Skip to main content
SpringerLink
Log in

An efficient in vitro propagation methodology for Annatto (Bixa orellana L.)

  • Research Article
  • Published:
Physiology and Molecular Biology of Plants Aims and scope Submit manuscript

Abstract

Efficient methods were developed for both in vitro seed germination and micropropagation of an economically important dye yielding multipurpose tree, Bixa orellana L. Mature seeds were inoculated onto Murashige and Skoog (MS) medium supplemented with different concentrations of gibberellic acid (GA3). Highest frequency of germination (93.3 %) was recorded on medium supplemented with 3 μM GA3 against 13.33 % in control. Nodal explants cultured on MS medium fortified with 5 μM isopentanyl adenine (2-iP) produced maximum explants response (93.3 %) and highest number of shoots (35.71). Addition of relatively higher concentration (15 μM) of benzyl adenine (BA) resulted in the production of significantly (P < 0.05) reduced number of shoots (12.66). Sucrose at 87.6 mM was found to be the best carbohydrate source for multiple shoot induction compared to glucose and fructose. Regenerated shoots (3–4 cm) were rooted (95.5 %) on agar gelled MS medium supplemented with 10 μM indole-3-butyric acid (IBA). In vitro developed plantlets with well-developed roots were potted and acclimatized initially in the growth chamber and then moved to a green house with 83.3 % survival. The present protocol avoids the use of auxins in shoot multiplication medium, which will lower the cost, avoid callus formation and thus reduces the possibility of somaclonal variation in the regenerated plants. The method is efficient to produce over 32,000 hardened plants within a 10-month culture period starting from a single nodal explant.

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.

Institutional subscriptions

Fig. 1

Similar content being viewed by others

References

  • Almeida JL, Almeida FCG, Nunes RDEP, Almeida FAG (1996) Bud initiation in leaf explants of annatto seedlings in different cytokinins. Cinencia Rural 26:45–49

    Google Scholar 

  • Aparnathi K, Lata R, Sharma R (2003) Annatto (Bixa orellana L.): its cultivation, preparation and usage. Int J Trop Agric 8:80–88

    Google Scholar 

  • Benson E, Danahar JE, Pimbley IM, Anderson CT, Wake JE, Daley S, Adams LK (2000) In vitro propagation of Primula scotica: a rare Scottish plant. Biodivers Conserv 9:711–726

    Article  Google Scholar 

  • Carvalho PR (1999) Technological advances and perspectives. Arch Latinoam Nutr 49:71–73

    Google Scholar 

  • Collins P, Hughes S (1991) Report in prospective in natural food symposium. Oversea food Ltd., England

    Google Scholar 

  • D’Souza MC, Sharon M (2001) In vitro clonal propagation of annatto (Bixa orellana L.). In Vitro Cell Dev Biol-Plant 37:168–172

    Article  Google Scholar 

  • De Klerk G, Brugge J, Marinova S (1997) Effectiveness of indole-acetic acid, indole butyric acid and naphthalene acetic acid during adventitious root formation in vitro in Malus Jork 9. Plant Cell Tiss Organ Cult 49:39–44

    Article  Google Scholar 

  • Debnath SC, McRae KB (2001) An efficient in vitro shoot propagation of cranberry (Vaccinium macrocarpon AIT.) by axillary bud proliferation. In Vitro Cell Dev Biol-Plant 37:243–249

    Article  CAS  Google Scholar 

  • George EF (1996) Plant propagation by tissue culture, Part 2: in practice. Exegetics Ltd, London, p 799

    Google Scholar 

  • Hartman HT, Kester KE (1983) Plant propagation and practices. Prentice Hall, India Pvt. Ltd, New Delhi

    Google Scholar 

  • Husain MK, Anis M, Shahzad A (2007) In vitro propagation of Indian kino (Pterocarpus marsupim Roxb.) using Thidiazuron. In Vitro Cell Dev Biol-Plant 43:59–64

    CAS  Google Scholar 

  • Joseph N, Siril EA, Nair GM (2010) Imbibition duration, seed treatment, seed mass and population influence germination of annatto (Bixa orellana L.) seeds. Seed Technol 32:37–45

    Google Scholar 

  • Khan PSSV, Prakash E, Rao KR (2002) Callus induction and plantlet regeneration in Bixa orellana L., an annatto-yielding tree. In Vitro Cell Dev Biol-Plant 38:186–190

    Article  CAS  Google Scholar 

  • Larkin PJ, Scowcroft WR (1981) Somaclonal variation- a novel source of variability from cell cultures for plant improvement. Theor Appl Genet 60:197–214

    Article  Google Scholar 

  • Ludwig-Muller J (2000) Indole-3-butyric acid in plant growth and development. Plant Growth Regul 32:219–230

    Article  CAS  Google Scholar 

  • Marcotrigiano M, McGlew SP (1991) A two-stage micropropagation system for cranberries. J Am Soc Hort Sci 116:911–916

    CAS  Google Scholar 

  • Marks TM, Simpson SE (2000) Interaction of explants type and indole-3-butyric acid during rooting in vitro in a range of difficult and easy-to-root woody plants. Plant Cell Tiss Organ Cult 62:65–74

    Article  CAS  Google Scholar 

  • Mercier H, Kerbauy GB (1997) Micropropagation of ornamental bromeliads (Bromeliaceae). Biotechnol Agric For 40:43–57

    Google Scholar 

  • Nassar AH, Khalifa SF, Hammouda FM, Shams KA (2001) In vitro propagation of Bixa orellana L. (annatto) an economically important plant newly introduced to Egypt. Egyptian J Bot 41:241–253

    Google Scholar 

  • Neto VBP, de Botelho MN, Aguiar R, Silva EAM, Otoni WC (2003) Somatic embryogenesis from immature zygotic embryos of annatto (Bixa orellana L.). In Vitro Cell Dev Biol-Plant 39:629–634

    Article  Google Scholar 

  • Neto VPB, Reis LB, Finger FL, Barros RS, Carvalho CR, Otoni WC (2009) Involvement of ethylene in the rooting of seedling shoot cultures of Bixa orellana L. In Vitro Cell Dev Biol-Plant 45:693–700

    Article  Google Scholar 

  • Parimalan R, Giridhar P, Gururaj HB, Ravishankar GA (2007) Organogenesis from cotyledon and hypocotyls derived explants of japhara (Bixa orellana L.). Act Bot Croat 66:153–160

    Google Scholar 

  • Parimalan R, Giridhar P, Gururaj HB, Ravishankar GA (2008) Mass multiplication of Bixa orellana L. through tissue culture for commercial propagation. Ind Crops and Prod 28:122–127

    Article  CAS  Google Scholar 

  • Parimalan R, Giridhar P, Gururaj HB, Ravishankar GA (2009) Micropropagation of Bixa orellana using phytohormones and triacontanol. Biol Plant 53:347–350

    Article  CAS  Google Scholar 

  • Parimalan R, Giridhar P, Ravishankar GA (2010) Enhanced shoot organogenesis in Bixa orellana L. in the presence of putrescine and silver nitrate. Plant Cell Tiss Organ Cult 105:285–290

    Article  Google Scholar 

  • Ramamurthy N, Savithramma N, Usha R, Swamy PM (1999) Multiple shoot induction and regeneration of japhara (Bixa orellana L.) through axillary bud derived callus cultures. J Plant Biol 26:231–235

    Google Scholar 

  • Satyanarayana A, Rao PG, Rao DG (2003) Chemistry, processing and toxicology of annatto (Bixa orellana L.). J Food Sci Technol 40:131–141

    CAS  Google Scholar 

  • Sharon M, D’Souza MC (2000) In vitro clonal propagation of annatto (Bixa orellana L.). Curr Sci 78:1532–1535

    Google Scholar 

  • Siril EA, Dhar U (1997) Micropropagation of mature Chinese tallow tree (Sapium sebiferum Roxb.). Plant Cell Rep 16:637–640

    Article  CAS  Google Scholar 

  • Snedecor GW, Cochran WG (1962) Statistical methods, 4th edn. The Iowa State University Press, Iowa

    Google Scholar 

  • Tiwari SK, Tiwari KP, Siril EA (2002) An improved micropropagation protocol for teak. Plant Cell Tiss Organ Cult 71:1–6

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Dr. Ashalatha S Nair, Professor and Head, Department of Botany for providing the facilities and Kerala State Council for Science, Technology and Environment (KSCSTE) Govt. of Kerala, Thiruvananthapuram, India, for the financial support (No.028/SRSLS/2007/CSTE).

Author information

Authors and Affiliations

  1. Department of Botany, University of Kerala, Kariavattom, Thiruvananthapuram, Kerala, 695 581, India

    Nisha Joseph, E. A. Siril & G. M. Nair

Authors
  1. Nisha Joseph
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E. A. Siril
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. G. M. Nair
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to E. A. Siril.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Joseph, N., Siril, E.A. & Nair, G.M. An efficient in vitro propagation methodology for Annatto (Bixa orellana L.). Physiol Mol Biol Plants 17, 263–270 (2011). https://doi.org/10.1007/s12298-011-0073-7

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12298-011-0073-7

Keywords

Search

Navigation