DOI QR코드

DOI QR Code

Overexpression of ginseng UGT72AL1 causes organ fusion in the axillary leaf branch of Arabidopsis

  • Nguyen, Ngoc Quy (Department of Plant Biotechnology, College of Agriculture and Life Science, Chonnam National University) ;
  • Lee, Ok Ran (Department of Plant Biotechnology, College of Agriculture and Life Science, Chonnam National University)
  • Received : 2016.11.18
  • Accepted : 2017.03.14
  • Published : 2017.07.15

Abstract

Background: Glycosylation of natural compounds increases the diversity of secondary metabolites. Glycosylation steps are implicated not only in plant growth and development, but also in plant defense responses. Although the activities of uridine-dependent glycosyltransferases (UGTs) have long been recognized, and genes encoding them in several higher plants have been identified, the specific functions of UGTs in planta remain largely unknown. Methods: Spatial and temporal patterns of gene expression were analyzed by quantitative reverse transcription (qRT)-polymerase chain reaction (PCR) and GUS histochemical assay. In planta transformation in heterologous Arabidopsis was generated by floral dipping using Agrobacterium tumefaciens (C58C1). Protein localization was analyzed by confocal microscopy via fluorescent protein tagging. Results: PgUGT72AL1 was highly expressed in the rhizome, upper root, and youngest leaf compared with the other organs. GUS staining of the promoter: GUS fusion revealed high expression in different organs, including axillary leaf branch. Overexpression of PgUGT72AL1 resulted in a fused organ in the axillary leaf branch. Conclusion: PgUGT72AL1, which is phylogenetically close to PgUGT71A27, is involved in the production of ginsenoside compound K. Considering that compound K is not reported in raw ginseng material, further characterization of this gene may shed light on the biological function of ginsenosides in ginseng plant growth and development. The organ fusion phenotype could be caused by the defective growth of cells in the boundary region, commonly regulated by phytohormones such as auxins or brassinosteroids, and requires further analysis.

Keywords

References

  1. Gershenzon J, Engelberth J. Secondary metabolites and plant defense. In: Taiz L, Zeiger E, editors. Plant physiology. 5th ed. Sunderland, MA: Sinauer Associates; 2010. p. 369-400.
  2. Lairson LL, Henrissat B, Davies GJ, Withers SG. Glycosyltransferases: structures, functions, and mechanisms. Annu Rev Biochem 2008;77:521-55. https://doi.org/10.1146/annurev.biochem.76.061005.092322
  3. Campbell JA, Davies GJ, Bulone V, Henrissat B. A classification of nucleotidediphospho-sugar glycosyltransferases based on amino acid sequence similarities. Biochem J 1997;326:929-39. https://doi.org/10.1042/bj3260929u
  4. Coutinho PM, Deleury E, Davies GJ, Henrissat B. An evolving hierarchical family classification for glycosyltransferases. J Mol Biol 2003;328:307-17. https://doi.org/10.1016/S0022-2836(03)00307-3
  5. Mackenzie PI, Owens IS, Burchell B, Bock KW, Bairoch A, Belanger A, Fournel-Gigleux S, Green M, Hum DW, Iyanagi T, et al. The UDP glycosyltransferase gene superfamily: recommended nomenclature update based on evolutionary divergence. Pharmacogenetics 1997;7:255-69. https://doi.org/10.1097/00008571-199708000-00001
  6. Lim EK, Bowles DJ. A class of plant glycosyltransferases involved in cellular homeostasis. EMBO J 2004;23:2915-22. https://doi.org/10.1038/sj.emboj.7600295
  7. Gachon CM, Langlois-Meurinne M, Saindrenan P. Plant secondary metabolism glycosyltransferases: the emerging functional analysis. Trends Plant Sci 2005;10:542-9. https://doi.org/10.1016/j.tplants.2005.09.007
  8. Roberts SC. Production and engineering of terpenoids in plant cell culture. Nat Chem Biol 2007;3:387-95. https://doi.org/10.1038/nchembio.2007.8
  9. Wee JJ, Park KM, Chung AS. Biological activities of ginseng and its application to human health. In: Benzie IFF, Wachtel-Galor S, editors. Herbal medicine: biomolecular and clinical aspects. 2nd ed. Boca Raton, FL: CRC Press/Taylor & Francis; 2011. p. 157-74.
  10. Luo H, Sun C, Sun Y, Wu Q, Li Y, Song J, Niu J, Cheng X, Xu H, Li C, et al. Analysis of the transcriptome of Panax notoginseng root uncovers putative triterpene saponin-biosynthetic genes and genetic markers. BMC Genomics 2011;12:S5.
  11. Kim YJ, Lee OR, Oh J, Jang MG, Yang DC. Functional analysis of 3-hydroxy-3-methylglutaryl coenzyme A reductase encoding genes in triterpene saponin-producing ginseng. Plant Physiol 2014;165:373-87. https://doi.org/10.1104/pp.113.222596
  12. Osbourn AE. Saponins in cereals. Phytochemistry 2003;62:1-4. https://doi.org/10.1016/S0031-9422(02)00393-X
  13. Yue CJ, Zhong JJ. Purification and characterization of UDPG: ginsenoside Rd glucosyltransferase from suspended cells of Panax notoginseng. Process Biochem 2005;40:3742-8. https://doi.org/10.1016/j.procbio.2005.05.001
  14. He YP, Yue CJ. Establishment of measurement system of ginsenoside Rh2 glycosyltransferase activity. Med Plant 2010;1:58-60.
  15. Jung SC, Kim W, Park SC, Jeong J, Park MK, Lim S, Lee Y, Im WT, Lee JH, Choi G, et al. Two ginseng UDP glycosyltransferases synthesize ginsenoside Rg3 and Rd. Plant Cell Physiol 2014;55:2177-88. https://doi.org/10.1093/pcp/pcu147
  16. Yan X, Fan Y, Wei W, Wang P, Liu Q, Wei Y, Zhang L, Zhao G, Yue J, Zhou Z. Production of bioactive ginsenoside compound K in metabolically engineered yeast. Cell Res 2014;24:770-3. https://doi.org/10.1038/cr.2014.28
  17. Bechtold N, Pelletier G. In planta Agrobacterium-mediated transformation of adult Arabidopsis thaliana plants by vacuum infiltration. In: Martinez-Zapater JM, Salinas J, editors. Arabidopsis protocols. Totowa, NJ: Humana Press; 1998. p. 259-66.
  18. Kim MK, Lee BS, In JG, Sun H, Yoon JH, Yang DC. Comparative analysis of expressed sequence tags (ESTs) of ginseng leaf. Plant Cell Rep 2006;25:599-606. https://doi.org/10.1007/s00299-005-0095-0
  19. Khorolragchaa A, Kim YJ, Rahimi S, Sukweenadhi J, Jang MG, Yang DC. Grouping and characterization of putative glycosyltransferase genes from Panax ginseng Meyer. Gene 2014;536:186-92. https://doi.org/10.1016/j.gene.2013.07.077
  20. Paquette S, Moller BL, Bak S. On the origin of family 1 plant glycosyltransferases. Phytochemistry 2003;62:399-413. https://doi.org/10.1016/S0031-9422(02)00558-7
  21. Wang X. Structure, mechanism and engineering of plant natural product glycosyltransferases. FEBS Lett 2009;583:3303-9. https://doi.org/10.1016/j.febslet.2009.09.042
  22. Langlois-Meurinne M, Gachon CM, Saindrenan P. Pathogen-responsive expression of glycosyltransferase genes UGT73B3 and UGT73B5 is necessary for resistance to Pseudomonas syringae pv tomato in Arabidopsis. Plant Physiol 2005;139:1890-901. https://doi.org/10.1104/pp.105.067223
  23. Song JT, Koo YJ, Seo HS, Kim MC, Choi YD, Kim JH. Overexpression of AtSGT1, an Arabidopsis salicylic acid glucosyltransferase, leads to increased susceptibility to Pseudomonas syringae. Phytochemistry 2008;69:1128-34. https://doi.org/10.1016/j.phytochem.2007.12.010
  24. Clay NK, Adio AM, Denoux C, Jander G, Ausubel FM. Glucosinolate metabolites required for an Arabidopsis innate immune response. Science 2009;323:95-101. https://doi.org/10.1126/science.1164627
  25. Grubb CD, Zipp BJ, Ludwig-Muller J, Masuno MN, Molinshi TF, Abel S. Arabidopsis glucosyltransferase UGT74B1 functions in glucosinolate biosynthesis and auxin homeostasis. Plant J 2004;40:893-908. https://doi.org/10.1111/j.1365-313X.2004.02261.x
  26. Tanaka K, Hayashi K, Natsume M, Kamiya Y, Sakakibara H, Kawaide H, Kasahara H. UGT74D1 catalyzes the glycosylation of 2-oxindole-3-acetic acid in the auxin metabolic pathway in Arabidopsis. Plant Cell Physiol 2014;55:218-28. https://doi.org/10.1093/pcp/pct173
  27. Lee OR, Kim SJ, Kim HJ, Hong JK, Ryu SB, Lee SH, Ganguly A, Cho HT. Phospholipase A2 is required for PIN-FORMED protein trafficking to the plasma membrane in the Arabidopsis root. Plant Cell 2010;22:1812-25. https://doi.org/10.1105/tpc.110.074211
  28. Kramer CM, Prata RT, Willits MG, De Luca V, Steffens JC, Graser G. Cloning and regiospecificity studies of two flavonoid glucosyltransferases from Allium cepa. Phytochemistry 2003;64:1069-76. https://doi.org/10.1016/S0031-9422(03)00507-7
  29. Bowles D, Isayenkova J, Lim EK, Poppenberger B. Glycosyltransferases: managers of small molecules. Curr Opin Plant Biol 2005;8:254-63. https://doi.org/10.1016/j.pbi.2005.03.007
  30. Lim EK, Doucet CJ, Li Y, Elias L, Worrall D, Spencer SP, Ross J, Bowles DJ. The activity of Arabidopsis glycosyltransferases toward salicylic acid, 4-hydroxybenzoic acid, and other benzoates. J Biol Chem 2002;277:586-92. https://doi.org/10.1074/jbc.M109287200
  31. Lim EK, Ashford DA, Hou B, Jackson RG, Bowles DJ. Arabidopsis glycosyltransferases as biocatalysts in fermentation for regioselective synthesis of diverse quercetin glucosides. Biotechnol Bioeng 2004;87:623-31. https://doi.org/10.1002/bit.20154
  32. Caputi L, Malnoy M, Goremykin V, Nikiforova S, Martens S. A genome-wide phylogenetic reconstruction of family 1 UDP-glycosyltransferases revealed the expansion of the family during the adaptation of plants to life on land. Plant J 2012;69:1030-42. https://doi.org/10.1111/j.1365-313X.2011.04853.x
  33. Dong T, Xu ZY, Park Y, Kim DH, Lee Y, Hwang I. Abscisic acid uridine diphosphate glucosyltransferases play a crucial role in abscisic acid homeostasis in Arabidopsis. Plant Physiol 2014;165:277-89. https://doi.org/10.1104/pp.114.239210
  34. Hou B, Lim EK, Higgins GS, Bowles DJ. N-glucosylation of cytokinins by glycosyltransferases of Arabidopsis thaliana. J Biol Chem 2004;279:47822-32. https://doi.org/10.1074/jbc.M409569200
  35. Husar S, Berthiller F, Fujioka S, Rozhon W, Khan M, Kalaivanan F, Elias L, Higgins GS, Li Y, Schuhmacher R, et al. Overexpression of the UGT73C6 alters brassinosteroid glucoside formation in Arabidopsis thaliana. BMC Plant Biol 2011;24:11-51.
  36. Hasegawa H. Proof of the mysterious efficacy of ginseng: basic and clinical trials: metabolic activation of ginsenoside: deglycosylation by intestinal bacteria and esterification with fatty acid. J Pharmacol Sci 2004;95:153-7. https://doi.org/10.1254/jphs.FMJ04001X4

Cited by

  1. Comparative transcriptome analysis of rhizome nodes and internodes in Panax. japonicus var. major reveals candidate genes involved in the biosynthesis of triterpenoid saponins vol.112, pp.2, 2017, https://doi.org/10.1016/j.ygeno.2019.06.025
  2. PgCYP76B93 docks on phenylurea herbicides and its expression enhances chlorotoluron tolerance in Arabidopsis vol.63, pp.1, 2017, https://doi.org/10.1186/s13765-020-00498-x
  3. Endophytes, biotransforming microorganisms, and engineering microbial factories for triterpenoid saponins production vol.41, pp.2, 2017, https://doi.org/10.1080/07388551.2020.1869691