Comparative Analysis of Qualitative and Bioactive Compounds of Whole and Refined Flours in Durum Wheat Grains with Different Year of Release and Yield Potential.

Durum wheat varieties are important sources of nutrients and provide remarkable amounts of phytochemicals. Especially, phenolics, which are mostly located in external layers of grains, have recently gained increased interest due to their high antioxidant power. This study aimed to evaluate the differences in the quality traits and phenolic compounds' concentration (e.g., phenolic acids) of different durum wheat genotypes, namely four Italian durum wheat cultivars and a USA elite variety, in relation to their yield potential and year of release. Phenolic acids were extracted both from wholemeal flour and semolina and analysed through HPLC-DAD analysis. Ferulic acid was the most represented phenolic acid, both in the wholemeal flour (438.3 µg g-1 dry matter) and in semolina (57.6 µg g-1 dry matter) across all cultivars, followed by p-coumaric acid, sinapic acid, vanillin, vanillic acid, syringic acid, and p-hydroxybenzoic acid. Among the cultivars, Cappelli showed the highest phenolic acid content, whilst Kronos had the lowest one. Negative correlations occurred between some phenolic acids and morphological and yield-related traits, especially for Nadif and Sfinge varieties. On the contrary, durum wheat genotypes with low yield potential such as Cappelli accumulated higher concentrations of phenolic acids under the same growing conditions, thereby significantly contributing to the health-promoting purposes.

Genome-Wide Identification, Characterization and Expression Pattern Analysis of the γ-Gliadin Gene Family in the Durum Wheat (Triticum durum Desf.) Cultivar Svevo.

Very recently, the genome of the modern durum wheat cv. Svevo was fully sequenced, and its assembly is publicly available. So, we exploited the opportunity to carry out an in-depth study for the systematic characterization of the γ-gliadin gene family in the cv. Svevo by combining a bioinformatic approach with transcript and protein analysis. We found that the γ-gliadin family consists of nine genes that include seven functional genes and two pseudogenes. Three genes, Gli-γ1a, Gli-γ3a and Gli-γ4a, and the pseudogene Gli-γ2a* mapped on the A genome, whereas the remaining four genes, Gli-γ1b, Gli-γ2b, Gli-γ3b and Gli-γ5b, and the pseudogene Gli-γ4b* mapped on the B genome. The functional γ-gliadins presented all six domains and eight-cysteine residues typical of γ-gliadins. The Gli-γ1b also presented an additional cysteine that could possibly have a role in the formation of the gluten network through binding to HMW glutenins. The γ-gliadins from the A and B genome differed in their celiac disease (CD) epitope content and composition, with the γ-gliadins from the B genome showing the highest frequency of CD epitopes. In all the cases, almost all the CD epitopes clustered in the central region of the γ-gliadin proteins. Transcript analysis during seed development revealed that all the functional γ-gliadin genes were expressed with a similar pattern, although significant differences in the transcript levels were observed among individual genes that were sometimes more than 60-fold. A progressive accumulation of the γ-gliadin fraction was observed in the ripening seeds that reached 34% of the total gliadin fraction at harvest maturity. We believe that the insights generated in the present study could aid further studies on gliadin protein functions and future breeding programs aimed at the selection of new healthier durum wheat genotypes.

Chitosan-Induced Activation of the Antioxidant Defense System Counteracts the Adverse Effects of Salinity in Durum Wheat.

The present study was carried out with the aim of (i) evaluating the effect of chitosan (CTS) on the growth of durum wheat under salinity and (ii) examining CTS-regulated mechanisms of salinity tolerance associated with the antioxidant defense system. To achieve these goals, durum wheat seedlings were treated with CTS at different molecular weight, low (L-CTS, 50-190 kDa), medium (M-CTS, 190-310 kDa) and high (H-CTS, 310-375 kDa). The results obtained show that exposure to 200 mM NaCl reduced the shoot and the root dried biomass by 38% and 59%, respectively. The growth impairment induced by salinity was strongly correlated with an increase in the superoxide anion production (5-fold), hydrogen peroxide content (2-fold) and malondialdehyde (MDA) content (4-fold). Seedlings responded to the oxidative stress triggered by salinity with an increase in the total phenolic content (TPC), total flavonoid content (TFC) and total antioxidant activity (TAA) by 67%, 51% and 32%, respectively. A salt-induced increase in the activity of the antioxidant enzymes superoxide dismutase and catalase (CAT) of 89% and 86%, respectively, was also observed. Treatment of salt-stressed seedlings with exogenous CTS significantly promoted seedling growth, with the strongest effects observed for L-CTS and M-CTS, which increased the shoot biomass of stressed seedlings by 32% and 44%, respectively, whereas the root dried biomass increased by 87% and 64%, respectively. L-CTS and M-CTS treatments also decreased the superoxide anion production (57% and 59%, respectively), the hydrogen peroxide content (35% and 38%, respectively) and the MDA content (48% and 56%, respectively) and increased the TPC (23% and 14%, respectively), the TFC (19% and 10%, respectively), the TAA (up to 10% and 7%, respectively) and the CAT activity (29% and 20%, respectively). Overall, our findings indicate that CTS exerts its protective role against the oxidative damages induced by salinity by enhancing the antioxidant defense system. L-CTS and M-CTS were the most effective in alleviating the adverse effect of NaCl, thus demonstrating that the CTS action is strictly related to its molecular weight.

Use of purple durum wheat to produce naturally functional fresh and dry pasta.

In this study, the effects of different milling procedures (roller-milling vs. stone-milling) and pasta processing (fresh vs. dried spaghetti), and cooking on the antioxidant components and sensory properties of purple durum wheat were investigated. Milling and pasta processing were performed using one purple and one conventional non-pigmented durum wheat genotypes, and the end-products were compared with commercial pasta. The results show that the stone milling process preserved more compounds with high health value (total fibre and carotenoids, and in the purple genotype, also anthocyanins) compared to roller-milling. The drying process significantly (p<0.05) reduced the content of anthocyanins (21.42 µg/g vs. 46.32 µg/g) and carotenoids (3.77 µg/g vs. 4.04 µg/g) with respect to the pasteurisation process involved in fresh pasta production. The sensory properties of pasta from the purple genotype did not significantly differ from commercial wholemeal pasta, and its in vitro glycemic index was even lower. Thus, it is possible to consider this genetic material as a good ingredient for the production of functional foods from cereals naturally rich in bioactive compounds.

Genetic variability in anthocyanin composition and nutritional properties of blue, purple, and red bread (Triticum aestivum L.) and durum (Triticum turgidum L. ssp. turgidum convar. durum) wheats.

Renewed interest in breeding for high anthocyanins in wheat (Triticum ssp.) is due to their antioxidant potential. A collection of different pigmented wheats was used to investigate the stability of anthocyanins over three crop years. The data show higher anthocyanins in blue-aleurone bread wheat (Triticum aestivum L.), followed by purple- and red-pericarp durum wheat (Triticum turgidum L. ssp. turgidum convar. durum), using cyanidin 3-O-glucoside as standard. HPLC of the anthocyanin components shows five to eight major anthocyanins for blue wheat extracts, compared to three anthocyanins for purple and red wheats. Delphinidin 3-O-rutinoside, delphinidin 3-O-glucoside, and malvidin 3-O-glucoside are predominant in blue wheat, with cyanidin 3-O-glucoside, peonidin 3-O-galactoside, and malvidin 3-O-glucoside in purple wheat. Of the total anthocyanins, 40-70% remain to be structurally identified. The findings confirm the high heritability for anthocyanins, with small genotype × year effects, which will be useful for breeding purposes, to improve the antioxidant potential of cereal-based foods.

Stay-green trait-antioxidant status interrelationship in durum wheat (Triticum durum) flag leaf during post-flowering.

Three independent durum wheat mutant lines that show delayed leaf senescence or stay-green (SG) phenotype, SG196, SG310 and SG504, were compared to the parental genotype, cv. Trinakria, with respect to the photosynthetic parameters and the cellular redox state of the flag leaf in the period from flowering to senescence. The SG mutants maintained their chlorophyll content and net photosynthetic rate for longer than Trinakria, thus revealing a functional SG phenotype. They also showed a better redox state as demonstrated by: (1) a lower rate of superoxide anion production due to generally higher activity of the antioxidant enzymes superoxide dismutase and catalase in all of the SG mutants and also of the total peroxidase in SG196; (2) a higher thiol content that can be ascribed to a higher activity of the NADPH-providing enzyme glucose-6-phosphate dehydrogenase in all of the SG mutants and also of the NADP(+)-dependent malic enzyme in SG196; (3) a lower pro-oxidant activity of lipoxygenase that characterises SG196 and SG504 mutants close to leaf senescence. Overall, these results show a general relationship in durum wheat between the SG phenotype and a better redox state. This relationship differs across the different SG mutants, probably as a consequence of the different set of altered genes underlying the SG trait in these independent mutant lines.

Insight into durum wheat Lpx-B1: a small gene family coding for the lipoxygenase responsible for carotenoid bleaching in mature grains.

BACKGROUND: The yellow colour of pasta products is one of the main criteria used by consumers to assess pasta quality. This character is due to the presence of carotenoid pigments in semolina. During pasta processing, oxidative degradation of carotenoid pigments occurs mainly due to lipoxygenase (LOX). In durum wheat (Triticum durum Desf.), two Lpx-1 genes have been identified on chromosome 4B, Lpx-B1.1 and Lpx-B1.2, and evidences have been reported that the deletion of Lpx-B1.1 is associated with a strong reduction in LOX activity in semolina. In the present study, we characterised the Lpx-B1 gene family identified in a durum wheat germplasm collection and related the distribution and expression of the Lpx-B1 genes and alleles to variations in LOX activity in the mature grains. RESULTS: In addition to the already known Lpx-B1.1 and Lpx-B1.2 genes, a new gene was identified, Lpx-B1.3, along with three different Lpx-B1.1 alleles, Lpx-B1.1a, Lpx-B1.1b and the partially deleted Lpx-B1.1c. Screening of the germplasm collection showed that all of the genotypes have one of the three Lpx-B1.1 alleles, associated with either Lpx-B1.2 or Lpx-B1.3, thus showing that in this collection the two genes are alternatives. Therefore, based on Lpx-B1 distribution, three different haplotypes were distinguished: haplotype I, carrying Lpx-B1.3 and the Lpx-B1.1b allele; haplotype II carrying Lpx-B1.2 and the Lpx-B1.1a allele; and haplotype III carrying Lpx-B1.2 and the Lpx-B1.1c allele. Determination of Lpx-B1 transcript abundance and total LOX activity in mature grains revealed differences among these three haplotypes: haplotypes I, II and III showed high, intermediate and low levels, respectively, of functional Lpx-B1 transcripts and enzymatic activity. CONCLUSIONS: In this germplasm collection, the Lpx-B1 gene family accounts for most of the total LOX activity in the mature grains. Information on these Lpx-B1 haplotypes provides significant improvement for prediction of LOX-1 activity levels in mature grains, and will therefore help in breeding programmes aimed at selection of new durum wheat genotypes with higher carotenoid contents in their end products.

The transcript levels of two plant mitochondrial uncoupling protein (pUCP)-related genes are not affected by hyperosmotic stress in durum wheat seedlings showing an increased level of pUCP activity.

Etiolated early seedlings of durum wheat submitted to moderate and severe salt (NaCl) and osmotic (mannitol) stress showed no relevant increase of both transcript levels of two plant uncoupling protein (pUCP)-related genes and maximal pUCP activity in purified mitochondria (which estimates protein level); contrarily, pUCP functioning due to endogenous free fatty acids strongly increased. These results show that pUCP activation under hyperosmotic stress may be due to modulation of pUCP reaction rather than to an increased protein synthesis. Finally, a properly developed method, based on a single membrane potential measurement, to evaluate both pUCP maximal activity and functioning, is reported.