Symbiotic fungi from a wild grass (Celtica gigantea) increase the growth, grain yield and quality of tritordeum under field conditions.

Plants function in symbiosis with numerous microorganisms, which might contribute to their adaptation and performance. In this study, we tested whether fungal strains in symbiotic interaction with roots of Celtica gigantea, a wild grass adapted to nutrient-poor soils in semiarid habitats, could improve the field performance of the agricultural cereal tritordeum (Triticum durum × Hordeum chilense). Seedlings of tritordeum were inoculated with 12 different fungal strains isolated from roots of Celtica gigantea that were first proved to promote the growth of tritordeum plants under greenhouse conditions. The inoculated seedlings were transplanted to field plots at two locations belonging to different climatic zones in terms of mean temperatures and precipitation in the Iberian Peninsula. Only one strain, Diaporthe iberica T6, had a significant effect on plant height, number of tillers and grain yield in one location. This result showed a substantial divergence between the results of greenhouse and field tests. In terms of grain nutritional quality, several parameters were differentially affected at both locations: Diaporthe T6, Pleosporales T7, Zygomycota T29 and Zygomycota T80 increased the content of total carotenoids, mainly lutein, in the colder location; whereas gluten proteins increased with several treatments in the warmer location. In conclusion, early inoculation of tritordeum plants with fungal symbionts had substantial beneficial effects on subsequent plant growth and development in the field. Regarding grain nutritional quality, the effect of inoculation was affected by the agroclimatic differences between both field locations.

Tritordeum: Promising Cultivars to Improve Health.

Tritordeum is an amphiploides species resulting from the hybridization between durum wheat (T. durum) and wild barley (H. chilense). This new cereal is considered a natural crop as it is obtained by traditional breeding techniques. Given its appreciable organoleptic characteristics, agronomic features, presence of interesting components, and good technological properties, Tritordeum is of promising interest for the development of health-oriented foods. In this study, we evaluated two registered Tritordeum cultivars, Bulel and Aucan. T. durum (Provenzal) was employed as the positive control. The extracted proteins were digested by gastric/pancreatic proteases, and their biological effects on Caco-2 differentiated on transwell inserts were determined. Changes in cell viability, monolayer permeability, organization of F-actin microfilaments, and ER stress triggered by protein-digested samples (DPs) were inspected. Our results showed that exposure to Provenzal-DPs promptly disrupted the tight junction barrier. Conversely, Aucan-DPs did not enhance monolayer permeability, whereas Bulel-DPs exerted only slight effects. Provental-DPs-induced toxicity was also confirmed by changes in cell viability and by the deep reorganization of the enterocyte cytoskeleton. In contrast, Aucan-DPs and Bulel-DPs did not affect monolayer viability and cytoskeleton structure. Overall, our findings suggest that both Tritordeum cultivars could be potential candidates for mitigating the toxicity of wheat flour.

Mycotoxin occurrence in kernels and straws of wheat, barley, and tritordeum.

Thirty-two varieties of common and durum wheat, hordeum, barley, and tritordeum collected over two harvesting years (2020 and 2021) were investigated for the presence of multiple Fusarium-related mycotoxins in asymptomatic plants. DON, 3-AcDON, 15-AcDON, T-2, HT-2, and ZEN together with the emerging mycotoxin ENN B and the major modified form of DON, namely DON3Glc, were quantified by means of UHPLC-MS/MS. Overall, DON and ENN B were the most frequently detected mycotoxins, albeit large inter-year variability was observed and related to different climate and weather conditions. Straws had higher mycotoxin contents than kernels and regarding DON occurrence tritordeum was found to be the most contaminated group on average for both harvesting years, while barley was the less contaminated one. Emerging mycotoxin ENN B showed comparable contents in kernels compared to straw, with a ratio close to 1 for tritordeum and barley. Regarding the occurrence of the other evaluated mycotoxins, T-2 and HT-2 toxins have been spotted in a few tritordeum samples, while ZEN has been frequently found only in straw from the harvesting year 2020. The data collected confirms the occurrence of multiple Fusarium mycotoxins in straws also from asymptomatic plants, highlighting concerns related to feed safety and animal health. The susceptibility of Tritordeum, hereby reported for the first time, suggests that careful measures in terms of monitoring, breeding, and cultural choices should be applied when dealing with his emerging crop.

In vitro faecal fermentation of Tritordeum breads and its effect on the human gut health.

Spontaneous fermentation of Tritordeum flour enhances the nutritional potential of this hybrid cereal. However, the effect of consumption of Tritordeum sourdough bread (SDB) on gut health remains to be elucidated. This study investigated the effect of in vitro digestion and faecal fermentation of SDB compared to that of traditional baker's yeast (BYB) Tritordeum bread. After 24-h anaerobic faecal fermentation, both SDB and BYB (1% w/v) induced an increase in the relative abundances of Bifidobacterium, Megasphaera, Mitsuokella, and Phascolarctobacterium genera compared to baseline, while concentrations of acetate and butyrate were significantly higher at 24 h for SDB compared to those for BYB. Integrity of intestinal epithelium, as assessed through in vitro trans-epithelial electrical resistance (TEER) assay, was slightly increased after incubation with SDB fermentation supernatants, but not after incubation with BYB fermentation supernatants. The SDB stimulated in vitro mucosal immune response by inducing early secretion of inflammatory cytokines, IL-6 and TNF-α, followed by downregulation of the inflammatory trigger through induction of anti-inflammatory IL-10 expression. Overall, our findings suggest that Tritordeum sourdough can modulate gut microbiota fermentation activity and positively impact the gut health.

Influence of Agronomic Practices on the Antioxidant Compounds of Pigmented Wheat (Triticum aestivum spp. aestivum L.) and Tritordeum (× Tritordeum martinii A. Pujadas, nothosp. nov.) Genotypes.

Twelve pigmented wheat genotypes, one tritordeum, and one common wheat were grown in three field experiments under varying nitrogen (N) fertilization rates to investigate the contributions of genotype, environment, and fertilization on the levels of phenolic acids, anthocyanins, carotenoids and antioxidant capacity of the grains. Soluble phenolic acids increased significantly (+16%) in the environment with high soil N content, while bound phenolic acids and anthocyanins decreased (-16 and -57%). N fertilization affected the agronomic and qualitative traits but had limited effects on some bioactive compounds (bound phenolic acids and anthocyanins). The greatest differences appeared among the color groups and within the same color types, with the black group showing the most anthocyanins and phenolic acids (34.4 and 1207 mg·kg-1) and the highest antioxidant capacity. Some of the cultivars could be promising for the development of innovative supply chains and the production of functional foods, as they showed good yield and quality performances, and good antioxidant features.

Corrigendum: Managing symptom profile of IBS-D patients with Tritordeum-based foods: results from a pilot study.

[This corrects the article DOI: 10.3389/fnut.2022.797192.].

Comparative compositions of grain of tritordeum, durum wheat and bread wheat grown in multi-environment trials.

Three genotypes each of bread wheat, durum wheat and tritordeum were grown in randomized replicated field trials in Andalusia (Spain) for two years and wholemeal flours analysed for a range of components to identify differences in composition. The contents of all components that were determined varied widely between grain samples of the individual species and in most cases also overlapped between the three species. Nevertheless, statistically significant differences between the compositions of the three species were observed. Notably, tritordeum had significantly higher contents of protein, some minerals (magnesium and iron), total phenolics and methyl donors. Tritordeum also had higher levels of total amino acids (but not asparagine) and total sugars, including raffinose. By contrast, bread wheat and tritordeum had similar contents of the two major dietary fibre components in white flour, arabinoxylan and ß-glucan, with significantly lower contents in durum wheat.

A Tritordeum-Based Diet for Female Patients with Diarrhea-Predominant Irritable Bowel Syndrome: Effects on Abdominal Bloating and Psychological Symptoms.

Most female patients with irritable bowel syndrome (IBS) complain of abdominal bloating rather than abdominal pain and diarrhea. The higher incidence in women could be due to the so-called dysfunctional gas handling. Since diet seems the most effective and durable strategy for managing IBS symptoms, we aimed to evaluate the effects of a 12 week diet based on a relatively new cereal, Tritordeum (TBD), on gastrointestinal (GI) symptoms, anthropometric and bioelectrical impedance parameters, and psychological profiles in 18 diarrhea-predominant IBS (IBS-D) female patients with abdominal bloating as the dominant symptom. The IBS Severity Scoring System (IBS-SSS), the Symptom Checklist-90 Revised, the Italian version of the 36-Item Short-Form Health Survey, and the IBS-Quality of Life questionnaire were administered. The TBD reduces the IBS-SSS "Intensity of abdominal bloating" with a concomitant improvement in the anthropometric profile. No correlation was found between "Intensity of abdominal bloating" and "Abdominal circumference". Anxiety, depression, somatization, interpersonal sensitivity, and phobic and avoidance manifestations were significantly reduced after TBD. Lastly, anxiety was correlated with "Intensity of abdominal bloating". Overall, these results suggest the possibility of lowering abdominal bloating and improving the psychological profile of female IBS-D patients using a diet based on an alternative grain such as Tritordeum.

Evaluation of the Effects of the Tritordeum-Based Diet Compared to the Low-FODMAPs Diet on the Fecal Metabolome of IBS-D Patients: A Preliminary Investigation.

Since irritable bowel syndrome (IBS)-a common gastrointestinal (GI) disorder-still lacks effective therapy, a nutritional approach may represent a practical alternative. Different reports demonstrated that a low-fermentable oligosaccharides, disaccharides, monosaccharides and polyols (FODMAPs) diet (LFD) reduces symptoms in IBS with diarrhea (IBS-D) patients, also inducing beneficial pathophysiological and biochemical modifications. More recently, diets with alternative cereals having a different gluten composition, such as tritordeum, have also been considered (TBD). We investigated the impact of TBD and LFD on the fecal metabolome composition in 38 IBS-D patients randomly allocated to the two diets for 12 weeks. Summarily, at baseline, the profile of fecal volatile organic compounds (VOCs) of IBS-D patients was not significantly different in the two groups. After treatment, significant changes were observed in the two groups regarding the VOCs content since some of them increased in the TBD group (namely, decanoic acid), whereas others (i.e., nonanal and ethanol) increased in the LFD one. Further, at baseline, short-chain fatty acids were positively related to inflammation and showed a significant decreasing trend after both diets compared to baseline values (namely, acetic and propanoic acid). Preliminary results from this pilot study suggest a potential positive intervention of TBD and LFD affecting the fecal metabolome composition in IBS-D patients.

Consumption of Tritordeum Bread Reduces Immunogenic Gluten Intake without Altering the Gut Microbiota.

Gluten proteins are responsible for the wheat breadmaking quality. However, gluten is also related to human pathologies for which the only treatment is a gluten-free diet (GFD). GFD has gained popularity among individuals who want to reduce their gluten intake. Tritordeum is a cereal species that originated after crossing durum wheat with wild barley and differs from bread wheat in its gluten composition. In this work, we have characterized the immunogenic epitopes of tritordeum bread and results from a four-phase study with healthy adults for preferences of bread and alterations in the gut microbiota after consuming wheat bread, gluten-free bread, and tritordeum bread are reported. Tritordeum presented fewer peptides related to gluten proteins, CD-epitopes, and IgE binding sites than bread wheat. Participants rated tritordeum bread higher than gluten-free bread. Gut microbiota analysis revealed that the adherence to a strict GFD involves some minor changes, especially altering the species producing short-chain fatty acids. However, the short-term consumption of tritordeum bread does not induce significant changes in the diversity or community composition of the intestinal microbiota in healthy individuals. Therefore, tritordeum bread could be an alternative for healthy individuals without wheat-related pathologies who want to reduce their gluten consumption without harming their gut health.

A Comparison of the Low-FODMAPs Diet and a Tritordeum-Based Diet on the Gastrointestinal Symptom Profile of Patients Suffering from Irritable Bowel Syndrome-Diarrhea Variant (IBS-D): A Randomized Controlled Trial.

The dietary approach low in oligosaccharides, disaccharides, monosaccharides, and fermentable polyols (FODMAPs-LFD) is a good strategy for treating irritable bowel syndrome (IBS). Beyond the LFD, other dietary approaches with beneficial effects may be hypothesized. Among them, consumption of Tritordeum-based foods (TBD, bread, bakery products, and pasta) in substitution of other cereals seem to achieve promising results. In a randomized controlled trial, we compared the effects of 12 weeks of LFD to TBD in improving the symptom profile of IBS-diarrhea (IBS-D) patients. The two diets equally improved gastrointestinal symptoms and QoL, measured by the IBS Severity Scoring System (IBS-SSS) questionnaire, reducing the total score after four weeks and maintaining this range until the end of treatment (IBS-SSS total score change: −132.1; 95% CI: −74.9 to −189.4 and −130.5; 95% CI: −73.2 to −187.7; p < 0.0001 after LFD and TBD, respectively). The two diets did not modify the micronutrients content when extended for 12 weeks. LFD could be regarded as a first-line dietary approach for IBS-D patients. However, TBD may represent a valid alternative, with high palatability, especially among Italian patients, for whom pasta is considered one of the main assets of dietetic culture, and would be easier to manage in their daily habits.

Managing Symptom Profile of IBS-D Patients With Tritordeum-Based Foods: Results From a Pilot Study.

In the past few years, increasing attention has been given to the pathologic role of specific foods in IBS, like wheat and other cereals. Recent literature describes IBS patients who may experience gastrointestinal (GI) and extra-GI symptoms precipitated by the ingestion of cereals. Tritordeum is a cereal of Spanish origin derived from the hybridization of durum wheat and wild barley. It is different from classic wheat for its gluten protein composition, with fewer carbohydrates and fructans and a higher content of proteins, dietary fibers, and antioxidants. This pilot study aimed to investigate the effects of a 12-week diet with Tritordeum-based foods in substitution of other cereals on the profile of GI symptoms (evaluated by appropriate questionnaire) and the health of the GI barrier (assessed by sugar absorption test and different markers of integrity and functions) in 16 diarrhea-predominant IBS (IBS-D) patients. The diet with Tritordeum-based foods (bread, bakery products, and pasta) significantly reduced IBS-D patients' symptoms. This amelioration appears to occur through an overall improvement of the GI barrier, as demonstrated by the reduced intestinal permeability and the decreased levels of markers of intestinal mucosal integrity, mucosal inflammation, and fermentative dysbiosis.

Tritordeum as an Innovative Alternative to Wheat: A Comparative Digestion Study on Bread.

Tritordeum results from the crossbreeding of a wild barley (Hordeum chilense) species with durum wheat (Triticum turgidum spp. turgidum). This hexaploid crop exhibits agronomic and rheological characteristics like soft wheat, resulting in an innovative raw material to produce baked goods. We applied a gel-based proteomic approach on refined flours to evaluate protein expression differences among two widespread tritordeum cultivars (Aucan and Bulel) taking as the reference semolina and flour derived from a durum and a soft wheat cvs, respectively. The products of in vitro digestion of model breads were analyzed to compare bio-accessibility of nutrients and mapping tritordeum bread resistant peptides. Significant differences among the protein profiles of the four flours were highlighted by electrophoresis. The amino acid bio-accessibility and the reducing sugars of tritordeum and wheat breads were comparable. Tritordeum cvs had about 15% higher alpha-amino nitrogen released at the end of the duodenal simulated digestion than soft wheat (p < 0.05). Bulel tritordeum flour, bread and digested bread had about 55% less R5-epitopes compared to the soft wheat. Differences in protein expression found between the two tritordeum cvs reflected in diverse digestion products and allergenic and celiacogenic potential of the duodenal peptides. Proteomic studies of a larger number of tritordeum cvs may be successful in selecting those with good agronomical performances and nutritional advantages.

5-n-Alkylresorcinol Profiles in Different Cultivars of Einkorn, Emmer, Spelt, Common Wheat, and Tritordeum.

5-n-Alkylresorcinols (AR) are bioactive compounds found in the edible parts of many cereals. Here, saturated and unsaturated homologues, including the oxidized forms 5-(2'-oxo) AR and their plant metabolites, were profiled by ultrahigh-performance liquid chromatography-ion mobility separation-high-resolution mass spectrometry in 18 cultivars of einkorn, emmer, spelt, common wheat, and tritordeum, cultivated in two consecutive years under uniform agronomic conditions. The average content of AR ranged between 672.5 ± 129.8 and 1408.9 ± 528.0 mg/kg, exceeding 2380 mg/kg in some samples and highlighting a superior content in tritordeum and in modern cultivars with respect to old wheat genotypes. By evaluating the effect of environmental and agronomic factors on the different variables, the harvest year resulted to be always significant, while location and variety influenced AR abundance only for some homologues. Furthermore, the spatial distribution of AR was investigated by mass spectrometry imaging using transversal cross sections of wheat kernels. Our results show that AR homologues are mainly localized in the testa and in the outer pericarp of wheat kernels.

The effect of nitrogen fertilization on the expression of protein in wheat and tritordeum varieties using a proteomic approach.

Wheat, an essential ingredient for several bakery preparations, is also responsible for gluten-related diseases in sensitive subjects. The effect of the N fertilization rate (80 vs 160 kg N ha-1) on gluten protein expression profile has been evaluated considering two soft wheats (landrace and modern) and one tritordeum cultivar (cv), grown in the same experimental field in North Italy. The proteins of refined flour were characterized through advanced proteomic approaches, including chromatography (RP-HPLC) and electrophoresis. A static model system was used to simulate in vitro digestion and the digestome peptides were examined by mass spectrometry and in silico approaches, to investigate the celiac and allergenic sequences. The CD-toxic epitopes in the digested samples were quantified by means of a R5 ELISA assay. The N fertilization rate increased the grain protein content, but it did not lead to any difference in gluten composition, with exception of glu/glia ratio in the modern wheat cv. Moreover, the gluten composition and the occurrence of toxic/allergenic epitopes varied to a great extent, according mostly to the genotype. A lower immunoreactivity, determined using R5 ELISA, was detected for the digested tritordeum flours than for the landrace (-51%) or modern (-58%) cvs, while no significant difference was observed for the N rates between each genotype. In silico analysis showed that tritordeum has fewer CD epitopes belonging to the ω-gliadins and a lower LMW-GS than the landrace or modern cv. Tritordeum presented fewer α-gliadin allergenic epitopes than the modern wheat cv. The lower frequency of celiac epitopes in tritordeum, compared to the old and the modern wheat, is probably due to the absence of a D genome.

Tritordeum: Creating a New Crop Species-The Successful Use of Plant Genetic Resources.

Hexaploid tritordeum is the amphiploid derived from the cross between the wild barley Hordeum chilense and durum wheat. This paper reviews the main advances and achievements in the last two decades that led to the successful development of tritordeum as a new crop. In particular, we summarize the progress in breeding for agronomic performance, including the potential of tritordeum as a genetic bridge for wheat breeding; the impact of molecular markers in genetic studies and breeding; and the progress in quality and development of innovative food products. The success of tritordeum as a crop shows the importance of the effective utilization of plant genetic resources for the development of new innovative products for agriculture and industry. Considering that wild plant genetic resources have made possible the development of this new crop, the huge potential of more accessible resources, such as landraces conserved in gene banks, goes beyond being sources of resistance to biotic and abiotic stresses. In addition, the positive result of tritordeum also shows the importance of adequate commercialization strategies and demonstrative experiences aimed to integrate the whole food chain, from producers to end-point sellers, in order to develop new products for consumers.

Using Einkorn and Tritordeum Brewers' Spent Grain to Increase the Nutritional Potential of Durum Wheat Pasta.

Brewers' spent grain (BSG), the major by-product of the brewing industry, can be used as a functional ingredient to increase the nutritional value of cereal-based products. In this work, micronized BSG from the einkorn and tritordeum brewing processes were characterized and used to produce four macaroni pasta formulations enriched with BSG at ratios of 5 g and 10 g/100 g of semolina. Einkorn BSG showed the highest values for all the parameters analyzed-proteins, total dietary fiber (TDF) and total antioxidant capacity (TAC)-except for ß-glucan. TDF increased up to 42 and 68% in pasta samples enriched with 10% of BSG from tritordeum and einkorn, respectively. The replacement of 10% of semolina with BSG from both cereals significantly increased the ß-glucan content and TAC values. Finally, the addition of BSG from einkorn and tritordeum affected to a minimal extent the sensory properties of cooked pasta, which showed higher values of optimal cooking time and cooking loss, but lower total organic matter compared to semolina pasta. Results from the sensorial judgment fell in the good quality ranges for durum wheat pasta; the incorporation of 10% of einkorn BSG resulted in the best compromise in terms of technological, nutritional and sensorial aspects of enriched pasta.

Tritordeum breads are well tolerated with preference over gluten-free breads in non-celiac wheat-sensitive patients and its consumption induce changes in gut bacteria.

BACKGROUND: The ingestion of wheat and other cereals are related to several gut disorders. The specific components responsible for non-celiac wheat-sensitivity (NCWS) may include gluten and other compounds. Tritordeum is a new cereal derived from crossing durum wheat with a wild barley species, which differs from bread wheat in its gluten composition. In the present work, we examined the response of NCWS patients to tritordeum bread Gastrointestinal symptoms as well as tritordeum acceptability, gluten immunogenic peptides excretion, and the composition and structure of the intestinal microbiota were evaluated. RESULTS: Gastrointestinal symptoms of the subjects showed no significant change between the gluten-free bread and the tritordeum bread. Participating subjects rated tritordeum bread higher than the gluten-free bread. Analysis of the bacterial gut microbiota indicated that tritordeum consumption does not alter the global structure and composition of the intestinal microbiota, and only a few changes in some butyrate-producing bacteria were observed. CONCLUSIONS: All the results derived from acceptability, biochemical and microbiological tests suggest that tritordeum may be tolerated by a sub-set of NCWS sufferers who do not require strict exclusion of gluten from their diet. © 2020 Society of Chemical Industry.

Carotenoid content in tritordeum is not primarily associated with esterification during grain development.

Tritordeums show a significant proportion of lutein esters which increases carotenoid stability and retention throughout the food chain. Esterification is a common means of carotenoid sequestration. A putative association between lutein esters formation acting as a metabolic sink during early stages of grain development and the high carotenoid content of tritordeums is analyzed in this work. Compared to wheat, tritordeums accumulated significantly higher lutein contents from 20 days post anthesis (dpa) but lutein esters were not detected until 36 dpa. Thus esterification is not acting as a metabolific sink before 36 dpa. The presence of lutein esters at late stages of grain development may have a complementary role in carotenoid accumulation by reducing and/or counteracting their catabolism. The differences for lutein esterification among tritordeums suggest the existence of diversity for xanthophyll acyl transferases that could be exploited to increase lutein retention in this cereal and through the food chain.

Mediation of a GDSL Esterase/Lipase in Carotenoid Esterification in Tritordeum Suggests a Common Mechanism of Carotenoid Esterification in Triticeae Species.

Carotenoids are essential in human diet, so that the development of programs toward carotenoid enhancement has been promoted in several crops. The cereal tritordeum, the amphiploid derived from the cross between Hordeum chilense Roem. et Schulz. and durum wheat has a remarkable carotenoid content in the endosperm. Besides, a high proportion of these carotenoids are esterified with fatty acids. The identification of the gene(s) responsible for xanthophyll esterification would be useful for breeding as esterified carotenoids show an increased ability to accumulate within plant cells and have a higher stability during post-harvest storage. In this work, we analyzed five genes identified as candidates for coding the xanthophyll acyltransferase (XAT) enzyme responsible for lutein esterification in H. chilense genome. All these genes were expressed during grain development in tritordeum, but only HORCH7HG021460 was highly upregulated. Sequence analysis of HORCH7HG021460 revealed a G-to-T transversion, causing a Glycine to Cysteine substitution in the protein of H290 (the only accession not producing quantifiable amounts of lutein esters, hereinafter referred as zero-ester) of H. chilense compared to the esterifying genotypes. An allele-specific marker was designed for the SNP detection in the H. chilense diversity panel. From the 93 accessions, only H290 showed the T allele and the zero-ester phenotype. Furthermore, HORCH7HG021460 is the orthologue of XAT-7D, which encodes a XAT enzyme responsible for carotenoid esterification in wheat. Thus, HORCH7HG021460 (XAT-7Hch) is a strong candidate for lutein esterification in H. chilense and tritordeum, suggesting a common mechanism of carotenoid esterification in Triticeae species. The transference of XAT-7Hch to wheat may be useful for the enhancement of lutein esters in biofortification programs.