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Differences in specificity and compensatory functions among three major starch synthases determine the structure of amylopectin in rice endosperm

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Abstract

The lengths of amylopectin-branched chains are precise and influence the physicochemical properties of starch, which determine starch functionality. Three major isozymes of starch synthases (SSs), SSI, SSII(a), and SSIII(a), are primarily responsible for amylopectin chain elongation in the storage tissues of plants. To date, the majority of reported rice mutants were generated using japonica cultivars, which have almost inactive SSIIa. Although three SSs share some overlapping chain length preferences, whether they complement each other remains unknown due to the absence of suitable genetic combinations of materials. In this study, rice ss1/SS2a/SS3a and SS1/SS2a/ss3a were newly generated, and the chain length distribution patterns of all the possible combinations of presence and absence of SSI, SSIIa, and SSIIIa activities were compared. This study demonstrated that SSIIa can complement most SSI functions that use glucan chains with DP 6–7 to generate DP 8–12 chains but cannot fully compensate for the elongation of DP 16–19 chains. This suggests that SSIIa preferentially elongates outer but not inner chains of amylopectin. In addition, the results revealed that neither SSI nor SSIIIa compensate for SSIIa. Neither SSI nor SSIIa compensate for elongation of DP >30 by SSIIIa. SSIIa could not resolve the pleiotropic increase of SSI caused by the absence of SSIIIa; instead, SSIIa further elongated those branches elongated by SSI. These results revealed compensatory differences among three major SS isozymes responsible for lengths of amylopectin branches.

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References

  • Abe N, Nakamura Y, Fujita N (2013) Thermal properties, morphology of starch granules, and crystallinity of endosperm starch in SSI and BE isozyme double mutant lines. J Appl Glycosci 60:171–176

    Article  CAS  Google Scholar 

  • Abe N, Asai H, Yago H, Oitome NF, Itoh R, Crofts N, Nakamura Y, Fujita N (2014) Relationships between starch synthase I and branching enzyme isozymes determined using double mutant rice lines. BMC Plant Biol 14:80

    Article  PubMed  PubMed Central  Google Scholar 

  • Akoh CC, Chang SW, Lee GC, Shaw JF (2008) Biocatalysis for the production of industrial products and functional foods from rice and other agricultural produce. J Agric Food Chem 26:10445–10451

    Article  Google Scholar 

  • Asai H, Abe N, Matsushima R, Crofts N, Oitome NF, Nakamura Y, Fujita N (2014) Deficiencies in both starch synthase IIIa and branching enzyme IIb lead to a significant increase in amylose in SSIIa-inactive japonica rice seeds. J Exp Bot 65:5497–5507

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bai X, Luo L, Yan W, Kovi MR, Zhan W, Xing Y (2010) Genetic dissection of rice grain shape using a recombinant inbred line population derived from two contrasting parents and fine mapping a pleiotropic quantitative trait locus qGL7. BMC Genet 11:16

    Article  PubMed  PubMed Central  Google Scholar 

  • Bao J, Xiao P, Hiratsuka M, Sun M, Umemoto T (2009) Granule-bound SSIIa protein content and its relationship with amylopectin structure and gelatinization temperature of rice starch. Starch 61:431–437

    Article  CAS  Google Scholar 

  • Benmoussa M, Moldenhauer KA, Hamaker BR (2007) Rice amylopectin fine structure variability affects starch digestion properties. J Agric Food Chem 55:1475–1479

    Article  CAS  PubMed  Google Scholar 

  • Brust H, Lehmann T, D’Hulst C, Fettke J (2014) Analysis of the functional interaction of Arabidopsis starch synthase and branching enzyme isoforms reveals that the cooperative action of SSI and BEs results in glucans with polymodal chain length distribution similar to amylopectin. PLoS ONE 9:e102364

    Article  PubMed  PubMed Central  Google Scholar 

  • Chen Y, Bao J (2016) Underlying mechanisms of zymographic diversity in starch synthase I and pullulanase in rice-developing endosperm. J Agric Food Chem 64:2030–2037

    Article  CAS  PubMed  Google Scholar 

  • Cheng A, Ismail I, Osman M, Hashim H (2012) Simple and rapid molecular techniques for identification of amylose levels in rice varieties. Int J Mol Sci 13:6156–6166

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Crofts N, Abe K, Aihara S, Itoh R, Nakamura Y, Itoh K, Fujita N (2012) Lack of starch synthase IIIa and high expression of granule-bound starch synthase I synergistically increase the apparent amylose content in rice endosperm. Plant Sci 193–194:62–69

    Article  PubMed  Google Scholar 

  • Crofts N, Abe N, Oitome NF, Matsushima R, Hayashi M, Tetlow IJ, Emes MJ, Nakamura Y, Fujita N (2015) Amylopectin biosynthetic enzymes from developing rice seed form enzymatically active protein complexes. J Exp Bot 66:4469–4482

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Delvallé D, Dumez S, Wattebled F, Roldán I, Planchot V, Berbezy P, Colonna P, Vyas D, Chatterjee M, Ball S, Mérida A, D’Hulst C (2005) Soluble starch synthase I: a major determinant for the synthesis of amylopectin in Arabidopsis thaliana leaves. Plant J 3:398–412

    Article  Google Scholar 

  • Fujita N (2014) Starch biosynthesis in rice endosperm. AGri-Biosci Monogr 4:1–18

    Article  Google Scholar 

  • Fujita N, Nakamura Y (2012) Distinct and overlapping functions of starch synthase isoforms. In: Tetlow IJ (ed) Essential reviews in experimental biology, vol 5: the synthesis and breakdown of starch. The society for experimental biology, London, pp 115–140

    Google Scholar 

  • Fujita N, Kubo A, Francisco PB Jr, Nakakita M, Harada K, Minaka N, Nakamura Y (1999) Purification, characterization, and cDNA structure of isoamylase from developing endosperm of rice. Planta 208:283–293

    Article  CAS  PubMed  Google Scholar 

  • Fujita N, Hasegawa H, Taira T (2001) The isolation and characterization of a waxy mutant of diploid wheat (Triticum monococcum L.). Plant Sci 160:595–602

    Article  CAS  PubMed  Google Scholar 

  • Fujita N, Yoshida M, Asakura N, Ohdan T, Miyao A, Hirochika H, Nakamura Y (2006) Function and characterization of starch synthase I using mutants in rice. Plant Physiol 140:1070–1084

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fujita N, Yoshida M, Kondo T, Saito K, Utsumi Y, Tokunaga T, Nishi A, Satoh H, Park JH, Jane JL, Miyao A, Hirochika H, Nakamura Y (2007) Characterization of SSIIIa-deficient mutants of rice: the function of SSIIIa and pleiotropic effects by SSIIIa deficiency in the rice endosperm. Plant Physiol 144:2009–2023

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fujita N, Toyosawa Y, Utsumi Y, Higuchi T, Hanashiro I, Ikegami A, Akuzawa S, Yoshida M, Mori A, Inomata K, Itoh R, Miyao A, Hirochika H, Satoh H, Nakamura Y (2009) Characterization of pullulanase (PUL)-deficient mutants of rice (Oryza sativa L.) and the function of PUL on starch biosynthesis in the developing rice endosperm. J Exp Bot 60:1009–1023

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fujita N, Satoh R, Hayashi A, Kodama M, Itoh R, Aihara S, Nakamura Y (2011) Starch biosynthesis in rice endosperm requires the presence of either starch synthase I or IIIa. J Exp Bot 62:4819–4831

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fujita N, Hanashiro I, Suzuki S, Higuchi T, Toyosawa Y, Utsumi Y, Itoh R, Aihara S, Nakamura Y (2012) Elongated phytoglycogen chain length in transgenic rice endosperm expressing active starch synthase IIa affects the altered solubility and crystallinity of the storage α-glucan. J Exp Bot 63:5859–5872

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fujita N, Hanashiro I, Toyosawa Y, Nakamura Y (2013) Functional study of rice starch synthase I (SSI) by using double mutant with lowered activities of SSI and isoalmylase1. J Appl Glycosci 60:45–51

    Article  CAS  Google Scholar 

  • Hanashiro I, Higuchi T, Aihara S, Nakamura Y, Fujita N (2011) Structures of starches from rice mutants deficient in the starch synthase isozyme SSI or SSIIIa. Biomacromolecules 12:1621–1628

    Article  CAS  PubMed  Google Scholar 

  • Hirano HY, Eiguchi M, Sano Y (1998) A single base change altered the regulation of the Waxy gene at the posttranscriptional level during the domestication of rice. Mol Biol Evol 15:978–987

    Article  CAS  PubMed  Google Scholar 

  • Hiratsuka M, Umemoto T, Aok i N, Katsuta M (2009) Development of SNP markers of starch synthase IIa (alk) and haplotype distribution in Rice Core Collections. Rice Genet Newslett 25: 80–82

    Google Scholar 

  • Hirose T, Terao T (2004) A comprehensive expression analysis of the starch synthase gene family in rice (Oryza sativa L.). Planta 220:9–16

    Article  CAS  PubMed  Google Scholar 

  • Hizukuri S (1985) Relationship between the distribution of the chain length of amylopectin and the crystalline structure of starch granules. Carbohydr Res 141:295–306

    Article  CAS  Google Scholar 

  • Hogg AC, Gause K, Hpfer P, Martin JM, Graybosch RA, Hansen LE, Giroux MJ (2013) Creation of a high-amylose durum wheat through mutagenesis of starch synthase II (SSIIa). J Cereal Sci 579:377–383

    Article  Google Scholar 

  • Huang B, Keeling PL, Hennen-Bierwagen TA, Myers AM (2016) Comparative in vitro analyses of recombinant maize starch synthases SSI, SSIIa, and SSIII reveal direct regulatory interactions and thermosensitivity. Arch Biochem Biophys 596:63–72

    Article  CAS  PubMed  Google Scholar 

  • Isshiki M, Morino K, Nakajima M, Okagaki RJ, Wessler SR, Izawa T, Shimamoto K (1998) A naturally occurring functional allele of the rice waxy locus has a GT to TT mutation at the 5′ splice site of the first intron. Plant J 15:133–138

    Article  CAS  PubMed  Google Scholar 

  • Jane J, Chen Y, Lee LF, McPherson AE, Wong KS, Radosavljevic M, Kasemsuwan T (1999) Effects of amylopectin branch chain length and amylose content on the gelatinization and pasting properties of starch. Cereal Chem 76:629–637

    Article  CAS  Google Scholar 

  • Jeon JS, Ryoo N, Hahn TR, Walia H, Nakamura Y (2010) Starch biosynthesis in cereal endosperm. Plant Physiol Biochem 48:383–392

    Article  CAS  PubMed  Google Scholar 

  • Jobling S (2004) Improving starch for food and industrial applications. Curr Opin Plant Biol 7:210–218

    Article  CAS  PubMed  Google Scholar 

  • Juliano BO, Perez CM, Blakeney AB, Castillo T, Kongseree N, Laignelet B, Lapis ET, Murty VVS, Paule CM, Webb BD (1981) International cooperative testing on the amylose content of milled rice. Starch 33:157–162

    Article  CAS  Google Scholar 

  • Lee SK, Eom JS, Hwang SK, Shin D, An G, Okita TW, Jeon JS (2016) Plastidic phosphoglucomutase and ADP-glucose pyrophosphorylase mutants impair starch synthesis in rice pollen grains and cause male sterility. J Exp Bot 67:5557–5569

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lii C, Lai VMF, Shen MC (2004) Changes in retrogradation properties of rice starches with amylose content and molecular properties. Cereal Chem 81:392–398

    Article  CAS  Google Scholar 

  • Lin Q, Huang B, Zhang M, Zhang X, Rivenbark J, Lappe RL, James MG, Myers AM, Hennen-Bierwagen TA (2012) Functional interactions between starch synthase III and isoamylase-type starch-debranching enzyme in maize endosperm. Plant Physiol 158:679–692

    Article  CAS  PubMed  Google Scholar 

  • Liu L, Ma X, Liu S, Zhu C, Jiang L, Wang Y, Shen Y, Ren Y, Dong H, Chen L, Liu X, Zhao Z, Zhai H, Wan J (2009) Identification and characterization of a novel Waxy allele from a Yunnan rice landrace. Plant Mol Biol 71:609–626

    Article  CAS  PubMed  Google Scholar 

  • Liu F, Romanova N, Lee EA, Ahmed R, Evans M, Gilbert EP, Morell MK, Emes MJ, Tetlow IJ (2012) Glucan affinity of starch synthase IIa determines binding of starch synthase I and starch-branching enzyme IIb to starch granules. Biochem J 448:373–387

    Article  CAS  PubMed  Google Scholar 

  • Luo J, Ahmed R, Kosar-Hashemi B, Larroque O, Butardo VM Jr, Tanner GJ, Colgrave ML, Upadhyaya NM, Tetlow IJ, Emes MJ, Millar A, Jobling SA, Morell MK, Li Z (2015) The different effects of starch synthase IIa mutations or variation on endosperm amylose content of barley, wheat and rice are determined by the distribution of starch synthase I and starch branching enzyme IIb between the starch granule and amyloplast stroma. Theor Appl Genet 128:1407–1419

    Article  CAS  PubMed  Google Scholar 

  • Matsuoka M, Ashikari M (2007) A quantitative trait locus regulating rice grain width. Nat Genet 39:583–584

    Article  CAS  PubMed  Google Scholar 

  • McMaugh SJ, Thistleton JL, Anschaw E, Luo J, Konik-Rose C, Wang H, Huang M, Larroque O, Regina A, Jobling SA, Morell MK, Li Z (2014) Suppression of starch synthase I expression affects the granule morphology and granule size and fine structure of starch in wheat endosperm. J Exp Bot 8:2189–2201

    Article  Google Scholar 

  • 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 2:173–185

    Article  Google Scholar 

  • Nakamura Y (2002) Towards a better understanding of the metabolic system for amylopectin biosynthesis in plants: rice endosperm as a model tissue. Plant Cell Physiol 43:718–725

    Article  CAS  PubMed  Google Scholar 

  • Nakamura Y (2015) Biosynthesis of reserve starch. In: Nakamura Y (ed) Starch. Springer, Japan, pp 161–209

    Chapter  Google Scholar 

  • Nakamura Y, Francisco PB Jr, Hosaka Y, Sato A, Sawada T, Kubo A, Fujita N (2005) Essential amino acids of starch synthase IIa differentiate amylopectin structure and starch quality between japonica and indica rice varieties. Plant Mol Biol 58:213–227

    Article  CAS  PubMed  Google Scholar 

  • Nakamura Y, Umemoto T, Ogata N, Kuboki Y, Yano M, Sasaki T (1996) Starch debranching enzyme (R-enzyme or pullulanase) from developing rice endosperm: purification, cDNA and chromosomal localization of the gene. Planta 199:209–218

    Article  CAS  PubMed  Google Scholar 

  • Nakamura Y, Kubo A, Shimamune T, Matsuda T, Harada K, Satoh H (1997) Correlation between activities of starch debranching enzyme and α-polyglucan structure in endosperms of sugary-1 mutants of rice. Plant J 12:143–153

    Article  CAS  Google Scholar 

  • Nakamura Y, Sakurai A, Inaba Y, Kimura K, Iwasawa N, Nagamine T (2002) The fine structure of amylopectin in endosperm from Asian cultivated rice can be largely classified into two classes. Starch 54:117–131

    Article  CAS  Google Scholar 

  • Nakamura Y, Ono M, Utsumi C, Steup M (2012) Functional interaction between plastidial starch phosphorylase and starch branching enzymes from rice during the synthesis of branched maltodextrins. Plant Cell Physiol 53:869–878

    Article  CAS  PubMed  Google Scholar 

  • Nakamura Y, Aihara S, Crofts N, Sawada T, Fujita N (2014) In vitro studies of enzymatic properties of starch synthases and interactions between starch synthase I and starch branching enzymes from rice. Plant Sci 224:1–8

    Article  CAS  PubMed  Google Scholar 

  • Nishi A, Nakamura Y, Tanaka N, Satoh H (2001) Biochemical and genetic analysis of the effects of amylose-extender mutation in rice endosperm. Plant Physiol 127:459–472

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • O’Shea MG, Morell MK (1996) High resolution slab gel electrophoresis of 8-amino-1,3,6-pyrenetrisulfonic acid (APTS) tagged oligosaccharides using a DNA sequencer. Electrophoresis 17:681–686

    Article  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Perez CM, Palmiano EP, Baun LC, Juliano BO (1973) Starch metabolism in the leaf sheaths and culm of rice. Plant Physiol 47:404–408

    Article  Google Scholar 

  • Pfister B, Lu KJ, Eicke S, Feil R, Lunn JE, Streb S, Zeeman SC (2014) Genetic evidence that chain length and branch point distributions are linked determinants of starch granule formation in Arabidopsis. Plant Physiol 165:1457–1474

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ryoo N, Yu C, Park CS, Baik MY, Park IM, Cho MH, Bhoo SH, An G, Hahn TR, Jeon JS (2007) Knockout of a starch synthase gene OsSSIIIa/Flo5 causes white-core floury endosperm in rice (Oryza sativa L.). Plant Cell Rep 26:1083–1095

    Article  CAS  PubMed  Google Scholar 

  • Sano Y (1984) Differential regulation of waxy gene expression in rice endosperm. Theor Appl Genet 68:467–473

    Article  CAS  PubMed  Google Scholar 

  • Sato Y, Takehisa H, Kamatsuki K, Minami H, Namiki N, Ikawa H, Ohyanagi H, Sugimoto K, Antonio BA, Nagamura Y (2013) RiceXPro version 3.0: expanding the informatics resource for rice transcriptome. Nucl Acids Res 41:D1206–D1213

    Article  CAS  PubMed  Google Scholar 

  • Satoh H, Nishi A, Yamashita K, Takemoto Y, Tanaka Y, Hosaka Y, Sakurai A, Fujita N, Nakamura Y (2003) Starch-branching enzyme I-deficient mutation specifically affects the structure and properties of starch in rice endosperm. Plant Physiol 133:1111–1121

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Satoh H, Shibahara K, Tokunaga T, Nishi A, Tasaki M, Hwang SK, Okita TW, Kaneko N, Fujita N, Yoshida M, Hosaka Y, Sato A, Utsumi Y, Ohdan T, Nakamura Y (2008) Mutation of the plastidial alpha-glucan phosphorylase gene in rice affects the synthesis and structure of starch in the endosperm. Plant Cell 20:1833–1849

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Seung D, Soyk S, Coiro M, Maier BA, Eicke S, Zeeman SC (2015) PROTEIN TARGETING TO STARCH is required for localising GRANULE-BOUND STARCH SYNTHASE to starch granules and for normal amylose synthesis in Arabidopsis. PLoS Biol 13:e1002080

    Article  PubMed  PubMed Central  Google Scholar 

  • Singletary GW, Banisadr R, Keeling PL (1997) Influence of gene dosage on carbohydrate synthesis and enzymatic activities in endosperm of starch-deficient mutants of maize. Plant Physiol 113:293–304

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Song XJ, Kuroha T, Ayano M, Furuta T, Nagai K, Komeda N, Segami S, Miura K, Ogawa D, Kamura T, Suzuki T, Higashiyama T, Yamasaki M, Mori H, Inukai Y, Wu J, Kitano H, Sakakibara H, Jacobsen SE, Ashikari M (2015) Rare allele of a previously unidentified histone H4 acetyltransferase enhances grain weight, yield, and plant biomass in rice. Proc Natl Acad Sci USA 112:76–81

    Article  CAS  PubMed  Google Scholar 

  • Sparla F, Falini G, Botticella E, Pirone C, Talamè V, Bovina R, Salvi S, Tuberosa R, Sestili F, Trost P (2014) New starch phenotypes produced by TILLING in barley. PLoS ONE 10:e107779

    Article  Google Scholar 

  • Szydlowski N, Ragel P, Hennen-Bierwagen TA, Planchot V, Myers AM, Mérida A, d’Hulst C, Wattebled F (2011) Integrated functions among multiple starch synthases determine both amylopectin chain length and branch linkage location in Arabidopsis leaf starch. J Exp Bot 62:4547–4559

    Article  CAS  PubMed  Google Scholar 

  • Takemoto-Kuno Y, Suzuki K, Nakamura S, Satoh H, Ohtsubo K (2006) Soluble starch synthase I effects differences in amylopectin structure between indica and japonica rice varieties. J Agric Food Chem 54:9234–9240

    Article  CAS  PubMed  Google Scholar 

  • Tanaka K, Ohnishi S, Kishimoto N, Kawasaki T, Baba T (1995) Structure, organization, and chromosomal location of the gene encoding a form of rice soluble starch synthase. Plant Physiol 108:677–683

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Toyosawa Y, Kawagoe Y, Matsushima R, Crofts N, Ogawa M, Fukuda M, Kumamaru T, Okazaki Y, Kusano M, Saito K, Toyooka K, Sato M, Ai Y, Jane JL, Nakamura Y, Fujita N (2016) Deficiency of starch synthase IIIa and IVb alters starch granule morphology from polyhedral to spherical in rice endosperm. Plant Physiol 170:1255–1270

    CAS  PubMed  PubMed Central  Google Scholar 

  • Umemoto T, Terashima K (2002) Activity of granule-bound starch synthase is an important determinant of amylose content in rice endosperm. Funct Plant Biol 29:1121–1124

    Article  CAS  Google Scholar 

  • Umemoto T, Yano M, Satoh H, Shomura A, Nakamura Y (2002) Mapping of a gene responsible for the difference in amylopectin structure between japonica-type and indica-type rice varieties. Theor Appl Genet 104:1–8

    Article  CAS  PubMed  Google Scholar 

  • Wang ZY, Zheng FQ, Shen GZ, Gao JP, Snustad DP, Li MG, Zhang JL, Hong MM (1995) The amylose content in rice endosperm is related to the post-transcriptional regulation of the waxy gene. Plant J 7:613–622

    Article  CAS  PubMed  Google Scholar 

  • Wu JL, Wu C, Lei C, Baraoidan M, Bordeos A, Madamba MR, Ramos-Pamplona M, Mauleon R, Portugal A, Ulat VJ, Bruskiewich R, Wang G, Leach J, Khush G, Leung H (2005) Chemical- and irradiation-induced mutants of indica rice IR64 for forward and reverse genetics. Plant Mol Biol 59:85–97

    Article  CAS  PubMed  Google Scholar 

  • Yamanaka S, Nakamura I, Watanabe KN, Sato Y (2004) Identification of SNPs in the waxy gene among glutinous rice cultivars and their evolutionary significance during the domestication process of rice. Theor Appl Genet 108:1200–1204

    Article  CAS  PubMed  Google Scholar 

  • Yamanouchi H, Nakamura Y (1992) Organ specificity of isoforms of starch branching enzyme (Qenzyme) in rice. Plant Cell Physiol 33: 985–991

    CAS  Google Scholar 

  • Zhang X, Szydlowski N, Delvallé D, D’Hulst C, James MG, Myers AM (2008) Overlapping functions of the starch synthases SSII and SSIII in amylopectin biosynthesis in Arabidopsis. BMC Plant Biol 23:96

    Article  Google Scholar 

  • Zhou H, Wang L, Liu G, Meng X, Jing Y, Shu X, Kong X, Sun J, Yu H, Smith SM, Wu D, Li J (2016) Critical roles of soluble starch synthase SSIIIa and granule-bound starch synthase Waxy in synthesizing resistant starch in rice. Proc Natl Acad Sci USA 113:12844–12849

    Article  CAS  PubMed Central  Google Scholar 

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Acknowledgements

The authors would like to thank Ms. Yuko Nakaizumi, Ms. Satoko Miura, and Ms. Misato Abe for taking care of the rice plants. The authors also would like to thank Enago for the English language review. This research project was partially supported by (1) The Science and Technology Research Promotion Program for Agriculture, Forestry and Fisheries and Food Industry (25033AB and 28029C; Naoko Fujita); (2) President’s Funds of Akita Prefectural University (Naoko Crofts and Dr. Naoko Fujita); (3) the Grant-in-Aid for JSPS fellows from Japan Society for the Promotion of Science (#15J40176; Naoko Crofts); and (4) The Japan Society for the Promotion of Science (#16K18571; Naoko Crofts).

Author contributions

NC designed, and both NC and KS performed the experiments. NFO provided technical assistance to KS. NC, YN and NF wrote the manuscript. The original research plan was conceived and supervised by NF.  

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Authors and Affiliations

  1. Department of Biological Production, Akita Prefectural University, 241-438 Shimoshinjo Nakano Kaidobata-Nishi, Akita City, Akita, 010-0195, Japan

    Naoko Crofts, Kyohei Sugimoto, Naoko F. Oitome, Yasunori Nakamura & Naoko Fujita

  2. Japan Society for the Promotion of Science (N.C.), Tokyo, Japan

    Naoko Crofts

  3. Akita Natural Science Laboratory, 25-44 Oiwake-Nishi, Tenno, Katagami City, Akita, 010-0101, Japan

    Yasunori Nakamura

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  1. Naoko Crofts
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  2. Kyohei Sugimoto
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Correspondence to Naoko Fujita.

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11103_2017_614_MOESM1_ESM.pdf

Supplemental Fig. 1 Replicative experiments for western blotting of the total and the starch granule-bound proteins demonstrating that the protein expression patterns agree with their genotypes (Fig. 3). (a) Western blotting of the total protein using the indicated antibodies. (b) Western blotting of the starch granule-bound proteins using the indicated antibodies, demonstrating that the active starch synthase (SS) IIa binds to the starch granules. Supplemental Fig. 2 Starch synthase (SS) activity staining without substrate adenosine diphosphate (ADP)-glucose. Gray arrowheads indicate outcomes of non-specific glucan hydrolysis activities. Supplemental Fig. 3 Chain length distribution analyses of ss1/SS2a/SS3a/Wx b-2 and average of segregates. (a) Chain length distribution patterns of starch synthases (SS)1/SS2a/SS3a (solid black), ss1/SS2a/SS3a-2 (green), SS1/ss2a/SS3a (pink), and ss1/ss2a/SS3a (dotted black) are compared to show the effect of with or without SSI and/or active SSIIa. (b) The differences in fine structure of amylopectin are shown as ΔMolar %. (ss1/SS2a/SS3a-1) – (SS1/SS2a/SS3a) is in green and (ss1/ss2a/SS3a) – (SS1/ss2a/SS3a) is dotted line. Statistical analyses were performed by averaging the values from four seeds of (data from individual ss1/SS2a/SS3a-1 seed) – (data from average of three SS1/SS2a/SS3a seeds). The bars represent standard error. (c) (ss1/SS2a/SS3a-2) – (SS1/SS2a/SS3a) is in light green. Statistical analyses were performed by averaging the values from four seeds of (data from individual ss1/SS2a/SS3a-2 seed) – (data from average of three SS1/SS2a/SS3a seeds). The bars represent standard error. (d) (SS1/SS2a/ss3a) – (SS1/SS2a/SS3a) is in blue and (SS1/ss2a/ss3a) – (SS1/ss2a/SS3a) is in gray. Statistical analyses were performed by averaging the values from four seeds of (data from individual SS1/SS2a/ss3a seed) – (data from average of three SS1/SS2a/SS3a seeds). The bars represent standard error. (PDF 428 KB)

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Crofts, N., Sugimoto, K., Oitome, N.F. et al. Differences in specificity and compensatory functions among three major starch synthases determine the structure of amylopectin in rice endosperm. Plant Mol Biol 94, 399–417 (2017). https://doi.org/10.1007/s11103-017-0614-8

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