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Aleurone Layer

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Gerard Branlard – 1st expert on this subject based on the ideXlab platform

  • proteome evolution of wheat triticum aestivum l Aleurone Layer at fifteen stages of grain development
    Journal of Proteomics, 2015
    Co-Authors: I. Nadaud, Christophe Chambon, Ayesha Tasleemtahir, Annelaure Chateignerboutin, Didier Viala, Gerard Branlard

    Abstract:

    Abstract The Aleurone Layer (AL) is the grain peripheral tissue; it is rich in micronutrients, vitamins, antioxidants, and essential amino acids. This highly nutritive part of the grain has been less studied partly because its isolation is so laborious. In the present study, the ALs of Triticum aestivum (variety Recital) were separated manually at 15 stages of grain development. A total of 327 proteins were identified using 2-DE LC-MS/MS. They were classified in six main groups and 26 sub-groups according to their biochemical function. Proteomic analysis revealed seven different profiles distributed among three main development stages: (i) early AL development, with proteins involved in intense metabolic activities in the growth and development of the cell wall compounds; (ii) the intermediate stage, characterized by oxidative stress and defense proteins (65%) linked with loss of water in peripheral Layers during grain filling; and (iii) AL maturation, involving the production of amino acids and the control of reactive oxidative species to enable the accumulation and maturation of globulins within the AL. The present study provides the first insights into developing proteome in the AL. We describe the numerous AL enzymes involved in the accumulation of storage protein and in the protection of the endosperm over time. Biological significance The hand dissection of wheat Aleurone Layer (AL) was carried in this study for the first time on fifteen developmental stages from cell differentiation to grain maturity. Three major phases were revealed over AL development: cell division activities, globulins storage, and grain protection. Enzymes related to metabolites and vitamins were abundantly expressed during the two first phases. In parallel to the progressive globulins accumulation, the final phase was characterized by key enzyme synthesis involved in energy production, amino-acids and antioxidant synthesis plus others to face hypoxia and dehydration of grain tissues.

  • proteomic analysis of the mature kernel Aleurone Layer in common and durum wheat
    Journal of Cereal Science, 2012
    Co-Authors: Smail Meziani, I. Nadaud, Christophe Chambon, Marzouk Benali, Brigitte Gaillardmartinie, Gerard Branlard

    Abstract:

    Abstract The Aleurone Layer (AL) is one of inner tissues removed from the grain with the wheat bran. It is the main source of vitamins, minerals and antioxidants of potential nutritional value in the wheat kernel. The AL of three varieties of each of the two main species of wheat, Triticum aestivum (ABD) and Triticum durum (AB), were manually dissected and analysed using two-dimensional gel-based proteomics. A total of 1258 and 1109 Coomassie-stained spots were detected in the AL of representatives of the ABD and AB genomes. In two varieties ( T. aestivum Chinese Spring and T. durum Bidi17), grown in two different years with full fungicide protection, no quantitative or qualitative (presence/absence) differences in spots were detected, suggesting that AL proteome is strongly genetically controlled. Comparison within and between species revealed a total of 339 AL significant protein spots. Among these spots, 30.8% differed within T. aestivum and 56.5% within T. durum varieties, whereas only 12.7% differed between the two species. Among the 142 AL proteins identified using MALDI-TOF and LC-MS/MS, 57% were globulin type storage proteins (Glo-3, Glo-3B, Glo-3C, Glo-2), 16.2% were involved in carbohydrate metabolism and 17.6% in defence/stress pathways. These variations in AL proteome are discussed.

  • Proteomic analysis of the mature kernel Aleurone Layer in common and durum wheat
    Journal of Cereal Science, 2012
    Co-Authors: Smail Meziani, I. Nadaud, Brigitte Gaillard-martinie, Christophe Chambon, Marzouk Benali, Gerard Branlard

    Abstract:

    The Aleurone Layer (AL) is one of inner tissues removed from the grain with the wheat bran. It is the main source of vitamins, minerals and antioxidants of potential nutritional value in the wheat kernel. The AL of three varieties of each of the two main species of wheat, Triticum aestivum (ABD) and Triticum durum (AB), were manually dissected and analysed using two-dimensional gel-based proteomics. A total of 1258 and 1109 Coomassie-stained spots were detected in the AL of representatives of the ABD and AB genomes. In two varieties (T. aestivum Chinese Spring and T. durum Bidi17), grown in two different years with full fungicide protection, no quantitative or qualitative (presence/absence) differences in spots were detected, suggesting that AL proteome is strongly genetically controlled. Comparison within and between species revealed a total of 339 AL significant protein spots. Among these spots, 30.8% differed within T. aestivum and 56.5% within T. durum varieties, whereas only 12.7% differed between the two species. Among the 142 AL proteins identified using MALDI-TOF and LC-MS/MS, 57% were globulin type storage proteins (Glo-3, Glo-3B, Glo-3C, Glo-2), 16.2% were involved in carbohydrate metabolism and 17.6% in defence/stress pathways. These variations in AL proteome are discussed. © 2012 Elsevier Ltd.

Christine Finnie – 2nd expert on this subject based on the ideXlab platform

  • Immobilisation of barley Aleurone Layers enables parallelisation of assays and analysis of transient gene expression in single cells.
    Plant Physiology and Biochemistry, 2017
    Co-Authors: Christina Mark, Arto Heiskanen, Claus Krogh Madsen, Martin Dufva, Jenny Emnéus, Henrik Brinch-pedersen, Christine Finnie

    Abstract:

    Abstract The barley Aleurone Layer is an established model system for studying phytohormone signalling, enzyme secretion and programmed cell death during seed germination. Most analyses performed on the Aleurone Layer are end-point assays based on cell extracts, meaning each sample is only analysed at a single time point. By immobilising barley Aleurone Layer tissue on polydimethylsiloxane pillars in the lid of a multiwell plate, continuous monitoring of living tissue is enabled using multiple non-destructive assays in parallel. Cell viability and menadione reducing capacity were monitored in the same Aleurone Layer samples over time, in the presence or absence of plant hormones and other effectors. The system is also amenable to transient gene expression by particle bombardment, with simultaneous monitoring of cell death. In conclusion, the easy to handle and efficient experimental setup developed here enables continuous monitoring of tissue samples, parallelisation of assays and single cell analysis, with potential for time course studies using any plant tissue that can be immobilised, for example leaves or epidermal peels.

  • spatio temporal appearance of α amylase and limit dextrinase in barley Aleurone Layer in response to gibberellic acid abscisic acid and salicylic acid
    Journal of the Science of Food and Agriculture, 2015
    Co-Authors: Azar Shahpiri, Nasim Talaei, Christine Finnie

    Abstract:

    BACKGROUND

    Cereal seed germination involves mobilization of storage reserves in the starchy endosperm to support seedling growth. In response to gibberellin produced by the embryo the Aleurone Layer synthesizes hydrolases that are secreted to the endosperm for degradation of storage products. In this study analysis of intracellular protein accumulation and release from barley Aleurone Layers is presented for the important enzymes in starch degradation: α-amylase and limit dextrinase (LD).

    RESULTS

    Proteins were visualized by immunoblotting in Aleurone Layers and culture supernatants from dissected Aleurone Layers incubated up to 72 h with either gibberellic acid (GA), abscisic acid (ABA) or salicylic acid (SA). The results show that α-amylase is secreted from Aleurone Layer treated with GA soon after synthesis but the release of LD to culture supernatants was significantly delayed and coincided with a general loss of proteins from Aleurone Layers.

    CONCLUSIONS

    Release of LD was found to differ from that of amylase and was suggested to depend on programmed cell death (PCD). Despite detection of intracellular amylase in untreated Aleurone Layers or Aleurone Layers treated with ABA or SA, α-amylase was not released from these samples. Nevertheless, the release of α-amylase was observed from Aleurone Layers treated with GA+ABA or GA+SA. © 2014 Society of Chemical Industry

  • Spatio‐temporal appearance of α‐amylase and limit dextrinase in barley Aleurone Layer in response to gibberellic acid, abscisic acid and salicylic acid
    Journal of the Science of Food and Agriculture, 2014
    Co-Authors: Azar Shahpiri, Nasim Talaei, Christine Finnie

    Abstract:

    BACKGROUND

    Cereal seed germination involves mobilization of storage reserves in the starchy endosperm to support seedling growth. In response to gibberellin produced by the embryo the Aleurone Layer synthesizes hydrolases that are secreted to the endosperm for degradation of storage products. In this study analysis of intracellular protein accumulation and release from barley Aleurone Layers is presented for the important enzymes in starch degradation: α-amylase and limit dextrinase (LD).

    RESULTS

    Proteins were visualized by immunoblotting in Aleurone Layers and culture supernatants from dissected Aleurone Layers incubated up to 72 h with either gibberellic acid (GA), abscisic acid (ABA) or salicylic acid (SA). The results show that α-amylase is secreted from Aleurone Layer treated with GA soon after synthesis but the release of LD to culture supernatants was significantly delayed and coincided with a general loss of proteins from Aleurone Layers.

    CONCLUSIONS

    Release of LD was found to differ from that of amylase and was suggested to depend on programmed cell death (PCD). Despite detection of intracellular amylase in untreated Aleurone Layers or Aleurone Layers treated with ABA or SA, α-amylase was not released from these samples. Nevertheless, the release of α-amylase was observed from Aleurone Layers treated with GA+ABA or GA+SA. © 2014 Society of Chemical Industry

I. Nadaud – 3rd expert on this subject based on the ideXlab platform

  • proteome evolution of wheat triticum aestivum l Aleurone Layer at fifteen stages of grain development
    Journal of Proteomics, 2015
    Co-Authors: I. Nadaud, Christophe Chambon, Ayesha Tasleemtahir, Annelaure Chateignerboutin, Didier Viala, Gerard Branlard

    Abstract:

    Abstract The Aleurone Layer (AL) is the grain peripheral tissue; it is rich in micronutrients, vitamins, antioxidants, and essential amino acids. This highly nutritive part of the grain has been less studied partly because its isolation is so laborious. In the present study, the ALs of Triticum aestivum (variety Recital) were separated manually at 15 stages of grain development. A total of 327 proteins were identified using 2-DE LC-MS/MS. They were classified in six main groups and 26 sub-groups according to their biochemical function. Proteomic analysis revealed seven different profiles distributed among three main development stages: (i) early AL development, with proteins involved in intense metabolic activities in the growth and development of the cell wall compounds; (ii) the intermediate stage, characterized by oxidative stress and defense proteins (65%) linked with loss of water in peripheral Layers during grain filling; and (iii) AL maturation, involving the production of amino acids and the control of reactive oxidative species to enable the accumulation and maturation of globulins within the AL. The present study provides the first insights into developing proteome in the AL. We describe the numerous AL enzymes involved in the accumulation of storage protein and in the protection of the endosperm over time. Biological significance The hand dissection of wheat Aleurone Layer (AL) was carried in this study for the first time on fifteen developmental stages from cell differentiation to grain maturity. Three major phases were revealed over AL development: cell division activities, globulins storage, and grain protection. Enzymes related to metabolites and vitamins were abundantly expressed during the two first phases. In parallel to the progressive globulins accumulation, the final phase was characterized by key enzyme synthesis involved in energy production, amino-acids and antioxidant synthesis plus others to face hypoxia and dehydration of grain tissues.

  • proteomic analysis of the mature kernel Aleurone Layer in common and durum wheat
    Journal of Cereal Science, 2012
    Co-Authors: Smail Meziani, I. Nadaud, Christophe Chambon, Marzouk Benali, Brigitte Gaillardmartinie, Gerard Branlard

    Abstract:

    Abstract The Aleurone Layer (AL) is one of inner tissues removed from the grain with the wheat bran. It is the main source of vitamins, minerals and antioxidants of potential nutritional value in the wheat kernel. The AL of three varieties of each of the two main species of wheat, Triticum aestivum (ABD) and Triticum durum (AB), were manually dissected and analysed using two-dimensional gel-based proteomics. A total of 1258 and 1109 Coomassie-stained spots were detected in the AL of representatives of the ABD and AB genomes. In two varieties ( T. aestivum Chinese Spring and T. durum Bidi17), grown in two different years with full fungicide protection, no quantitative or qualitative (presence/absence) differences in spots were detected, suggesting that AL proteome is strongly genetically controlled. Comparison within and between species revealed a total of 339 AL significant protein spots. Among these spots, 30.8% differed within T. aestivum and 56.5% within T. durum varieties, whereas only 12.7% differed between the two species. Among the 142 AL proteins identified using MALDI-TOF and LC-MS/MS, 57% were globulin type storage proteins (Glo-3, Glo-3B, Glo-3C, Glo-2), 16.2% were involved in carbohydrate metabolism and 17.6% in defence/stress pathways. These variations in AL proteome are discussed.

  • Proteomic analysis of the mature kernel Aleurone Layer in common and durum wheat
    Journal of Cereal Science, 2012
    Co-Authors: Smail Meziani, I. Nadaud, Brigitte Gaillard-martinie, Christophe Chambon, Marzouk Benali, Gerard Branlard

    Abstract:

    The Aleurone Layer (AL) is one of inner tissues removed from the grain with the wheat bran. It is the main source of vitamins, minerals and antioxidants of potential nutritional value in the wheat kernel. The AL of three varieties of each of the two main species of wheat, Triticum aestivum (ABD) and Triticum durum (AB), were manually dissected and analysed using two-dimensional gel-based proteomics. A total of 1258 and 1109 Coomassie-stained spots were detected in the AL of representatives of the ABD and AB genomes. In two varieties (T. aestivum Chinese Spring and T. durum Bidi17), grown in two different years with full fungicide protection, no quantitative or qualitative (presence/absence) differences in spots were detected, suggesting that AL proteome is strongly genetically controlled. Comparison within and between species revealed a total of 339 AL significant protein spots. Among these spots, 30.8% differed within T. aestivum and 56.5% within T. durum varieties, whereas only 12.7% differed between the two species. Among the 142 AL proteins identified using MALDI-TOF and LC-MS/MS, 57% were globulin type storage proteins (Glo-3, Glo-3B, Glo-3C, Glo-2), 16.2% were involved in carbohydrate metabolism and 17.6% in defence/stress pathways. These variations in AL proteome are discussed. © 2012 Elsevier Ltd.