A method for cell suspension culture and plant regeneration of Angelica sinensis (Oliv.) Diels

Abstract

Angelica sinensis (Oliv.) Diels is an edible and medicinal plant honored as “female ginseng”. It is widely used as valuable traditional Chinese medicine for over 2000 years. In the present study, an efficient procedure has been developed for establishment of cell suspension culture and plant regeneration of Angelica sinensis (Oliv.) Diels. Friable Callus derived from hypocotyl was used to establish cell suspension culture. Dynamics of biomass, FA content and medium conductivity have been investigated during the growth cycle of the suspension culture. The maximum content of FA and biomass were both observed on the 16th day of the cultivation cycle. The effect of basic medium, inoculum density, initial pH, illumination and various concentrations of sucrose have been analyzed, to optimize biomass growth and FA production. The maximum biomass was obtained in ½MS medium. In response to various sucrose concentrations, cell cultures with 40 g/L sucrose produced the highest dry biomass, but the highest production of FA was obtained in medium with 30 g/L sucrose. The optimal inoculum density was 10.3925 g/L. Initial medium pH 5.0 was beneficial to biomass and FA accumulation. Illumination had no effect on biomass, but it was favorable for FA synthesis. Cells cultured over a year were transferred to ½MS medium supplemented with 10% coconut water showed embryoid formation at a frequency of 100%. The research results could provide technical support for large-scale production of cells and seedlings, genetic transformation and somatic hybridization in A. sinensis studies.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3

References

  1. Abbas MS, El-Shabrawi HM, Soliman AS, Mai AS (2018) Optimization of germination, callus induction, and cell suspension culture of African locust beans Parkia biglobosa (Jacq.) Benth. J Genetic Biotechnol 16:191–201

    Article  Google Scholar 

  2. Ali M, Abbasi BH, Ahmad N, Ali SS, Ali S, Ali GS (2016) Sucrose-enhanced biosynthesis of medicinally important antioxidant secondary metabolites in cell suspension cultures of Artemisia absinthium L. Bioprocess Biosyst Eng 39:1–10

    Article  CAS  Google Scholar 

  3. Arias JP, Zapata K, Rojano B, Arias M (2016) Effect of light wavelength on cell growth, content of phenolic compounds and antioxidant activity in cell suspension cultures of Thevetia peruviana. J Photochem Photobiol B Biol 163:87–91

    Article  CAS  Google Scholar 

  4. Baque MA, Lee EJ, Paek KY (2012) Sucrose regulated enhanced induction of anthraquinone, phenolics, flavonoids biosynthesis and activities of antioxidant enzymes in adventitious root suspension cultures of Morinda citrifolia (L.). Acta Physiol Plant 34:417–417

    Article  CAS  Google Scholar 

  5. Bioprocess (2016) Sucrose-enhanced biosynthesis of medicinally important antioxidant secondary metabolites in cell suspension cultures of Artemisia absinthium L Biosys Eng 39:1–10

    Google Scholar 

  6. Cetin ES, Baydar NG (2016) Elicitor applications to cell suspension culture for production of phenolic compounds in grapevine. Tarim Bilimleri Dergisi 22:42–53

    Article  Google Scholar 

  7. Chen XP, Li W, Xiao XF, Zhang LL, Liu CX (2013) Phytochemical and pharmacological studies on radix angelica sinensis. Chin J Nat Med 11:577–587

    Article  CAS  PubMed  Google Scholar 

  8. China Pharmacopoeia Commission (2015) Chinese Pharmacopoeia. Chemical Industry Press, China

    Google Scholar 

  9. Ganapathi S, Karg F (1990) Recent advances in indole alkaloid production by Catharanthus raseus (Periwinkle). J Exp Bot 41:259–267

    Article  CAS  Google Scholar 

  10. Hattori T, Nakagawa S, Nakamura K (1990) High-level expression of tuberous root storage protein genes of sweet potato in stems of plantlets grown in vitro on sucrose medium. Plant Mol Biol 14:595–604

    Article  CAS  PubMed  Google Scholar 

  11. Hippolyte I, Marin B, Baccou JC, Jonard R (1992) Growth and rosmarinic acid production in cell suspension cultures of Salvia officinalis L Plant Cell Rep 11:109–112

    Article  CAS  PubMed  Google Scholar 

  12. Hook IL (2014) Danggui to Angelica sinensis root: are potential benefits to European women lost in translation? a review. J Ethnopharmacol 152:1–13

    Article  PubMed  Google Scholar 

  13. Huang B, Han L, Li S, Yan C (2015) Optimization of induction, subculture conditions, and growth kinetics of Angelica sinensis (Oliv.) Diels callus. Pharmacogn Mag 11:574–578

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Karwasara VS, Dixit VK (2013) Culture medium optimization for camptothecin production in cell suspension cultures of Nothapodytes nimmoniana (J. Grah.) Mabberley. Plant Biotechnol Rep 7:357–369

    Article  Google Scholar 

  15. Kashyap S, Kapoor N, Kale RD (2016) Coscinium fenestratum: callus and suspension cell culture of the endangered medicinal plant using vermicompost extract and coelomic fluid as plant tissue culture media. Am J Plant Sci 7:899–906

    Article  Google Scholar 

  16. Kühn C, Hajirezaei MR, Fernie AR, Roessnertunali U, Czechowski T, Hirner B et al (2003) The sucrose transporter stsut1 localizes to sieve elements in potato tuber phloem and influences tuber physiology and development. Plant Physiol 131:102–113

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Kurkdjian A, Morot-Gaudry JF, Wuilleme S, Lamant A, Jolivet E, Guern J (1981) Evidence for an action of fusicoccin on the vacuolar pH of Acer pseudoplatanus cells in suspension culture. Plant Sci Lett 23:233–243

    Article  CAS  Google Scholar 

  18. Li Q, Tang M, Tan Y, Ma D, Wang Y, Zhang H (2016) Improved production of chlorogenic acid from cell suspension cultures of Lonicera macranthoids. Trop J Pharm Res 15:919–927

    Article  CAS  Google Scholar 

  19. Lian ML, Chakrabarty D, Paek KY (2002) Effect of plant growth regulators and medium composition on cell growth and saponin production during cell-suspension culture of mountain ginseng (Panax ginseng, C. A. Mayer). J Plant Biol 45:201–206

    Article  CAS  Google Scholar 

  20. Liu X, Yan Y, Liu Y, Mo T, Wang X, Song Y et al (2018) Cell culture establishment and regulation of two phenylethanoid glycosides accumulation in cell suspension culture of desert plant Cistanche tubulosa. Plant Cell Tissue Organ Cult 134:1–12

    Article  CAS  Google Scholar 

  21. Martinez BC, Park CH (1993) Characteristics of batch suspension cultures of preconditioned Coleus blumei cells: sucrose effect. Biotechnol Prog 9:97–100

    Article  CAS  Google Scholar 

  22. Modarres M, Esmaeilzadeh SB, Taghavizadeh MY (2018) Enhanced production of phenolic acids in cell suspension culture of Salvia leriifolia Benth. using growth regulators and sucrose. Cytotechnology 70:1–10

    Article  CAS  Google Scholar 

  23. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  24. Nagella P, Murthy HN (2010) Establishment of cell suspension cultures of Withania somnifera for the production of withanolide A. Bioresour Technol 101:6735–6739

    Article  CAS  Google Scholar 

  25. Nalawade SM, Sagare AP, Lee CY, Kao CL, Tsay HS (2003) Studies on tissue culture of Chinese medicinal plant resources in Taiwan and their sustainable utilization. Bot Bull Acad Sin Taipei 44:79–98

    Google Scholar 

  26. Pacheco G, Garcia R, Lugato D, Vianna M, Mansur E (2012) Plant regeneration, callus induction and establishment of cell suspension cultures of Passiflora alata Curtis. Sci Hortic 144:42–47

    Article  CAS  Google Scholar 

  27. Qi XJ, Chen RY, Wang W (2010) Cell suspension culture of Gentiana macrophylla (I). Chin Trad Herb Drugs 41:636–638

    CAS  Google Scholar 

  28. Rao SR, Ravishankar GA (2003) Plant cell cultures: chemical factories of secondary metabolites. Biotechnol Adv 20:101–153

    Google Scholar 

  29. Ryu DDY, Lee SO, Romani RJ (1990) Determination of growth rate for plant cell cultures: comparative studies. Biotechnol Bioeng 35:305–311

    Article  CAS  PubMed  Google Scholar 

  30. Sathish S, Venkatesh R, Safia N, Sathishkumar R (2018) Studies on growth dynamics of embryogenic cell suspension cultures of commercially important Indica rice cultivars ASD16 and Pusa basmati. Biotech 8:1–9

    Google Scholar 

  31. Sharma N, Gauchan DP, Dhakal A, Luitel A, Shakya S (2015) Establishment of regenerative callus, suspension system and molecular characterization of Stevia Rebaudiana Bertoni for the production of stevioside in In-vitro. Int J Res Appl Sci Eng Technol 3:133–143

    Google Scholar 

  32. Tsay HS, Huang HL (1998) Somatic embryo formation and germination from immature embryo-derived suspension-cultured cells of Angelica sinensis (Oliv.) Diels. Plant Cell Rep 17:670–674

    Article  CAS  PubMed  Google Scholar 

  33. Vasil V, Vasil IK (1982) Characterization of an embryogenic cell suspension culture derived from cultured inflorescences of Pennisetum americanum (Pearl millet, Gramineae). Am J Bot 69:1441–1449

    Article  Google Scholar 

  34. Wang YC, Liu CZ, Zhao B, Ouyang F (2000) Kinetics and stoichiometry in suspension culture of Artemisia annua L. hairy root. Eng Chem Metall 21:42–46

    Google Scholar 

  35. Wang YQ, Ye QH, Zhu Y (2008) Preliminary study on the cell suspension culture of Eucommia ulmoides and secondary metabolite-chlorogenic acid. Guihaia 28:671–674

    CAS  Google Scholar 

  36. Wang J, Gao WY, Yin SS, Liu H, Wei CL (2012) Research progress in medicinal plant cell suspension culture. China J Chin Mater Med 37:3680–3683

    Google Scholar 

  37. Wu SJ, Ng LT, Lin CC (2004) Antioxidant activities of some common ingredients of traditional Chinese medicine, angelica sinensis, lycium barbarum and poria cocos. Phytother Res 18:1008–1012

    Article  PubMed  Google Scholar 

  38. Xu J, Liu C, Fang X, Feng P (1997) Effects of medium pH dectease on salidroside release and cell viability in cell suspension culture of Rhodiola sachalinensis. Acta Bot Sin 39:1022–1029

    Google Scholar 

  39. Zhang W, Seki M, Furusaki S, Middelberg APJ (1998) Anthocyanin synthesis, growth and nutrient uptake in suspension cultures of strawberry cells. J Ferment Bioeng 86:72–78

    Article  CAS  Google Scholar 

  40. Zhang HY, Bi WG, Yu Y, Liao WB (2012) Angelica sinensis (Oliv.) Diels in china: distribution, cultivation, utilization and variation. Genetic Resour Crop Evol 59:607–613

    Article  Google Scholar 

  41. Zhao Y, Sun J, Yu L, Chen P (2013) Chromatographic and mass spectrometric fingerprinting analyses of Angelica sinensis (Oliv.) Diels-derived dietary supplements. Anal Bioanal Chem 405:4477–4485

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This work was supported by funds of the Twelfth Five-Year National Science and Technology Support Project of China (No. 2011BAI05B02).

Author information

Affiliations

Authors

Corresponding author

Correspondence to C. Y. He.

Ethics declarations

Conflict of interest

We declare that this work was done by the authors named in this article and all liabilities pertaining to claims relating to the content of this article will be borne by the authors.

Additional information

Communicated by Jochen Kumlehn.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Zhang, Y.H., Lu, Y.Y., He, C.Y. et al. A method for cell suspension culture and plant regeneration of Angelica sinensis (Oliv.) Diels. Plant Cell Tiss Organ Cult 136, 313–322 (2019). https://doi.org/10.1007/s11240-018-1517-3

Download citation

Keywords

  • Angelica sinensis (Oliv.) Diels
  • Cell suspension culture
  • Ferulic acid
  • Sucrose
  • Inoculum density
  • Plant regeneration