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Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 136, Issue 2, pp 323–337 | Cite as

Plant-specific transcription factor LrTCP4 enhances secondary metabolite biosynthesis in Lycium ruthenicum hairy roots

  • Aysha Arif Chahel
  • Shaohua ZengEmail author
  • Zubaida YousafEmail author
  • Yinyin Liao
  • Ziyin Yang
  • Xiaoyi Wei
  • Wang YingEmail author
Original Article
  • 168 Downloads

Abstract

Lycium ruthenicum Murr. is an important medicinal plant from traditional Chinese medicine. It contains various biologically active compounds, such as phenolics and alkaloids. These secondary metabolites are used extensively in dietary food and pharmaceutical products. However, these phenolics occur at very low concentrations in the roots, and thus, it is expensive to commercially produce them. The present study was proposed to induce a hairy root culture system for the first time in L. ruthenicum to achieve a high concentration of phenolic polyamines and other non-targeted secondary metabolites. The over-expression sequence of the TCP4 gene was retrieved from the transcriptome data of L. ruthenicum (LrTCP4-OE), and a gene construct (with pCAMBIA 1307) was integrated into the genome of L. ruthenicum by Ri-mediated genetic transformation. A total of 21 metabolites were tentatively identified by using ultrahigh-performance liquid chromatography coupled to photodiode array detector/quadrupole time-of-flight mass spectrometry (UPLC-PDA-qTOF-MS). Transgenic hairy root clones had higher relative abundances of kukoamine A and 17 other secondary metabolites than did control-type hairy roots. After 1 month, high-growth transgenic and non-transgenic hairy root lines were subjected to UPLC analysis for absolute quantification (with an authentic standard) of total kukoamine A. Transgenic hairy root lines (LrTCP4-OE) showed higher kukoamine A accumulation (0.14%) than did control hairy roots (0.11%). This enhanced productivity correlated with increased TCP4-OE activity, validating the primary role that TCP4 plays in total kukoamine A synthesis and the efficiency of non-targeted metabolomic techniques in studying plant metabolites.

Keywords

Traditional Chinese medicine Hairy root culture system TCP4 over-expression UPLC-qTOF Kukoamine A 

Notes

Acknowledgements

This work is financially supported by the National Natural Science Foundation of China (31470391 and 31770334), Youth Innovation Promotion Association, CAS (2015286), and the Scientific Project of Ningxia Agriculture Comprehensive Development (znnfkj2015).

Supplementary material

11240_2018_1518_MOESM1_ESM.pptx (111 kb)
Supplementary material 1 (PPTX 111 KB)
11240_2018_1518_MOESM2_ESM.docx (31 kb)
Supplementary material 2 (DOCX 31 KB)

References

  1. Ali SS, Kasoju N, Luthra A, Singh A, Sharanabasava H, Sahu A (2008) Indian medicinal herbs as sources of antioxidants. Food Res Int 41:1–15CrossRefGoogle Scholar
  2. Altintas A, Kosar M, Kirimer N, Baser KHC, Demirci BC (2006) Composition of the essential oils of Lycium barbarum and L. ruthenicum fruits. Chem Nat Compd 42:24–25CrossRefGoogle Scholar
  3. Bassard JE, Ullmann P, Bernier F, Werck-Reichhart D (2010) Phenolamides: bridging polyamines to the phenolic metabolism. Phytochemistry 71:1808–1824CrossRefGoogle Scholar
  4. Bourgaud F, Gravot A, Milesi S, Gontier E (2001) Production of plant secondary metabolites: a historical perspective. Plant Sci 161:839–851CrossRefGoogle Scholar
  5. Chandra S, Chandra R (2011) Engineering secondary metabolite production in hairy roots. Phytochem Rev 10:371CrossRefGoogle Scholar
  6. Cheng M, Lowe BA, Spencer TM, Ye X, Armstrong CL (2004) Factors influencing Agrobacterium-mediated transformation of monocotyledonous species. In Vitro Cell & Dev Biol-Plant 40:31–45CrossRefGoogle Scholar
  7. Cohen FJ, Manni A, Glikman P, Bartholomew M, Demers L (1988) Involvement of the polyamine pathway in antiestrogeninduced growth inhibition of human breast cancer. Can Res 48:6819–6825Google Scholar
  8. Danisman S et al (2012) Arabidopsis class I and class II TCP transcription factors regulate jasmonic acid metabolism and leaf development antagonistically. Plant Physiol 159:1511–1523CrossRefGoogle Scholar
  9. DeBoer KD, Lye JC, Aitken CD, Su AK-K, Hamill JD (2009) The A622 gene in Nicotiana glauca (tree tobacco): evidence for a functional role in pyridine alkaloid synthesis. Plant Mol Biol 69:299CrossRefGoogle Scholar
  10. Fu X, Cheng S, Liao Y, Huang B, Du B, Zeng W, Jiang Y, Duan X, Yang Z (2018) Comparative analysis of pigments in red and yellow banana fruit. Food Chem 239:1009–1018CrossRefGoogle Scholar
  11. Funayama S, Yoshida K, Konno C, Hikino H (1980) Structure of kukoamine A, a hypotensive principle of Lycium chinense root barks1. Tetrahedron Lett 21:1355–1356CrossRefGoogle Scholar
  12. Funayama S, Zhang G-R, Nozoe S (1995) Kukoamine B, a spermine alkaloid from Lycium chinense. Phytochemistry 38:1529–1531CrossRefGoogle Scholar
  13. Gaudin V, Varin T, Jouanin L (1994) Bacterial genesmodifying hormonal balances in plants. Plant Physiol Biochem 32:11–29Google Scholar
  14. Gnonlonfin BGJ, Ambaliou S, Leon B (2012) Review scopoletin—a coumarin phytoalexin with medicinal properties. Crit Rev Plant Sci 31:47–56CrossRefGoogle Scholar
  15. Hadjipavlou-Litina D, Garnelis T, Athanassopoulos CM, Papaioannou D (2009) Kukoamine A analogs with lipoxygenase inhibitory activity. J Enzyme Inhibition Med Chem 24:1188–1193CrossRefGoogle Scholar
  16. Hao J (2012) GbTCP, a cotton TCP transcription factor, confers fibre elongation and root hair development by a complex regulating system. J Exp Bot 63:6267–6281CrossRefGoogle Scholar
  17. Hassan V, Fatemeh MK, Reza K, Mir Babak B, Mehdi MF (2014) Isolation and structure elucidation of coumarin and cinamate derivatives from Lycium ruthenicum. Q Iran Chem Commun 2:277–282Google Scholar
  18. Hu Z, Yang J, Guo G, Zheng G (2000) Establishment of transformed Lycium barbarum Line. mediated with Agrobacterium rhizogenes and factors affecting transformation. Acta Bot Boreali-Occidentalia Sinica 20:766–771Google Scholar
  19. Hu Z, Wang Y, Wu Y, Li W (2006) Effects of light and plant growth regulators on growth of normal and hairy root of Lycium barbarum in vitro China J Chin Mater Med 31:106–110Google Scholar
  20. Jacob A, Malpathak N (2005) Manipulation of MS and B5 components for enhancement of growth and solasodine production in hairy root cultures of Solanum khasianum Clarke. Plant Cell Tissue Org Cult 80:247–257CrossRefGoogle Scholar
  21. Jain A, Singh S (2015) Effect of growth regulators and elicitors for the enhanced production of solasodine in hairy root culture of Solanum melongena (L.) J Indian Bot Soc 94:23–39Google Scholar
  22. Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907CrossRefGoogle Scholar
  23. Jouhikainen K, Lindgren L, Jokelainen T, Hiltunen R, Teeri TH, Oksman-Caldentey K-M (1999) Enhancement of scopolamine production in Hyoscyamus muticus L. hairy root cultures by genetic engineering. Planta 208:545–551CrossRefGoogle Scholar
  24. Kai K, Shimizu B, Mizutani M, Watanabe K, Sakata K (2006) Accumulation of coumarins in Arabidopsis thaliana. Phytochemistry 67:379–386CrossRefGoogle Scholar
  25. Kajikawa M, Hirai N, Hashimoto T (2009) A PIP-family protein is required for biosynthesis of tobacco alkaloids. Plant Mol Biol 69:287CrossRefGoogle Scholar
  26. Kliebenstein DJ, Osbourn A (2012) Making new molecules—evolution of pathways for novel metabolites in plants. Curr Opin Plant Biol 15:415–423CrossRefGoogle Scholar
  27. Kosugi S, Ohashi Y (2002) DNA binding and dimerization specificity and potential targets for the TCP protein family. Plant J 30:337–348CrossRefGoogle Scholar
  28. Li C, Potuschak T, Colón-Carmona A, Gutiérrez RA, Doerner P (2005) Arabidopsis TCP20 links regulation of growth and cell division control pathways. Proc Natl Acad Sci USA 102:12978–12983CrossRefGoogle Scholar
  29. Li F, Jin Z, Zhao D, Cheng L, Fu C, Ma F (2006) Overexpression of the Saussurea medusa chalcone isomerase gene in S. involucrata hairy root cultures enhances their biosynthesis of apigenin. Phytochemistry 67:553–560CrossRefGoogle Scholar
  30. Li Y-Y, Wang H, Zhao C, Huang Y-Q, Tang X, Cheung H-Y (2015) Identification and characterization of kukoamine metabolites by multiple ion monitoring triggered enhanced product ion scan method with a triple-quadruple linear ion trap mass spectrometer. J Agric Food Chem 63:10785–10790CrossRefGoogle Scholar
  31. Liu X et al (2011a) Dual targets guided screening and isolation of kukoamine B as a novel natural anti-sepsis agent from traditional Chinese herb Cortex lycii. Int Immunopharmacol 11:110–120CrossRefGoogle Scholar
  32. Liu X et al (2011b) Kukoamine B, a novel dual inhibitor of LPS and CpG DNA, is a potential candidate for sepsis treatment. Br J Pharmacol 162:1274–1290CrossRefGoogle Scholar
  33. López-Cobo A, Gómez-Caravaca AM, Cerretani L, Segura-Carretero A, Fernández-Gutiérrez A (2014) Distribution of phenolic compounds and other polar compounds in the tuber of Solanum tuberosum L. by HPLC-DAD-q-TOF and study of their antioxidant activity. J Food Compos Anal 36:1–11CrossRefGoogle Scholar
  34. Mehrotra S, Srivastava V, Rahman L, Kukreja LK (2013) Overexpression of a Catharanthus tryptophan decarboxylase (tdc) gene leads to enhanced terpenoid indole alkaloid (TIA) production in transgenic hairy root lines of Rauwolfia serpentina Plant Cell Tissue Org Cult 115:377–384CrossRefGoogle Scholar
  35. Moyano E, Fornalé S, Palazón J, Cusidó RM, Bagni N, Piñol MT (2002) Alkaloid production in Duboisia hybrid hairy root cultures overexpressing the. pmt gene. Phytochemistry 59:697–702CrossRefGoogle Scholar
  36. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497.  https://doi.org/10.1111/j.1399-3054.1962.tb08052.x CrossRefGoogle Scholar
  37. Nagella P, Thiruvengadam M, Jung SJ, Murthy HN, Chung IM (2013) Establishment of Gymnema sylvestre hairy root cultures for the production of gymnemic acid. Acta Physiol Plant 35:3067–3073CrossRefGoogle Scholar
  38. Ogawa T et al (2017) Seed metabolome analysis of a transgenic rice line expressing cholera toxin B-subunit. Sci Rep 7:5196  https://doi.org/10.1038/s41598-017-04701-w CrossRefGoogle Scholar
  39. Parapunova V et al (2014) Identification, cloning and characterization of the tomato TCP transcription factor family. BMC Plant Biol 14:157CrossRefGoogle Scholar
  40. Parr AJ, Mellon FA, Colquhoun IJ, Davies HV (2005) Dihydrocaffeoyl polyamines (kukoamine and allies) in potato (Solanum tuberosum) tubers detected during metabolite profiling. J Agric Food Chem 53:5461–5466CrossRefGoogle Scholar
  41. Peng Q, Liu H, Lei HJ, Wang XQ (2016) Relationship between structure and immunological activity of an arabinogalactan from Lycium ruthenicum. Food Chem 194:595–600CrossRefGoogle Scholar
  42. Ponasik JA, Strickland C, Faerman C, Savvides S, Karplus PA, Ganem B (1995) Kukoamine A and other hydrophobic acylpolyamines: potent and selective inhibitors of Crithidia fasciculate trypanothione reductase. Biochem J 311:371–375CrossRefGoogle Scholar
  43. Praveen N, Murthya HN (2012) Synthesis of withanolide A depends on carbon source and medium pH in hairy root cultures of Withania somnifera. Ind Crops Prod 35:241–243CrossRefGoogle Scholar
  44. Qiu J et al (2012) Screening natural antioxidants in peanut shell using DPPH–HPLC–DAD–TOF/MS methods. Food Chem 135:2366–2371CrossRefGoogle Scholar
  45. Rogoza LN, Salakhutdinov NF, Tolstikov GA (2005) Plant alkaloids of the polymethyleneamine series. Russ Chem Rev 74:381–396CrossRefGoogle Scholar
  46. Sarvepalli K, Nath U (2011) Hyper-activation of the TCP4 transcription factor in Arabidopsis thaliana accelerates multiple aspects of plant maturation. Plant J 67:595–607CrossRefGoogle Scholar
  47. Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative CT method. Nat Protoc 3:1101CrossRefGoogle Scholar
  48. Schoch G, Goepfert S, Morant M, Hehn A, Meyer D, Ullmann P, Werck-Reichhart D (2001) CYP98A3 from Arabidopsis thaliana is a 3′-hydroxylase of phenolic esters, a missing link in the phenylpropanoid pathway. J Biol Chem 276:36566–36574CrossRefGoogle Scholar
  49. Sevón N, Dräger B, Hiltunen R, Oksman-Caldentey K-M (1997) Characterization of transgenic plants derived from hairy roots of Hyoscyamus muticus. Plant Cell Rep 16:605–611CrossRefGoogle Scholar
  50. Talhaoui N, Gómez-Caravaca AM, León L, De la Rosa R, Segura-Carretero A, Fernández-Gutiérrez A (2014) Determination of phenolic compounds of ‘Sikitita’olive leaves by HPLC-DAD-TOF-MS. Comparison with its parents ‘Arbequina’and ‘Picual’olive leaves. LWT-Food Sci Technol 58:28–34CrossRefGoogle Scholar
  51. Thwe A, Arasu MV, Li X, Park CH, Kim SJ, Al-Dhabi NA, Park SU (2016) Effect of different Agrobacterium rhizogenes strains on hairy root induction and phenylpropanoid biosynthesis in tartary buckwheat (Fagopyrum tataricum Gaertn). Front Microbiol 7:318CrossRefGoogle Scholar
  52. Tzfira T, Vaidya M, Citovsky V (2004) Involvement of targeted proteolysis in plant genetic transformation by Agrobacterium. Nature 431:87–92CrossRefGoogle Scholar
  53. Vanhala L, Hiltunen R, Oksman-Caldentey K-M (1995) Virulence of different Argrobacterium strains on hairy root formation of Hyoscyamus muticus. Plant Cell Rep 14:236–240CrossRefGoogle Scholar
  54. Wang CJ et al (2003) Defining the molecular requirements for the selective delivery of polyamine conjugates into cells containing active polyamine transporters. J Med Chem 46:5129–5138CrossRefGoogle Scholar
  55. Weigel D, Glazebrook J (2006) Transformation of Agrobacterium using electroporation. Cold Spring Harb Protoc.  https://doi.org/10.1101/pdb.prot4665 Google Scholar
  56. Zeng S, Liu Y, Wu M, Liu X, Shen X, Liu C, Wang Y (2014) Identification and validation of reference genes for quantitative real-time PCR normalization and its applications in Lycium. PLoS ONE 9:e97039.  https://doi.org/10.1371/journal.pone.0097039 CrossRefGoogle Scholar
  57. Zheng J et al (2011) Anthocyanins composition and antioxidant activity of wild Lycium ruthenicum Murr. from Qinghai-Tibet Plateau. Food Chem 126:859–865CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Key Laboratory of South China Agricultural Plant Molecular Analysis and Genetic Improvement, Provincial Key Laboratory of Applied Botany, South China Botanical GardenChinese Academy of SciencesGuangzhouPeople’s Republic of China
  2. 2.Plant Molecular Taxonomy, Department of BotanyLahore College for Women UniversityLahorePakistan

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