Abstract
The barley shrunken grain mutant M292 has a novel high-amylose starch phenotype caused by a mutation in the starch synthase IIa gene (SsIIa) located at the starch excess-6 (sex6) locus on chromosome 7H of barley. The loss of SSIIa enzyme activity leads to a decrease in amylopectin synthesis to less than 20% of the levels found in wild-type grains. Detailed composition analysis indicates that the contents of protein, non-starch polysaccharides, lipid, sucrose and hexoses, and fructo-oligosaccharides are increased in mature M292 grain compared to wild type. Using a microarray analysis, we characterize the differences between the transcription profiles of wild-type and mutant barley endosperms at mid-grain fill. The expression changes include genes involved in carbon storage, stress-related genes, and a number of transcripts with unassigned function. The changes in gene expression are discussed in terms of the altered grain composition of the mutant seed.
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References
Andon NL, Hollingworth S, Koller A, Greenland AJ, Yates JR, Haynes PA (2002) Proteomic characterization of wheat amyloplasts using identification of proteins by tandem mass spectrometry. Proteomics 2:1156–1168
Andrawis A, Solomon M, Delmer DP (1993) Cotton fiber annexins: a potential role in the regulation of callose synthase. Plant J 3:763–772
Appels R, Dvorak J (1982) The wheat ribosomal DNA spacer region: its structure and variation in populations and among species. Theor Appl Genet 63:337–348
Ball SG, Morell MK (2003) From bacterial glycogen to starch: understanding the biogenesis of the plant starch granule. Annu Rev Plant Biol 54:207–233
Balmer Y, Vensel WH, DuPont FM, Buchanan BB, Hurkman WJ (2006) Proteome of amyloplasts isolated from develo** wheat endosperm presents evidence of broad metabolic capability. J Exp Bot 57:1591–1602
Bernfeld P (1955) Amylases alpha and beta. In: Colowick S, Kaplan N (eds) Methods in enzymology. Academic, NY, p 149
Bird AR, Flory C, Davies DA, Usher S, Top** DL (2004a) A novel barley cultivar (Himalaya 292) with a specific gene mutation in starch synthase IIa raises large bowel starch and short chain fatty acids in rats. J Nutr 134:831–835
Bird AR, Jackson M, King RA, Davies DA, Usher S, Top** DL (2004b) A novel high amylose barley cultivar (Hordeum vulgare var. Himalaya 292) lowers plasma cholesterol and alters indices of large-bowel fermentation in pigs. Br J Nutr 92:607–615
Boston RS, Viitanen PV, Vierling E (1996) Molecular chaperones and protein folding in plants. Plant Mol Biol l32:191–222
Campbell JA, Hansen RW, Wilson JR (1999) Cost-effective colorimetric microtitre plate enzymatic assays for sucrose, glucose and fructose in sugarcane tissue extracts. J Sci Food Agric 79:232–236
Clarke B, Rahman S (2005) A microarray analysis of wheat grain hardness. Theor Appl Genet 110:1259–1267
Clarke BC, Hobbs M, Skylas D, Appels R (2000) Genes active in develo** wheat endosperm. Funct Integr Genomics 1:44–55
da Silva PMFR, Eastmond PJ, Hill LM, Smith AM, Rawsthorne S (1997) Starch metabolism in develo** embryos of oilseed rape. Planta 203:480–487
Davies C, Robinson SP (2000) Differential screening indicates a dramatic change in mRNA profiles during grape berry ripening. Cloning and characterization of cDNAs encoding putative cell wall and stress response proteins. Plant Physiol 122:803–812
Doehlert DC, Kuo TM (1994) Gene expression in develo** kernels of some endosperm mutants of maize. Plant Cell Physiol 35:411–418
Fincher GB, Stone BA (1986) Cell walls and their components in cereal grain technology. Adv Cereal Sci Technol 8:207–295
Franco OL, Rigden DJ, Melo FR, Bloch JC, Silva CP, Grossi de Sa MF (2000) Activity of wheat alpha-amylase inhibitors towards bruchid alpha-amylases and structural explanation of observed specificities. Eur J Biochem 267:2166–2173
Giroux MJ, Boyer C, Feix G, Hannah LC (1994) Coordinated transcriptional regulation of storage product genes in the maize endosperm. Plant Physiol 106:713–722
Guerin JR, Lance RCM, Wallace W (1992) Release and activation of barley beta-amylase by malt endopeptidases. J Cereal Sci 15:5–14
Harvey AJ, Hrmova M, Fincher GB (2001) Regulation of genes encoding β-D-gucan glucohydrolases in barley (Hordeum vulgare). Physiol Plant 113:108–120
Hrmova M, Fincher GB (2001) Structure–function relationships and β-d-glucan endo- and exohydrolases from higher plants. Plant Mol Biol 47:73–91
James MG, Denyer K, Myers AM (2003) Starch synthesis in the cereal endosperm. Curr Opin Plant Biol 6:215–222
Konarev AV (1996) Interaction of insect digestive enzymes with plant protein inhibitors and host-parasite co-evolution. Euphytica 92:89–94
Kossmann J, Lloyd J (2000) Understanding and influencing starch biochemistry. Crit Rev Plant Sci 19:171–226
Kumar A, Taylor MA, Mad Arif SA, Davies HV (1996) Potato plants expressing antisense and sense S-adenosylmethionine decarboxylase (SAMDC) transgenes show altered levels of polyamines and ethylene: antisense plants display abnormal phenotypes. Plant J 9:147–158
Laudencia-Chingcuanco DL, Stamova BS, You FM, Lazo GR, Beckles DM, Anderson OD (2007) Transcriptional profiling of wheat caryopsis development using cDNA microarrays. Plant Mol Biol 63:651–668
Lunn JE, Hatch MD (1995) Primary partitioning and storage of photosynthate in sucrose and starch in leaves of C4 plants. Planta 197:385–391
Merendino N, D’Aquino M, Molinari R, De Gara L, D’Egidio MG, Paradiso A, Cecchini C, Corradini C, Tomassi G (2006) Chemical characterization and biological effects of immature durum wheat in rats. J Cereal Sci 43:129–136
Millar HA, Sweetlove LJ, Giege P, Leaver CJ (2001) Analysis of the Arabidopsis mitochondrial proteome. Plant Physiol 127:1711–1727
Morell MK, Rahman S, Regina A, Appels R, Li Z (2001) Wheat starch biosynthesis. Euphytica 119:55–58
Morell MK, Kosar-Hashemi B, Cmiel M, Samuel MS, Chandler P, Rahman S, Buleon A, Batey IL, Li Z (2003) Barley sex6 mutants lack starch synthase IIa activity and contain a starch with novel properties. Plant J 34:173–185
Morell MK, Li Z, Regina A, Rahman S, d’Hulst C, Ball SG (2006) Control of starch biosynthesis in vascular plants and algae. In: Plaxton WC, McManus MT (eds) Control of primary metabolism in plants. Annual Plant Reviews 22:258–289. Blackwell Publishing, Oxford, UK
Myers AM, Morell MK, James MG, Ball SG (2000) Recent progress towards understanding the amylopectin crystal. Plant Physiol 122:989–997
Nijtmans LGJ, de Jong L, Sanz MA, Coates PJ, Berden JA, Back JW, Muijsers AO, van der Spek H, Grivell LA (2000) Prohibitins act as a membrane-bound chaperone for the stabilization of the mitochondrial proteins. EMBO J 19:2444–2451
Ohdan T, Francisco PB Jr, Sawada T, Hirose T, Terao T, Satoh H, Nakamura Y (2005) Expression profiling of genes involved in starch synthesis in sink and source organs of rice. J Exp Bot 56:3229–3244
Østergaard H, Rasmussen SK, Roberts TH, Hejgaard J (2000) Inhibitory serpins from wheat with reactive centers resembling glutamine-rich repeats of prolamin storage proteins. J Biol Chem 275:33272–33279
Poole R, Barker G, Wilson ID, Coghill JA, Edwards KJ (2007) Measuring global gene expression in polyploidy; a cautionary note from allohexaploid wheat. Funct Integr Genomics 7:207–219
Prosky L, Asp N-G, Furda I, DeVries JW, Schweizer TF, Harland BF (1985) Determination of total dietary fibre in foods and food product: collaborative study. J Assoc Off Agric Chem 68:677
RACI (2003) Official testing methods of the Cereal Chemistry Division of the Royal Australian Chemical Institute, Method 02-03. Dumas (combustion) total nitrogen determination
Rahman S, Li Z, Batey I, Cochrane MP, Appels R, Morell MK (2000) Genetic alteration of starch functionality in wheat. J Cereal Sci 31:91–110
Regina A, Bird A, Top** D, Bowden S, Freeman J, Barsby T, Kosar-Hashemi B, Li Z, Rahman S, Morell M (2006) High-amylose wheat generated by RNA interference improves indices of large-bowel health in rats. Proc Natl Acad Sci USA 103:3546–3551
Ruuska S, Rebetzke G, van Herwaarden A, Richards RA, Fettell N, Tabe L, Jenkins CLD (2006) Genotypic variation in water-soluble carbohydrate accumulation in wheat. Funct Plant Biol 33:799–809
Schulman AH, Tomooka S, Suzuki A, Myllarinen P, Hizukuri S (1995) Structural analysis of starch from normal and shx (shrunken endosperm) barley (Hordeum vulgare L.). Carbohydr Res 275:361–369
Shin H, Brown RM Jr (1999) GTPase Activity and biochemical characterization of a recombinant cotton fiber annexin. Plant Physiol 119:925–934
Smith AM (2001) The biosynthesis of starch granules. Biomacromolecules 2:335–341
Tetlow IJ, Morell MK, Emes MJ (2004) Recent developments in understanding the regulation of starch metabolism in higher plants. J Exp Bot 55:2131–2145
Theander O, Aman P, Westerlund E, Andersson R, Pettersson D (1995) Total dietary fibre determined as neutral sugar residues, uronic acid residues and Klason lignin (the Uppsala method). J AOAC Int 78:1030–1044
Top** DL, Morell KM, King RA, Li Z, Bird AR, Noakes M (2003) Resistant starch and health—Himalaya 292, a novel barley cultivar to deliver benefits to consumers. Starke 55:539–545
Tsuchiya K, Urahara T, Konishi T, Kotake T, Tohno-oka T, Komae K, Kawada N, Tsumuraya Y (2005) Biosynthesis of (1-3),(1-4)-β-glucan in develo** endosperms of barley (Hordeum vulgare). Physiol Plant 125:181–191
Whetten R, Sun Y-H, Zhang E, Sederoff R (2001) Functional genomics and cell wall biosynthesis in loblolly pine. Plant Mol Biol 47:275–291
Wilson DL, Buckley MJ, Helliwell CA, Wilson IW (2003) New normalisation methods for cDNA microarray data. Bioinformatics 19:1325–1332
Winter H, Huber SC (2000) Regulation of sucrose metabolism in higher plants: localization and regulation of activity of key enzymes. Crit Rev Plant Sci 19:31–67
Wobus U, Sreenivasulu N, Borisjuk L, Rolletschek H, Panitz R, Gubatz S, Weschke W (2004) Molecular physiology and genomics of develo** barley grains. Recent Res Dev Plant Mol Biol 2:1–29
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Clarke, B., Liang, R., Morell, M.K. et al. Gene expression in a starch synthase IIa mutant of barley: changes in the level of gene transcription and grain composition. Funct Integr Genomics 8, 211–221 (2008). https://doi.org/10.1007/s10142-007-0070-7
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DOI: https://doi.org/10.1007/s10142-007-0070-7