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.

As the number falls, alternatives to the Hagberg-Perten falling number method: A review.

This review examines the application, limitations, and potential alternatives to the Hagberg-Perten falling number (FN) method used in the global wheat industry for detecting the risk of poor end-product quality mainly due to starch degradation by the enzyme α-amylase. By viscometry, the FN test indirectly detects the presence of α-amylase, the primary enzyme that digests starch. Elevated α-amylase results in low FN and damages wheat product quality resulting in cakes that fall, and sticky bread and noodles. Low FN can occur from preharvest sprouting (PHS) and late maturity α-amylase (LMA). Moist or rainy conditions before harvest cause PHS on the mother plant. Continuously cool or fluctuating temperatures during the grain filling stage cause LMA. Due to the expression of additional hydrolytic enzymes, PHS has a stronger negative impact than LMA. Wheat grain with low FN/high α-amylase results in serious losses for farmers, traders, millers, and bakers worldwide. Although blending of low FN grain with sound wheat may be used as a means of moving affected grain through the marketplace, care must be taken to avoid grain lots from falling below contract-specified FN. A large amount of sound wheat can be ruined if mixed with a small amount of sprouted wheat. The FN method is widely employed to detect α-amylase after harvest. However, it has several limitations, including sampling variability, high cost, labor intensiveness, the destructive nature of the test, and an inability to differentiate between LMA and PHS. Faster, cheaper, and more accurate alternatives could improve breeding for resistance to PHS and LMA and could preserve the value of wheat grain by avoiding inadvertent mixing of high- and low-FN grain by enabling testing at more stages of the value stream including at harvest, delivery, transport, storage, and milling. Alternatives to the FN method explored here include the Rapid Visco Analyzer, enzyme assays, immunoassays, near-infrared spectroscopy, and hyperspectral imaging.

Genome-wide association mapping of the 'super-soft' kernel texture in white winter wheat.

KEY MESSAGE: The novel super-soft kernel phenotype has the potential to improve wheat processing and flour quality. We identified genomic regions associated with this kernel texture in white winter wheat. Grain hardness is a key determinant of wheat milling and baking quality. The recently discovered 'super-soft' kernel phenotype has the potential to improve wheat processing and flour quality. However, the genetic basis underlying the super-soft trait in wheat is not yet well understood. In this study, we investigated the phenotypic and genotypic structure of the super-soft trait in a collection of 172 advanced soft white winter wheat breeding lines and cultivars adapted to the Pacific Northwest region of the USA. This collection had a continuous distribution for grain hardness index (single-kernel characterization system). Ten super-soft genotypes showed hardness index ≤ 12 including the cultivar Jasper. Over 98,000 SNP markers from genotyping-by-sequencing were used for association mapping (GWAS). The GWAS identified 20 significant markers associated with grain hardness. These significant SNPs corresponded to seven QTL on chromosomes 2B, 3A, 3B, 5A, 6B,7A, and one unaligned chromosome. Two of these QTL, QSKhard.wql-3A and QSKhard.wql-5A, had large effects and distinguished between the normal soft and the super-soft classes. QSKhard.wql-3A and QSKhard.wql-5A reduced the hardness index by 11.7 and 13.1 on average, respectively. The remaining QTL had small effects and reduced grain hardness within the normal soft range. QSKhard.wql-2B, QSKhard.wql-3A, QSKhard.wql-3B, and QSKhard.wql-6B were not previously reported to be in genomic regions of grain hardness-related genes/QTL. The identified super-soft genotypes as well as the SNPs associated with lower grain hardness will be useful to assist breeding for this grain texture trait.

The antimicrobial properties of the puroindolines, a review.

Antimicrobial proteins, and especially antimicrobial peptides (AMPs) hold great promise in the control of animal and plant diseases with low risk of pathogen resistance. The two puroindolines, a and b, from wheat control endosperm softness of the wheat caryopsis (grain), but have also been shown to inhibit the growth and kill various bacteria and fungi, while showing little toxicity to erythrocytes. Puroindolines are small (~ 13 kDa) amphipathic proteins with a characteristic tryptophan-rich domain (TRD) that is part of an 18 or 19 amino acid residue loop subtended by a disulfide bond. This review presents a brief history of the puroindolines, their physical-chemical characteristics, their interaction with lipids and membranes, and their activity as antimicrobial proteins and AMPs. In this latter context, the use of the TRDs of puroindoline a and b in puroindoline AMP function is reviewed. The activity of puroindoline a and b and their AMPs appear to act through similar but somewhat different modes, which may involve membrane binding, membrane disruption and ion channel formation, and intra-cellular nucleic acid binding and metabolic disruption. Natural and synthetic mutants have identified key elements of the puroindolines for antimicrobial activity.

Determinants of wheat noodle color.

Noodles are a leading food in the world, and color is a key determinant of consumer acceptance. In this review the two prominent forms of wheat noodles are considered: white salted and alkaline. Many of the preparation and evaluation strategies are the same for both, with prominence placed on 'brightness' (L*) or a lack of discoloration (ΔL*), and the absence of 'specks.' All raw noodles darken over time. Increasing the protein content of flours almost always translates into darker noodles. Greater discoloration is also associated with higher flour extraction rates, higher ash contents, and higher starch damage. Increasing storage time, dough water absorption, and temperature all often lead to greater discoloration. There is a large range in noodle color variation, and much of this variation is associated with genetics. Consequently, much research has been devoted to methods of screening germplasm, either as whole seeds, meals, flours, or noodle sheets. Polyphenol oxidase (PPO) is a primary culprit in noodle discoloration and has guided much of the research on noodle color. It is now possible to select germplasm with very low levels of PPO through the use of efficacious phenotype screens and the use of molecular markers. The success of this research has provided the opportunity to select wheat breeding lines with nil PPO activity, and to combine favorable alleles at multiple PPO loci. Yet, when noodles are prepared, we continue to observe discoloration. As our ability to minimize PPO activity increases, this 'non-PPO' discoloration has become more important. Perhaps the 'holy grail' is a noodle that never discolors, and has the 'perfect' level of a* (redness, zero?) and b* (yellowness/creaminess). Published 2018. This article is a U.S. Government work and is in the public domain in the USA.

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.

A review of the occurrence of Grain softness protein-1 genes in wheat (Triticum aestivum L.).

Grain softness protein-1 (Gsp-1) is a small, 495-bp intronless gene found throughout the Triticeae tribe at the distal end of group 5 chromosomes. With the Puroindolines, it constitutes a key component of the Hardness locus. Gsp-1 likely plays little role in grain hardness, but has direct interest due to its utility in phylogeny and its role in arabinogalactan peptides. Further role(s) remain to be identified. In the polyploid wheats, Triticum aestivum and T. turgidum, the gene is present in a homoeologous series. Since its discovery, there have been conflicting reports and data as to the number of Gsp-1 genes and the level of sequence polymorphism. Little is known about allelic variation within a species. In the simplest model, a single Gsp-1 gene is present in each wheat and Aegilops tauschii genome. The present review critically re-examines the published and some unpublished data (sequence available in the NCBI nucleotide and MIPS Wheat Genome Databases). A number of testable hypotheses are identified, and include the level of polymorphism that may represent (and define) different Gsp-1 alleles, the existence of a fourth Gsp-1 gene, and the apparent, at times, high level of naturally-occurring or artifactual gene chimeras. In summary, the present data provide firm evidence for at most, three Gsp-1 genes in wheat, although there are numerous data that suggest a more complex model.

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.

The distal portion of the short arm of wheat (Triticum aestivum L.) chromosome 5D controls endosperm vitreosity and grain hardness.

Kernel vitreosity is an important trait of wheat grain, but its developmental control is not completely known. We developed back-cross seven (BC(7)) near-isogenic lines in the soft white spring wheat cultivar Alpowa that lack the distal portion of chromosome 5D short arm. From the final back-cross, 46 BC(7)F(2) plants were isolated. These plants exhibited a complete and perfect association between kernel vitreosity (i.e. vitreous, non-vitreous or mixed) and Single Kernel Characterization System (SKCS) hardness. Observed segregation of 10:28:7 fit a 1:2:1 Chi-square. BC(7)F(2) plants classified as heterozygous for both SKCS hardness and kernel vitreosity (n = 29) were selected and a single vitreous and non-vitreous kernel were selected, and grown to maturity and subjected to SKCS analysis. The resultant phenotypic ratios were, from non-vitreous kernels, 23:6:0, and from vitreous kernels, 0:1:28, soft:heterozygous:hard, respectively. Three of these BC(7)F(2) heterozygous plants were selected and 40 kernels each drawn at random, grown to maturity and subjected to SKCS analysis. Phenotypic segregation ratios were 7:27:6, 11:20:9, and 3:28:9, soft:heterozygous:hard. Chi-square analysis supported a 1:2:1 segregation for one plant but not the other two, in which cases the two homozygous classes were under-represented. Twenty-two paired BC(7)F(2):F(3) full sibs were compared for kernel hardness, weight, size, density and protein content. SKCS hardness index differed markedly, 29.4 for the lines with a complete 5DS, and 88.6 for the lines possessing the deletion. The soft non-vitreous kernels were on average significantly heavier, by nearly 20%, and were slightly larger. Density and protein contents were similar, however. The results provide strong genetic evidence that gene(s) on distal 5DS control not only kernel hardness but also the manner in which the endosperm develops, viz. whether it is vitreous or non-vitreous.

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.

Increasing the Versatility of Durum Wheat through Modifications of Protein and Starch Composition and Grain Hardness.

Although durum wheat (Triticum durum L. ssp. durum Desf.) has traditionally been used to make a range of food products, its use has been restricted due to the absence of the D-genome glutenin proteins, the relatively low variability in starch composition, and its very hard grain texture. This review focuses on the manipulation of the starch and protein composition and modification of the hardness of durum wheat in order to improve its technological and nutritional value and expand its utilization for application to a wider number of end products. Starch is composed of amylopectin and amylose in a 3:1 ratio, and their manipulation has been explored for achieving starch with modified composition. In particular, silencing of the genes involved in amylose and amylopectin synthesis has made it possible to isolate durum wheat lines with amylose content varying from 2-3% up to 75%. This has created opportunities for new products with different properties and enhanced nutritional value. Durum-made bread has generally inferior quality to bread made from common wheat. Attempts to introduce the Glu-D1 subunits 1Dx5 + 1Dy10 and 1Dx2 + 1Dy12 produced stronger dough, but the former produced excessively strong, inelastic doughs, and loaf volume was either inferior or not affected. In contrast, the 1Dx2 + 1Dy12 sometimes improved bread loaf volume (LV) depending on the glutenin subunit background of the genotype receiving these genes. Further breeding and selection are needed to improve the dough extensibility to allow higher LV and better texture. The versatility of durum wheat has been greatly expanded with the creation of soft-textured durum via non-GMO introgression means. This soft durum mills like soft hexaploid wheat and has similar baking properties. The pasta quality is also not diminished by the soft-textured kernels. The Glu-D1 locus containing the subunits 1Dx2 + 1Dy12 has also been introgressed to create higher quality soft durum bread.

Genomic Selection for End-Use Quality and Processing Traits in Soft White Winter Wheat Breeding Program with Machine and Deep Learning Models.

Breeding for grain yield, biotic and abiotic stress resistance, and end-use quality are important goals of wheat breeding programs. Screening for end-use quality traits is usually secondary to grain yield due to high labor needs, cost of testing, and large seed requirements for phenotyping. Genomic selection provides an alternative to predict performance using genome-wide markers under forward and across location predictions, where a previous year's dataset can be used to build the models. Due to large datasets in breeding programs, we explored the potential of the machine and deep learning models to predict fourteen end-use quality traits in a winter wheat breeding program. The population used consisted of 666 wheat genotypes screened for five years (2015-19) at two locations (Pullman and Lind, WA, USA). Nine different models, including two machine learning (random forest and support vector machine) and two deep learning models (convolutional neural network and multilayer perceptron) were explored for cross-validation, forward, and across locations predictions. The prediction accuracies for different traits varied from 0.45-0.81, 0.29-0.55, and 0.27-0.50 under cross-validation, forward, and across location predictions. In general, forward prediction accuracies kept increasing over time due to increments in training data size and was more evident for machine and deep learning models. Deep learning models were superior over the traditional ridge regression best linear unbiased prediction (RRBLUP) and Bayesian models under all prediction scenarios. The high accuracy observed for end-use quality traits in this study support predicting them in early generations, leading to the advancement of superior genotypes to more extensive grain yield trails. Furthermore, the superior performance of machine and deep learning models strengthens the idea to include them in large scale breeding programs for predicting complex traits.

Environment characterization and genomic prediction for end-use quality traits in soft white winter wheat.

End-use quality phenotyping is laborious and expensive, thus, testing may not occur until later generations in wheat breeding programs. We investigated the pattern of genotype × environment (G × E) interaction for end-use quality traits in soft white wheat (Triticum aestivum L.) and tested the effectiveness of implementing genomic selection to optimize breeding for these traits. We used a multi-environment unbalanced dataset comprised of 672 breeding lines and cultivars adapted to the Pacific Northwest region of the United States, which were evaluated for 14 end-use quality traits. Genetic correlations between environments based on factor analytic models showed low-to-moderate G × E interaction for most traits but high G × E interaction for grain and flour protein. A total of 40,518 single-nucleotide polymorphism markers were used for genomic prediction. Genomic prediction accuracies were high for most traits thereby justifying the use of genomic selection to assist breeding for superior end-use quality in soft white wheat. Excluding outlier environments based on genetic correlations between environments was more effective in increasing genomic prediction accuracies compared with that based on environment clustering analysis. For kernel size, kernel weight, milling score, ash, and flour swelling volume, excluding outlier environments increased prediction accuracies by 1-11%. However, for grain and flour protein, flour yield, and cookie diameter, excluding outlier environments did not improve genomic prediction performance.

Inhibitory Effect of Puroindoline Peptides on Campylobacter jejuni Growth and Biofilm Formation.

Puroindolines are small, amphipathic, wheat proteins that determine the hardness of the wheat kernel and protect crops from different pathogens. Puroindoline A (PinA) and puroindoline B (PinB) are two major isoforms of puroindolines. These proteins have antibacterial and antifungal properties mainly attributed to their characteristic tryptophan-rich domains (TRDs). In this in vitro study, we investigated the antimicrobial effect of PinA and PinB synthetic peptides against the growth and biofilm formation of Campylobacter jejuni. C. jejuni is an important microaerobic, foodborne pathogen that causes gastrointestinal and neurological diseases in humans. Our results showed that: (1) PinA, but not PinB, has strong antimicrobial activity against C. jejuni clinical strains 81-176 and F38011, Escherichia coli O157:H7, methicillin-resistant Staphylococcus aureus, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes; (2) The substitution of two tryptophan residues to glycine (W→G) in the TRD of PinA abolishes its antimicrobial activity against these microorganisms; (3) PinA functions additively with two common antibiotics (ciprofloxacin and erythromycin) to inhibit or inactivate C. jejuni strains; (4) PinA damages the C. jejuni cellular membrane, (5) PinA is cytotoxic to human INT 407 cells at high concentrations; and (6) PinA inhibits C. jejuni biofilm formation. In summary, this study demonstrates the antimicrobial activity of PinA against C. jejuni growth and biofilm formation and further confirms the potential use of PinA as a therapeutic agent in health care or as preservatives in the agri-food industry.

Physical mapping and a new variant of Puroindoline b-2 genes in wheat.

Puroindoline a and b proteins soften the endosperm of wheat kernels. When the underlying puroindoline genes are altered by mutation or are deleted, kernels become harder. Thus, puroindoline a and b (Pina and Pinb) play an important role in wheat quality and utilization. Recently, additional Pinb genes have been reported. In the present report, we provide corroborating coding and additional 5' and 3' flanking sequence for three Pinb variants: Pinb-2v1, Pinb-2v2, and Pinb-2v3. Additionally, a new Pinb variant, Pinb-2v4, is reported. All four variants were physically mapped using Chinese Spring (CS) diteolosomics, nullisomic-tetrasomics, and CS-Cheyenne disomic substitution lines. Results place Pinb-2v1 on 7DL, Pinb-2v2 on 7BL, Pinb-2v3 on 7B, and Pinb-2v4 on 7AL. Pinb-2v1 and Pinb-2v4 were present in all cvs. examined: CS, Cheyenne, Recital, Wichita and Winsome. Pinb-2v2 was present in CS and Recital; Pinb-2v3 was present in Cheyenne, Wichita, and Winsome. These results are not wholly consistent with prior research and additional studies will be required to reconcile discrepancies. The discovery of Pinb-2v4 and the mapping of all four variants will contribute to a better understanding of gene duplication events in wheat and their bearing on wheat kernel texture and grain utilization.

Seed-specific expression of the wheat puroindoline genes improves maize wet milling yields.

The texture of maize (Zea mays L.) seeds is important to seed processing properties, and soft dent maize is preferred for both wet-milling and livestock feed applications. The puroindoline genes (Pina and Pinb) are the functional components of the wheat (Triticum aestivum L.) Hardness locus and together function to create soft grain texture in wheat. The PINs (PINA and PINB) are believed to act by binding to lipids on the surface of starch granules, preventing tight adhesion between starch granules and the surrounding protein matrix during seed maturation. Here, maize kernel structure and wet milling properties were successfully modified by the endosperm-specific expression of wheat Pins (Pina and Pinb). Pins were introduced into maize under the control of a maize gamma-Zein promoter. Three Pina/Pinb expression positive transgenic lines were evaluated over two growing seasons. Textural analysis of the maize seeds indicated that the expression of PINs decreased adhesion between starch and protein matrix and reduced maize grain hardness significantly. Reduction in pressure required to fracture kernels ranged from 15.65% to 36.86% compared with control seeds. Further, the PINs transgenic maize seeds had increased levels of extractable starch as characterized by a small scale wet milling method. Starch yield was increased by 4.86% on average without negatively impacting starch purity. The development of softer maize hybrids with higher starch extractability would be of value to maize processors.

Physical Mapping of Peroxidase Genes and Development of Functional Markers for TaPod-D1 on Bread Wheat Chromosome 7D.

Peroxidase (POD) activity in wheat (Triticum aestivum L.) grain influences natural carotenoid pigment content and is associated with the color of flour, and processing and product quality. Here, we report the molecular characterization and physical mapping of POD genes in bread wheat. The complete genomic DNA (gDNA) sequence of two POD genes (TaPod-A2 and TaPod-D1), and the partial gDNA sequence of two additional POD genes (TaPod-A3 and TaPod-B1) from wheat were characterized using in silico cloning and validated through laboratory experiments. Using a set of 21 nullisomic-tetrasomic (NT) lines, six group-7 ditelosomic (Dt) lines, and 38 group-7 deletion (Del) lines of Chinese Spring (CS), TaPod-A2 and TaPod-D1 were found to be physically located on 0.73-0.83 and on the most distal 0.39 fraction arm length (FL) of 7AS and 7DS in cv. CS, respectively; whereas, TaPod-A3 and TaPod-B1 were assigned to the 0.40-0.49 and 0.40-0.48 FL of 7AL and 7BL, respectively. Based on single nucleotide polymorphisms (SNPs) of two alleles at the TaPod-D1 locus, two functional markers POD-7D1 and POD-7D6 were developed, amplifying 540- and 640-bp, fragments in varieties with higher and lower POD activities, respectively. A total of 224 wheat varieties were analyzed and showed a significant association between the polymorphic fragments and POD activity using POD-7D1 and POD-7D6 markers. The analysis of variance (ANOVA) indicated the average POD activities of 115 varieties with TaPod-D1a were significantly lower than 109 varieties with TaPod-D1b (P < 0.01). This study provides useful information of the POD genes in bread wheat, insight into wheat genome synteny and structure, gene-specific markers, and contributes a valuable resource for quality improvement in wheat breeding programs.

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.

Structural consequences of the interaction of puroindolines with gluten proteins.

The effect of puroindolines (PINs) on structural characteristics of wheat proteins was investigated in Triticum turgidum ssp. durum (cv. Svevo) and Triticum aestivum (cv. Alpowa) and in their respective derivatives in which PIN genes were expressed (Soft Svevo) or the distal end of the short arm of chromosome 5D was deleted and PINs were not expressed (Hard Alpowa). The presence of PINs decreased the amount of cold-SDS extractable proteins and the accessibility of protein thiols to specific reagents, but resulted in facilitated solvation of gluten proteins, as detected by tryptophan fluorescence measurements carried out on minimally mixed flour/water mixtures. We propose that PINs and gluten proteins are interacting in the grain or flour prior to mixing. Hydrophobic interactions between PINs and some of the gluten proteins modify the pattern of interactions among gluten proteins, thus providing an additional mechanistic rationale for the effects of PINs on kernel hardness.

Genetic Dissection of End-Use Quality Traits in Adapted Soft White Winter Wheat.

Soft white wheat is used in domestic and foreign markets for various end products requiring specific quality profiles. Phenotyping for end-use quality traits can be costly, time-consuming and destructive in nature, so it is advantageous to use molecular markers to select experimental lines with superior traits. An association mapping panel of 469 soft white winter wheat cultivars and advanced generation breeding lines was developed from regional breeding programs in the U.S. Pacific Northwest. This panel was genotyped on a wheat-specific 90 K iSelect single nucleotide polymorphism (SNP) chip. A total of 15,229 high quality SNPs were selected and combined with best linear unbiased predictions (BLUPs) from historical phenotypic data of the genotypes in the panel. Genome-wide association mapping was conducted using the Fixed and random model Circulating Probability Unification (FarmCPU). A total of 105 significant marker-trait associations were detected across 19 chromosomes. Potentially new loci for total flour yield, lactic acid solvent retention capacity, flour sodium dodecyl sulfate sedimentation and flour swelling volume were also detected. Better understanding of the genetic factors impacting end-use quality enable breeders to more effectively discard poor quality germplasm and increase frequencies of favorable end-use quality alleles in their breeding populations.