Abstract
Key message
High levels of two major starch synthases, SSIIa and GBSSI, in ss3a ss4b double mutant rice alter the starch structure but fail to recover the polygonal starch granule morphology.
Abstract
The endosperm starch granule is polygonal in wild-type rice but spherical in double mutant japonica rice lacking genes encoding two of the five major Starch synthase (SS) isozymes expressed in endosperm, SSIIIa and SSIVb. Japonica rice naturally has low levels of SSIIa and Granule-bound SSI (GBSSI). Therefore, introduction of active SSIIa allele and/or high-expressing GBSSI allele from indica rice into the japonica rice mutant lacking SS isozymes can help elucidate the compensatory roles of SS isozymes in starch biosynthesis. In this study, we crossed the ss3a ss4a double mutant japonica rice with the indica rice to generate three new rice lines with high and/or low SSIIa and GBSSI levels, and examined their starch structure, physicochemical properties, and levels of other starch biosynthetic enzymes. Lines with high SSIIa levels showed more SSI and SSIIa bound to starch granule, reduced levels of short amylopectin chains (7 ≤ DP ≤ 12), increased levels of amylopectin chains with DP > 13, and consequently higher gelatinization temperature. Lines with high GBSSI levels showed an increase in amylose content. The ADP-glucose content of the crude extract was high in lines with low or high SSIIa and low GBSSI levels, but was low in lines with high GBSSI. Addition of high SSIIa and GBSSI altered the starch structure and physicochemical properties but did not affect the starch granule morphology, confirming that SSIIIa and SSIVb are key enzymes affecting starch granule morphology in rice. The relationship among SS isozymes and its effect on the amount of substrate (ADP-glucose) is discussed.
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References
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. https://doi.org/10.1093/jxb/eru310
Bahaji A, Baroja-Fernández E, Sánchez-López AM, Muñoz FJ, Li J, Almagro G, Montero M, Pujol P, Galarza R, Kaneko K, Oikawa K, Wada K, Mitsui T, Pozueta-Romero J (2014) HPLC-MS/MS analyses show that the near-Starchless aps1 and pgm leaves accumulate wild type levels of ADPglucose: further evidence for the occurrence of important ADPglucose biosynthetic pathway(s) alternative to the pPGI-pPGM-AGP pathway. PLoS ONE 9:e104997. https://doi.org/10.1371/journal.pone.0104997
Bertoft E (2013) On the building block and backbone concepts of amylopectin structure. Cereal Chem 90:294–311. https://doi.org/10.1094/CCHEM-01-13-0004-FI
Burton RA, Jenner H, Carrangis L, Fahy B, Fincher GB, Hylton C, Laurie DA, Parker M, Waite D, Van Wegen S, Verhoeven T, Denyer K (2002) Starch granule initiation and growth are altered in barley mutants that lack isoamylase activity. Plant J 31:97–112. https://doi.org/10.1046/j.1365-313x.2002.01339.x
Cakir B, Shiraishi S, Tuncel A, Matsusaka H, Satoh R, Singh S, Crofts N, Hosaka Y, Fujita N, Hwang SK, Satoh H, Okita TW (2016) Analysis of the rice ADP-glucose transporter (OsBT1) indicates the presence of regulatory processes in the amyloplast stroma that control ADP-glucose flux into starch. Plant Physiol 170:1271–1283. https://doi.org/10.1104/pp.15.01911
Cakir B, Tian L, Crofts N, Chou HL, Koper K, Ng CY, Tuncel A, Gargouri M, Hwang SK, Fujita N, Okita TW (2019) Re-programming of gene expression in the CS8 rice line over-expressing ADPglucose pyrophosphorylase induces a suppressor of starch biosynthesis. Plant J 97:1073–1088. https://doi.org/10.1111/tpj.14180
Chen GX, Zhou JW, Liu YL, Lu XB, Han CX, Zhang WY, Xu YH, Yan YM (2016) Biosynthesis and regulation of wheat amylose and amylopectin from proteomic and phosphoproteomic characterization of granule-binding proteins. Sci Rep 6:33111. https://doi.org/10.1038/srep33111
Chen GX, Zhen SM, Liu YL, Yan X, Zhang M, Yan YM (2017) In vivo phosphoproteome characterization reveals key starch granule-binding phosphoproteins involved in wheat water-deficit response. BMC Plant Biol 17. https://doi.org/10.1186/s12870-017-1118-z
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. https://doi.org/10.3390/ijms13056156
Clarke BR, Denyer K, Jenner CF, Smith AM (1999) The relationship between the rate of starch synthesis the adenosine 5′-diphosphoglucose concentration and the amylose content of starch in developing pea embryos. Planta 209:324–329. https://doi.org/10.1007/s004250050639
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. https://doi.org/10.1016/j.plantsci.2012.05.006
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. https://doi.org/10.1093/jxb/erv212
Crofts N, Sugimoto K, Oitome NF, Nakamura Y, Fujita N (2017) 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. https://doi.org/10.1007/s11103-017-0614-8
Crofts N, Itoh A, Abe M, Miura S, Oitome NF, Bao J, Fujita N (2019) Three major nucleotide polymorphisms in the Waxy gene correlated with the amounts of extra-long chains of amylopectin in rice cultivars with S or L-type amylopectin. J Appl Glycosci 66:37–460. https://doi.org/10.5458/jag.jag.JAG-2018_005
Crumpton-Taylor M, Pike M, Lu KJ, Hylton CM, Feil R, Eicke S, Lunn JE, Zeeman SC, Smith AM (2013) Starch synthase 4 is essential for coordination of starch granule formation with chloroplast division during Arabidopsis leaf expansion. New Phytol 200:1064–1075. https://doi.org/10.1111/nph.12455
Cuesta-Seijo JA, Nielsen MM, Ruzanski C, Krucewicz K, Beeren SR, Rydhal MG, Yoshimura Y, Striebeck A, Motawia MS, Willats WGT, Palcic MM (2016) In vitro biochemical characterization of all barley endosperm starch synthases. Front Plant Sci 6:1265. https://doi.org/10.3389/fpls.2015.01265
Fujita N (2014) Starch biosynthesis in rice endosperm. Agri-Biosci Monogr. https://doi.org/10.5047/agbm.2014.00401.0001
Fujita N (2015) Manipulation of rice starch properties for application. In: Nakamura Y (ed) Starch. Springer, Japan, pp 335–369
Fujita N, Nakamura Y (2012) Distinct and overlapping functions of starch synthase isoforms. In: IJ Tetlow (ed) Essential reviews in experimental biology, Vol. 5. The Synthesis and Breakdown of Starch, The Society for Experimental Biology, London, pp. 115–140
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. https://doi.org/10.1007/s004250050560
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. https://doi.org/10.1016/S0168-9452(00)00408-8
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 40:1707–1084. https://doi.org/10.1104/pp.105.071845
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. https://doi.org/10.1104/pp.107.102533
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. https://doi.org/10.1093/jxb/err125
Gámez-Arjona FM, Li J, Raynaud S, Barija-Fernández E, Muñoz FJ, Ovecka M, Ragel P, Bahaji A, Pozueta-Romero J, Mérida A (2011) Enhancing the expression of starch synthase class IV results in increased levels of both transitory and long-term storage starch. Plant Biotechnol J 9:1049–1060. https://doi.org/10.1111/j.1467-7652.2011.00626.x
Gao M, Wanat J, Stinard PS, James MG, Myers AM (1998) Characterization of dull1, a maize gene coding for a novel starch synthase. Plant Cell 10:399–412. https://doi.org/10.1105/tpc.10.3.399
Grimaud F, Rogniaux H, James MG, Myers AM, Planchot V (2008) Proteome and phosphoproteome analysis of starch granule-associated proteins from normal maize and mutants affected in starch biosynthesis. J Exp Bot 59:3395–3406. https://doi.org/10.1093/jxb/ern198
Guo H, Liu L, Li X, Yan Z, Xie Y, Xiong H, Zhao L, Gu J, Zhao S, Liu L (2017) Novel mutant alleles of the starch synthesis gene TaSSIVb-D result in the reduction of starch granule number per chloroplast in wheat. BMC Genom 18. https://doi.org/10.1186/s12864-017-3724-4
Hanashiro I, Itoh K, Kuratomi Y, Yamazaki M, Igarashi T, Matsugasako JI, Takeda Y (2008) Granule-bound starch synthase I is responsible for biosynthesis of extra-Long unit chains of amylopectin in rice. Plant and Cell Physiol 49: 925–933. https://doi.org/10.1093/pcp/pcn066
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. Biomacromol 12:1621–1628. https://doi.org/10.1021/bm200019q
Hawkins E, Chen J, Watson‐Lazowski A, Ahn‐Jarvis J, Barclay JE, Fahy B, Hartley M, Warren FJ, Seung D (2021) STARCH SYNTHASE 4 is required for normal starch granule initiation in amyloplasts of wheat endosperm. New Phytol 230:2371–2386. https://doi.org/10.1111/nph.17342
Hiratsuka M, Umemoto T, Aoki N, Katsuta M (2010) Development of SNP markers of starch synthase IIa (alk) and haplotype distribution in rice core collections. Rice genet Newslet 25:80–82
Hirose T, Terao T (2004) A comprehensive expression analysis of the starch synthase gene family in rice (Oryza sativa L.). Planta 220:9–16. https://doi.org/10.1007/s00425-004-1314-6
Hayashi M, Kodama M, Nakamura Y, Fujita N (2015) Thermal and pasting properties, morphology of starch granules, and crystallinity of endosperm starch in the rice SSI and SSIIIa double-mutant. J Appl Glycosci 62:81–86. https://doi.org/10.5458/jag.jag.JAG-2015_007
Hayashi M, Crofts N, Oitome NF, Fujita N (2018) Analyses of starch biosynthetic protein complexes and starch properties from developing mutant rice seeds with minimal starch synthase activities. BMC Plant Biol 18:59. https://doi.org/10.1186/s12870-018-1270-0
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. https://doi.org/10.1093/oxfordjournals.molbev.a026013
Hirose T, Terao T (2004) A comprehensive expression analysis of the starch synthase gene family in rice (Oryza sativa L.). Planta 220:9–16. https://doi.org/10.1007/s00425-004-1314-6
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. https://doi.org/10.1016/S0008-6215(00)90461-0
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. https://doi.org/10.1046/j.1365-313x.1998.00189.x
Itoh Y, Crofts N, Abe M, Hosaka Y, Fujita N (2017) Characterization of the endosperm starch and the pleiotropic effects of biosynthetic enzymes on their properties in novel mutant rice lines with high resistant starch and amylose content. Plant Sci 258:52–60. https://doi.org/10.1016/j.plantsci.2017.02.002
Kaneko K, Inomata T, Masui T, Koshu T, Umezawa Y, Itoh K, Pozueta-Romero J, Mitsui T (2014) Nucleotide pyrophosphatase/phosphodiesterase 1 exerts a negative effect on starch accumulation and growth in rice seedlings under high temperature and CO2 concentration conditions. Plant Cell Physiol 55:320–332. https://doi.org/10.1093/pcp/pct139
Liu DR, Huang WX, Cai XL (2013) Oligomerization of rice granule-bound starch synthase 1 modulates its activity regulation. Plant Sci 210:141–210. https://doi.org/10.1016/j.plantsci.2013.05.019
Mehrpouyan S, Menon U, Tetlow IJ, Emes MJ (2021) Protein phosphorylation regulates maize endosperm starch synthase IIa activity and protein-protein interactions. Plant J 105:1098–1112. https://doi.org/10.1111/tpj.15094
Miura S, Crofts N, Saito Y, Hosaka Y, Oitome NF, Watanabe T, Kumamaru T, Fujita N (2018) Starch synthase IIa-deficient mutant rice line produces endosperm starch with lower gelatinization temperature than japonica rice cultivars. Front Plant Sci 9:645. https://doi.org/10.3389/fpls.2018.00645
Morita R, Crofts N, Shibatani N, Miura S, Hosaka Y, Oitome NF, Ikeda KI, Fujita N, Fukayama H (2019) CO2-responsive CCT protein stimulates the ectopic expression of particular starch biosynthesis-related enzymes, which markedly change the structure of starch in the leaf sheaths of rice. Plant Cell Physiol 60:961–972. https://doi.org/10.1093/pcp/pcz008
Nakamura Y (2015) Biosynthesis of reserve starch. In: Nakamura Y (ed) Starch. Springer, Japan, pp 161–209
Nakamura Y, Yuki K, Park SY, Ohya T (1989) Carbohydrate metabolism in the developing endosperm of rice grains. Plant Cell Physiol 30:833–839. https://doi.org/10.1093/oxfordjournals.pcp.a077813
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. https://doi.org/10.1002/1521-379X(200204)54:3/4%3c117::AID-STAR117%3e3.0.CO;2-2
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. https://doi.org/10.1007/s11103-005-6507-2
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. https://doi.org/10.1016/j.plantsci.2014.03.021
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. https://doi.org/10.1104/pp.010127
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. https://doi.org/10.1093/jxb/eri292
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. https://doi.org/10.1002/elps.1150170410
Radchuk VV, Borisjuk L, Sreenivasulu N, Merx K, Mock HP, Rolletschek H, Wobus U, Weschke W (2009) Spatiotemporal profiling of starch biosynthesis and degradation in the developing barley grain. Plant Physiol 150:190–204. https://doi.org/10.1104/pp.108.133520
Ragel P, Streb S, Feil R, Sahrawy M, Annunziata MG, Lunn JE, Zeeman S, Mérida Á (2013) Loss of starch granule initiation has a deleterious effect on the growth of Arabidopsis plants due to an accumulation of ADP-glucose. Plant Physiol 163:75–85. https://doi.org/10.1104/pp.113.223420
Roldán I, Wattebled F, Lucas MM, Delvallé D, Planchot V, Jimenéz S, Pérez R, Ball S, D’Hulst C, Mérida A (2007) The phenotype of soluble starch synthase IV defective mutants of Arabidopsis thaliana suggests a novel function of elongation enzymes in the control of starch granule formation. Plant J 49:492–504. https://doi.org/10.1104/pp.108.133520
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. https://doi.org/10.1007/s00299-007-0309-8
Sano Y (1984) Differential regulation of waxy gene expression in rice endosperm. Theor Appl Genet 68:467–473. https://doi.org/10.1007/bf00254822
Seung D, Smith AM (2019) Starch granule initiation and morphogenesis-progress in Arabidopsis and cereals. J Exp Bot 70:771–784. https://doi.org/10.1093/jxb/ery412
Seung D, Lu KJ, Stettler M, Streb S, Zeeman SC (2016) Degradation of glucan primers in the absence of starch synthase 4 disrupts starch granule initiation in Arabidopsis. J Biol Chem 291:20718–20728. https://doi.org/10.1074/jbc.M116.730648
Sikka VM, Choi SB, Kavakli H, Sakulsingharoj C, Gupta S, Ito H, Okita TW (2001) Subcellular compartmentation and allosteric regulation of the rice endosperm ADPglucose pyrophosphorylase. Plant Sci 161:461–468. https://doi.org/10.1016/S0168-9452(01)00431-9
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. https://doi.org/10.1104/pp.113.1.293
Stamova BS, Laudencia-Chingcuanco D, Beckles DM (2009) Transcriptomic analysis of starch biosynthesis in the developing grain of hexaploid wheat. Int J Plant Genomics 2009:407426. https://doi.org/10.1155/2009/407426
Szydlowski N, Ragel P, Raynaud S, Lucas MM, Roldán I, Montero M, Muñoz FJ, Ovecka M, Bahaji A, Planchot V, Pozueta-Romero J, D’Hulst C, Mérida A (2009) Starch granule initiation in Arabidopsis requires the presence of either class IV or class III starch synthases. Plant Cell 21:2443–2457. https://doi.org/10.1105/tpc.109.066522
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. https://doi.org/10.1093/jxb/err172
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. https://doi.org/10.1021/jf061200i
Takeda Y, Hizukuri S, Juliano BO (1987) Structures of rice amylopectins with low and high affinities for iodine. Carbohydr Res 168:79–88. https://doi.org/10.1016/0008-6215(87)80008-3
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. https://doi.org/10.1104/pp.15.01232
Tuncel A, Kawaguchi J, Ihara Y, Matsusaka H, Nishi A, Nakamura T, Kuhara S, Hirakawa H, Nakamura Y, Cakir B, Nagamine A, Okita TW, Hwang SK, Satoh H (2014) The rice endosperm ADP-glucose pyrophosphorylase large subunit is essential for optimal catalysis and allosteric regulation of the heterotetrameric enzyme. Plant Cell Physiol 55:1169–1183. https://doi.org/10.1093/pcp/pcu057
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. https://doi.org/10.1071/PP01145
Umemoto T, Terashima K, Nakamura Y, Satoh H (1999) Differences in amylopectin structure between two rice varieties in relation to the effects of temperature during grain-filling. Starch 51:58–62. https://doi.org/10.1002/(SICI)1521-379X(199903)51:2%3c58::AID-STAR58%3e3.0.CO;2-J
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 Apply Genet 104:1–8. https://doi.org/10.1007/s001220200000
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. https://doi.org/10.1046/j.1365-313x.1995.7040613.x
Xu L, You H, Zhang O, Xiang X (2020) Genetic effects of soluble starch synthase IV-2 and it with ADPglucose pyrophorylase large unit and pullulanase on rice qualities. Rice 13. https://doi.org/10.1186/s12284-020-00409-0
Yamanouchi H, Nakamura Y (1992) Organ specificity of isoforms of starch branching enzyme (Qenzyme) in rice. Plant Cell Physiol 33:985–991. https://doi.org/10.1093/oxfordjournals.pcp.a078351
Yamamoto K, Sawada S, Ogino I (1973) Properties of rice starch prepared by alkali merhod with various condition. J Jap Soc Starch Sci 20:99–104
Yamamoto K, Sawada S, Ogino I (1981) Effects of quality and quantity of alkali solution on the properties of rice starch. J Jap Soc Starch sci 28:241–244
Acknowledgements
We thank Yuko Nakaizumi (Akita Prefectural University) for growing the rice plants and Bioedit for English language review. Pseudomonas isoamylase used for debranching amylopectin was a kind gift from Hayashibara Co., Ltd. We also thank Prof. Toshiaki Mitsui and Dr. Kentaro Kaneko (University of Niigata) for detailed suggestions on the ADP-glucose measurement.
Funding
This work was supported by the Science and Technology Research Promotion Program for Agriculture, Forestry and Fisheries and Food Industry (25033AB and 28029C to N.F.); the President’s Funds of Akita Prefectural University (N.F. and N.C.); Grant-in-Aid for the JSPS fellows from Japan Society for the Promotion of Science (#15J40176, JP18J40020, 20K05961 to N.C., and 19H01608 to N.F.); and the Japan Society for the Promotion of Science (#16K18571 and JP18K14438 to N.C.).
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N.C., A.D., M.S., Y.H., N.F.O., and N.F. generated new rice lines; A.D. performed protein analyses; A.D., N.F.O., Y.H., and N.F. performed starch analyses; N.C. and K.N. analyzed the ADP-glucose content; and N.C. and N.F. wrote the manuscript. All authors read and approved the final manuscript.
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Crofts, N., Domon, A., Miura, S. et al. Starch synthases SSIIa and GBSSI control starch structure but do not determine starch granule morphology in the absence of SSIIIa and SSIVb. Plant Mol Biol 108, 379–398 (2022). https://doi.org/10.1007/s11103-021-01197-x
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DOI: https://doi.org/10.1007/s11103-021-01197-x