Rye-durum wheat 1BL.1RS translocation: implications for drought tolerance and nutritional status.

The translocation of the short chromosome arm 1RS of rye onto the 1B chromosome of common wheat has been shown to improve resistance to stress and yield. Here, translocation was operated in durum wheat and its effects on drought tolerance were evaluated. Both the 1BL.1RS translocation line (Svevo 1BL.1RS) and the corresponding Svevo control were exposed to drought for 7 days. Significant differences were found in root morphology between Svevo and Svevo 1BL.1RS under control and drought conditions. Although Svevo 1BL.1RS experienced more severe growth inhibition due to drought than Svevo, it exhibited greater resilience to oxidative stress. Furthermore, several drought-responsive genes were upregulated in both shoots and roots only in the translocation line. Notably, in roots of Svevo 1BL.1RS, the expression of these genes was also higher in the control condition compared to Svevo, suggesting that these genes could be constitutively expressed at higher levels in the translocation line. Moreover, the 1BL.1RS translocation had a significant impact on the plant's ability to accumulate nutrients under drought. Overall, the impact on sulfate accumulation and the expression of genes associated with its assimilation pathways are particularly noteworthy, highlighting the involvement of sulfur in the plant response to water stress. Additionally, the genetic characterization of Svevo 1BL.1RS revealed variants extending beyond the translocation, located in drought stress-responsive genes.

The High-Molecular-Weight Glutenin Subunits of the T. timopheevii (AuAuGG) Group.

Polyploid wheats include a group of tetraploids known as Timopheevii (AuAuGG), which are represented by two subspecies: Triticum timopheevii ssp. timopheevii (cultivated) and Triticum timopheevii ssp. araraticum (wild). The combined use of electrophoretic (SDS-PAGE) and chromatographic (RP-HPLC) techniques carried out on high-molecular-weight glutenin subunits (HMW-GSs) permitted the association of different x- and y-type subunits to the A and G genomes and the assessment of allelic variation present at corresponding loci. The results also revealed that in both subspecies, accessions are present that possess expressed y-type subunits at the Glu-A1 locus. Genes corresponding to these subunits were amplified and amplicons corresponding to x- and y-type genes associated with the A genome were detected in all accessions, including those without expressed x- and y-type subunits. The comparison with genes of polyploid wheats confirmed the structural characteristics of typical y-type genes, with the presence of seven cysteine residues and with hexapeptide and nonapeptide repeat motifs. The identification of wild and cultivated T. timopheevii with both x- and y-type glutenin subunits at the Glu-A1 and Glu-G1 loci represents a useful source for the modification of the allelic composition of HMW-GSs in cultivated wheats with the ultimate objective of improving technological properties.

Corrigendum: Characterization of Triticum turgidum sspp. durum, turanicum and polonicum grown in Central Italy in relation to technological and nutritional aspects.

[This corrects the article DOI: 10.3389/fpls.2023.1269212.].

Mutations in starch biosynthesis genes affect chloroplast development in wheat pericarp.

Starch bioengineering in cereals has produced a plethora of genotypes with new nutritional and technological functionalities. Modulation of amylose content from 0 to 100% was inversely correlated with starch digestibility and promoted a lower glycemic index in food products. In wheat, starch mutants have been reported to exhibit various side effects, mainly related to the seed phenotype. However, little is known about the impact of altered amylose content and starch structure on plant metabolism. Here, three bread wheat starch mutant lines with extreme phenotypes in starch branching and amylose content were used to study plant responses to starch structural changes. Omics profiling of gene expression and metabolic patterns supported changes, confirmed by ultrastructural analysis in the chloroplast of the immature seeds. In detail, the identification of differentially expressed genes belonging to functional categories related to photosynthesis, chloroplast and thylakoid (e.g. CURT1), the alteration in the accumulation of photosynthesis-related compounds, and the chloroplast alterations (aberrant shape, grana stacking alteration, and increased number of plastoglobules) suggested that the modification of starch structure greatly affects starch turnover in the chloroplast, triggering oxidative stress (ROS accumulation) and premature tissue senescence. In conclusion, this study highlighted a correlation between starch structure and chloroplast functionality in the wheat kernel.

Characterization of Triticum turgidum sspp. durum, turanicum, and polonicum grown in Central Italy in relation to technological and nutritional aspects.

Introduction: Wheat is a staple food, with the two most common species being Triticum aestivum and Triticum turgidum ssp. durum. Moreover, the latter, T. turgidum, includes other tetraploid subspecies, among which the sspp. turanicum (Khorasan wheat) and polonicum (Polish wheat), whose importance has increased in the last decades, representing alternative crops for marginal areas, in addition to being a source of genetic diversity. Methods: In this work, different accessions of these three subspecies of T. turgidum have been grown in 2 years in the same environment and have been characterized for technological properties and factors affecting nutritional quality, such as fiber amount and the content of micro- and macro-nutrients in grains, and for root morphological traits. Results: These analyses allowed the identification, in particular, of a Polish wheat accession showing better technological performances, a higher amount of positive micro- and macro-elements, and a lower amount of toxic cadmium. The modern variety Svevo and the Polish Pol2 showed the lowest and the highest shoot:root ratio, respectively. The high shoot:root ratio in Pol2 was mainly attributable to the decrease in root growth. Although Pol2 had a lower root biomass, its particular root morphology made it more efficient for nutrient uptake, as evident from the greater accumulation of micro- and macro-nutrients. Discussion: These results underline that it is not possible to draw general conclusions about the difference between primitive and modern wheats, but rather a case-by-case approach should be chosen.

Impact of Variation in Amylose Content on Durum Wheat cv. Svevo Technological and Starch Properties.

Reserve starch, the main component of durum wheat semolina, is constituted of two glucan homopolymers (amylose and amylopectin) that differ in their chemical structure. Amylose is mainly a linear structure formed of α-1,4-linked glucose units, with a lower polymerization degree, whereas amylopectin is a highly branched structure of α-1,4-chains linked by α-1,6-bonds. Variation of the amylose/amylopectin ratio has a profound effect on the starch properties which may impact the wheat technological and nutritional characteristics and their possible use in the food and non-food sector. In this work a set of genotypes, with a range of amylose from 14.9 to 57.8%, derived from the durum wheat cv. Svevo was characterised at biochemical and rheological level and used to produce pasta to better understand the role of amylose content in a common genetic background. A negative correlation was observed between amylose content and semolina swelling power, starch peak viscosity, and pasta stickiness. A worsening of the firmness was observed in the low amylose pasta compared to the control (cv. Svevo), whereas no difference was highlighted in the high amylose samples. The resistant starch was higher in the high amylose (HA) pasta compared to the control and low amylose (LA) pasta. Noteworthy, the extent of starch digestion was reduced in the HA pasta while the LA genotypes offered a higher starch digestion, suggesting other possible applications.

The functional implications of high-amylose wholegrain wheat flours: An in vitro digestion and fermentation approach combined with metabolomics.

Wheat flour is one of the most prevalent foodstuffs for human consumption, and novel strategies are underway to enhance its nutritional properties. This work evaluated wholegrain flours from bread wheat lines with different amylose/amylopectin ratios through in vitro starch digestion and large intestine fermentation. High-amylose flours presented a higher resistant starch content and lower starch hydrolysis index. Moreover, UHPLC-HRMS metabolomics was carried out to determine the profile of the resulting in vitro fermentates. The multivariate analysis highlighted distinctive profiles between the flours derived from the different lines compared to the wild type. Peptides, glycerophospholipids, polyphenols, and terpenoids were identified as the main markers of the discrimination. The high-amylose flour fermentates showed the richest bioactive profile, containing stilbenes, carotenoids, and saponins. Present findings pave the way toward applying high-amylose flours to design novel functional foods.

Exploring Variability of Free Asparagine Content in the Grain of Bread Wheat (Triticum aestivum L.) Varieties Cultivated in Italy to Reduce Acrylamide-Forming Potential.

Acrylamide, a suspected human carcinogen, is generated during food processing at high temperatures in the Maillard reaction, which involves reducing sugars and free asparagine. In wheat derivatives, free asparagine represents a key factor in acrylamide formation. Free asparagine levels in the grain of different wheat genotypes has been investigated in recent studies, but little is known about elite varieties that are cultivated in Italy. Here, we analysed the accumulation of free asparagine in a total of 54 bread wheat cultivars that are relevant for the Italian market. Six field trials in three Italian locations over two years were considered. Wholemeal flours obtained from harvested seeds were analysed using an enzymatic method. Free asparagine content ranged from 0.99 to 2.82 mmol/kg dry matter in the first year, and from 0.55 to 2.84 mmol/kg dry matter in the second year. Considering the 18 genotypes that were present in all the field trials, we evaluated possible environment and genetic influences for this trait. Some cultivars seemed to be highly affected by environment, whereas others showed a relative stability in free asparagine content across years and locations. Finally, we identified two varieties showing the highest free asparagine levels in our analysis, representing potential useful materials for genotype x environment interaction studies. Two other varieties, which were characterized by low amounts of free asparagine in the considered samples, may be useful for the food industry and for future breeding programs aimed to reduce acrylamide-forming potential in bread wheat.

Effect of Three Bakery Products Formulated with High-Amylose Wheat Flour on Post-Prandial Glycaemia in Healthy Volunteers.

Both Glycaemic index (GI) and Glycaemic Load (GL) were introduced to measure the impact of a carbohydrate-containing food on blood glucose. From this perspective, high-amylose (HA) flours, with a higher percentage of resistant starch (RS), may represent a suitable raw material to improve the glycaemic response. The present work aims to investigate the GI of HA bakery products (biscuits, taralli and bread) compared to products obtained from conventional flour. Ten healthy volunteers were enrolled and their capillary blood glucose was measured every 15 min for 2 h after the consumption of HA and control products containing 50 g of available carbohydrates. On average, in the three bakery products, the amount of total starch replaced by RS was equal to 12%. HA biscuits and HA bread showed significantly lower GI than their control counterparts (p = 0.0116 and p = 0.011, respectively) and better glycaemic control. From the survey to assess liking and willingness to pay on HA snacks, HA packages received an average premium of €0.66 compared to control products. Although HA flour results in lower GI in both biscuits and bread, further studies are needed to evaluate the correct composition of HA products to have beneficial effects on post-prandial glycaemia.

Molecular insights into the role of amylose/amylopectin ratio on gluten protein organization.

Waxy (WX) and high-amylose (HA) wheat flours have interesting functional and/or nutritional characteristics, but low technological properties compared to regular wheat. Here a set of three wheat lines, having different amylose content but sharing the same varietal background, were compared to shed light on the role of the amylose/amylopectin ratio on the protein conformational changes that lead to gluten formation. Despite the absence of differences in their protein profile, as also confirmed by thiolomic approaches, both WX and HA lines developed a weaker gluten than the control sample. The altered amylose/amylopectin ratio exerts a matrix effect establishing a competition for water with proteins, leading to a different protein structure and three-dimensional organization of the gluten network. These results add a piece to the understanding of the molecular aspects that oversee matrix effects on gluten formation in wheat, which description can be helpful for a rational optimization of the transformation process.

Whole-exome sequencing of selected bread wheat recombinant inbred lines as a useful resource for allele mining and bulked segregant analysis.

Although wheat (Triticum aestivum L.) is the main staple crop in the world and a major source of carbohydrates and proteins, functional genomics and allele mining are still big challenges. Given the advances in next-generation sequencing (NGS) technologies, the identification of causal variants associated with a target phenotype has become feasible. For these reasons, here, by combining sequence capture and target-enrichment methods with high-throughput NGS re-sequencing, we were able to scan at exome-wide level 46 randomly selected bread wheat individuals from a recombinant inbred line population and to identify and classify a large number of single nucleotide polymorphisms (SNPs). For technical validation of results, eight randomly selected SNPs were converted into Kompetitive Allele-Specific PCR (KASP) markers. This resource was established as an accessible and reusable molecular toolkit for allele data mining. The dataset we are making available could be exploited for novel studies on bread wheat genetics and as a foundation for starting breeding programs aimed at improving different key agronomic traits.

Characterization of Fresh Pasta Made of Common and High-Amylose Wheat Flour Mixtures.

This study aims to assess the main biochemical, technological, and nutritional properties of a few samples of fresh pasta composed of commercial common wheat flour blended with increasing percentages, ranging from 0 to 100%, of high-amylose wheat flour. Although the technological parameters of such samples remained practically constant, fresh pasta samples including 50 to 100% of high-amylose wheat flour were classifiable as foods with a low in vitro glycemic index of about 43%. However, only fresh pasta made of 100% high-amylose wheat flour exhibited a resistant starch-to-total starch ratio greater than 14% and was therefore eligible to claim a physiological effect of improved glucose metabolism after a meal, as according to EU Regulation 432/2012.

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.

Enrichment of provitamin A content in durum wheat grain by suppressing ß-carotene hydroxylase 1 genes with a TILLING approach.

KEY MESSAGE: The suppression of the HYD-1 gene by a TILLING approach increases the amount of ß-carotene in durum wheat kernel. Vitamin A deficiency is a major public health problem that affects numerous countries in the world. As humans are not able to synthesize vitamin A, it must be daily assimilated along with other micro- and macronutrients through the diet. Durum wheat is an important crop for Mediterranean countries and provides a discrete amount of nutrients, such as carbohydrates and proteins, but it is deficient in some essential micronutrients, including provitamin A. In the present work, a targeting induced local lesions in genomes strategy has been undertaken to obtain durum wheat genotypes biofortified in provitamin A. In detail, we focused on the suppression of the ß-carotene hydroxylase 1 (HYD1) genes, encoding enzymes involved in the redirection of ß-carotene toward the synthesis of the downstream xanthophylls (neoxanthin, violaxanthin and zeaxanthin). Expression analysis of genes involved in carotenoid biosynthesis revealed a reduction of the abundance of HYD1 transcripts greater than 50% in mutant grain compared to the control. The biochemical profiling of carotenoid in the wheat mutant genotypes highlighted a significant increase of more than 70% of ß-carotene compared to the wild-type sibling lines, with no change in lutein, α-carotene and zeaxanthin content. This study sheds new light on the molecular mechanism governing carotenoid biosynthesis in durum wheat and provides new genotypes that represent a good genetic resource for future breeding programs focused on the provitamin A biofortification through non-transgenic approaches.

QTL Analysis of Five Morpho-Physiological Traits in Bread Wheat Using Two Mapping Populations Derived from Common Parents.

Traits such as plant height (PH), juvenile growth habit (GH), heading date (HD), and tiller number are important for both increasing yield potential and improving crop adaptation to climate change. In the present study, these traits were investigated by using the same bi-parental population at early (F2 and F2-derived F3 families) and late (F6 and F7, recombinant inbred lines, RILs) generations to detect quantitative trait loci (QTLs) and search for candidate genes. A total of 176 and 178 lines were genotyped by the wheat Illumina 25K Infinium SNP array. The two genetic maps spanned 2486.97 cM and 3732.84 cM in length, for the F2 and RILs, respectively. QTLs explaining the highest phenotypic variation were found on chromosomes 2B, 2D, 5A, and 7D for HD and GH, whereas those for PH were found on chromosomes 4B and 4D. Several QTL detected in the early generations (i.e., PH and tiller number) were not detected in the late generations as they were due to dominance effects. Some of the identified QTLs co-mapped to well-known adaptive genes (i.e., Ppd-1, Vrn-1, and Rht-1). Other putative candidate genes were identified for each trait, of which PINE1 and PIF4 may be considered new for GH and TTN in wheat. The use of a large F2 mapping population combined with NGS-based genotyping techniques could improve map resolution and allow closer QTL tagging.

Contribution to Breadmaking Performance of Two Different HMW Glutenin 1Ay Alleles Expressed in Hexaploid Wheat.

Two expressed alleles of the 1Ay high-molecular-weight glutenin subunit (HMW-GS), 1Ay21* and 1AyT1, previously introduced in durum and bread wheat, were separately introgressed into the Australian bread wheat (Triticum aestivum L.) cv. Livingston. The developed lines had different allelic compositions compared to that of the parental cultivar (1Ax1), having either 1Ax21+1Ay21* or 1Ax1+1AyT1 at the Glu-A1 locus. Since 1Ax21 and 1Ax1 are known to have the same effects on quality, differences observed between the two sets of the developed lines are attributed to the two introgressed Ay genes. Yield and agronomic performance of the lines were evaluated in the field, and the protein, dough, and baking quality attributes were evaluated by large-scale quality testing. Results demonstrated that the subunit 1Ay21* increased unextractable polymeric protein by up to 14.3% and improved bread loaf volume by up to 9.2%. On the other hand, subunit 1AyT1 increased total grain protein by up to 9% along with dough elasticity. Furthermore, milling extraction was higher, and flour ash was lower in the 1Ay21* lines compared to the lines integrating 1AyT1. Both sets of the 1Ay introgression lines reduced dough-mixing time compared to the recurrent parent Livingston. The results also showed that 1Ay21* had a higher potential to improve the baking quality than 1AyT1 under the Livingston genetic background. Both alleles showed the potential to be utilized in breeding programs to improve the breadmaking quality.

A Cross between Bread Wheat and a 2D(2R) Disomic Substitution Triticale Line Leads to the Formation of a Novel Disomic Addition Line and Provides Information of the Role of Rye Secalins on Breadmaking Characteristics.

A bread wheat line (N11) and a disomic 2D(2R) substitution triticale line were crossed and backrossed four times. At each step electrophoretic selection for the seeds that possessed, simultaneously, the complete set of high molecular weight glutenin subunits of N11 and the two high molecular weight secalins of rye, present in the 2D(2R) line, was carried out. Molecular cytogenetic analyses of the BC4F8 generation revealed that the selection carried out produced a disomic addition line (2n = 44). The pair of additional chromosomes consisted of the long arm of chromosome 1R (1RL) from rye fused with the satellite body of the wheat chromosome 6B. Rheological analyses revealed that the dough obtained by the new addition line had higher quality characteristics when compared with the two parents. The role of the two additional high molecular weight secalins, present in the disomic addition line, in influencing improved dough characteristics is discussed.

Reduction of Allergenic Potential in Bread Wheat RNAi Transgenic Lines Silenced for CM3, CM16 and 0.28 ATI Genes.

Although wheat is used worldwide as a staple food, it can give rise to adverse reactions, for which the triggering factors have not been identified yet. These reactions can be caused mainly by kernel proteins, both gluten and non-gluten proteins. Among these latter proteins, α-amylase/trypsin inhibitors (ATI) are involved in baker's asthma and realistically in Non Celiac Wheat Sensitivity (NCWS). In this paper, we report characterization of three transgenic lines obtained from the bread wheat cultivar Bobwhite silenced by RNAi in the three ATI genes CM3, CM16 and 0.28. We have obtained transgenic lines showing an effective decrease in the activity of target genes that, although showing a higher trypsin inhibition as a pleiotropic effect, generate a lower reaction when tested with sera of patients allergic to wheat, accounting for the important role of the three target proteins in wheat allergies. Finally, these lines show unintended differences in high molecular weight glutenin subunits (HMW-GS) accumulation, involved in technological performances, but do not show differences in terms of yield. The development of new genotypes accumulating a lower amount of proteins potentially or effectively involved in allergies to wheat and NCWS, not only offers the possibility to use them as a basis for the production of varieties with a lower impact on adverse reaction, but also to test if these proteins are actually implicated in those pathologies for which the triggering factor has not been established yet.

Noodles Made from High Amylose Wheat Flour Attenuate Postprandial Glycaemia in Healthy Adults.

Previous research has not considered the effect of high amylose wheat noodles on postprandial glycaemia. The aim of the study is to investigate the effect of consumption of high amylose noodles on postprandial glycaemia over 2-h periods by monitoring changes in blood glucose concentration and calculating the total area under the blood glucose concentration curve. Twelve healthy young adults were recruited to a repeated measure randomised, single-blinded crossover trial to compare the effect of consuming noodles (180 g) containing 15%, 20% and 45% amylose on postprandial glycaemia. Fasting blood glucose concentrations were taken via finger-prick blood samples. Postprandial blood glucose concentrations were taken at 15, 30, 45, 60, 90 and 120 min. Subjects consuming high amylose noodles made with flour containing 45% amylose had significantly lower blood glucose concentration at 15, 30 and 45 min (5.5 ± 0.11, 6.1 ± 0.11 and 5.6 ± 0.11 mmol/L; p = 0.01) compared to subjects consuming low amylose noodles with 15% amylose (5.8 ± 0.12, 6.6 ± 0.12 and 5.9 ± 0.12 mmol/L). The total area under the blood glucose concentration curve after consumption of high amylose noodles with 45% amylose was 640.4 ± 9.49 mmol/L/min, 3.4% lower than consumption of low amylose noodles with 15% amylose (662.9 ± 9.49 mmol/L/min), p = 0.021. Noodles made from high amylose wheat flour attenuate postprandial glycaemia in healthy young adults, as characterised by the significantly lower blood glucose concentration and a 3.4% reduction in glycaemic response.

Can Manipulation of Durum Wheat Amylose Content Reduce the Glycaemic Index of Spaghetti?

Resistant starch (RS) in foods has positive benefits for potentially alleviating lifestyle diseases. RS is correlated positively with starch amylose content. This study aimed to see what level of amylose in durum wheat is needed to lower pasta GI. The silencing of starch synthases IIa (SSIIa) and starch branching enzymes IIa (SBEIIa), key genes involved in starch biosynthesis, in durum wheat cultivar Svevo was performed and spaghetti was prepared and evaluated. The SSIIa and SBEIIa mutants have a 28% and 74% increase in amylose and a 2.8- and 35-fold increase in RS, respectively. Cooked pasta was softer, with higher cooking loss but lower stickiness compared to Svevo spaghetti, and with acceptable appearance and colour. In vitro starch digestion extent (area under the digestion curve) was decreased in both mutants, but much more in SBEIIa, while in vivo GI was only significantly reduced from 50 to 38 in SBEIIa. This is the first study of the glycaemic response of spaghetti prepared from SBEIIa and SSIIa durum wheat mutants. Overall pasta quality was acceptable in both mutants but the SBEIIa mutation provides a clear glycaemic benefit and would be much more appealing than wholemeal spaghetti. We suggest a minimum RS content in spaghetti of ~7% is needed to lower GI which corresponded to an amylose content of ~58%.