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This review proposes a new model, "the two-step branching and improper branch clearing model", to explain how amylopectin is synthesised through the action of starch synthases and starch-branching enzymes coupled with the occasional use of a debranching enzyme to correct improper branch formation.
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The authors set out to design a freeze-thaw-stable starch from first principles. They reasoned that if they removed the amylose and decreased the chain length of amylopectin, then the resulting starch would be less likely to retrograde. An amylose-containing short-chain amylopectin starch had already been created by antisense downregulation of two soluble starch synthases [36]. Retransformation of this line with an antisense GBSS construct produced a starch that was amylose-free, had short-chain amylopectin and was incredibly freeze-thaw stable. The remarkable thing about this starch is that it is made by only one starch synthase gene (i.e. SSI) and there is no significant reduction in yield.
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Jobling S.A., Westcott R.J., Tayal A., Jeffcoat R., Schwall G.P. Production of a freeze-thaw-stable potato starch by antisense inhibition of three starch synthase genes. Nat Biotechnol. 20:2002;295-299 The authors set out to design a freeze-thaw-stable starch from first principles. They reasoned that if they removed the amylose and decreased the chain length of amylopectin, then the resulting starch would be less likely to retrograde. An amylose-containing short-chain amylopectin starch had already been created by antisense downregulation of two soluble starch synthases [36]. Retransformation of this line with an antisense GBSS construct produced a starch that was amylose-free, had short-chain amylopectin and was incredibly freeze-thaw stable. The remarkable thing about this starch is that it is made by only one starch synthase gene (i.e. SSI) and there is no significant reduction in yield.
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Jobling, S.A.1
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Role of granule-bound starch synthase in determination of amylopectin structure and starch granule morphology in potato
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This study showed that granule defects in modified starch lines that contained reduced levels of SSIII were corrected when the expression of GBSS was also reduced. The authors discuss the implications of the presence long glucan chains in amylopectin and amylose on the granule structure of these starches.
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Fulton D.C., Edwards A., Pilling E., Robinson H.L., Fahy B., Seale R., Kato L., Donald A.M., Geigenberger P., Martin C., Smith A.M. Role of granule-bound starch synthase in determination of amylopectin structure and starch granule morphology in potato. J Biol Chem. 277:2002;10834-10841 This study showed that granule defects in modified starch lines that contained reduced levels of SSIII were corrected when the expression of GBSS was also reduced. The authors discuss the implications of the presence long glucan chains in amylopectin and amylose on the granule structure of these starches.
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Bird A.R., Brown I.L., Topping D.L. Starches, resistant starches, the gut microflora and human health. Curr Issues Intest Microbiol. 1:2000;25-37.
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A minor form of starch branching enzyme in potato (Solanum tuberosum L.) tubers has a major effect on starch structure: Cloning and characterisation of multiple forms of SBE a
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Jobling S.A., Schwall G.P., Westcott R.J., Sidebottom C.M., Debet M., Gidley M.J., Jeffcoat R., Safford R. A minor form of starch branching enzyme in potato (Solanum tuberosum L.) tubers has a major effect on starch structure: cloning and characterisation of multiple forms of SBE A. Plant J. 18:1999;163-171.
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Schwall G.P., Safford R., Westcott R.J., Jeffcoat R., Tayal A., Shi Y.C., Gidley M.J., Jobling S.A. Production of very-high-amylose potato starch by inhibition of SBE A and B. Nat Biotechnol. 18:2000;551-554.
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22
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Immunomodulation of enzyme function in plants by single-domain antibody fragments
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A novel method of inhibiting gene function using a special form of antibody was used to create starches that have very high amylose levels. This was the first demonstration of immunomodulation in a transgenic organism, and opens up the possibility of inhibiting enzyme function or altering metabolism in ways that were not previously possible.
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Jobling S.A., Jarman C., Teh M.M., Holmberg N., Blake C., Verhoeyen M.E. Immunomodulation of enzyme function in plants by single-domain antibody fragments. Nat Biotechnol. 21:2003;77-80 A novel method of inhibiting gene function using a special form of antibody was used to create starches that have very high amylose levels. This was the first demonstration of immunomodulation in a transgenic organism, and opens up the possibility of inhibiting enzyme function or altering metabolism in ways that were not previously possible.
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The two isoforms of pea GBSS, one found predominantly in the leaf (GBSSIb) and one in the embryo (GBSSIa), produce different sized amyloses when transformed into the amylose-free mutant of potato.
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Edwards A., Vincken J.P., Suurs L.C., Visser R.G., Zeeman S., Smith A., Martin C. Discrete forms of amylose are synthesized by isoforms of GBSSI in pea. Plant Cell. 14:2002;1767-1785 The two isoforms of pea GBSS, one found predominantly in the leaf (GBSSIb) and one in the embryo (GBSSIa), produce different sized amyloses when transformed into the amylose-free mutant of potato.
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Edwards, A.1
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28
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0038372492
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Barley sex6 mutants lack starch synthase IIa activity and contain a starch with novel properties
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A novel high-amylose starch is discovered in the barley sex6 mutant. The amylopectin that remains in high-amylose starches normally has long branches, and these starches usually have high gelatinisation temperatures, with some very-high-amylose starches requiring autoclaving to be solubilised. The novel high-amylose starch, however, had amylopectin with relatively short branches and gelatinised at lower temperatures than normal starch. In addition to the loss of SSIIa, the granule distribution of several starch-synthesising enzymes was altered, suggesting that these enzymes may form complexes with SSIIa. Further molecular analysis is needed to determine the basis of the apparent high amylose levels in the sex6 mutant because SSIIa mutants in other plants do not show such severe phenotypes.
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Morell M.K., Kosar-Hashemi B., Cmiel M., Samuel M.S., Chandler P., Rahman S., Buleon A., Batey I.L., Li Z. Barley sex6 mutants lack starch synthase IIa activity and contain a starch with novel properties. Plant J. 34:2003;173-185 A novel high-amylose starch is discovered in the barley sex6 mutant. The amylopectin that remains in high-amylose starches normally has long branches, and these starches usually have high gelatinisation temperatures, with some very-high-amylose starches requiring autoclaving to be solubilised. The novel high-amylose starch, however, had amylopectin with relatively short branches and gelatinised at lower temperatures than normal starch. In addition to the loss of SSIIa, the granule distribution of several starch-synthesising enzymes was altered, suggesting that these enzymes may form complexes with SSIIa. Further molecular analysis is needed to determine the basis of the apparent high amylose levels in the sex6 mutant because SSIIa mutants in other plants do not show such severe phenotypes.
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Morell, M.K.1
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29
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Enhanced ADP-glucose pyrophosphorylase activity in wheat endosperm increases seed yield
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Smidansky E.D., Clancy M., Meyer F.D., Lanning S.P., Blake N.K., Talbert L.E., Giroux M.J. Enhanced ADP-glucose pyrophosphorylase activity in wheat endosperm increases seed yield. Proc Natl Acad Sci USA. 99:2002;1724-1729.
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Regierer B., Fernie A.R., Springer F., Perez-Melis A., Leisse A., Koehl K., Willmitzer L., Geigenberger P., Kossmann J. Starch content and yield increase as a result of altering adenylate pools in transgenic plants. Nat Biotechnol. 20:2002;1256-1260.
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From bacterial glycogen to starch: Understanding the biogenesis of the plant starch granule
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A useful review that contrasts the differences between synthesising glycogen, a soluble glucan polymer and a starch that forms the remarkable semicrystalline granule.
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Ball S.G., Morell M.K. From bacterial glycogen to starch: understanding the biogenesis of the plant starch granule. Annu Rev Plant Biol. 54: 2003;207-233 A useful review that contrasts the differences between synthesising glycogen, a soluble glucan polymer and a starch that forms the remarkable semicrystalline granule.
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The authors describe the real phenotype of a starch-branching enzyme I mutant for the first time. As with other SBEI mutants in maize or potato lines in which SEBI has been downregulated by antisense techniques, there was no change in amylose levels. In the rice mutant, however, the fine structure of amylopectin was slightly changed: there were more short chains and fewer long chains, although the changes were not large. These changes gave rise to a starch that was slightly more soluble in urea and had a slightly lower onset of gelatinisation than wildtype starches.
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Satoh H., Nishi A., Yamashita K., Takemoto Y., Tanaka Y., Hosaka Y., Sakurai A., Fujita N., Nakamura Y. Starch branching enzyme-I-deficient mutation specifically affects the structure and properties of starch in rice endosperm. Plant Physiol. 133:2003;1111-1121 The authors describe the real phenotype of a starch-branching enzyme I mutant for the first time. As with other SBEI mutants in maize or potato lines in which SEBI has been downregulated by antisense techniques, there was no change in amylose levels. In the rice mutant, however, the fine structure of amylopectin was slightly changed: there were more short chains and fewer long chains, although the changes were not large. These changes gave rise to a starch that was slightly more soluble in urea and had a slightly lower onset of gelatinisation than wildtype starches.
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Edwards A., Fulton D.C., Hylton C.M., Jobling S.A., Gidley M., Rossner U., Martin C., Smith A.M. A combined reduction in activity of starch synthases II and III of potato has novel effects on the starch of tubers. Plant J. 17:1999;251-261.
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The authors describe the development of a new sweet potato line whose starch has a remarkably low gelatinisation temperature, approximately 20°C lower than that of the wildtype. This starch has properties similar to those of a potato starch that was developed by antisense inhibition of two starch synthase genes [36]. The amylopectin structure of the two starches is similar in that they both have more short chains and fewer long chains than is normal. Because sweet potato is hexaploid, this mutation is likely to be dominant. It is possible that a genome rearrangement has occurred that created an antisense gene to a starch synthase gene, but more work is necessary to identify the gene(s) responsible.
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Katayama K., Komae K., Kohyama K., Kato T., Tamiya S., Komaki K. New sweet potato line having low gelatinization temperature and altered starch structure. Starch-Stärke. 54:2002;51-57 The authors describe the development of a new sweet potato line whose starch has a remarkably low gelatinisation temperature, approximately 20°C lower than that of the wildtype. This starch has properties similar to those of a potato starch that was developed by antisense inhibition of two starch synthase genes [36]. The amylopectin structure of the two starches is similar in that they both have more short chains and fewer long chains than is normal. Because sweet potato is hexaploid, this mutation is likely to be dominant. It is possible that a genome rearrangement has occurred that created an antisense gene to a starch synthase gene, but more work is necessary to identify the gene(s) responsible.
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Japonica rice varieties have better eating quality and their starch gelatinises at a lower temperature than indica varieties. The amylopectin of japonica varieties is enriched in short chains and depleted in intermediate-sized chains compared to indica varieties. The gene responsible for this difference, previously known as alk, is now known to encode SSIIa.
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Umemoto T., Yano M., Satoh H., Shomura A., Nakamura Y. Mapping of a gene responsible for the difference in amylopectin structure between japonica-type and indica-type rice varieties. Theor Appl Genet. 104:2002;1-8 Japonica rice varieties have better eating quality and their starch gelatinises at a lower temperature than indica varieties. The amylopectin of japonica varieties is enriched in short chains and depleted in intermediate-sized chains compared to indica varieties. The gene responsible for this difference, previously known as alk, is now known to encode SSIIa.
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The long-sort-after enzyme that phosphorylates starch is functionally characterised as an alpha-glucan water dikinase, providing a biochemical explanation of how the phosphate is incorporated into starch. Nevertheless, the target structures within starch that are recognised by this enzyme remain to be determined.
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Ritte G., Lloyd J.R., Eckermann N., Rottmann A., Kossmann J., Steup M. The starch-related R1 protein is an alpha-glucan, water dikinase. Proc Natl Acad Sci USA. 99:2002;7166-7171 The long-sort-after enzyme that phosphorylates starch is functionally characterised as an alpha-glucan water dikinase, providing a biochemical explanation of how the phosphate is incorporated into starch. Nevertheless, the target structures within starch that are recognised by this enzyme remain to be determined.
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A comprehensive microscopic examination of several highly modified starches, including antisense GBSS, SBEI and SBEII and the starch- phosphorylating enzyme GWD.
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This is the first functional study of the suitability of various potato starches (including one transgenic starch) that differ mainly in phosphate content for paper manufacture. The best correlation with paper filler retention was starch stickiness and those starches that performed best had rapid granule burst, which was related to their phosphate content.
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Blennow A., Bay-Smidt A., Leonhardt P., Bandsholm O., Madsen M.H. Starch paste stickiness is a relevant native starch selection criterion for wet-end paper manufacturing. Starch-Stärke. 55:2003;381-389 This is the first functional study of the suitability of various potato starches (including one transgenic starch) that differ mainly in phosphate content for paper manufacture. The best correlation with paper filler retention was starch stickiness and those starches that performed best had rapid granule burst, which was related to their phosphate content.
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Yu Abs S-M: Transgenic rice expressing a thermostable amylopullulanase in seeds leads to starch autohydrolysis and production of high-protein flour. American Society of Plant Biologists Meeting: 2003 July 25-30; Honlulu, Hawaii. 2003. [URL: http://abstracts.aspb.org/pb2003/public/P34/0355.html]. The processing of the grains of transgenic rice plants that express a thermostable amylopullulanase at temperatures of around 908C results in a remarkably efficient complete conversion of starch into soluble sugars. A signal peptide that directs the amylopullulanase to the apoplast was used to direct the overexpression of this protein. The amylopullulanase was apparently mistargetted to the plastid by an unknown mechanism, however, and this resulted in a slightly modified starch structure.
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Alternative splicing of the first two exons of a SBE gene generates one form with an amino-terminal extension that is found in both the soluble and granule-bound starch fractions. The form without this extension is exclusively soluble and has different kinetic properties.
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Hamada S., Ito H., Hiraga S., Inagaki K., Nozaki K., Isono N., Yoshimoto Y., Takeda Y., Matsui H. Differential characteristics and subcellular localization of two starch-branching enzyme isoforms encoded by a single gene in Phaseolus vulgaris L. J Biol Chem. 277:2002;16538-16546 Alternative splicing of the first two exons of a SBE gene generates one form with an amino-terminal extension that is found in both the soluble and granule-bound starch fractions. The form without this extension is exclusively soluble and has different kinetic properties.
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