Advertisement

Elicitation of Isatis tinctoria L. hairy root cultures by salicylic acid and methyl jasmonate for the enhanced production of pharmacologically active alkaloids and flavonoids

  • Qing-Yan Gai
  • Jiao JiaoEmail author
  • Xin Wang
  • Yu-Ping Zang
  • Li-Li Niu
  • Yu-Jie FuEmail author
Original Article
  • 64 Downloads

Abstract

Plant cell and organ cultures via the implementation of effective elicitation strategies can offer attractive biotechnological platforms for the enhanced production of phytochemicals of pharmaceutical interest. For the first time, the elicitation of exogenous signal molecules was conducted to enhance the production of pharmacologically active alkaloids and flavonoids in Isatis tinctoria L. hairy root cultures (ITHRCs). ITHRCs III and V correspondingly possessing high alkaloid and flavonoid productivity were adopted for elicitation treatments. The maximum accumulation of alkaloids in ITHRCs III elicited by 142.61 µM salicylic acid for 28.18 h and flavonoids in ITHRCs V elicited by 179.54 µM methyl jasmonate for 41.87 h increased 5.89- and 11.21-folds as compared with controls, respectively. Moreover, expressions of 11 genes involved in alkaloid and flavonoid biosynthetic pathways were significantly up-regulated following elicitation, among which YUCCA, CHI and F3′H genes might play a crucial role in the target phytochemical augmentation. Overall, two effective elicitation protocols were provided here to improve the yields of bioactive alkaloids and flavonoids in ITHRCs, which was useful for the scale-up production of these valuable compounds to meet the demands for natural bioactive ingredients by pharmaceutical industries.

Keywords

Elicitation Isatis tinctoria L. Hairy root cultures Bioactive phytochemicals Yield enhancement 

Notes

Acknowledgements

The authors gratefully acknowledge the financial supports by National Natural Science Foundation of China for Youths (31800492), Fundamental Research Funds for the Central Universities (2572018BU02 and 2572017DA04), Heilongjiang Province Science Foundation for Youths (QC2017012), National Key R&D Program of China (2017YFD0600205), Scientific Research Start-up Funds for Talents Introduction of Northeast Forestry University (YQ2017-03), and Fundamental Research Funds for the Central Universities (2572017AA08).

Author Contributions

QYG, JJ and YJF conceived and designed the experiments. QYG, JJ, and XW performed the experiments. GQY and JJ analyzed the data. GQY, XW, YPZ, and LLN contributed reagents/materials/analysis tools. All the authors contributed to writing and editing of the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11240_2018_1553_MOESM1_ESM.doc (97 kb)
Supplementary material 1 (DOC 97 KB)

References

  1. Ahuja I, Kissen R, Bones AM (2012) Phytoalexins in defense against pathogens. Trends Plant Sci 17:73–90CrossRefPubMedGoogle Scholar
  2. Chen J, Dong X, Li Q, Zhou X, Gao S, Chen R, Sun L, Zhang L, Chen W (2013) Biosynthesis of the active compounds of Isatis indigotica based on transcriptome sequencing and metabolites profiling. BMC Genom 14:857CrossRefGoogle Scholar
  3. Chen G, Wang S, Huang X, Hong J, Du L, Zhang L, Ye L (2015) Environmental factors affecting growth and development of Banlangen (Radix Isatidis) in China. African J Plant Sci 9:421–426CrossRefGoogle Scholar
  4. el Jaber-Vazdekis N, Barres ML, Ravelo ÁG, Zárate R (2008) Effects of elicitors on tropane alkaloids and gene expression in Atropa baetica transgenic hairy roots. J Nat Prod 71:2026–2031CrossRefPubMedGoogle Scholar
  5. Gai QY, Jiao J, Luo M, Wang W, Ma W, Zu YG, Fu YJ (2015a) Establishment of high-productive Isatis tinctoria L. hairy root cultures: a promising approach for efficient production of bioactive alkaloids. Biochem Eng J 95:37–47CrossRefGoogle Scholar
  6. Gai QY, Jiao J, Luo M, Wei ZF, Zu YG, Ma W, Fu YJ (2015b) Establishment of hairy root cultures by Agrobacterium rhizogenes mediated transformation of Isatis tinctoria L. for the efficient production of flavonoids and evaluation of antioxidant activities. PLoS ONE 10:e0119022CrossRefPubMedGoogle Scholar
  7. Giri CC, Zaheer M (2016) Chemical elicitors versus secondary metabolite production in vitro using plant cell, tissue and organ cultures: recent trends and a sky eye view appraisal. Plant Cell Tissue Organ Cult 126:1–18CrossRefGoogle Scholar
  8. Hamburger M (2002) Isatis tinctoria–From the rediscovery of an ancient medicinal plant towards a novel anti-inflammatory phytopharmaceutical. Phytochem Rev 1:333–344CrossRefGoogle Scholar
  9. Isah T, Umar S, Mujib A, Sharma MP, Rajasekharan PE, Zafar N, Frukh A (2018) Secondary metabolism of pharmaceuticals in the plant in vitro cultures: strategies, approaches, and limitations to achieving higher yield. Plant Cell Tissue Organ Cult 132:239–265CrossRefGoogle Scholar
  10. Kranner I, Minibayeva FV, Beckett RP, Seal CE (2010) What is stress? Concepts, definitions and applications in seed science. New Phytol 188:655–673CrossRefPubMedGoogle Scholar
  11. Lim W, Li J (2017) Co-expression of onion chalcone isomerase in Del/Ros1-expressing tomato enhances anthocyanin and flavonol production. Plant Cell Tissue Organ Cult 128:113–124CrossRefGoogle Scholar
  12. Lin CW, Tsai FJ, Tsai CH, Lai CC, Wan L, Ho TY, Hsieh CC, Lee Chao PD (2005) Anti-SARS coronavirus 3C-like protease effects of Isatis indigotica root and plant-derived phenolic compounds. Antiviral Res 68:36–42CrossRefPubMedGoogle Scholar
  13. Liu S, Ju J, Xia G (2014) Identification of the flavonoid 3′-hydroxylase and flavonoid 3′, 5′-hydroxylase genes from Antarctic moss and their regulation during abiotic stress. Gene 543:145–152CrossRefPubMedGoogle Scholar
  14. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2– ∆∆CT method. Methods 25:402–408CrossRefPubMedGoogle Scholar
  15. Malik S, Hossein Mirjalili M, Fett-Neto AG, Mazzafera P, Bonfil M (2013) Living between two worlds: two-phase culture systems for producing plant secondary metabolites. Crit Rev Biotechnol 33:1–22CrossRefPubMedGoogle Scholar
  16. Muir SR, Collins GJ, Robinson S, Hughes S, Bovy A, Ric De Vos CH, van Tunen AJ, Verhoeyen ME (2001) Overexpression of petunia chalcone isomerase in tomato results in fruit containing increased levels of flavonols. Nat Biotechnol 19:470–474CrossRefPubMedGoogle Scholar
  17. Murthy HN, Lee EJ, Paek KY (2014) Production of secondary metabolites from cell and organ cultures: strategies and approaches for biomass improvement and metabolite accumulation. Plant Cell Tissue Organ Cult 118:1–16CrossRefGoogle Scholar
  18. Narayani M, Srivastava S (2017) Elicitation: a stimulation of stress in in vitro plant cell/tissue cultures for enhancement of secondary metabolite production. Phytochem Rev 16:1227–1252CrossRefGoogle Scholar
  19. Nguyen TKO, Jamali A, Grand E, Morreel K, Marcelo P, Gontier E, Dauwe R (2017) Phenylpropanoid profiling reveals a class of hydroxycinnamoyl glucaric acid conjugates in Isatis tinctoria leaves. Phytochemistry 144:127–140CrossRefPubMedGoogle Scholar
  20. Ramirez-Estrada K, Vidal-Limon H, Hidalgo D, Moyano E, Golenioswki M, Cusidó RM, Palazon J (2016) Elicitation, an effective strategy for the biotechnological production of bioactive high-added value compounds in plant cell factories. Molecules 21:182CrossRefPubMedGoogle Scholar
  21. Schmitz C, Fritsch L, Fischer R, Schillberg S, Rasche S (2016) Statistical experimental designs for the production of secondary metabolites in plant cell suspension cultures. Biotechnol Lett 38:2007–2014CrossRefPubMedGoogle Scholar
  22. Suzuki M, Yamazaki C, Mitsui M, Kakei Y, Mitani Y, Nakamura A, Ishii T, Soeno K, Shimada Y (2015) Transcriptional feedback regulation of YUCCA genes in response to auxin levels in Arabidopsis. Plant Cell Rep 34:1343–1352CrossRefPubMedGoogle Scholar
  23. Tang X, Xiao Y, Lv T, Wang F, Zhu Q, Zheng T, Yang J (2014) High-throughput sequencing and de novo assembly of the Isatis indigotica transcriptome. PLoS ONE 9:e102963CrossRefPubMedGoogle Scholar
  24. Venugopalan A, Srivastava S (2015) Enhanced camptothecin production by ethanol addition in the suspension culture of the endophyte, Fusarium solani. Bioresour Technol 188:251–257CrossRefPubMedGoogle Scholar
  25. Wang YT, Yang ZF, Zhao SS, Qin S, Guan WD, Huang QD, Zhao YS, Lin Q, Mo ZY (2011) Screening of anti-H1N1 active constituents from Radix Isatidis. J Guangzhou Univ Tradit Chin Med 4:419–422Google Scholar
  26. Zhang HC, Liu JM, Lu HY, Gao SL (2009) Enhanced flavonoid production in hairy root cultures of Glycyrrhiza uralensis Fisch by combining the over-expression of chalcone isomerase gene with the elicitation treatment. Plant Cell Rep 28:1205–1213CrossRefPubMedGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Key Laboratory of Forest Plant Ecology, Ministry of EducationNortheast Forestry UniversityHarbinPeople’s Republic of China
  2. 2.Beijing Advanced Innovation Center for Tree Breeding by Molecular DesignBeijing Forestry UniversityBeijingPeople’s Republic of China

Personalised recommendations