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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.
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Secas , Regulação da Expressão Gênica de Plantas , Raízes de Plantas , Secale , Translocação Genética , Triticum , Triticum/genética , Triticum/fisiologia , Secale/genética , Secale/fisiologia , Raízes de Plantas/genética , Raízes de Plantas/fisiologia , Cromossomos de Plantas/genética , Estresse Fisiológico/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Resistência à SecaRESUMO
KEY MESSAGE: Two allelic variants of Pp-A3 and Pp-B1 were identified in purple durum wheat. Molecular markers at both loci were developed and validated on an independent panel, offering a breakthrough for wheat improvement. Purple wheats are a class of cereals with pigmented kernels of particular interest for their antioxidant and anti-inflammatory properties. Although two complementary loci (Pp-B1 and Pp-A3), responsible for purple pericarp have been pinpointed in bread wheat (Triticum aestivum L.), in durum wheat (Triticum durum Desf.) the causative genes along with functional and non-functional alleles are still unknown. Here, using a quantitative trait loci (QTL) mapping approach on a RIL population derived from purple and non-purple durum wheat genotypes, we identified three major regions on chromosomes 2A, 3A, and 7B explaining the highest phenotypic variation (> 50%). Taking advantage of the Svevo genome, a MYB was reannotated on chromosome 7B and reported as a candidate for Pp-B1. An insertion of ~ 1.6 kb within the first exon led to a non-functional allele (TdPpm1b), whereas the functional allele (TdPpm1a) was characterized and released for the first time in durum wheat. Pp-A3 was instead identified as a duplicated gene, of which only one was functional. The promoter sequencing of the functional allele (TdPpb1a) revealed six 261-bp tandem repeats in purple durum wheat, whereas one unit (TdPpb1b) was found in the yellow once. Functional molecular markers at both loci were developed to precisely discriminate purple and not purple genotypes, representing a valuable resource for selecting superior purple durum lines at early growth stages. Overall, our results expand the understanding of the function of MYB and bHLH activators in durum wheat, paving new ways to explore cis-regulatory elements at the promoter level.
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Alelos , Mapeamento Cromossômico , Genótipo , Fenótipo , Locos de Características Quantitativas , Triticum , Triticum/genética , Triticum/crescimento & desenvolvimento , Mapeamento Cromossômico/métodos , Marcadores Genéticos , Genes de Plantas , Pigmentação/genética , Cromossomos de Plantas/genéticaRESUMO
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.
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Glutens , Triticum , Glutens/genética , Glutens/química , Triticum/genética , Peso Molecular , Alelos , Poliploidia , Subunidades Proteicas/genética , Subunidades Proteicas/químicaRESUMO
Drought is a major challenge for the cultivation of durum wheat, a crucial crop for global food security. Plants respond to drought by adjusting their mineral nutrient profiles to cope with water scarcity, showing the importance of nutrient plasticity for plant acclimation and adaptation to diverse environments. Therefore, it is essential to understand the genetic basis of mineral nutrient profile plasticity in durum wheat under drought stress to select drought-tolerant varieties. The research study investigated the responses of different durum wheat genotypes to severe drought stress at the seedling stage. The study employed an ionomic, molecular, biochemical and physiological approach to shed light on distinct behaviors among different genotypes. The drought tolerance of SVEMS16, SVEVO, and BULEL was related to their capacity of maintaining or increasing nutrient's accumulation, while the limited nutrient acquisition capability of CRESO and S.CAP likely resulted in their susceptibility to drought. The study highlighted the importance of macronutrients such as SO42-, NO3-, PO43-, and K+ in stress resilience and identified variant-containing genes potentially influencing nutritional variations under drought. These findings provide valuable insights for further field studies to assess the drought tolerance of durum wheat genotypes across various growth stages, ultimately ensuring food security and sustainable production in the face of changing environmental conditions.
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Secas , Minerais , Triticum , Triticum/genética , Triticum/fisiologia , Triticum/metabolismo , Minerais/metabolismo , Genótipo , Adaptação Fisiológica/genética , Resistência à SecaRESUMO
An increasing number of individuals are eating out due to work and study commitments. This trend directly influences people's food choices, especially those who frequently rely on snacks and pre-packaged foods. Consuming these foods can lead to long-term health consequences. Adding functional foods to vending machines could lead to healthier choices. Our aim is to evaluate the acceptability and willingness to pay (WTP) of workers and students for a snack pack of novel functional biscuits (FBs) made with high amylose contents. We found that the experimental flour used is effective in preventing various non-communicable diseases; two phases of analysis were carried out on 209 participants. The participants blindly tested the products and only after the sensory evaluation were they informed about the biscuits' health contents. Firstly, the blind investigation highlighted the acceptability of the FBs compared to the conventional biscuits. Secondly, the finite mixture model on WTP revealed that some consumers are interested in the health benefits associated with high-amylose test blends and others are focused on hedonistic taste. The design of a communication strategy and industry approach should aim to assist consumers in comprehending the health benefits and sensory aspects of novel functional foods available on the market.
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The criteria of "Distinctness, Uniformity and Stability" as well as a high "overall quality index" are used to register the Italian modern varieties to the national register. Differently, local conservation varieties can be certified under different EU Directives that facilitate, as an overall objective, the preservation of biodiversity and the containment of genetic erosion. In recent years, products derived from ancient grains are perceived to be healthier and more sustainable by consumers, especially in Italy, with consequent higher market prices. The ancient tetraploid wheat varieties registered in the national register of conservation varieties amount to 28, 24 of which are Sicilian. They are supposed to have wide genetic variability compared to modern ones, making them vulnerable to fraud because they are difficult to trace. It is therefore important to have tools able to discriminate between autochthonous Sicilian varieties. This can be completed by gluten proteins composition, which also provides information on the technological properties of derived products. Fifty-one accessions belonging to twenty-two ancient varieties of Sicilian tetraploid (mostly durum) wheat were analyzed. Although wide intra-accession and intra-varietal variability measurements were assessed, the gliadin pattern of bulks of seeds belonging to each variety was discriminatory. Moreover, differences in technological attitudes were found between landraces. This paves the way to use gluten protein patterns for traceability, allowing local farmers and producers to valorize their products and assure consumers regarding the transparency of the entire supply chain.
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[This corrects the article DOI: 10.3389/fpls.2023.1269212.].
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BACKGROUND: In wheat-derived bakery products, the quantity of free asparagine (fAsn) has been identified as a key factor in acrylamide (AA) formation. Based on this assumption, four varieties of common wheat (Triticum aestivum L.), Stromboli, Montecarlo, Sothys and Cosmic, selected for their different fAsn content inside the grain, were studied to evaluate their potential in the production of pizza with reduced AA levels. To this purpose, wholemeal and refined flours were obtained from each variety. RESULTS: The fAsn content ranged from 0.25 to 3.30 mmol kg-1, with higher values for wholemeal flours which also showed greater amount of ash, fibre and damaged starch than refined wheat flours. All types of flours were separately used to produce wood oven baked pizza base, according to the Traditional Speciality Guaranteed EU Regulation (97/2010). AA reduction in the range 47-68% was found for all the selected wheat cultivars, compared with a commercial flour, with significantly lower values registered when refined flour was used. Moreover, refined leavened dough samples showed decreased levels of fAsn and reducing sugars due to the fermentation activity of yeasts. Furthermore, it was confirmed that pizza made with wholemeal flours exhibited lower rapidly digestible starch (RDS) and rapidly available glucose (RAG) values compared to that prepared with the refined flour. CONCLUSION: This study clearly shows that a reduced asparagine content in wheat flour is a key factor in the mitigation of AA formation in pizza base. Unfortunately, at the same time, it is highlighted how it is necessary to sacrifice the beneficial effects of fibre intake, such as lowering the glycaemic index, in order to reduce AA. © 2024 Society of Chemical Industry.
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Asparagina , Farinha , Asparagina/química , Amido , Triticum/química , Acrilamida/química , Madeira , PãoRESUMO
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.
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Amilose , Triticum , Amilose/metabolismo , Triticum/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Amido/metabolismo , Mutação/genética , Cloroplastos/genética , Cloroplastos/metabolismoRESUMO
Although the Med-Diet is a healthy diet model, it is affected by current dietary habits. Therefore, new foods with improved nutritional value should be developed to respond to the needs of people following the Med-Diet. This study was focused on developing high-ß-glucan flat bread (bazlama) with a relatively lower GI. A bread wheat (cv. Tosunbey) flour was enriched with the flour of a high-ß-glucan-content hull-less barley (cv. Chifaa) flour (15, 30, 45 and 60%) to develop a functional bazlama. The nutritional and technological properties of bazlama samples enriched with barley flour were compared with the ones produced from bread wheat. All of the barley flour-enriched bazlama samples had higher yellowness values (b*) than the control (both crumb and crust), which is generally preferred by the consumers. Texture results indicated that bazlama samples became harder with the increase in barley flour supplementation level. The results showed that 3 g of ß-glucan can be provided from the barley flour-enriched bazlama samples (at 45 and 60% levels), and this is the limit to carry health claims. The bazlama samples enriched with barley flour were richer in Mg, K, Mn, Fe, and Zn minerals than the control (100% Tosunbey flour). While the glycemic index (GI) of commercial bread wheat and Tosunbey bazlama samples were high (88.60% and 79.20%, respectively), GI values of the bazlama samples enriched with 60% (64.73) and 45% barley flour (68.65) were medium. The lower GI values of barley flour-enriched bazlama samples are probably due to the higher ß-glucan contents of the bazlama samples. Additionally, as the barley flour supplementation level of the bazlama samples increased, the phenolics and antioxidant capacities of free and bound extracts increased compared to bread wheat bazlama. The results indicated that hull-less barley (cv. Chifaa) with high ß-glucan content may be utilized at relatively higher levels (45 and 60%) to produce bazlama with improved nutritional properties.
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BACKGROUD: Pseudomonas syringae pv. actinidiae (Psa), P. syringae pv. tomato (Pst) and P. savastanoi pv. savastanoi (Psav) are bacterial plant pathogens with worldwide impact that are mainly managed by the preventive application of cupric salts. These are dangerous for ecosystems and have favoured the selection of resistant strains, so they are candidates to be replaced in the next few years. Thus, there is an urgent need to find efficient and bio-based solutions to mitigate these bacterial plant diseases. Nanotechnology could represent an innovative way to control plant diseases, providing alternative solutions to the agrochemicals traditionally employed, thanks to the formulation of the so-called third-generation and nanotechnology-based agrochemicals. RESULTS: In this work, a novel nanostructured formulation (NPF) composed of cellulose nanocrystals (CNC) as carrier, high amylose starch (HAS) as excipient, and chitosan (CH) and gallic acid (GA) as antimicrobials, was tested at 2% in vitro and in vivo with respect to the three different Pseudomonas plant pathogens. In vitro agar assays demonstrated that the NPF inhibited ≤80% Psa, Pst and Psav. Moreover, the NPF did not decrease biofilm synthesis and it did not influence bacterial cells flocculation and adhesion. On plants, the NPF displayed complete biocompatibility and boosted the transcript levels of the major systemic acquired resistance responsive genes in kiwifruit and olive plants. CONCLUSION: This works provides novel and valuable information regarding the several modes-of-action of the novel NPF, which could potentially be useful to mitigate Psa, Pst and Psav infections even in organic agriculture. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
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Quitosana , Olea , Pseudomonas syringae , Olea/microbiologia , Ecossistema , Antibacterianos , Agroquímicos , Doenças das Plantas/microbiologiaRESUMO
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.
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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.
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Selenium (Se) is an essential micronutrient of fundamental importance to human health and the main Se source is from plant-derived foods. Plants mainly take up Se as selenate (SeO42-), through the root sulfate transport system, because of their chemical similarity. The aims of this study were (1) to characterize the interaction between Se and S during the root uptake process, by measuring the expression of genes coding for high-affinity sulfate transporters and (2) to explore the possibility of increasing plant capability to take up Se by modulating S availability in the growth medium. We selected different tetraploid wheat genotypes as model plants, including a modern genotype, Svevo (Triticum turgidum ssp. durum), and three ancient Khorasan wheats, Kamut, Turanicum 21, and Etrusco (Triticum turgidum ssp. turanicum). The plants were cultivated hydroponically for 20 days in the presence of two sulfate levels, adequate (S = 1.2 mM) and limiting (L = 0.06 mM), and three selenate levels (0, 10, 50 µM). Our findings clearly showed the differential expression of genes encoding the two high-affinity transporters (TdSultr1.1 and TdSultr1.3), which are involved in the primary uptake of sulfate from the rhizosphere. Interestingly, Se accumulation in shoots was higher when S was limited in the nutrient solution.
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Selênio , Triticum , Humanos , Ácido Selênico , Triticum/metabolismo , Tetraploidia , Sulfatos/metabolismo , Selênio/metabolismo , GenótipoRESUMO
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.
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Amilose , Farinha , Humanos , Triticum/química , Fermentação , Amido/química , Metabolômica , DigestãoRESUMO
Fusarium head blight (FHB) and Fusarium crown rot (FCR) are managed by the application of imidazole fungicides, which will be strictly limited by 2030, as stated by the European Green Deal. Here, a novel and eco-sustainable nanostructured particle formulation (NPF) is presented by following the principles of the circular economy. Cellulose nanocrystals (CNC) and resistant starch were obtained from the bran of a high amylose (HA) bread wheat and employed as carrier and excipient, while chitosan and gallic acid were functionalized as antifungal and elicitor active principles. The NPF inhibited conidia germination and mycelium growth, and mechanically interacted with conidia. The NPF optimally reduced FHB and FCR symptoms in susceptible bread wheat genotypes while being biocompatible on plants. The expression level of 21 genes involved in the induction of innate immunity was investigated in Sumai3 (FHB resistant) Cadenza (susceptible) and Cadenza SBEIIa (a mutant characterized by high-amylose starch content) and most of them were up-regulated in Cadenza SBEIIa spikes treated with the NPF, indicating that this genotype may possess an interesting genomic background particularly responsive to elicitor-like molecules. Quantification of fungal biomass revealed that the NPF controlled FHB spread, while Cadenza SBEIIa was resistant to FCR fungal spread. The present research work highlights that the NPF is a powerful weapon for FHB sustainable management, while the genome of Cadenza SBEIIa should be investigated deeply as particularly responsive to elicitor-like molecules and resistant to FCR fungal spread.
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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.
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Micronutrient malnutrition affects more than half of the world population. Reduced bioavailability of microelements in the raw materials is considered one of the main causes of mineral deficiency in populations whose diet is largely based on the consumption of staple crops. In this context, the production of low phytic acid (lpa) cereals is a main goal of the breeding programs, as phytic acid (PA) binds essential mineral cations such as iron (Fe), zinc (Zn), manganese (Mn), potassium (K), calcium (Ca) and magnesium (Mg) precipitating in the form of phytate salts poorly digested by monogastric animals, including humans, due to the lack of phytases in the digestive tract. Since PA limits the bioavailability of microelements, it is widely recognized as an anti-nutritional compound. A Targeting Induced Local Lesions IN Genomes (TILLING) approach has been undertaken to silence the genes encoding the TdABCC13 proteins, known as Multidrug-Resistance associated Proteins 3 (TdMRP3), transporters involved in the accumulation of PA inside the vacuole in durum wheat. The TdMRP3 complete null genotypes showed a significant reduction in the content of PA and were able to accumulate a higher amount of essential micronutrients (Fe, Zn, Mn) compared to the control. The number of spikelets and seeds per spike, traits associated with the agronomic performances, were reduced compared to the control, but the negative effect was in part balanced by the increased grain weight. The TdMRP3 mutant lines showed morphological differences in the root apparatus such as a significant decrease in the number of root tips, root length, volume and surface area and an increase in root average diameter compared to the control plants. These materials represent a promising basis for obtaining new commercial durum wheats with higher nutritional value.
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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.