Skip to main content
SpringerLink
Log in

Optimization of a Liquid Culture System for Shoot Regeneration and Achieving an Enriched Level of Scopadulcic Acid B in the Leaf Organ Cultures of Scoparia dulcis L. by Response Surface Methodology

  • Plant Development/Regeneration
  • Published:
In Vitro Cellular & Developmental Biology - Plant Aims and scope Submit manuscript

Abstract

Response surface methodology (RSM) approach was utilized in the present investigation to optimize the constituents of Murashige and Skoog’s (MS) liquid medium to enrich the scopadulcic acid B (SDB) content in Scoparia dulcis L. RSM approach using the central composite design (CCD) was employed to identify the precise concentration of growth regulators of medium and substantiated in shake-flask cultivation. CCD-RSM model revealed a strong agreement, with a predicted coefficient of determination (R2) values of 0.881 and 0.872 for the shoot regeneration percentage (Y1) and the average number of shoots per explant (Y2), respectively. RSM predicted augmented conditions of MS liquid medium considerably influence the shoot proliferation (Y1 and Y2) in the leaf explants of S. duclis. The medium was fortified with 3.59 μM kinetin (KN; X1), 6.00 μM 6-benzylaminopurine (BAP; X2), 3.93 μM indole-3-acetic acid (IAA; X3), and 25.84 g L−1 of sucrose (X4). The present investigation conquered a maximal response of Y1 and Y2 with 91.28 ± 3.85 and 82.26 ± 2.13, respectively. The experimentally detected values are in close agreement with the predicted values of 90.07 and 79.70, respectively. This proposed that the developed design using CCD had efficacy in the optimization of medium components. The enhanced level of SDB, 9.89 ± 0.98 mg g−1 FW (ca. 5-fold), in the plantlets grown on liquid medium was significantly greater than those accomplished in the other tested plant tissues, comprises of field-grown parent plant leaves and in vitro developed callus and micropropagated plants on MS solid medium. For the first time, the methodology was developed effectively using RSM to promote the shoot proliferation and enhance the SDB production in the leaf organ culture of S. dulcis on MS liquid medium.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Abbasi Z, Hooshyar S, Bagherieh-Najjar MB (2016) Improvement of callus production and shoot regeneration using various organs of soybean (Glycine max L. Merr) by response surface methodology. In Vitro Cell Dev Biol-Plant 52:537–545

    Article  CAS  Google Scholar 

  • Aileni M, Rao Kokkirala V, Reddy Kota S, Umate P, Abbagani S (2008) Efficient in-vitro regeneration from mature leaf explants of Scoparia dulcis L., an ethnomedicinal plant. J Herbs Spices Med Plants 14:200–207

    Article  CAS  Google Scholar 

  • Bansal M, Sudhakara Reddy M, Kumar A (2017) Optimization of cell growth and bacoside-a production in suspension cultures of Bacopa monnieri (L.) Wettst. using response surface methodology. In Vitro Cell Dev Biol-Plant 53:527–537

    Article  CAS  Google Scholar 

  • Burkill HM (2000) The useful plants of west tropical Africa, 2nd Ed, 5, Royal Botanic Gardens, Kew, pp 648

  • Chakraborty D, Bandyopadhyay A, Bandopadhyay S, Gupta K, Chatterjee A (2010) Use of response surface methodology for optimization of a shoot regeneration protocol in Basilicum polystachyon. In Vitro Cell Dev Biol-Plant 46:451–459

    Article  Google Scholar 

  • Chong MN, Zhuc HY, Jin B (2010) Response surface optimization of photocatalytic process for degradation of Congo Red using H-titanate nanofiber catalyst. Chem Eng J 156:278–285

    Article  CAS  Google Scholar 

  • Davioud E, Kan C, Hamon J, Tempé J, Husson H-P (1989) Production of indole alkaloids by in vitro root cultures from Catharanthus trichophyllus. Phytochemistry 28:2675–2680

    Article  CAS  Google Scholar 

  • Elibol M (2004) Optimization of medium composition for actinorhodin production by Streptomyces coelicolor A3(2) with response surface methodology. Process Biochem 39:1057–1062

    Article  CAS  Google Scholar 

  • Hayashi T, Smith FT, Lee KH (1987) Antitumor agents. 89. Psychorubrin, a new cytotoxic naphthoquinone from Psychotria rubra and its structure-activity relationships. J Med Chem 30:2005–2008

    Article  CAS  Google Scholar 

  • Hayashi T, Okamura K, Kawasaki M, Morita N (1991) Two chemotypes of Scoparia dulcis in Paraguay. Phytochemistry 30:3617–3620

    Article  CAS  Google Scholar 

  • Hayashi T, Kawasaki M, Okamura K, Tamada Y, Morita N, Tezuka Y, Kikuchi T, Miwa Y, Taga T (1992) Scoparic acid A, a beta-glucuronidase inhibitor from Scoparia dulcis. J Nat Prod 55:1748–1755

  • Hayashi T, Okamura K, Kawasaki M, Morita N (1993a) Production of diterpenoids by cultured cells from two chemotypes of Scoparia dulcis. Phytochemistry 33:353–356

    Article  CAS  Google Scholar 

  • Hayashi T, Okamura K, Tamada Y, Iida A, Fujita T, Morita N (1993b) A new chemotype of Scoparia dulcis. Phytochemistry 33:349–352

    Article  Google Scholar 

  • Hayashi T, Kasahara K, Sankawa U (1997) Efficient production of biologically active diterpenoids by leaf organ culture of Scoparia dulcis. Phytochemistry 46:517–520

    Article  CAS  Google Scholar 

  • Hayashi T, Asai T, Sankawa U (1999) Mevalonate-independent biosynthesis of bicyclic and tetracyclic diterpenes of Scoparia dulcis L. Tetrahedron Lett 40:8239–8243

    Article  CAS  Google Scholar 

  • Hirata K, Yamanaka A, Kurano N, Miyamoto K, Miura Y (1987) Production of indole alkaloids in multiple shoot culture of Catharanthus roseus (L). G. Don. Agric Biol Chem 51:1311–1317

    CAS  Google Scholar 

  • Krishnan PN, Decruse SW, Radha RK (2011) Conservation of medicinal plants of Western Ghats, India and its sustainable utilization through in vitro technology. In Vitro Cell Dev Biol-Plant 47:110–122

    Article  Google Scholar 

  • Kuhnt S, Rudak N (2013) Simultaneous optimization of multiple responses with the R package JOP. J Stat Softw 54:1–23

    Article  Google Scholar 

  • Latha M, Pari L, Sitasawad S, Bhonde R (2004) Insulin-secretagogue activity and cytoprotective role of the traditional antidiabetic plant Scoparia dulcis (Sweet Broomweed). Life Sci 75:2003–2014

    Article  CAS  Google Scholar 

  • Lee CL, Wang WL (1997) Biological statistics. Science Press, Beijing, China

  • Lenth RV (2009) Response-surface methods in R, using rsm. J Stat Softw 32:1–17

    Article  Google Scholar 

  • Liu Q, Yang Q-M, Hu H-J, Yang L, Yang Y-B, Chou G-X, Wang Z-T (2014) Bioactive diterpenoids and flavonoids from the aerial parts of Scoparia dulcis. J Nat Prod 77:1594–1600

    Article  CAS  Google Scholar 

  • Majumder S, Rahman MM, Bhadra SK (2011) Micropropagation of Scoparia dulcis Linn. through induction of indirect organogenesis. Asia Pac J Mol Biol Biotechnol 1:11–17

  • Mukherjee PK (2003) Exploring botanicals in Indian system of medicine-regulatory perspectives. Clin Res Regul Aff 20:249–264

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Naveenchandra PM, Bhattacharya S, Ravishankar GA (2011) Culture media optimization through response surface methodology for in vitro shoot bud development of Solanum melongena L. for micropropagation. Int J Bioautomation 15:159–172

    CAS  Google Scholar 

  • Nkembo MK, Kurosaki F, Lee J-B, Hayashi T (2005a) Stimulation of calcium signal transduction results in enhancement of production of scopadulcic acid B by methyl jasmonate in the cultured tissues of Scoparia dulcis. Plant Biotechnol 22:333–337

    Article  CAS  Google Scholar 

  • Nkembo MK, Lee J-B, Hayashi T (2005b) Selective enhancement of scopadulcic acid B production in the cultured tissues of Scoparia dulcis by methyl jasmonate. Chem Pharm Bull 53:780–782

    Article  CAS  Google Scholar 

  • Osei-Safo D, Chama MA, Addae-Mensah I, Waibel R (2009) Hispidulin and other constituents of Scoparia dulcis Linn. J Sci Technol 29:7–15

    Google Scholar 

  • Pamunuwa G, Karunaratne N, Waisundara VY (2016) Antidiabetic properties, bioactive constituents, and other therapeutic effects of Scoparia dulcis. Evid Based Complement Alternat Med 8243215:1–11

    Article  Google Scholar 

  • Pari L, Venkateswaran S (2002) Hypoglycaemic activity of Scoparia dulcis L. extract in alloxan induced hyperglycaemic rats. Phytother Res 16:662–664

    Article  CAS  Google Scholar 

  • Premkumar G, Sankaranarayanan R, Jeeva S, Rajarathinam K (2011) Cytokinin induced shoot regeneration and flowering of Scoparia dulcis L. (Scrophulariaceae)-an ethnomedicinal herb. Asian Pac J Trop Biomed 1:169–172

    Article  CAS  Google Scholar 

  • R Core Team (2018) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. URL https://www.R-project.org/

  • Rana B, Awasthi P, Kumbhar BK (2012) Optimization of processing conditions for cyanide content reduction in fresh bamboo shoot during NaCl treatment by response surface methodology. J Food Sci Technol 49:103–109

    Article  CAS  Google Scholar 

  • Rao Y, Lu S, Liu B, Tzeng Y (2006) Enhanced production of an extracellular protease from Beauveria bassiana by optimization of cultivation processes. Biochem Eng J 28:57–66

    Article  CAS  Google Scholar 

  • Srivastava S, Srivastava AK (2012) Statistical medium optimization for enhanced azadirachtin production in hairy root culture of Azadirachta indica. In Vitro Cell Dev Biol-Plant 48:73–84

    Article  CAS  Google Scholar 

  • Tabata M, Yamamoto H, Hiraoka N, Konoshima M (1972) Organization and alkaloid production in tissue cultures of Scopolia parviflora. Phytochemistry 11:949–955

    Article  CAS  Google Scholar 

  • Taylor L (2006) Vassourinha monograph 4/Scoparia dulcis. In: The rainforest, pharmacy to the world. Raintree Nutrition, Inc., Carson City

  • Wang Y, Fang X, An F, Wang G, Zhang X (2011) Improvement of antibiotic activity of Xenorhabdus bovienii by medium optimization using response surface methodology. Microb Cell Fact 10:98

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors are grateful to Prof. Russell V. Lenth, Department of Statistics and Actuarial Science, The University of Iowa, USA, for assistance with R statistical programming. The authors also thank the anonymous reviewers for their constructive comments.

Funding

The financial support (grant reference No. MRP/3011/09) was from the University Grants Commission, New Delhi.

Author information

Author notes

  1. Gandhi Premkumar and Thirupathi Karuppanapandian contributed equally to this work.

Authors and Affiliations

  1. Department of Botany, V.H.N. Senthikumara Nadar College (Autonomous), Virudhunagar, 626001, India

    Gandhi Premkumar, Chandran Sureshpandian & Kaniappan Rajarathinam

  2. Department of Horticultural Science, Faculty of AgriSciences, Stellenbosch University, Private Bag X1, Stellenbosch, 7602, South Africa

    Thirupathi Karuppanapandian

  3. Department of Biotechnology, Pondicherry University, Pondicherry, 605014, India

    Neelakanda Arumugam

  4. Department of Mathematics, V.H.N. Senthikumara Nadar College (Autonomous), Virudhunagar, 626001, India

    Avadayappan Selvam

Authors
  1. Gandhi Premkumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Thirupathi Karuppanapandian
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Chandran Sureshpandian
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Neelakanda Arumugam
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Avadayappan Selvam
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Kaniappan Rajarathinam
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Gandhi Premkumar or Thirupathi Karuppanapandian.

Additional information

Editor: Randall Niedz

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Premkumar, G., Karuppanapandian, T., Sureshpandian, C. et al. Optimization of a Liquid Culture System for Shoot Regeneration and Achieving an Enriched Level of Scopadulcic Acid B in the Leaf Organ Cultures of Scoparia dulcis L. by Response Surface Methodology. In Vitro Cell.Dev.Biol.-Plant 56, 60–71 (2020). https://doi.org/10.1007/s11627-019-10037-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11627-019-10037-z

Keywords

Search

Navigation