Genetic architecture of end-use quality traits in soft white winter wheat.

BACKGROUND: Genetic improvement of end-use quality is an important objective in wheat breeding programs to meet the requirements of grain markets, millers, and bakers. However, end-use quality phenotyping is expensive and laborious thus, testing is often delayed until advanced generations. To better understand the underlying genetic architecture of end-use quality traits, we investigated the phenotypic and genotypic structure of 14 end-use quality traits in 672 advanced soft white winter wheat breeding lines and cultivars adapted to the Pacific Northwest region of the United States. RESULTS: This collection of germplasm had continuous distributions for the 14 end-use quality traits with industrially significant differences for all traits. The breeding lines and cultivars were genotyped using genotyping-by-sequencing and 40,518 SNP markers were used for association mapping (GWAS). The GWAS identified 178 marker-trait associations (MTAs) distributed across all wheat chromosomes. A total of 40 MTAs were positioned within genomic regions of previously discovered end-use quality genes/QTL. Among the identified MTAs, 12 markers had large effects and thus could be considered in the larger scheme of selecting and fixing favorable alleles in breeding for end-use quality in soft white wheat germplasm. We also identified 15 loci (two of them with large effects) that can be used for simultaneous breeding of more than a single end-use quality trait. The results highlight the complex nature of the genetic architecture of end-use quality, and the challenges of simultaneously selecting favorable genotypes for a large number of traits. This study also illustrates that some end-use quality traits were mainly controlled by a larger number of small-effect loci and may be more amenable to alternate selection strategies such as genomic selection. CONCLUSIONS: In conclusion, a breeder may be faced with the dilemma of balancing genotypic selection in early generation(s) versus costly phenotyping later on.

Late-maturity α-amylase (LMA): exploring the underlying mechanisms and end-use quality effects in wheat.

MAIN CONCLUSION: A comprehensive understanding of LMA from the underlying molecular aspects to the end-use quality effects will greatly benefit the global wheat industry and those whose livelihoods depend upon it. Late-maturity α-amylase (LMA) leads to the expression and protein accumulation of high pI α-amylases during late grain development. This α-amylase is maintained through harvest and leads to an unacceptable low falling number (FN), the wheat industry's standard measure for predicting end-use quality. Unfortunately, low FN leads to significant financial losses for growers. As a result, wheat researchers are working to understand and eliminate LMA from wheat breeding programs, with research aims that include unraveling the genetic, biochemical, and physiological mechanisms that lead to LMA expression. In addition, cereal chemists and quality scientists are working to determine if and how LMA-affected grain impacts end-use quality. This review is a comprehensive overview of studies focused on LMA and includes open questions and future directions.

Evidence of intralocus recombination at the Glu-3 loci in bread wheat (Triticum aestivum L.).

KEY MESSAGE: Recombination at the Glu-3 loci was identified, and strong genetic linkage was observed only between the amplicons representing i-type and s-type genes located, respectively, at the Glu-A3 and Glu-B3 loci. The low-molecular weight glutenin subunits (LMW-GSs) are one of the major components of wheat seed storage proteins and play a critical role in the determination of wheat end-use quality. The genes encoding this class of proteins are located at the orthologous Glu-3 loci (Glu-A3, Glu-B3, and Glu-D3). Due to the complexity of these chromosomal regions and the high sequence similarity between different LMW-GS genes, their organization and recombination characteristics are still incompletely understood. This study examined intralocus recombination at the Glu-3 loci in two recombinant inbred line (RIL) and one doubled haploid (DH) population, all segregating for the Glu-A3, Glu-B3, and Glu-D3 loci. The analysis was conducted using a gene marker system that consists of the amplification of the complete set of the LMW-GS genes and their visualization by capillary electrophoresis. Recombinant marker haplotypes were detected in all three populations with different recombination rates depending on the locus and the population. No recombination was observed between the amplicons representing i-type and s-type LMW-GS genes located, respectively, at the Glu-A3 and Glu-B3 loci, indicating tight linkage between these genes. Results of this study contribute to better understanding the genetic linkage and recombination between different LMW-GS genes, the structure of the Glu-3 loci, and the development of more specific molecular markers that better represent the genetic diversity of these loci. In this way, a more precise analysis of the contribution of various LMW-GSs to end-use quality of wheat may be achieved.

Selecting High-Performing and Stable Pea Genotypes in Multi-Environmental Trial (MET): Applying AMMI, GGE-Biplot, and BLUP Procedures.

A large amount of data on various traits is accumulated over the course of a breeding program and can be used to optimize various aspects of the crop improvement pipeline. We leveraged data from advanced yield trials (AYT) of three classes of peas (green, yellow, and winter peas) collected over ten years (2012-2021) to analyze and test key aspects fundamental to pea breeding. Six balanced datasets were used to test the predictive success of the BLUP and AMMI family models. Predictive assessment using cross-validation indicated that BLUP offered better predictive accuracy as compared to any AMMI family model. However, BLUP may not always identify the best genotype that performs well across environments. AMMI and GGE, two statistical tools used to exploit GE, could fill this gap and aid in understanding how genotypes perform across environments. AMMI's yield by environmental IPCA1, WAASB by yield plot, and GGE biplot were shown to be useful in identifying genotypes for specific or broad adaptability. When compared to the most favorable environment, we observed a yield reduction of 80-87% in the most unfavorable environment. The seed yield variability across environments was caused in part by weather variability. Hotter conditions in June and July as well as low precipitation in May and June affected seed yield negatively. In conclusion, the findings of this study are useful to breeders in the variety selection process and growers in pea production.

Quantitative proteomic analysis of super soft kernel texture in soft white spring wheat.

Super soft kernel texture is associated with superior milling and baking performance in soft wheat. To understand the mechanism underlying super soft kernel texture, we studied proteomic changes between a normal soft and a super soft during kernel development. The cultivar 'Alpowa', a soft white spring wheat, was crossed to a closely related super soft spring wheat line 'BC2SS163' to produce F6 recombinant inbred lines (RILs). Four normal soft RILs and four super soft RILs along with the parents were selected for proteomic analysis. Alpowa and the normal soft RILs showed hardness indices of 20 to 30, whereas BC2SS163 and the super soft RILs showed hardness indices of -2 to -6. Kernels were collected from normal soft and super soft genotypes at 7 days post anthesis (dpa), 14 dpa, 28 dpa, and maturity and were subject to quantitative proteomic analysis. Throughout kernel development, 175 differentially abundant proteins (DAPs) were identified. Most DAPs were observed at 7 dpa, 14 dpa, and 28 dpa. Of the 175 DAPs, 32 had higher abundance in normal soft wheat, whereas 143 DAPs had higher abundance in super soft wheat. A total of 18 DAPs were associated with carbohydrate metabolism and five DAPs were associated with lipids. The gene TraesCS4B02G091100.1 on chromosome arm 4BS, which encodes for sucrose-phosphate synthase, was identified as a candidate gene for super soft kernel texture in BC2SS163. This study enhanced our understanding of the mechanism underlying super soft kernel texture in soft white spring wheat.

Prevalence of Puroindoline D1 and Puroindoline b-2 variants in U.S. Pacific Northwest wheat breeding germplasm pools, and their association with kernel texture.

Kernel texture is a major factor influencing the classification and end use properties of wheat (Triticum aestivum L.), and is mainly controlled by the Puroindoline a (Pina) and Puroindoline b (Pinb) genes. Recently, a new puroindoline gene, Puroindoline b-2 (Pin b-2), was identified. In this study, 388 wheat cultivars and advanced breeding lines from the U.S. Pacific Northwest were investigated for frequencies of Puroindoline D1 alleles and Pinb-2 variants 2 and 3. Results indicated that Pinb-D1b (74.0%) was the predominant genotype among hard wheats (N = 196), the only other hard allele encountered was Pina-D1b (26.0%). Across all varieties, Pinb-2v3 was the predominant genotype (84.5%) compared with Pinb-2v2 (15.5%). However, among 240 winter wheat varieties (124 soft white, 15 club, 68 hard red and 33 hard white varieties), all carried Pinb-2v3. Among spring wheats, Pinb-2v2 and Pinb-2v3 frequencies were more variable (soft white 25.0:75.0, hard red 58.2:41.8 and hard white 40.0:60.0, respectively). Kernel texture variation was analyzed using 247 of the 388 wheat varieties grown in multi-location factorial trials in up to 7 crop years. The range of variety means among the four groups, soft winter, soft spring, hard winter and hard spring, was on the order of 15-25 single kernel characterization system (SKCS) Hardness Index. The least significant difference for each of these trials ranged from 2.8 to 5.6 SKCS Hardness Index. Observations lead to the conclusion that Pinb-2 variants do not exert a prominent effect on kernel texture, however, Pinb-2 variants do identify features of wheat germ plasm structure in the U.S. Pacific Northwest.

Prediction of Protein Concentration in Pea (Pisum sativum L.) Using Near-Infrared Spectroscopy (NIRS) Systems.

Breeding for increased protein concentration is a priority in field peas. Having a quick, accurate, and non-destructive protein quantification method is critical for screening breeding materials, which the near-infrared spectroscopy (NIRS) system can provide. Partial least square regression (PLSR) models to predict protein concentration were developed and compared for DA7250 and FT9700 NIRS systems. The reference protein data were accurate and exhibited a wider range of variation (15.3−29.8%). Spectral pre-treatments had no clear advantage over analyses based on raw spectral data. Due to the large number of samples used in this study, prediction accuracies remained similar across calibration sizes. The final PLSR models for the DA7250 and FT9700 systems required 10 and 13 latent variables, respectively, and performed well and were comparable (R2 = 0.72, RMSE = 1.22, and bias = 0.003 for DA7250; R2 = 0.79, RMSE = 1.23, and bias = 0.055 for FT9700). Considering three groupings for protein concentration (Low: <20%, Medium: ≥20%, but ≤25%, and High: >25%), none of the entries changed from low to high or vice versa between the observed and predicted values for the DA7250 system. Only a single entry moved from a low category in the observed data to a high category in the predicted data for the FT9700 system in the calibration set. Although the FT9700 system outperformed the DA7250 system by a small margin, both systems had the potential to predict protein concentration in pea seeds for breeding purposes. Wavelengths between 950 nm and 1650 nm accounted for most of the variation in pea protein concentration.

Serum Melatonin Values in Normal Dogs and Dogs with Seizures.

Epilepsy, or recurrent seizures, is reported to be the most common neurologic condition in dogs; 20-30% of epileptic dogs are considered to be pharmacoresistent to one medication. The hormone melatonin has been shown to have significant anticonvulsant effects; epileptic humans have lower serum melatonin levels than unaffected individuals. We hypothesize that serum levels of melatonin will be lower in dogs with seizures as compared with normal dogs. Sixty-two dogs were enrolled in the study: 29 normal dogs (Group 1) and 33 dogs with seizures (Group 2). Blood sampling was done at three separate time points (8:00 a.m., 12:00, and 4:00 p.m.). The majority of dogs in Groups 1 (69%) and 2 (76%) had serum melatonin levels <0.5 pg/mL as measured by radioimmunoassay. There were no significant differences in serum melatonin values between the groups or within groups when time of blood draw, length of sample freezing, time of day/year of sampling, or presence of anticonvulsant therapy were compared. There were no notable differences in daytime serum melatonin values in normal dogs versus dogs with seizures. The majority of daytime serum melatonin levels were <0.5 pg/mL in dogs with and without seizures.

Influence of Soft Kernel Texture on Fresh Durum Pasta.

This study examined the quality of fresh pasta made from 3 varieties of a new type of durum wheat possessing soft kernel texture, as compared to fresh pasta made from commercial samples of durum semolina, durum flour, and bread flour, each at 3 levels of hydration (28%, 30%, and 32%, respectively). Soft durum possesses a small part of chromosome 5D that carries the Hardness locus and puroindoline genes. The soft durum lines were derived from the durum varieties Svevo, Alzada, and Havasu. The soft durum pasta exhibited low cooking weight increase (water uptake) (115% to 122%), the lowest cooking loss (∼3% to 4%), high firmness (269.3, 265.8, and 297.9 g, Soft Svevo, Soft Havasu, and Soft Alzada, respectively, versus 239.7 and 273.6 g, durum flour and semolina, respectively), low stickiness (4.17 to 4.96 g·s for the soft durums compared with 5.04 for the semolina), and raw and cooked pasta color comparable to or superior to those exhibited by the durum semolina (high L* and b* ). The soft durum samples also exhibited pasta quality superior to both the durum flour and bread flour samples. These results challenge the long-standing view that high-quality pasta must be made from durum semolina. PRACTICAL APPLICATION: This study illustrates the quality and potential applications of soft durum wheat in pasta manufacturing. As a new type of wheat, understanding these properties is crucial for manufacturers and others who may be interested in utilizing soft durum.

Goniometry and Limb Girth in Miniature Dachshunds.

Purpose. To report the mean and median pelvic limb joint angles and girth measurements in miniature Dachshunds presenting with varying degrees of pelvic limb weakness secondary to thoracolumbar intervertebral disc extrusion. Methods. 15 miniature Dachshunds who presented to WSU-VTH for thoracolumbar disc extrusion. Dachshunds varied in neurologic status from ambulatory paraparetic to paraplegic at the time of measurements. Results. There were no significant differences in joint angles or girth among the three groups (ambulatory paraparetic, nonambulatory paraparetic, or paraplegic) (P > 0.05). When group was disregarded and values for extension, flexion, and girth combined, no differences existed. Conclusions. Goniometry and limb girth measurements can successfully be made in the miniature Dachshund; however, the shape of the Dachshund leg makes obtaining these values challenging. There were no differences in joint angle or girth measurements between dogs with varying neurologic dysfunction at the time of measurement.

Repeatability of Mice Consumption Discrimination of Wheat (Triticum aestivum L.) Varieties across Field Experiments and Mouse Cohorts.

UNLABELLED: Whole grain wheat (Triticum aestivum L.) foods can provide critical nutrients for health and nutrition in the human diet. Potential flavor differences among varieties can be examined using consumption discrimination of the house mouse (Mus musculus L.) as a model system. This study examines consistency and repeatability of the mouse model and potentially, wheat grain flavor. A single elimination tournament design was used to measure relative consumption preference for hard red spring and hard white spring varieties across all 3 experiments in combination with 2 mouse cohorts. Fifteen replicate mice were used in 24-h trials to examine differences in preference among paired wheat varieties until an overall "winner" was established as the most highly preferred variety of wheat. In all 3 experiment-cohort combinations, the same varieties were preferred as the "winner" of both the hard red spring and hard white spring wheat varieties, Hollis and BR 7030, respectively. Despite the consistent preference for these varieties across experiments, the degree (magnitude) to which the mice preferred these varieties varied across experiments. For the hard white spring wheat varieties, the small number of varieties and confounding effects of experiment and cohort limited our ability to accurately gauge repeatability. Conversely, for the hard red spring wheat varieties, consumption preferences were consistent across experiments and mice cohorts. The single-elimination tournament model was effective in providing repeatable results in an effort to more fully understand the mouse model system and possible flavor differences among wheat varieties. PRACTICAL APPLICATION: The mouse model system used here is effective in identifying wheat varieties that may be more or less desirable to humans in whole wheat foods. The system identifies consistent differences across different mouse cohorts and crop years.

Determination of optimal storage temperature and duration for analysis of total and isoenzyme lactate dehydrogenase activities in canine serum and cerebrospinal fluid.

BACKGROUND: Lactate dehydrogenase (LDH) exists as 5 isoenzymes in both serum and cerebrospinal fluid (CSF). Human studies have demonstrated that changes in LDH activity can be correlated with a particular disease. OBJECTIVES: Conflicting reports regarding the stability of LDH made it necessary to determine storage conditions before further study of the diagnostic power of this enzyme's activity can be pursued in dogs. The purpose of this study was to optimize measurement of LDH activity and analysis of its isoenzyme profile in canine serum and CSF through proper storage. METHODS: Serum and CSF were collected from 5 healthy dogs. Samples were stored at 22°C, 4°C, or -20°C for up to 2 months. Total LDH activity was measured spectrophotometrically. Isoenzyme profiles were determined using the QuickGel LDH Isoenzyme technique and densitometric scanning. Retention of > 70% LDH activity in stored samples was considered clinically acceptable. RESULTS: Serum and CSF stored at -20°C retained > 85% of the total LDH activity for 4 weeks, although CSF total LDH activity degraded by > 10% within 24 hours of storage. All serum LDH isoenzymes retained > 85% activity for up to 4 weeks at -20°C. CSF LDH isoenzyme activity degraded rapidly, therefore CSF LDH should be evaluated within 72 hours to assure > 75% of LDH isoenzyme activity. CONCLUSIONS: Proper storage at -20°C can optimize detection of total LDH activity and the LDH isoenzyme profile in canine serum and CSF. This information is important for evaluating the potential usefulness of LDH in veterinary medicine.

Effect of processing on phenolic composition of dough and bread fractions made from refined and whole wheat flour of three wheat varieties.

This study investigated the effect of breadmaking on the assay of phenolic acids from flour, dough, and bread fractions of three whole and refined wheat varieties. Comparison of the efficacy of two commonly used methods for hydrolysis and extraction of phenoilc acids showed that yields of total phenolic acids (TPA) were 5-17% higher among all varieties and flour types when samples were directly hydrolyzed in the presence of ascorbate and EDTA as compared to the method separating free, soluble conjugates and bound, insoluble phenolic acids. Ferulic acid (FA) was the predominant phenolic acid, accounting for means of 59 and 81% of TPA among all refined and whole wheat fractions, respectively. All phenolic acids measured were more abundant in whole wheat than in refined samples. Results indicated that the total quantified phenolic acids did not change significantly when breads were prepared from refined and whole wheat flour. Thus, the potential phytochemical health benefits of total phenolic acids appear to be preserved during bread baking.