A Corrigendum on Characterization of Triticum turgidum sspp. durum, turanicum, and polonicum grow... more A Corrigendum on 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 an... more 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.
Amylase/trypsin inhibitors (ATIs) are widely consumed in cereal-based foods and have been implica... more Amylase/trypsin inhibitors (ATIs) are widely consumed in cereal-based foods and have been implicated in adverse reactions to wheat exposure, such as respiratory and food allergy, and intestinal responses associated with coeliac disease and noncoeliac wheat sensitivity. ATIs occur in multiple isoforms which differ in the amounts present in different types of wheat (including ancient and modern ones). Measuring ATIs and their isoforms is an analytical challenge as is their isolation for use in studies addressing their potential effects on the human body. ATI isoforms differ in their spectrum of bioactive effects in the human gastrointestinal (GI), which may include enzyme inhibition, inflammation and immune responses and of which much is not known. Similarly, although modifications during food processing (exposure to heat, moisture, salt, acid, fermentation) may affect their structure and activity as shown in vitro, it is important to relate these changes to effects that may present in the GI tract. Finally, much of our knowledge of their potential biological effects is based on studies in vitro and in animal models. Validation by human studies using processed foods as commonly consumed is warranted. We conclude that more detailed understanding of these factors may allow the effects of ATIs on human health to be better understood and when possible, to be ameliorated, for example by innovative food processing. We therefore review in short our current knowledge of these proteins, focusing on features which relate to their biological activity and identifying gaps in our knowledge and research priorities.
The recently growing demand of gluten-reduced beer is leading to the development of diverse appro... more The recently growing demand of gluten-reduced beer is leading to the development of diverse approaches to be applied in brewing. The current work focuses on the development of an innovative and sustainable biocatalytic tool for the continuous production of gluten-reduced beer, based on the application of immobilized prolyl endopeptidase from Aspergillus niger (AN-PEP). This food-grade protease has been immobilized on A. niger chitosan beads and applied, for the first time, for the reduction of gluten in a commercial beer from barley malt. The immobilization procedure was optimized for maximizing the specific activity of the biocatalyst (0.016 I.U./ mg BSAeq) and the best performance was reached using an immobilization solution at an initial protein concentration of 0.3 mg BSAeq /mL. The immobilization increased the thermal stability of the protease, which showed similar catalytic properties in synthetic beer (toward the synthetic substrate Z-Gly-Pro-pNA) when it was applied at 20°C or at 50°C. The continuous treatment in fluidized bed reactor (FBR), containing 10 g of immobilized AN-PEP (corresponding to 0.0036 g BSAeq), was optimized varying the flow rate (Q v). The suitable conditions to achieve reduction of the intact gluten of authentic beer was Q v of 728 mL/min. The continuous treatment in FBR allowed us to reduce the initial gluten content (65 mg/kg) in the commercial beer from barley malt, reaching the concentration of 19 mg/ kg after 9 h and 15 mg/kg after 10 h of treatment.
... IN THE WHEAT ENDOSPERM AND BELONG TO THE GLUTENIN FRACTION P. Ferrante, C. Patacchini, S. Mas... more ... IN THE WHEAT ENDOSPERM AND BELONG TO THE GLUTENIN FRACTION P. Ferrante, C. Patacchini, S. Masci, R. D'Ovidio, D. Lafiandra ... This latter was perfomed by the Molecular Structure Facility at University of California, Davis by means of an Applied Biosystems Procise ...
Background: Among wheat gluten proteins, the α-type gliadins are the major responsible for celiac... more Background: Among wheat gluten proteins, the α-type gliadins are the major responsible for celiac disease, an autoimmune disorder that affects about 1% of the world population. In fact, these proteins contain several toxic and immunogenic epitopes that trigger the onset of the disease. The α-type gliadins are a multigene family, encoded by genes located at the complex Gli-2 loci. Results: Here, three bread wheat deletion lines (Gli-A2, Gli-D2 and Gli-A2/Gli-D2) at the Gli-2 loci were generated by the introgression in the bread wheat cultivar Pegaso of natural mutations, detected in different bread wheat cultivars. The molecular characterization of these lines allowed the isolation of 49 unique expressed genes coding α-type gliadins, that were assigned to each of the three Gli-2 loci. The number and the amount of α-type gliadin transcripts were drastically reduced in the deletion lines. In particular, the line Gli-A2/Gli-D2 contained only 12 active α-type gliadin genes (−75.6% respect to the cv. Pegaso) and a minor level of transcripts (−80% compared to cv. Pegaso). Compensatory pleiotropic effects were observed in the two other classes of gliadins (ω-and γ-gliadins) either at gene expression or protein levels. Although the comparative analysis of the deduced amino acid sequences highlighted the typical structural features of α-type gliadin proteins, substantial differences were displayed among the 49 proteins for the presence of toxic and immunogenic epitopes. Conclusion: The deletion line Gli-A2/Gli-D2 did not contain the 33-mer peptide, one of the major epitopes triggering the celiac disease, representing an interesting material to develop less "toxic" wheat varieties.
Con sideration s abou t th e effect of in corporation of two rare LMW-GS in du ru m wh eat in com... more Con sideration s abou t th e effect of in corporation of two rare LMW-GS in du ru m wh eat in comparison to bread wh eat dou gh s.
The Royal Society of Chemistry eBooks, May 19, 2004
... The uee of biotechnology to study wheat endosperm development and improve grain quality. D. L... more ... The uee of biotechnology to study wheat endosperm development and improve grain quality. D. Lafiandra, S. Masci and R. D'Ovidio Copyright Year:2004 ISBN: 978-1-84755-209-9 DOI:10.1039/9781847552099-00001. ...
dependent on the wheat species. The combination of manual curation of the MaxQuant search with Sk... more dependent on the wheat species. The combination of manual curation of the MaxQuant search with Skyline revealed a very good performance. The DIA approach with analytical flow found similar results compared to absolute quantitation except for some minor ATIs, which were not detected. Comparison of applied methods revealed that peptide selection is a crucial step for protein quantitation. Wheat proteomics faces challenges due to the high genetic complexity, the close relationship to other cereals and the incomplete, redundant protein database requiring sensitive, precise and accurate LC-MS/MS methods.
The Royal Society of Chemistry eBooks, May 19, 2004
... Bread Wheat Line Over-Expressing a LMW-GS. S. Masci, R. D'Ovidio, F. Scossa, C. ... more ... Bread Wheat Line Over-Expressing a LMW-GS. S. Masci, R. D'Ovidio, F. Scossa, C. Patacchini, D. Lafiandra, OD Anderson and AE Blechl Copyright Year:2004 ISBN: 978-1-84755-209-9 DOI:10.1039/9781847552099-00010. ...
Plant genetic transformation and regeneration has become a valuable research tool for functional ... more Plant genetic transformation and regeneration has become a valuable research tool for functional genomics. A successful transformation event involves the transfer of the target gene into a suitable explant, the integration and expression of the transgene into the host genome and the regeneration of the fertile transgenic plants from the transformed tissues. Wheat is considered as a recalcitrant species even if many efforts have been done in recent years to improve transformation efficiency. The transformation of its progenitors has never been attempted, even though the possibility to transform wild hemmer represents a valuable tool to evaluate structural and functional variability occurring in wild hemmer and explaining its higher adaptation to abiotic stresses. In this paper we report, as far as we know, for the first time, the microparticle transformation of immature embryos of the wild hemmer Triticum dicoccoides with the Tapgip1 gene. The transformation method was successfully transferred from durum wheat and several transgenic lines were obtained. Its application for the exploitation of wheat progenitors for molecular breeding is of great relevance for genomic and functional genomics studies. This result, indeed, opens new perspectives in complementation studies for the comprehension of durum and bread wheat adaptation mechanisms to stresses.
Frontiers in sustainable food systems, Jul 31, 2020
Wheat and its derived foods are widespread, representing one of the main food sources globally. D... more Wheat and its derived foods are widespread, representing one of the main food sources globally. During the last decades, the incidence of disorders related to wheat has become a global issue for the human population, probably linked to the spread of wheat-derived foods. It has been ascertained that structural and metabolic proteins, like α-amylase/trypsin inhibitors (ATI), are involved in the onset of wheat allergies (bakers' asthma) and probably Non-Coeliac Wheat Sensitivity (NCWS). The ATI are a group of exogenous protease inhibitors, which are encoded by a multigene family dispersed over several chromosomes in durum and bread wheat. WTAI-CM3 and WTAI-CM16 subunits are considered among the main proteins involved in the onset of bakers' asthma and probably NCWS. A CRISPR-Cas9 multiplexing strategy was used to edit the ATI subunits WTAI-CM3 and WTAI-CM16 in the grain of the Italian durum wheat cultivar Svevo with the aim to produce wheat lines with reduced amount of potential allergens involved in adverse reactions. Using a marker gene-free approach, whereby plants are regenerated without selection agents, homozygous mutant plants without the presence of CRISPR vectors were obtained directly from T 0 generation. This study demonstrates the capability of CRISPR technology to knock out immunogenic proteins in a reduced time compared to conventional breeding programmes. The editing of the two target genes was confirmed either at molecular (sequencing and gene expression study) or biochemical (immunologic test) level. Noteworthy, as a pleiotropic effect, is the activation of the ATI 0.28 pseudogene in the edited lines.
HAL (Le Centre pour la Communication Scientifique Directe), Feb 7, 2013
HAL is a multidisciplinary open access archive for the deposit and dissemination of scientific re... more HAL is a multidisciplinary open access archive for the deposit and dissemination of scientific research documents, whether they are published or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. L'archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d'enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.
A Corrigendum on Characterization of Triticum turgidum sspp. durum, turanicum, and polonicum grow... more A Corrigendum on 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 an... more 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.
Amylase/trypsin inhibitors (ATIs) are widely consumed in cereal-based foods and have been implica... more Amylase/trypsin inhibitors (ATIs) are widely consumed in cereal-based foods and have been implicated in adverse reactions to wheat exposure, such as respiratory and food allergy, and intestinal responses associated with coeliac disease and noncoeliac wheat sensitivity. ATIs occur in multiple isoforms which differ in the amounts present in different types of wheat (including ancient and modern ones). Measuring ATIs and their isoforms is an analytical challenge as is their isolation for use in studies addressing their potential effects on the human body. ATI isoforms differ in their spectrum of bioactive effects in the human gastrointestinal (GI), which may include enzyme inhibition, inflammation and immune responses and of which much is not known. Similarly, although modifications during food processing (exposure to heat, moisture, salt, acid, fermentation) may affect their structure and activity as shown in vitro, it is important to relate these changes to effects that may present in the GI tract. Finally, much of our knowledge of their potential biological effects is based on studies in vitro and in animal models. Validation by human studies using processed foods as commonly consumed is warranted. We conclude that more detailed understanding of these factors may allow the effects of ATIs on human health to be better understood and when possible, to be ameliorated, for example by innovative food processing. We therefore review in short our current knowledge of these proteins, focusing on features which relate to their biological activity and identifying gaps in our knowledge and research priorities.
The recently growing demand of gluten-reduced beer is leading to the development of diverse appro... more The recently growing demand of gluten-reduced beer is leading to the development of diverse approaches to be applied in brewing. The current work focuses on the development of an innovative and sustainable biocatalytic tool for the continuous production of gluten-reduced beer, based on the application of immobilized prolyl endopeptidase from Aspergillus niger (AN-PEP). This food-grade protease has been immobilized on A. niger chitosan beads and applied, for the first time, for the reduction of gluten in a commercial beer from barley malt. The immobilization procedure was optimized for maximizing the specific activity of the biocatalyst (0.016 I.U./ mg BSAeq) and the best performance was reached using an immobilization solution at an initial protein concentration of 0.3 mg BSAeq /mL. The immobilization increased the thermal stability of the protease, which showed similar catalytic properties in synthetic beer (toward the synthetic substrate Z-Gly-Pro-pNA) when it was applied at 20°C or at 50°C. The continuous treatment in fluidized bed reactor (FBR), containing 10 g of immobilized AN-PEP (corresponding to 0.0036 g BSAeq), was optimized varying the flow rate (Q v). The suitable conditions to achieve reduction of the intact gluten of authentic beer was Q v of 728 mL/min. The continuous treatment in FBR allowed us to reduce the initial gluten content (65 mg/kg) in the commercial beer from barley malt, reaching the concentration of 19 mg/ kg after 9 h and 15 mg/kg after 10 h of treatment.
... IN THE WHEAT ENDOSPERM AND BELONG TO THE GLUTENIN FRACTION P. Ferrante, C. Patacchini, S. Mas... more ... IN THE WHEAT ENDOSPERM AND BELONG TO THE GLUTENIN FRACTION P. Ferrante, C. Patacchini, S. Masci, R. D'Ovidio, D. Lafiandra ... This latter was perfomed by the Molecular Structure Facility at University of California, Davis by means of an Applied Biosystems Procise ...
Background: Among wheat gluten proteins, the α-type gliadins are the major responsible for celiac... more Background: Among wheat gluten proteins, the α-type gliadins are the major responsible for celiac disease, an autoimmune disorder that affects about 1% of the world population. In fact, these proteins contain several toxic and immunogenic epitopes that trigger the onset of the disease. The α-type gliadins are a multigene family, encoded by genes located at the complex Gli-2 loci. Results: Here, three bread wheat deletion lines (Gli-A2, Gli-D2 and Gli-A2/Gli-D2) at the Gli-2 loci were generated by the introgression in the bread wheat cultivar Pegaso of natural mutations, detected in different bread wheat cultivars. The molecular characterization of these lines allowed the isolation of 49 unique expressed genes coding α-type gliadins, that were assigned to each of the three Gli-2 loci. The number and the amount of α-type gliadin transcripts were drastically reduced in the deletion lines. In particular, the line Gli-A2/Gli-D2 contained only 12 active α-type gliadin genes (−75.6% respect to the cv. Pegaso) and a minor level of transcripts (−80% compared to cv. Pegaso). Compensatory pleiotropic effects were observed in the two other classes of gliadins (ω-and γ-gliadins) either at gene expression or protein levels. Although the comparative analysis of the deduced amino acid sequences highlighted the typical structural features of α-type gliadin proteins, substantial differences were displayed among the 49 proteins for the presence of toxic and immunogenic epitopes. Conclusion: The deletion line Gli-A2/Gli-D2 did not contain the 33-mer peptide, one of the major epitopes triggering the celiac disease, representing an interesting material to develop less "toxic" wheat varieties.
Con sideration s abou t th e effect of in corporation of two rare LMW-GS in du ru m wh eat in com... more Con sideration s abou t th e effect of in corporation of two rare LMW-GS in du ru m wh eat in comparison to bread wh eat dou gh s.
The Royal Society of Chemistry eBooks, May 19, 2004
... The uee of biotechnology to study wheat endosperm development and improve grain quality. D. L... more ... The uee of biotechnology to study wheat endosperm development and improve grain quality. D. Lafiandra, S. Masci and R. D'Ovidio Copyright Year:2004 ISBN: 978-1-84755-209-9 DOI:10.1039/9781847552099-00001. ...
dependent on the wheat species. The combination of manual curation of the MaxQuant search with Sk... more dependent on the wheat species. The combination of manual curation of the MaxQuant search with Skyline revealed a very good performance. The DIA approach with analytical flow found similar results compared to absolute quantitation except for some minor ATIs, which were not detected. Comparison of applied methods revealed that peptide selection is a crucial step for protein quantitation. Wheat proteomics faces challenges due to the high genetic complexity, the close relationship to other cereals and the incomplete, redundant protein database requiring sensitive, precise and accurate LC-MS/MS methods.
The Royal Society of Chemistry eBooks, May 19, 2004
... Bread Wheat Line Over-Expressing a LMW-GS. S. Masci, R. D'Ovidio, F. Scossa, C. ... more ... Bread Wheat Line Over-Expressing a LMW-GS. S. Masci, R. D'Ovidio, F. Scossa, C. Patacchini, D. Lafiandra, OD Anderson and AE Blechl Copyright Year:2004 ISBN: 978-1-84755-209-9 DOI:10.1039/9781847552099-00010. ...
Plant genetic transformation and regeneration has become a valuable research tool for functional ... more Plant genetic transformation and regeneration has become a valuable research tool for functional genomics. A successful transformation event involves the transfer of the target gene into a suitable explant, the integration and expression of the transgene into the host genome and the regeneration of the fertile transgenic plants from the transformed tissues. Wheat is considered as a recalcitrant species even if many efforts have been done in recent years to improve transformation efficiency. The transformation of its progenitors has never been attempted, even though the possibility to transform wild hemmer represents a valuable tool to evaluate structural and functional variability occurring in wild hemmer and explaining its higher adaptation to abiotic stresses. In this paper we report, as far as we know, for the first time, the microparticle transformation of immature embryos of the wild hemmer Triticum dicoccoides with the Tapgip1 gene. The transformation method was successfully transferred from durum wheat and several transgenic lines were obtained. Its application for the exploitation of wheat progenitors for molecular breeding is of great relevance for genomic and functional genomics studies. This result, indeed, opens new perspectives in complementation studies for the comprehension of durum and bread wheat adaptation mechanisms to stresses.
Frontiers in sustainable food systems, Jul 31, 2020
Wheat and its derived foods are widespread, representing one of the main food sources globally. D... more Wheat and its derived foods are widespread, representing one of the main food sources globally. During the last decades, the incidence of disorders related to wheat has become a global issue for the human population, probably linked to the spread of wheat-derived foods. It has been ascertained that structural and metabolic proteins, like α-amylase/trypsin inhibitors (ATI), are involved in the onset of wheat allergies (bakers' asthma) and probably Non-Coeliac Wheat Sensitivity (NCWS). The ATI are a group of exogenous protease inhibitors, which are encoded by a multigene family dispersed over several chromosomes in durum and bread wheat. WTAI-CM3 and WTAI-CM16 subunits are considered among the main proteins involved in the onset of bakers' asthma and probably NCWS. A CRISPR-Cas9 multiplexing strategy was used to edit the ATI subunits WTAI-CM3 and WTAI-CM16 in the grain of the Italian durum wheat cultivar Svevo with the aim to produce wheat lines with reduced amount of potential allergens involved in adverse reactions. Using a marker gene-free approach, whereby plants are regenerated without selection agents, homozygous mutant plants without the presence of CRISPR vectors were obtained directly from T 0 generation. This study demonstrates the capability of CRISPR technology to knock out immunogenic proteins in a reduced time compared to conventional breeding programmes. The editing of the two target genes was confirmed either at molecular (sequencing and gene expression study) or biochemical (immunologic test) level. Noteworthy, as a pleiotropic effect, is the activation of the ATI 0.28 pseudogene in the edited lines.
HAL (Le Centre pour la Communication Scientifique Directe), Feb 7, 2013
HAL is a multidisciplinary open access archive for the deposit and dissemination of scientific re... more HAL is a multidisciplinary open access archive for the deposit and dissemination of scientific research documents, whether they are published or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. L'archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d'enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.
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