Plant Molecular Biology Reporter

, Volume 30, Issue 5, pp 1210–1217 | Cite as

Cloning and Characterization of the Wx Gene Encoding a Granule-Bound Starch Synthase in Lotus (Nelumbo nucifera Gaertn)

  • Ye Lu
  • Liangjun Li
  • Yong Zhou
  • Qingsong Gao
  • Guohua Liang
  • Xuehao Chen
  • Xiaohua Qi
Original Paper


The granule-bound starch synthase (GBSS) proteins were widely considered as one of the most important enzymes in plant amylose synthesis. However, understanding of the molecular basis of the GBSS protein in lotus remains fragmented. In this work, a lotus Wx gene, encoding a GBSS (GenBank accession no. EU938541), was isolated and characterized. This gene comprises 13 exons and 12 introns and covers 4152 bp (GenBank accession no. FJ602702). The exons of Wx gene have similar lengths, while the introns vary greatly. Phylogenetic tree indicated that the lotus GBSS protein belongs to a GBSS I subgroup. The expression of the Wx gene varies in different organs of the lotus during its development process and is also expressed differently in different cultivars. The Wx gene is expressed at a higher level in the rhizomes of cultivar Meirenhong than in those of cultivar Elian 4. This study elucidates more molecular information about the Wx gene in lotus and provides a theoretical foundation for the genes regulation and the modification of starch quality.


Lotus (Nelumbo nucifera Gaertn) Wx gene Granule-bound starch synthase Cloning Expression analysis 



This work was supported by National Natural Science Foundation of China (31071795) and Special Fund for Agro-scientific Research in the Public Interest of China (200903017-02).


  1. Anja K, Evert J, Richard V (1994) Formation and deposition of amylose in the potato tuber starch granule are affected by the reduction of granule-bound starch synthase gene expression. Plant Cell 6:43–52Google Scholar
  2. Ball S, Guan HP, James M, Myers A, Keeling P, Mouille G, Buleon A, Colonna P, Preiss J (1996) From glycogen to amylopectin: a model for the biogenesis of the plant starch granule. Cell 86:349–352PubMedCrossRefGoogle Scholar
  3. Ball SG, Marion HBJ, Richard GF (1998) Progress in understanding the biosynthesis of amylose. Trends Plant Sci 3:462–467Google Scholar
  4. Bochanikova I, Wasserman LA, Krivandin AV, Fornal J, Blaszczak W, Chernykh V (2003) Structure and thermodynamic melting parameters of wheat starches with different amylose content. J Ther Anal Calor 74:681–695CrossRefGoogle Scholar
  5. Buleon A, Colonna P, Planchot V, Ball S (1998) Starch granules: structure and biosynthesis. Int J Biol Macromol 23:85–112PubMedCrossRefGoogle Scholar
  6. Cai XL, Wang ZY, Xing YY, Zhang JL, Hong MM (1998) Aberrant splicing of intron1 leads to the heterogeneous 5ʹUTR and decreased expression of waxy gene in rice cultivars of intermediate amylose content. The Plant J 14:459–465CrossRefGoogle Scholar
  7. Clarka JR, Robertson M, Anisworth CC (1991) Nucleotide sequence of a wheat (Triticum aestivum L.) cDNA clone encoding the waxy protein. Plant Mol Biol 16:1099–1101CrossRefGoogle Scholar
  8. Denyer K (2006) The isolation and characterisation of novel low amylose mutants of Pisum sativum L. Plant Cell Environ 18:1019–1026Google Scholar
  9. Du HX. Zhao JS, Park JY, Kim SH, Chang KJ (2010) Antioxidant and hepatic protective effects of lotus root hot water extract with taurine supplementation in rats fed a high fat diet. J Biomed Sci 17 Suppl 1:S39Google Scholar
  10. Emanuelsson O, Nielsen H, Brunak S, von Heijne G (2000) Predicting subcellular localization of proteins based on their N-terminal amino acid sequence. J Mol Biol 300:1005–1016PubMedCrossRefGoogle Scholar
  11. Finn RD, Mistry J, Tate J, Coggill P, Heger A, Pollington JE, Gavin OL, Gunasekaran P, Ceric G, Forslund K, Holm L, Sonnhammer ELL, Eddy SR, Bateman A (2010) The Pfam protein families’ database. Nucl Acids Res 38:211–222CrossRefGoogle Scholar
  12. Garrow JS, James WPT, Ralph A (2000) Human nutrition and dietetics.Google Scholar
  13. Han YP, Xu ML, Liu X, Yan CJ, Korban SS, Chen X, Gu MH (2004) Genes coding for starch branching enzymes are major contributors to viscosity characteristics in waxy rice (Oryza sativa L.). Plant Sci 166:357–364CrossRefGoogle Scholar
  14. 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 15:978–987PubMedCrossRefGoogle Scholar
  15. Hizukuri S, Takeda Y, Maruta N, Juliano BO (1989) Molecular structural characteristics of starch. Carbohydrate Res 189:227–235CrossRefGoogle Scholar
  16. Horton P, Park JY, Obayashi T, Fujita N, Harada H, Adams-Collier CJ, Nakai K (2007) WoLF PSORT: protein localization predictor. Nucl Acids Res 35:585–587CrossRefGoogle Scholar
  17. Hovenkamp-Hermelink JHM, Jacobsen E, Ponstein AS, Visser RGF, Vos-Scheperkeuter GH, Bijmolt EW, Vries JN, Witholt B, Feenstra WJ (1987) Isolation of an amylose-free mutant of the potato (Solanum tuberosum L.). Theor Appl Genet 75:217–221CrossRefGoogle Scholar
  18. Huang GW, Ma JH, Han YZ, Chen XJ, Fu YF (2011) Cloning and expression analysis of the soybean CO-like gene GmCOL9. Plant Mol Biol Rep 29:352–359CrossRefGoogle Scholar
  19. Jeon JS, Ryoo N, Hahn TR, Walia H, Nakamura Y (2010) Starch biosynthesis in cereal endosperm. Plant Physiol Biochem 48:383–392PubMedCrossRefGoogle Scholar
  20. Li YZ, Ma HM, Zhang JL, Wang ZY, Hong MM (1995) Effects of the first intron of rice waxy gene on the expression of foreign genes in rice and tobacco protoplasts. Plant Sci 108:181–190CrossRefGoogle Scholar
  21. Li LJ, Lin HM, Cao PS (2003) The problems of pollution-free production of Lotus root in Jiangsu Province. Chin Agric Sci Bull 19:156–158Google Scholar
  22. Li LJ, Zhang XD, Pan EC, Sun L, Xie K, Gu L, Cao PS (2006a) Relationship of starch synthesis with its related enzymes’ activities during rhizome development. Sci Agric Sin 39:2307–2312Google Scholar
  23. Li LJ, Zhang XD, Shen XP, Lei S, Xie K, Gu L, Cao PS (2006b) Studies on starch RVA profile and starch granule shape in rhizome of Nelumbo nucifera Gaertn. Acta Hort Sin 33:534–538Google Scholar
  24. Li LJ, Zhang XD, Xie K, Sun L, Gu L, Cao PS (2006c) Study on the carbohydrate metabolism in Lotus Root during storage in field through the winter. Chin Veg 4:11–13Google Scholar
  25. Liu J, Zhang M, Wang S (2010a) Processing characteristics and flavour of full lotus root powder beverage. J Sci Food Agric 90:2482–2489PubMedCrossRefGoogle Scholar
  26. Liu XM, Anderson JM, Pijut PM (2010b) Cloning and characterization of Prunus serotina AGAMOUS, a putative flower homeotic gene. Plant Mol Biol Rep 28:193–203CrossRefGoogle Scholar
  27. Morrison TB, Weis JJ, Wittwer CT (1998) Quantification of low-copy transcripts by continuous SYBR Green I monitoring during amplification. Biotechnology 24:954–958Google Scholar
  28. Murata T, Sugiyama T, Akazawa T (1965) Enzymic mechanism of starch synthesis in glutinous rice grains. Biochem Biophys Res Commun 18:371–376PubMedCrossRefGoogle Scholar
  29. Nakamura T (1995) Production of waxy (amylose-free) wheats. Mol Gen Genet 248:253–259PubMedCrossRefGoogle Scholar
  30. Nakamura T, Vrinten P, Hayakawa K, Ikeda J (1998) Characterization of a granule-bound starch synthase isoform found in the pericarp of wheat. Plant Physiol 118:451–459PubMedCrossRefGoogle Scholar
  31. Nielsen H, Engelbrecht J, Brunak S, Von Heijne G (1997) Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. Prot Eng 10:1–6CrossRefGoogle Scholar
  32. Okaqaki RJ, Wessler SR (1988) Comparison of non-mutant and mutant waxy gene in rice and maize. Genetics 120:1137–1143Google Scholar
  33. Orlando C, Pinzaniand P, Pazzagli M (1998) Developments in quantitative PCR. Clin Chem Lab Med 36:255–269PubMedCrossRefGoogle Scholar
  34. Sun MM, Abdula SE, Lee HJ, Cho YC, Han LZ, Koh HJ (2011) Molecular aspect of good eating quality formation in japonica rice. PLoS One 6:1–12CrossRefGoogle Scholar
  35. Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599PubMedCrossRefGoogle Scholar
  36. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucl Acids Res 25:4876–4882PubMedCrossRefGoogle Scholar
  37. Tian ZX, Qian Q, Liu QQ, Yan MX, Liu XF, Yan CJ, Liu GF, Gao ZY, Tang SZ, Zeng DL, Wang YH, Yu JM, Gu MH, Li JY (2009) Allelic diversities in rice starch biosynthesis lead to a diverse array of rice eating and cooking qualities. PNAS 106:21760–21765PubMedCrossRefGoogle Scholar
  38. Venu RC, Sreerekha MV, Nobuta K, Beló A, Ning Y, An G, Meyers BC, Wang GL (2011) Deep sequencing reveals the complex and coordinated transcriptional regulation of genes related to grain quality in rice cultivars. BMC Genomics 12:190–202PubMedCrossRefGoogle Scholar
  39. Wang ZY, Zheng FQ, Shen GZ, Gao JP, Snustad DP, Zhang JL, Hong MM (1995) The amylose content in rice endosperm is related to the post-transcriptional regulation of the waxy gene. Plant J 7:613–622PubMedCrossRefGoogle Scholar
  40. Weatherwax P (1922) A rare carbohydrate in waxy maize. Genetics 7:568–572PubMedGoogle Scholar
  41. Xiong GY, Liu DB, Chen F, Zhang JM, Fan XP (2009) Study of the combined effect of N, P, Kand Zn fertilizers on lotus root. Soils Fert Sci China 1:31–34Google Scholar
  42. Xiong GY, Tong J, Liu DB, Deng YG, Wang JM, Fang JK, Cao GS, Wang BW, Ding HH (2011) Present status investigation of production and fertilization on rhizome Lotus in Hubei province. Hubei Agric Sci 50:3934–3938Google Scholar
  43. Xu CJ, Chen KS, Zhang B, Wang QJ, Ye WJ (2004) A study on methods of RNA extraction from citrus tissues. J Fruit Sci 21:136–140Google Scholar
  44. Yang C, Zhao TJ, Yu DY, Cai JY (2011) Isolation and functional characterization of a SERK Gene from Soybean (Glycine max (L.) Merr.). Plant Mol Biol Rep 29:334–344CrossRefGoogle Scholar
  45. Zdobnov EM, Apweiler R (2001) InterProScan: an integration platform for the signature-recognition methods in InterPro. Bioinformatics 17:847–848PubMedCrossRefGoogle Scholar
  46. Zhang YJ, Chen HC, Jiang JX, Liu FL, Liu JF (2008) Establishment of NIR models for components determination in fresh Lotus Roots. J Chin Inst Food Sci Technol 8:122–127Google Scholar
  47. Zhao YW (1999) Chinese aquatic vegetables. China Agriculture Press, BeijingGoogle Scholar
  48. Zhu LJ, Liu QQ, Sang YJ, Gu MH, Shi YC (2010) Underlying reasons for waxy rice flours having different pasting properties. Food Chem 120:94–100CrossRefGoogle Scholar
  49. Zhuang YL, Ren GJ, He CM, Li XY, Meng QM, Zhu CF, Wang RC, Zhang JR (2010) Cloning and characterization of a maize cDNA encoding glutamate decarboxylase. Plant Mol Biol Rep 28:620–626CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Ye Lu
    • 1
    • 2
  • Liangjun Li
    • 1
    • 2
  • Yong Zhou
    • 1
  • Qingsong Gao
    • 1
  • Guohua Liang
    • 1
  • Xuehao Chen
    • 2
  • Xiaohua Qi
    • 2
  1. 1.Key Laboratory of the Ministry of Education for Plant Functional GenomicsYangzhou UniversityYangzhouChina
  2. 2.Laboratory of Aquatic VegetablesYangzhou UniversityYangzhouChina

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