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Figure S4: Monosaccharide content in wheat dry grain, flour, isolated outer pericarp and intermediate layers (var. Recital.) Material and methods: Samples:

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Presentation on theme: "Figure S4: Monosaccharide content in wheat dry grain, flour, isolated outer pericarp and intermediate layers (var. Recital.) Material and methods: Samples:"— Presentation transcript:

1 Figure S4: Monosaccharide content in wheat dry grain, flour, isolated outer pericarp and intermediate layers (var. Recital.) Material and methods: Samples: flour (type 55) from milled Recital grains (800 mg) dry whole grain grinded in liquid nitrogen (100 mg) manually dissected outer pericarp grinded in liquid nitrogen (100 mg) manually dissected intermediate layers (cross cells tube cells, testa and nucellar epidermis )(100 mg) grinded in liquid nitrogen (100 mg) Preparation of alcohol-insoluble residue (AIR) Small sugars (glucose, sucrose, etc.) were removed using 80% ethanol for 10 min; the insoluble residue was isolated by centrifugation (6300g; 10 min). The procedure was repeated once and the residue was washed with 95% ethanol and again isolated by centrifugation (6300g; 10 min). The supernatant was discarded and the residue was dried in two steps: first in an oven at 40 °C for 24 h and then with P2O5 in a vacuum oven at 40 °C for 48h. Individual neutral sugar content was determined following hydrolysis and alditol acetate derivatisation as described in Dervilly et al. (2000) according to the method of Englyst and Cummings (1988). Uronic acids were assayed by an automated m-hydroxydipenyl method (Blumenktrantz & Asboe- Hansen, 1973; Thibault 1979). Galacturonic acid was estimated taking advantage of the differential response of glucuronic acid and galacturonic acid in the presence of sodium tetraborate (Thibault 1979). Results (mean of two measurements) In wheat grain : Rhamnose indicates the presence of rhamnogalacturonans Arabinose mainly comes from arabinoxylans, also from arabinogalactan proteins Xylose mainly comes from arabinoxylans Mannose comes from mannans Glucose comes mainly from mixed-linked glucans and cellulose and from starch in flour and whole grain Galactose mainly comes from arabinogalactan proteins and possibly from heteroxylans Galacturonic acid indicates the presence of pectic homogalacturonans and/or rhamnogalacturonans % of AIRwhole grainouter pericarpinterm layersflour Rhamnose (Rha)0.280.00 0.33 Arabinose (Ara)2.7227.1010.750.89 Xylose (Xyl)4.5224.1232.11.54 Mannose (Man)0.250.830.000.23 Galactose (Gal)0.411.541.050.25 Glucose (Glc)75.9327.5119.6291.00 Galacturonic acid0.170.310.700.03 Glucuronic acid0.333.172.640.14

2 Grain Standards Rha Fuc Ara Xyl Glc Inositol Man Gal Inositol Intermediate layers Rha Ara Xyl Glc Man Gal Ara Xyl Glc Gal mV Gas-liquid chromatograms from dry wheat grain and intermediate layers. Dervilly et al. (2000) Isolation of homogeneous fractions from wheat water-soluble arabinoxylans. J. Agric. Food Chem. 48, 270-278. Englyst HN and Cummings JH (1988) Improved method of measurement of dietary fiber as non-starch polysaccharides in plant foods. J. Assoc. Off. Anal. Chem. 71, 808-814. Blumenkrantz N and Asboe-Hansen (1973) New method for quantitative determination of uronic acids. Analytical Biochemistry 54, 484-489. Thibault J-F (1979) Automatisation du dosage des substances pectiques par la méthode au méta-hydroxydipenyl. Lebensmitell Wissenschaft und Technologie (LWT) 12, 247-251

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