Plant Molecular Biology

, Volume 84, Issue 3, pp 359–369 | Cite as

Novel role of ZmaNAC36 in co-expression of starch synthetic genes in maize endosperm

  • Junjie Zhang
  • Jiang Chen
  • Qiang Yi
  • Yufeng Hu
  • Hanmei Liu
  • Yinghong Liu
  • Yubi HuangEmail author


Starch is an essential commodity that is widely used as food, feed, fuel and in industry. However, its mechanism of synthesis is not fully understood, especially in terms of the expression and regulation of the starch synthetic genes. It was reported that the starch synthetic genes were co-expressed during maize endosperm development; however, the mechanism of the co-expression was not reported. In this paper, the ZmaNAC36 gene was amplified by homology-based cloning, and its expression vector was constructed for transient expression. The nuclear localization, transcriptional activation and target sites of the ZmaNAC36 protein were identified. The expression profile of ZmaNAC36 showed that it was strongly expressed in the maize endosperm and was co-expressed with most of the starch synthetic genes. Moreover, the expressions of many starch synthesis genes in the endosperm were upregulated when ZmaNAC36 was transiently overexpressed. All our results indicated that NAC36 might be a transcription factor and play a potential role in the co-expression of starch synthetic genes in the maize endosperm.


Starch synthesis Gene regulation Transcription factor NAC Maize endosperm 



ADP-glucose pyrophosphorylase


Adenosine diphosphate glucose


Arabidopsis thaliana transcription activation factor


Cup-shaped cotyledon


Day after pollination


Debranching enzymes


Granule-bound starch synthase


Isopropyl β-d-1-thiogalactopyranoside


Starch-branching enzymes


Soluble granule-bound starch synthase


Starch phosphorylase



This work was supported by the National Key Basic Research Program of China (No: 2014CB138205), cultivating fund of excellent master degree theses of Sichuan Agriculture University, and the Preferentially Financing projects of scientific and technological activities of overseas students in Sichuan province.

Supplementary material

11103_2013_153_MOESM1_ESM.doc (284 kb)
Supplementary material 1 (DOC 284 kb)


  1. Ball SG, Morell MK (2003) From bacterial glycogen to starch: understanding the biogenesis of the plant starch granule. Annu Rev Plant Biol 54:207–233PubMedCrossRefGoogle Scholar
  2. Chen J, Huang B, Li Y-P, Du H, Gu Y, Liu H-M, Zhang J-J, Huang Y-B (2011) Synergistic influence of sucrose and abscisic acid on the genes involved in starch synthesis in maize endosperm. Carbohydr Res 346(13):1684–1691PubMedCrossRefGoogle Scholar
  3. Du H, Zhang L, Liu L, Tang X-F, Yang W-J, Wu Y-M, Huang Y-B, Tang Y-X (2009) Biochemical and molecular characterization of plant MYB transcription factor family. Biochemistry (Mosc) 74(1):1–11CrossRefGoogle Scholar
  4. Ferreira S, Senning M, Sonnewald S, Keßling PM, Goldstein R, Sonnewald U (2010) Comparative transcriptome analysis coupled to X-ray CT reveals sucrose supply and growth velocity as major determinants of potato tuber starch biosynthesis. BMC Genomics 11:93–110PubMedCentralPubMedCrossRefGoogle Scholar
  5. Fu F-F, Xue H-W (2010) Coexpression analysis identifies Rice Starch Regulator1, a rice AP2/EREBP family transcription factor, as a novel rice starch biosynthesis regulator. Plant Physiol 154(2):927–938PubMedCentralPubMedCrossRefGoogle Scholar
  6. Fujita M, Fujita Y, Maruyama K, Seki M, Hiratsu K, Ohme-Takagi M, Tran LSP, Yamaguchi-Shinozaki K, Shinozaki K (2004) A dehydration induced NAC protein, RD26, is involved in a novel ABA dependent stress signaling pathway. Plant J 39(6):863–876PubMedCrossRefGoogle Scholar
  7. Giraud E, Van Aken O, Ho LHM, Whelan J (2009) The transcription factor ABI4 is a regulator of mitochondrial retrograde expression of ALTERNATIVE OXIDASE1a. Plant Physiol 150(3):1286–1296PubMedCentralPubMedCrossRefGoogle Scholar
  8. Giroux MJ, Boyer C, Feix G, Hannah LC (1994) Coordinated transcriptional regulation of storage product genes in the maize endosperm. Plant Physiol 106(2):713–722PubMedCentralPubMedGoogle Scholar
  9. Hannah LC (2005) Starch synthesis in the maize endosperm. Maydica 50:497–506Google Scholar
  10. Hengen PN (1995) Purification of His-Tag fusion proteins from Escherichia coli. Trends Biochem Sci 20(7):285–286PubMedCrossRefGoogle Scholar
  11. Hennen-Bierwagen TA, Liu F, Marsh RS, Kim S, Gan Q, Tetlow IJ, Emes MJ, James MG, Myers AM (2008) Starch biosynthetic enzymes from developing maize endosperm associate in multisubunit complexes. Plant Physiol 146(4):1892–1908PubMedCentralPubMedCrossRefGoogle Scholar
  12. Hennen-Bierwagen TA, Lin Q, Grimaud F, Planchot V, Keeling PL, James MG, Myers AM (2009) Proteins from multiple metabolic pathways associate with starch biosynthetic enzymes in high molecular weight complexes: a model for regulation of carbon allocation in maize amyloplasts. Plant Physiol 149(3):1541–1559PubMedCentralPubMedCrossRefGoogle Scholar
  13. Hu Y-F, Li Y, Zhang J-J, Liu H-M, Chen Z, Huang Y-B (2011) PzsS3a, a novel endosperm specific promoter from maize (Zea mays L.) induced by ABA. Biotechnol Lett 33(7):1465–1471PubMedCrossRefGoogle Scholar
  14. Hu Y-F, Li Y-P, Zhang J-J, Liu H-M, Tian M-L, Huang Y-B (2012) Binding of ABI4 to a CACCG motif mediates the ABA-induced expression of the ZmSSI gene in maize (Zea mays L.) endosperm. J Exp Bot 63(16):5979–5989PubMedCrossRefGoogle Scholar
  15. Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6(13):3901–3907PubMedGoogle Scholar
  16. Keeling PL (1999) From enzyme activity to flux control: a quest to understand starch deposition in developing cereal grains. In: Bryant JA, Burrell MM, Kruger NJ (eds) Plant carbohydrate biochemistry. BIOS Scientific Publishers, Oxford, pp 91–103Google Scholar
  17. Kim SY, Seo PJ, Bae M, Yoon HK, Park CM (2007) Exploring membrane-associated NAC transcription factors in Arabidopsis: implications for membrane biology in genome regulation. Nucleic Acids Res 35(1):203–213PubMedCentralPubMedCrossRefGoogle Scholar
  18. 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(2):399–410PubMedCentralPubMedCrossRefGoogle Scholar
  19. Li L, Ilarslan H, James MG, Myers AM, Wurtele ES (2007) Genome wide co-expression among the starch debranching enzyme genes AtISA1, AtISA2, and AtISA3 in Arabidopsis thaliana. J Exp Bot 58(12):3323–3342PubMedCrossRefGoogle Scholar
  20. Myers N, Mittermeier RA, Mittermeier CG, Da Fonseca GAB, Kent J (2000) Biodiversity hotspots for conservation priorities. Nature 403(6772):853–858PubMedCrossRefGoogle Scholar
  21. 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(5):869–878PubMedCrossRefGoogle Scholar
  22. Ohdan T, Francisco PB, 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(422):3229–3244PubMedCrossRefGoogle Scholar
  23. Olsen AN, Ernst HA, Leggio LL, Skriver K (2005a) DNA-binding specificity and molecular functions of NAC transcription factors. Plant Sci 169:785–797CrossRefGoogle Scholar
  24. Olsen AN, Ernst HA, Leggio LL, Skriver K (2005b) NAC transcription factors: structurally distinct, functionally diverse. Trends Plant Sci 10(2):79–87PubMedCrossRefGoogle Scholar
  25. Ren T, Qu F, Morris T-J (2000) HRT gene function requires interaction between a NAC protein and viral capsid protein to confer resistance to turnip crinkle virus. Plant Cell 12(10):1917–1926PubMedCentralPubMedGoogle Scholar
  26. Rushton PJ, Somssich IE, Ringler P, Shen Q-J (2010) WRKY transcription factors. Trends Plant Sci 15(5):247–258PubMedCrossRefGoogle Scholar
  27. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, New York, pp 1486–1493 (Chinese version)Google Scholar
  28. Schmidt RJ, Burr FA, Aukerman MJ, Burr B (1990) Maize regulatory gene opaque-2 encodes a protein with a “leucine-zipper” motif that binds to zein DNA. Proc Natl Acad Sci USA 87(1):46–50PubMedCrossRefGoogle Scholar
  29. Schmidt RJ, Ketudat M, Aukerman MJ, Hoschek G (1992) Opaque-2 is a transcriptional activator that recognizes a specific target site in 22-kD zein genes. Plant Cell 4(6):689–700PubMedCentralPubMedGoogle Scholar
  30. Stower H (2011) Gene regulation: resolving transcription factor binding. Nat Rev Genet 13(2):71PubMedGoogle Scholar
  31. Sun C, Palmqvist S, Olsson H, Borén M, Ahlandsberg S, Jansson C (2003) A novel WRKY transcription factor, SUSIBA2, participates in sugar signaling in barley by binding to the sugar-responsive elements of the iso1 promoter. Plant Cell 15(9):2076–2092PubMedCentralPubMedCrossRefGoogle Scholar
  32. Tetlow IJ, Wait R, Lu Z, Akkasaeng R, Bowsher CG, Esposito S, Kosar-Hashemi B, Morell MK, Emes MJ (2004) Protein phosphorylation in amyloplasts regulates starch branching enzyme activity and protein–protein interactions. Plant Cell 16(3):694–708PubMedCentralPubMedCrossRefGoogle Scholar
  33. Tetlow IJ, Beisel KG, Cameron S, Makhmoudova A, Liu F, Bresolin NS, Wait R, Morell MK, Emes MJ (2008) Analysis of protein complexes in wheat amyloplasts reveals functional interactions among starch biosynthetic enzymes. Plant Physiol 146(4):1878–1891PubMedCentralPubMedCrossRefGoogle Scholar
  34. Tickle P, Burrell MM, Coates SA, Emes MJ, Tetlow IJ, Bowsher CG (2009) Characterization of plastidial starch phosphorylase in Triticum aestivum L. endosperm. J Plant Physiol 166(14):1465–1478PubMedCrossRefGoogle Scholar
  35. Tsai HL, Lue W-L, Lu K-J, Hsieh M-H, Wang SM, Chen J (2009) Starch synthesis in Arabidopsis is achieved by spatial cotranscription of core starch metabolism genes. Plant Physiol 151(3):1582–1595PubMedCentralPubMedCrossRefGoogle Scholar
  36. Xie Q, Frugis G, Colgan D, Chua N-H (2000) Arabidopsis NAC1 transduces auxin signal downstream of TIR1 to promote lateral root development. Genes Dev 14(23):3024–3036PubMedCrossRefGoogle Scholar
  37. Zeeman SC, Thorneycroft D, Schupp N, Chapple A, Weck M, Dunstan H, Haldimann P, Bechtold N, Smith AM, Smith SM (2004) Plastidial α-glucan phosphorylase is not required for starch degradation in Arabidopsis leaves but has a role in the tolerance of abiotic stress. Plant Physiol 135(2):849–858PubMedCentralPubMedCrossRefGoogle Scholar
  38. Zhang J-J, Hu Y-F, Huang Y-B (2008) Relationship between activities of key enzymes involved in starch synthesis and accumulation in maize inbred lines during grain filling. Russ J Plant Physiol 55(2):249–255CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Junjie Zhang
    • 1
    • 2
  • Jiang Chen
    • 1
  • Qiang Yi
    • 1
  • Yufeng Hu
    • 3
  • Hanmei Liu
    • 2
  • Yinghong Liu
    • 1
  • Yubi Huang
    • 1
    • 3
    Email author
  1. 1.Maize Research InstituteSichuan Agricultural UniversityChengduChina
  2. 2.College of Life ScienceSichuan Agricultural UniversityYa’anChina
  3. 3.College of AgricultureSichuan Agricultural UniversityChengduChina

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