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FLOURY ENDOSPERM12 Encoding Alanine Aminotransferase 1 Regulates Carbon and Nitrogen Metabolism in Rice

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Abstract

Starch is a major storage substance in cereal grains, and starch biosynthesis is a complex process. In order to elucidate regulation of the starch biosynthesis pathway, we screened a series of rice (Oryza sativa L.) endosperm mutants. In this study, we identified a floury white-core endosperm mutant named floury endosperm12 (flo12). The flo12 mutant exhibited loosely packed starch granules and a lower thousand kernel weight compared to wild type. Semithin sections revealed that compound starch grains (SG) in flo12 interior endosperm cells were developed abnormally. Furthermore, amylose content was decreased, while total protein content was significantly increased in flo12 grains. Map-based cloning showed that FLO12 encodes rice alanine aminotransferase 1 (OsAlaAT1). OsAlaAT1 is highly expressed in developing endosperm. Subcellular localization showed that OsAlaAT1 is localized in the cytosol. Moreover, the expression of most starch synthesis-related genes was decreased, while most of the storage protein coding genes had elevated expression levels in the flo12 mutant. In addition, overexpression of the OsAlaAT1 gene increased grain weight. In brief, we demonstrated that OsAlaAT1 regulates carbon and nitrogen metabolism, which provides a new insight for the improvement of rice quality and yield.

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References

  • Cai Y, Li S, Jiao G, Sheng Z, Wu Y, Shao G, Xie L, Peng C, Xu J, Tang S, Wei X, Hu P (2018) OsPK2 encodes a plastidic pyruvate kinase involved in rice endosperm starch synthesis, compound granule formation and grain filling. Plant Biotechnol J 16:1878–1891

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chen S, Tao L, Zeng L, Vega-Sanchez ME, Umemura K, Wang GL (2006) A highly efficient transient protoplast system for analyzing defence gene expression and protein-protein interactions in rice. Mol Plant Pathol 7:417–427

    Article  CAS  PubMed  Google Scholar 

  • Chiu WL, Niwa Y, Zeng W, Hirano T, Kobayashi H, Sheen J (1996) Engineered GFP as a vital reporter in plants. Curr Biol 6:325–330

    Article  CAS  PubMed  Google Scholar 

  • Farre EM, Geigenberger P, Willmitzer L, Trethewey RN (2000) A possible role for pyrophosphate in the coordination of cytosolic and plastidial carbon metabolism within the potato tuber. Plant Physiol 123:681–688

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fu FF, Xue HW (2010) Coexpression analysis identifies Rice Starch Regulator1, a rice AP2/EREBP family transcription factor, as a novel rice starch biosynthesis regulator. Plant Physiol 154:927–938

    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, 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 

  • Good AG, Johnson SJ, De Pauw M, Carroll RT, Savidov N, Vidmar J, Lu Z, Taylor G, Stroeher V (2007) Engineering nitrogen use efficiency with alanine aminotransferase. Can J Bot 85:252–262

    Article  CAS  Google Scholar 

  • Han X, Wang Y, Liu X, Jiang L, Ren Y, Liu F, Peng C, Li J, Jin X, Wu F, Wang J, Guo X, Zhang X, Cheng Z, Wan J (2012) The failure to express a protein disulphide isomerase-like protein results in a floury endosperm and an endoplasmic reticulum stress response in rice. J Exp Bot 63:121–130

    Article  CAS  PubMed  Google Scholar 

  • Hiei Y, Ohta S, Komari T, Kumashiro T (1994) Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J 6:271–282

    Article  CAS  PubMed  Google Scholar 

  • Igarashi D, Miwa T, Seki M, Kobayashi M, Kato T, Tabata S, Shinozaki K, Ohsumi C (2003) Identification of photorespiratory glutamate:glyoxylate aminotransferase (GGAT) gene in Arabidopsis. Plant J 33:975–987

    Article  CAS  PubMed  Google Scholar 

  • Ishiyama K, Inoue E, Tabuchi M, Yamaya T, Takahashi H (2004) Biochemical background and compartmentalized functions of cytosolic glutamine synthetase for active ammonium assimilation in rice roots. Plant Cell Physiol 45:1640–1647

    Article  CAS  PubMed  Google Scholar 

  • James MG, Robertson DS, Myers AM (1995) Characterization of the maize gene sugary1, a determinant of starch composition in kernels. Plant Cell 7:417–429

    Article  CAS  PubMed  PubMed Central  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 

  • Kang HG, Park S, Matsuoka M, An G (2005) White-core endosperm floury endosperm-4 in rice is generated by knockout mutations in the C-type pyruvate orthophosphate dikinase gene (OsPPDKB). Plant J 42:901–911

    Article  CAS  PubMed  Google Scholar 

  • Kikuchi H, Hirose S, Toki S, Akama K, Takaiwa F (1999) Molecular characterization of a gene for alanine aminotransferase from rice (Oryza sativa). Plant Mol Biol 39:149–159

    Article  CAS  PubMed  Google Scholar 

  • Kubo A, Fujita N, Harada K, Matsuda T, Satoh H, Nakamura Y (1999) The starch-debranching enzymes isoamylase and pullulanase are both involved in amylopectin biosynthesis in rice endosperm. Plant Physiol 121:399–409

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lee SK, Hwang SK, Han M, Eom JS, Kang HG, Han Y, Choi SB, Cho MH, Bhoo SH, An G, Hahn TR, Okita TW, Jeon JS (2007) Identification of the ADP-glucose pyrophosphorylase isoforms essential for starch synthesis in the leaf and seed endosperm of rice (Oryza sativa L.). Plant Mol Biol 65:531–546

    Article  CAS  PubMed  Google Scholar 

  • Liepman AH, Olsen LJ (2003) Alanine aminotransferase homologs catalyze the glutamate:glyoxylate aminotransferase reaction in peroxisomes of Arabidopsis. Plant Physiol 131:215–227

    Article  CAS  PubMed  PubMed Central  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 

  • Long W, Wang Y, Zhu S, Jing W, Wang Y, Ren Y, Tian Y, Liu S, Liu X, Chen L, Wang D, Zhong M, Zhang Y, Hu T, Zhu J, Hao Y, Zhu X, Zhang W, Wang C, Zhang W, Wan J (2018) FLOURY SHRUNKEN ENDOSPERM1 Connects Phospholipid Metabolism and Amyloplast Development in Rice. Plant Physiol 177:698–712

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Martin C, Smith AM (1995) Starch biosynthesis. Plant Cell 7:971–985

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Matsushima R, Maekawa M, Kusano M, Tomita K, Kondo H, Nishimura H, Crofts N, Fujita N, Sakamoto W (2016) Amyloplast Membrane Protein SUBSTANDARD STARCH GRAIN6 Controls Starch Grain Size in Rice Endosperm. Plant Physiol 170:1445–1459

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • McAllister CH, Facette M, Holt A, Good AG (2013) Analysis of the enzymatic properties of a broad family of alanine aminotransferases. PLoS One 8:e55032

    Google Scholar 

  • Mikami I, Aikawa M, Hirano HY, Sano Y (1999) Altered tissuespecific expression at the Wx gene of the opaque mutants in rice. Euphytica 105:91–97

    Article  CAS  Google Scholar 

  • Miyashita Y, Dolferus R, Ismond KP, Good AG (2007) Alanine aminotransferase catalyses the breakdown of alanine after hypoxia in Arabidopsis thaliana. Plant J 49:1108–1121

    Article  CAS  PubMed  Google Scholar 

  • Nelson OE, Rines HW (1962) The enzymatic deficiency in the waxy mutant of maize. Biochem Biophys Res Commun 9:297–300

    Article  CAS  PubMed  Google Scholar 

  • Niessen M, Krause K, Horst I, Staebler N, Klaus S, Gaertner S, Kebeish R, Araujo WL, Fernie AR, Peterhansel C (2012) Two alanine aminotranferases link mitochondrial glycolate oxidation to the major photorespiratory pathway in Arabidopsis and rice. J Exp Bot 63:2705–2716

    Article  CAS  PubMed  PubMed Central  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 

  • Peng C, Wang Y, Liu F, Ren Y, Zhou K, Lv J, Zheng M, Zhao S, Zhang L, Wang C, Jiang L, Zhang X, Guo X, Bao Y, Wan J (2014) FLOURY ENDOSPERM6 encodes a CBM48 domaincontaining protein involved in compound granule formation and starch synthesis in rice endosperm. Plant J 77:917–930

    Article  CAS  PubMed  Google Scholar 

  • Qu L, Takaiwa F (2004) Evaluation of tissue specificity and expression strength of rice seed component gene promoters in transgenic rice. Plant Biotechnol J 2:113–125

    Article  CAS  Google Scholar 

  • Rocha M, Sodek L, Licausi F, Hameed MW, Dornelas MC, van Dongen JT (2010) Analysis of alanine aminotransferase in various organs of soybean (Glycine max) and in dependence of different nitrogen fertilisers during hypoxic stress. Amino Acids 39:1043–1053

    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 

  • She KC, Kusano H, Koizumi K, Yamakawa H, Hakata M, Imamura T, Fukuda M, Naito N, Tsurumaki Y, Yaeshima M, Tsuge T, Matsumoto K, Kudoh M, Itoh E, Kikuchi S, Kishimoto N, Yazaki J, Ando T, Yano M, Aoyama T, Sasaki T, Satoh H, Shimada H (2010) A novel factor FLOURY ENDOSPERM2 is involved in regulation of rice grain size and starch quality. Plant Cell 22:3280–3294

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shrawat AK, Carroll RT, DePauw M, Taylor GJ, Good AG (2008) Genetic engineering of improved nitrogen use efficiency in rice by the tissue-specific expression of alanine aminotransferase. Plant Biotechnol J 6:722–732

    Article  CAS  PubMed  Google Scholar 

  • Son D, Sugiyama T (1992) Molecular cloning of an alanine aminotransferase from NAD-malic enzyme type C4 plant Panicum miliaceum. Plant Mol Biol 20:705–713

    Article  CAS  PubMed  Google Scholar 

  • Sonoda Y, Ikeda A, Saiki S, von Wiren N, Yamaya T, Yamaguchi J (2003) Distinct expression and function of three ammonium transporter genes (OsAMT1;1-1;3) in rice. Plant Cell Physiol 44:726–734

    Article  CAS  PubMed  Google Scholar 

  • Sun H, Qian Q, Wu K, Luo J, Wang S, Zhang C, Ma Y, Liu Q, Huang X, Yuan Q, Han R, Zhao M, Dong G, Guo L, Zhu X, Gou Z, Wang W, Wu Y, Lin H, Fu X (2014) Heterotrimeric G proteins regulate nitrogen-use efficiency in rice. Nat Genet 46:652–656

    Article  CAS  PubMed  Google Scholar 

  • Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5:Molecular Evolutionary Genetics Analysis Using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods. Mol Biol Evol 28:2731–2739

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tamura W, Hidaka Y, Tabuchi M, Kojima S, Hayakawa T, Sato T, Obara M, Kojima M, Sakakibara H, Yamaya T (2010) Reverse genetics approach to characterize a function of NADH-glutamate synthase1 in rice plants. Amino Acids 39:1003–1012

    Article  CAS  PubMed  Google Scholar 

  • Tamura W, Kojima S, Toyokawa A, Watanabe H, Tabuchi-Kobayashi M, Hayakawa T, Yamaya T (2011) Disruption of a Novel NADH-Glutamate Synthase2 Gene Caused Marked Reduction in Spikelet Number of Rice. Front Plant Sci 2:57

    Article  PubMed  PubMed Central  Google Scholar 

  • Wang JC, Xu H, Zhu Y, Liu QQ, Cai XL (2013) OsbZIP58, a basic leucine zipper transcription factor, regulates starch biosynthesis in rice endosperm. J Exp Bot 11:3453–3466

    Article  CAS  Google Scholar 

  • Wang YH, Ren YL, Liu X, Jiang L, Chen LM, Han XH, Jin MN, Liu SJ, Liu F, Lv J, Zhou KN, Su N, Bao YQ, Wan JM (2010) OsRab5a regulates endomembrane organization and storage protein trafficking in rice endosperm cells. Plant J 64:812–824

    Article  CAS  PubMed  Google Scholar 

  • Yang J, Kim SR, Lee SK, Choi H, Jeon JS, An G (2015) Alanine aminotransferase 1 (OsAlaAT1) plays an essential role in the regulation of starch storage in rice endosperm. Plant Sci 240:79–89

    Article  CAS  PubMed  Google Scholar 

  • Zhang L, Ren Y, Lu B, Yang C, Feng Z, Liu Z, Chen J, Ma W, Wang Y, Yu X, Wang Y, Zhang W, Wang Y, Liu S, Wu F, Zhang X, Guo X, Bao Y, Jiang L, Wan J (2016) FLOURY ENDOSPERM7 encodes a regulator of starch synthesis and amyloplast development essential for peripheral endosperm development in rice. J Exp Bot 67:633–647

    Article  CAS  PubMed  Google Scholar 

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  1. These authors contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory for Crop Genetics and Germplasm Enhancement, Jiangsu Plant Gene Engineering Research Center, Nanjing Agricultural University, Nanjing, 210095, China

    Mingsheng Zhong, Xi Liu, Yunlong Wang, Jianping Zhu, Xuan Teng, Erchao Duan, Fan Wang, Huan Zhang, Mingming Wu, Yuanyuan Hao, Xiaopin Zhu, Ruonan Jing, Xiuping Guo, Ling Jiang, Yihua Wang & Jianmin Wan

  2. National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

    Yulong Ren & Jianmin Wan

  3. College of Life Sciences, Nanjing Agricultural University, Nanjing, 210095, China

    Feng Liu

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  1. Mingsheng Zhong
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  2. Xi Liu
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  3. Feng Liu
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  5. Yunlong Wang
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Correspondence to Yihua Wang or Jianmin Wan.

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Zhong, M., Liu, X., Liu, F. et al. FLOURY ENDOSPERM12 Encoding Alanine Aminotransferase 1 Regulates Carbon and Nitrogen Metabolism in Rice. J. Plant Biol. 62, 61–73 (2019). https://doi.org/10.1007/s12374-018-0288-z

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  • DOI: https://doi.org/10.1007/s12374-018-0288-z

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