Biologia plantarum 61:147-154, 2017 | DOI: 10.1007/s10535-016-0645-z

Cloning cDNA and functional characterization of UDP-glucose pyrophosphorylase in Dendrobium officinale

R.-L. Wan1,2,3, J. Sun1,3, T. He1, Y.-D. Hu1, Y. Zhao2, Y. Wu1, Z. Chun1,*
1 Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, P.R. China
2 College of Life Sciences, Sichuan University, Chengdu, P.R. China
3 Graduate University of the Chinese Academy of Sciences, Beijing, P.R. China

Dendrobium officinale is a traditional Chinese medicinal herb that produces promising bioactive polysaccharides. However, the biosynthetic pathway of polysaccharides in this herb remains to be elucidated. The uridine diphosphate glucose pyrophosphorylase (UGPase) is a key enzyme for the production of uridine diphosphate glucose, which is a major glycosyl donor for synthesis of polysaccharides. This study identified a novel UGPase gene from D. officinale termed as DoUGP. Bioinformatics and subcellular-localization of the DoUGP protein indicate that it belongs to the UGPase-A type and was localized in cytoplasm. The DoUGP was revealed to be constitutively expressed in all organs, and the highest mRNA content was detected in stems, the organs with the highest polysaccharide content. Furthermore, sucrose feeding experiments in D. officinale demonstrate that sucrose addition could increase DoUGP transcription significantly and enhance polysaccharide accumulation accordingly. Together, we conclude that DoUGP probably plays an important role in polysaccharide biosynthesis of D. officinale and is a potential target for quality breeding of this orchid.

Keywords: gene expression; polysaccharide synthesis; subcellular localization
Subjects: UDP-glucose pyrophosphorylase; polysaccharide synthesis; gene expression; phylogenetic tree; sucrose

Received: December 4, 2015; Revised: March 28, 2016; Accepted: March 31, 2016; Published: January 1, 2017Show citation

ACS AIP APA ASA Harvard Chicago IEEE ISO690 MLA NLM Turabian Vancouver
Wan, R.-L., Sun, J., He, T., Hu, Y.-D., Zhao, Y., Wu, Y., & Chun, Z. (2017). Cloning cDNA and functional characterization of UDP-glucose pyrophosphorylase in Dendrobium officinale. Biologia plantarum, 61(1), 147-154. doi: 10.1007/s10535-016-0645-z.
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    Supplementary files

    Download filebpl-201701-0015_S1.pdf

    File size: 71.03 kB

    References

    1. Abe, T., Niiyama, H., and Sasahara, T.: Cloning of cDNA for UDP-glucose pyrophosphorylase and the expression of mRNA in rice endosperm. - Theor. appl. Genet. 105: 216-221, 2002. Go to original source...
    2. Chen, R., Zhao, X., Shao, Z., Zhu, L., He, G.: Multiple isoforms of UDP-glucose pyrophosphorylase in rice. - Physiol. Plant. 129: 725-736, 2007. Go to original source...
    3. Chen, Y.S., Liu, W.H.: [Optimal extraction procedure of polysaccharides from Dendrobium candidum and effects of polysaccharides on overexpression of NF-B agent in vascular endothelial cells induced by high glucose.] - J. Shanxi Coll. Tradit. Chin. Med. 12: 28-31, 2011. [In Chin.]
    4. Ciereszko, I., Johansson, H., Kleczkowski, L.: Sucrose and light regulation of a cold-inducible UDP-glucose pyrophosphorylase gene via a hexokinase-independent and abscisic acidinsensitive pathway in Arabidopsis. - Biochem. J. 354: 67-72, 2001. Go to original source...
    5. Delmer, D.P., Amor, Y.: Cellulose biosynthesis. - Plant Cell 7: 987-1000, 1995. Go to original source...
    6. Dubois, M., Gilles, K.A., Hamilton J.K., Rebers P.A., Smith F.: Colorimetric method for determination of sugars and related substances. - Anal. Chem. 28: 350-356, 1956. Go to original source...
    7. Eimert, K., Villand, P., Kilian, A., Kleczkowski, L.A.: Cloning and characterization of several cDNAs for UDP-glucose pyrophosphorylase from barley (1) tissues. - Gene 170: 227-232, 1996. Go to original source...
    8. Geisler, M., Wilczynska, M., Karpinski, S., Kleczkowski, L.A.: Toward a blueprint for UDP-glucose pyrophosphorylase structure/function properties: homology-modeling analyses. - Plant mol. Biol. 56: 783-794, 2004. Go to original source...
    9. He, T.G., Yang, L.T., Li, Y.R., Wang, C.Q., Su, J.: [Effect of sucrose on growth of protocorms of Dendrobium candidum and accumulation of polysaccharide.] - J. Anhui Agr. Sci. 35: 3817-3819, 2007b. [In Chin.]
    10. Huang, M., Cai, T., Liu, Q.: [Effects of polysaccharides from Dendrobium candidum on white blood cells and lymph cell moving inhibition factor of mice.] - Nat. Prod. Res. Dev. 8: 39-41, 1996. [In Chin.]
    11. Jin, Q., Yao, Y., Cai, Y., Lin, Y.: Molecular cloning and sequence analysis of a phenylalanine ammonia-lyase gene from Dendrobium. - PLoS ONE 8: e62352, 2013. Go to original source...
    12. Johansson, H.: Gene Regulation of UDP-glucose Synthesis and Metabolism in Plants. - Thesis. Umeå University, Umeå, 2003.
    13. Katsube, T., Kazuta, Y., Tanizawa, K., Fukui, T.: Expression in Escherichia coli of UDP-glucose pyrophosphorylase cDNA from potato tuber and functional assessment of the five lysyl residues located at the substrate-binding site. - Biochemistry 30: 8546-8551, 1991. Go to original source...
    14. Kleczkowski, L.A.: Glucose activation and metabolism through UDP-glucose pyrophosphorylase in plants. - Phytochemistry 37: 1507-1515, 1994. Go to original source...
    15. Kleczkowski, L.A., Decker, D.: Sugar activation for production of nucleotide sugars as substrates for glycosyltransferases in plants. - J. appl. Glycosci. 62: 25-36, 2015. Go to original source...
    16. Kleczkowski, L.A., Geisler, M., Ciereszko, I., Johansson, H.: UDP-glucose pyrophosphorylase. An old protein with new tricks. - Plant Physiol. 134: 912-918, 2004. Go to original source...
    17. Kleczkowski, L.A., Geisler, M., Fitzek, E., Wilczynska, M.: A common structural blueprint for plant UDP-sugar-producing pyrophosphorylases. - Biochem. J. 439: 375-379, 2011. Go to original source...
    18. Kleczkowski, L.A., Kunz, S., Wilczynska, M.: Mechanisms of UDP-glucose synthesis in plants. - Crit. Rev. Plant Sci. 29: 191-203, 2010. Go to original source...
    19. Litterer, L.A., Plaisance, K.L., Schnurr, J.A., Storey, K.K., Jung, H.J.G., Gronwald, J.W., Somers, D.A.: Biosynthesis of UDP-glucuronic acid in developing soybean embryos: possible role of UDP-sugar pyrophosphorylase. - Physiol. Plant. 128: 200-211, 2006a. Go to original source...
    20. Litterer, L.A., Schnurr, J.A., Plaisance, K.L., Storey, K.K., Gronwald, J.W., Somers, D.A.: Characterization and expression of Arabidopsis UDP-sugar pyrophosphorylase. - Plant Physiol. Biochem. 44: 171-180, 2006b. Go to original source...
    21. Luo, A.X., Ge, Z., Fan, Y.J., Luo, A.S., Chun, Z., He, X.J.: in vitro and in vivo antioxidant activity of a water-soluble polysaccharide from Dendrobium denneanum. - Molecules 16: 1579-1592, 2011. Go to original source...
    22. Martz, F., Wilczynska, M., Kleczkowski, L.: Oligomerization status, with the monomer as active species, defines catalytic efficiency of UDP-glucose pyrophosphorylase. - Biochem. J. 367: 295-300, 2002. Go to original source...
    23. McCoy, J.G., Bitto, E., Bingman, C.A., Wesenberg, G.E., Bannen, R.M., Kondrashov, D.A., Phillips, G.N.: Structure and dynamics of UDP-glucose pyrophosphorylase from Arabidopsis thaliana with bound UDP-glucose and UTP. - J. mol. Biol. 366: 830-841, 2007. Go to original source...
    24. Meng, M., Geisler, M., Johansson, H., Mellerowicz, E.J., Karpinski, S., Kleczkowski, L.A.: Differential tissue/organdependent expression of two sucrose-and cold-responsive genes for UDP-glucose pyrophosphorylase in Populus. - Gene 389: 186-195, 2007. Go to original source...
    25. Meng, M., Wilczynska, M., Kleczkowski L.A.: Molecular and kinetic characterization of two UDP-glucose pyrophosphorylases, products of distinct genes, from Arabidopsis. - Biochim. biophys. Acta 1784: 967-972, 2008. Go to original source...
    26. Murashige, T., Skoog, F.: A revised medium for rapid growth and bioassays with tobacco tissue cultures. - Physiol. Plant. 15: 473-497, 1962. Go to original source...
    27. Okazaki, Y., Shimojima, M., Sawada, Y., Toyooka, K., Narisawa, T., Mochida, K., Tanaka, H., Matsuda, F., Hirai, A., Hirai, M.Y.: A chloroplastic UDP-glucose pyrophosphorylase from Arabidopsis is the committed enzyme for the first step of sulfolipid biosynthesis. - Plant Cell 21: 892-909, 2009. Go to original source...
    28. Pfaffl, M.W.: A new mathematical model for relative quantification in real-time RT-PCR. - Nucl. Acids Res. 29: e45-e45, 2001. Go to original source...
    29. Pua, E.C., Lim, S.S.-W., Liu, P., Liu, J.Z.: Expression of a UDPglucose pyrophosphorylase cDNA during fruit ripening of banana (1). - Funct. Plant Biol. 27: 1151-1159, 2000. Go to original source...
    30. Sparkes, I.A., Runions, J., Kearns, A., Hawes, C.: Rapid, transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants. - Nat. Protoc. 4: 2019-2025, 2006. Go to original source...
    31. Spychalla, J.P., Scheffler, B.E., Sowokinos, J.R., Bevan, M.W.: Cloning, antisense RNA inhibition, and the coordinated expression of UDP-glucose pyrophosphorylase with starch biosynthetic genes in potato-tubers. - J. Plant Physiol. 144: 444-453, 1994. Go to original source...
    32. Tomoda, M., Takahashi, M., Nakatsuka, S.: Water-soluble carbohydrates of Zizyphi Fructus(2): isolation of two polysaccharides and structure of an arabinan. - Chem. Pharm. Bull. 21: 707-711, 1973. Go to original source...
    33. Tsuboi, K.K., Fukunaga, K., Petricciani, J.C.: Purification and specific kinetic properties of erythrocyte uridine diphosphate glucose pyrophosphorylase. - J. biol. Chem. 244: 1008-1015, 1969.
    34. Xia, L., Liu, X., Guo, H., Zhang, H., Zhu, J., Ren, F.: Partial characterization and immunomodulatory activity of polysaccharides from the stem of Dendrobium officinale (1) in vitro. - J. Funct. Foods 4: 294-301, 2012. Go to original source...
    35. Xiao, J.H., Chen, D.X., Xiao, Y., Liu, J.W., Liu, Z.L., Wan, W.H., Fang, N., Tan, B.B., Liang, Z.Q., Liu, A.Y.: Optimization of submerged culture conditions for mycelial polysaccharide production in Cordyceps pruinosa. - Process Biochem. 39: 2241-2247, 2004. Go to original source...
    36. Xing, X.H., Cui, S.W., Nie, S.P., Phillips, G.O., Douglas Goff, H., Wang, Q.: A review of isolation process, structural characteristics, and bioactivities of water-soluble polysaccharides from Dendrobium plants. - Bioact. Carbohydr. Diet. Fibre 1: 131-147, 2013. Go to original source...
    37. Yang, H., Wang, S.C., Wang, Z.T., Hu, Z.B.: Structural analysis of polysaccharides from Dendroblum candidum. - Chin. Pharm. J. 40: 254-255, 2004.
    38. Yang, J., Meng, H.L., Yang, S.C., Zhang, W., Zha, Y.H., Wen, G.S.: [Correlation between soluble polysaacharide and sucrose metabolic enzymes in Dendrobium officinale.] - J. West Chin. Forest. Sci. 41: 62-67, 2012. [In Chin.]
    39. Yang, J., Sun, H.: [Effect of different elicitors on accumulation of astragalus polysaccharides in the suspension culture cell of Astragalus membranaceus.] - J. Anhui Agr. Sci. 42: 8954-8956, 8959, 2014. [In Chin.]
    40. Yoo, S.D., Cho, Y.H., Sheen, J.: Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. - Nat. Protoc. 2: 1565-1572, 2007. Go to original source...
    41. Wang, Q., Zhang, X., Li, F., Hou, Y., Liu, X., Zhang, X.: Identification of a UDP-glucose pyrophosphorylase from cotton (Gossypium hirsutum L.) involved in cellulose biosynthesis in Arabidopsis thaliana. - Plant Cell Rep. 30: 1303-1312, 2011. Go to original source...
    42. Wu, X., Liu, D., Hu, Z.: [Study on Relationship between uridine diphosphate glucose pyrophosphorylase activity and polysaccharide content in hairy root of Astragalus membranaceus.] - Acta Univ. Tradit. Med. Sin. Pharm. Shanghai 14: 53-55, 2000. [In Chin.]
    43. Wu, X.J., Du, M., Weng, Y.Q., Liu, D., Hu, Z.B.: UGPase of Astragalus membranaceus: cDNA cloning, analyzing and expressing in Escherichia coli. - J. Integr. Plant Biol. 6: 009, 2002.
    44. Zha, X.Q., Luo, J.P., Jiang, S.T., Wang, J.H.: Enhancement of polysaccharide production in suspension cultures of protocorm-like bodies from Dendrobium huoshanense by optimization of medium compositions and feeding of sucrose. - Process Biochem. 42: 344-351, 2007. Go to original source...

    Return to the content