The method of drying greatly affected cellular ultrastructure. The most pronounced change upon air-drying of developing samples was disappearance of individual protein bodies and conversion of the cytoplasm into a matrix-like material similar in appearance to storage protein matrix found in mature wheat endosperm. Freeze-dried wheats maintained nearly natural ultrastructure but exhibited various amounts of freeze d...

Effect of Soft Kernel Texture on the Milling Properties of Soft Durum Wheat

Worldwide, nearly 20 times more common wheat (Triticum aestivum) is produced than durum wheat (T. turgidum subsp. durum). Durum wheat is predominately milled into coarse semolina owing to the extreme hardness of the kernels. Semolina, lacking the versatility of traditional flour, is used primarily in the production of pasta. The puroindoline genes, responsible for kernel softness in wheat, have been introduced into durum via homoeologous recombination. The objective of this study was to determine what impact the introgression of the puroindoline genes, and subsequent expression of the soft kernel phenotype, had on the milling properties and flour characteristics of durum wheat. Three grain lots of Soft Svevo and one of Soft Alzada, two soft-kernel back-cross derived durum varieties, were milled into flour on the modified Quadrumat Senior laboratory mill at 13, 14, and 16% temper levels. Samples of Svevo (a durum wheat and recurrent parent of Soft Svevo), Xerpha (a soft white winter wheat), and Expresso (a...