Skip to main content

Establishment of anthocyanin-producing cell suspension cultures of Cleome rosea Vahl ex DC. (Capparaceae)

Plant Cell, Tissue and Organ Culture (PCTOC) volume 106, pages 537–545 (2011)Cite this article

Abstract

The effects of different levels of Murashige and Skoog (MS) basal medium, 2,4-dichlorophenoxyacetic acid (2,4-D), and sucrose on anthocyanin production and biomass accumulation of cell suspension cultures of Cleome rosea were investigated. Cultures were established in liquid MS medium containing 30 g l−1 sucrose and supplemented with 0.90 μM 2,4-D. Proliferating cell suspension cultures achieved the highest growth capacity, a fourfold increase in biomass accumulation, following subculture at the exponential growth phase, 14–18 days of culture. Moreover, the presence of 2,4-D was essential for anthocyanin production and biomass accumulation. On the other hand, increasing levels of sucrose above 30 g l−1 resulted in a drastic reduction in biomass accumulation. Anthocyanin production was highest in cell suspension cultures grown on half-strength MS medium (1/2 MS), 30 g l−1 sucrose, and 0.45 μM 2,4-D. These cell suspension cultures were mainly composed of small aggregates of spherical cells with similar morphology observed in anthocyanin-producing and non-producing cultures. Moreover, microscopic analysis of anthocyanin-producing cultures showed the presence of mixtures of non-pigmented, low-pigmented, and high-pigmented cells.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

39,95 €

Price includes VAT (Finland)

Fig. 1
Fig. 2
Fig. 3

Abbreviations

CV:

Color value

2,4-D:

2,4-dichlorophenoxyacetic acid

DW:

Dry weight

FW:

Fresh weight

References

  • Bagchi D, Sen CK, Bagchi M, Atalay M (2004) Anti-angiogenic, antioxidant and anti-carcinogenic properties of a novel anthocyanin-rich berry extract formula. Biochemistry 69:75–78

    PubMed  CAS  Google Scholar 

  • Barker AV, Pilbeam DJ (2007) Handbook of plant nutrition. CRC Press, Boca Raton

    Google Scholar 

  • Bernabé-Antonio A, Estrada-Zúñiga ME, Buendía-González L, Reyes-Chilpa R, Chávez-Ávila VM, Cruz-Sosa F (2010) Production of anti-HIV-1 calanolides in a callus culture of Calophyllum brasiliense (Cambes). Plant Cell Tiss Org Cult 103:33–40

    Article  Google Scholar 

  • Bonfill M, Mangas S, Moyano E, Cusido RM, Palazón J (2011) Production of centellosides and phytosterols in cell suspension cultures of Centella asiatica. Plant Cell Tiss Org Cult 104:61–67

    Article  CAS  Google Scholar 

  • Boonsnongcheep P, Korsangruang S, Soonthornchareonnon N, Chintapakorn Y, Saralamp P, Prathanturarug S (2010) Growth and isoflavonoid accumulation of Pueraria candollei var. candollei and P. candollei var. mirifica cell suspension cultures. Plant Cell Tiss Org Cult 101:119–126

    Article  CAS  Google Scholar 

  • Bourgaud F, Gravot A, Milesi S, Gontier E (2001) Production of plant secondary metabolites: a historical perspective. Plant Sci 161:839–851

    Article  CAS  Google Scholar 

  • Brindle P, Timberlake CF (1996) Anthocyanins as natural food colors: selected aspects. Food Chem 58:103–109

    Article  Google Scholar 

  • Callebaut A, Terahara N, de Haan M, Decleire M (1997) Stability of anthocyanin composition in Ajuga reptans callus and cell suspension cultures. Plant Cell Tiss Org Cult 50:195–201

    Article  CAS  Google Scholar 

  • Do CB, Cormier F (1990) Accumulation of anthocyanins enhanced by a high osmotic potential in grape (Vitis vinifera L.) cell-suspensions. Plant Cell Rep 9:143–146

    Article  CAS  Google Scholar 

  • Edahiro JI, Seki M (2006) Phenylpropanoid metabolite supports cell aggregate formation in strawberry cell suspension culture. J Biosci Bioeng 102:8–13

    Article  PubMed  CAS  Google Scholar 

  • Estrada-Zúñiga ME, Cruz-Sosa F, Rodríguez-Monroy M, Verde-Calvo JR, Vernon-Carter EJ (2009) Phenylpropanoid production in callus and cell suspension cultures of Buddleja cordata Kunth. Plant Cell Tiss Org Cult 97:39–47

    Article  Google Scholar 

  • Filippini R, Caniato R, Piovan A, Cappelletti EM (2003) Production of anthocyanins by Catharanthus roseus. Fitoterapia 74:62–67

    Article  PubMed  CAS  Google Scholar 

  • Gould K, Davies K, Winefield C (2008) Anthocyanins: biosynthesis, function and applications. Springer, New York

    Google Scholar 

  • Hirasuma TJ, Shuler ML, Lackney VK, Spanswick RM (1991) Enhance anthocyanin production in grape cell culture. Plant Sci 78:107–120

    Article  Google Scholar 

  • Kolewe ME, Gaurav V, Roberts SC (2008) Pharmaceutically active natural product synthesis and supply via plant cell culture technology. Mol Pharm 5:243–256

    Article  PubMed  CAS  Google Scholar 

  • Konczac-Islam I, Yoshinaga M, Nakatami M, Terahara N, Yamakawa O (2000) Establishment and characteristic of an anthocyanin-producing cell line from sweet potato storage root. Plant Cell Rep 19:472–477

    Article  Google Scholar 

  • Madhusudhan R, Ravishankar GA (1996) Gradient of anthocyanin in cell aggregates of Daucus carota in suspension cultures. Biotechnol Lett 18:1253–1256

    Article  CAS  Google Scholar 

  • Meyer HJ, van Staden J (1995) The in vitro production of an anthocyanin from callus culture of Oxalis linearis. Plant Cell Tiss Org Cult 40:55–58

    Article  CAS  Google Scholar 

  • Meyer JE, Pépin MF, Smith MAL (2002) Anthocyanin production from Vaccinium pahalae: limitations of the physical microenvironment. J Biotechnol 93:45–57

    Article  PubMed  CAS  Google Scholar 

  • 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 

  • Nagella P, Murthy HN (2011) Effects of macroelements and nitrogen source on biomass accumulation and withanolide-A production from cell suspension cultures of Withania somnifera (L.) Dunal. Plant Cell Tiss Org Cult 104:119–124

    Article  CAS  Google Scholar 

  • Nakamura M, Seki M, Furusaki S (1998) Enhanced anthocyanin methylation by growth limitation in strawberry suspension culture. Enzyme Microb Technol 22:404–408

    Article  CAS  Google Scholar 

  • Narayan MS, Venkataraman LV (2000) Characterisation of anthocyanins derived from carrot (Daucus carota) cell culture. Food Chem 70:361–363

    Article  CAS  Google Scholar 

  • Narayan MS, Thimmaraju R, Bhagyalakshmi B (2005) Interplay of growth regulators during solid-state and liquid-state batch cultivation of anthocyanin producing cell line of Daucus carota. Process Biochem 40:351–358

    Article  CAS  Google Scholar 

  • Nawa Y, Asano S, Motoori S, Ohtani T (1993) Production of anthocyanins, carotenoids and proanthocyanidins by culture cells of rabbiteye blueberry (Vaccinium ashei Reade). Biosci Biotech Biochem 57:770–774

    Article  CAS  Google Scholar 

  • Pèpin MF, Smith MAL, Reid JF (1999) Application of imaging tools to plant cell culture: relationship between plant cell aggregation and flavonoid production. In Vitro Cell Dev Biol Plant 35:290–295

    Article  Google Scholar 

  • Qu J, Zhang W, Yu X, Jin M (2005) Instability of anthocyanin accumulation in Vitis vinifera L. var. Gamay Fréaux suspension cultures. Biotechnol Bioprocess Eng 10:155–161

    Article  CAS  Google Scholar 

  • Rao SR, Ravishankar GA (2002) Plant cell cultures: chemical factories of secondary metabolites. Biotechnol Adv 20:101–153

    Article  PubMed  CAS  Google Scholar 

  • Saito K, Mizukami H (2002) Plant cell cultures as producers of secondary metabolites. In: Oksman-Caldentey K-M, Barz WH (eds) Plant biotechnology and trangenic plants. Marcel Dekker, New York

    Google Scholar 

  • Sakamoto K, Iida K, Sawamura K, Hajiro K, Asada Y, Yoshikawa T, Furuya T (1993) Effects of nutrients on anthocyanin production in cultured cells of Aralia cordata. Phytochemistry 33:357–360

    Article  CAS  Google Scholar 

  • Simões C, Santos AS, Albarello N, Figueiredo SFL (2004) Shoot organogenesis and plantlet regeneration from stem explants of Cleome rosea Vahl (Capparaceae). J Plant Biotech 6:199–204

    Google Scholar 

  • Simões C, Mattos JCP, Sabino KCC, Caldeira-de-Araújo A, Coelho MGP, Albarello N, Figueiredo SFL (2006) Medicinal potential from in vivo and acclimatized plants of Cleome rosea Vahl ex DC. (Capparaceae). Fitoterapia 77:94–99

    Article  PubMed  Google Scholar 

  • Simões C, Albarello N, Callado CH, Castro TC, Mansur E (2009a) New approaches for shoot production and in vitro root cultures of Cleome rosea Vahl. Plant Cell Tiss Org Cult 98:79–86

    Article  Google Scholar 

  • Simões C, Bizarri CHB, Castro TC, Coutada LCM, Silva AJR, Albarello N, Mansur E (2009b) Anthocyanin production in callus cultures of Cleome rosea: modulation by culture conditions and characterization of pigments by means of HPLC-DAD/ESIMS. Plant Physiol Biochem 47:895–903

    Article  PubMed  Google Scholar 

  • Vogelien DL, Hrazdina G, Reevers S, Dougall DK (1990) Phenotypic differences in anthocyanin accumulation among clonally related cultured cells of carrot. Plant Cell Tiss Org Cult 22:213–222

    Article  CAS  Google Scholar 

  • Wai-Leng L, Lai-Keng C (2004) Establishment of Orthosiphon stamineus cell suspension culture for cell growth. Plant Cell Tiss Org Cult 78:101–106

    Article  CAS  Google Scholar 

  • Wang J, Mazza G (2002) Inhibitory effects of anthocyanins and other phenolic compounds on nitric oxide production in LPS/IFN-γ-active RAW 2647 macrophages. J Agric Food Chem 50:850–857

    Article  PubMed  CAS  Google Scholar 

  • Yoshinaga M, Yamakawa O, Nakatani M (1999) Genotypic diversity of anthocyanin content and composition in purple-fleshed sweet potato (Ipomoea batatas (L.) Lam. Breeding Sci 49:43–47

    CAS  Google Scholar 

  • Zamith LR, Scarano FR (2006) Restoration of a restinga sandy coastal plain in Brazil: survival and growth of planted woody species. Restoration Ecol 14:87–94

    Article  Google Scholar 

  • Zhang W, Curtin C, Kikuchi M, Franco C (2002) Integration of jasmonic acid and light irradiation for enhancement of anthocyanin biosynthesis in Vitis vinifera suspension cultures. Plant Sci 162:459–468

    Article  Google Scholar 

  • Zhang W, Franco C, Curtin C, Conn S (2004) To stretch the bondary of secondary metabolite production in plant cell-based bioprocessing: anthocyanin as a case study. J Biomed Biotechnol 5:264–271

    Article  CAS  Google Scholar 

  • Zhao D, Huang Y, Jin Z, Qu W, Lu D (2003) Effect of aggregate size in cell cultures of Saussurea medusa on cell growth and jaceosidin production. Plant Cell Rep 21:1129–1133

    Article  PubMed  CAS  Google Scholar 

  • Zhong JJ, Yoshida M, Fujiyama K, Seki T, Yoshida T (1993) Enhancement of anthocyanin production by Perilla frutescens cells in a stirred bioreactor with internal light irradiation. J Ferment Bioeng 75:299–303

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors are grateful to Maria Francisca Santoro Assunção for the valuable technical assistance. This work was supported by Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).

Author information

Authors and Affiliations

  1. Núcleo de Biotecnologia Vegetal, Universidade do Estado do Rio de Janeiro (UERJ), Rua São Francisco Xavier, 524, PHLC, sala 509, Maracanã, Rio de Janeiro, RJ, CEP 20550-013, Brazil

    Claudia Simões-Gurgel, Lívia da Silva Cordeiro, Tatiana Carvalho de Castro, Norma Albarello & Elisabeth Mansur

  2. Laboratório de Anatomia Vegetal, Universidade do Estado do Rio de Janeiro (UERJ), Rio de Janeiro, Brazil

    Cátia Henriques Callado

Authors
  1. Claudia Simões-Gurgel
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lívia da Silva Cordeiro
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tatiana Carvalho de Castro
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Cátia Henriques Callado
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Norma Albarello
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Elisabeth Mansur
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Claudia Simões-Gurgel.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Simões-Gurgel, C., Cordeiro, L.d.S., de Castro, T.C. et al. Establishment of anthocyanin-producing cell suspension cultures of Cleome rosea Vahl ex DC. (Capparaceae). Plant Cell Tiss Organ Cult 106, 537–545 (2011). https://doi.org/10.1007/s11240-011-9945-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-011-9945-3

Keywords

  • Cell aggregate
  • Color value
  • 2,4-dichlorophenoxyacetic acid
  • Salt concentration
  • Sucrose