Skip to main content
SpringerLink
Log in

Direct shoot organogenesis on hypocotyl explants with collar region from in vitro seedlings of Coffea canephora Pierre ex. Frohner cv. C × R and Agrobacterium tumefaciens-mediated transformation

  • Original Paper
  • Published:
Plant Cell, Tissue and Organ Culture (PCTOC) Aims and scope Submit manuscript

Abstract

Direct differentiation of shoot buds from the collar region of hypocotyl segments of Coffea canephora was obtained on Murashige and Skoog (MS) medium supplemented with 40 μM silver nitrate (AgNO3) and growth regulators indole-3-acetic acid (IAA) and N6 benzyladenine (BA). The highest response to shoot differentiation of 60% frequency and the maximum number of multiple shoots (2–3) per explant were obtained on MS medium containing 8.87 μM BA and 2.85 μM IAA. Apart from this, 70% of hypocotyl explants produced yellow friable embryogenic callus and also globular primary somatic embryos. Subsequent transfer onto the same medium induced secondary somatic embryogenesis. The micro-shoots, upon transfer to the same medium, in the following 6 weeks developed into 4-cm-long shoots with a single root. Further subculturing onto the same medium induced 4–5 roots in a 4-week period. The resulting plantlets were hardened and transferred to micro-pots containing sand:compost mixture (1:2), where 65% of them survived and resumed growth. By using optimal levels of AgNO3, it was possible to obtain effective direct organogenesis and embryogenesis. This system was used for genetic transformation using Agrobacterium tumefaciens. A stable transformation frequency of 2–5% was obtained when both types of explants, i.e., hypocotyl explants with collar region or hypocotyl explants without collar region, were co-cultivated with A. tumefaciens GV 3101 harboring pCAMBIA 1305.2 binary vector. This is the first report of a hypocotyl collar region-based Agrobacterium-mediated transformation protocol for the economically important tropical plant C. canephora.

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. 1A–I
Fig. 2A–F
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

AgNO3 :

Silver nitrate

BA:

Benzyladenine

IAA:

Indole-3-aceticacid

MS:

Murashige and Skoog

NAA:

α-naphthaleneacetic acid

References

  • Beyer EM (1976) A potent inhibitor of ethylene action in plants. Plant Physiol 58:268–271

    Article  CAS  PubMed  Google Scholar 

  • Beyer EM (1979) Effect of silver ion, carbon dioxide, and oxygen on ethylene action and metabolism. Plant Physiol 63:163–173

    Article  Google Scholar 

  • Broothaerts W, Mitchell HJ, Weir B, Kaines S, Smith LMA, Yang W, Mayer JE, Roa-Rodríguez C, Jefferson RA (2005) Gene transfer to plants by diverse species of bacteria. Nature 433:629–633

    Article  CAS  PubMed  Google Scholar 

  • Ducos JP, Labbe G, Lambot C, Pétiard V (2007) Pilot scale process for the production of pre-germinated somatic embryos of selected robusta (Coffea canephora) clones. In Vitro Cell Dev Biology Plant 43:652–659

    Article  Google Scholar 

  • Duncan DR, Williams ME, Zehr BE, Widholm JM (1985) The production of callus capable of plant regeneration from immature embryos of numerous Zea mays genotypes. Planta 165:322–332

    Article  CAS  Google Scholar 

  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158

    Article  CAS  PubMed  Google Scholar 

  • Giridhar P, Reddy BO, Ravishankar GA (2001) Silver nitrate influences in vitro shoot multiplication and root formation in Vanilla planifolia Andr. Current Sci 81:1166–1170

    CAS  Google Scholar 

  • Giridhar P, Indu EP, Vijaya Ramu D, Ravishankar GA (2003) Effect of silver nitrate on in vitro shoot growth of coffee. Tropical Sci 43:144–146

    Article  Google Scholar 

  • Giridhar P, Indu EP, Vinod K, Chandrashekar A, Ravishankar GA (2004) Direct somatic embryogenesis from Coffea arabica L. and Coffea canephora P ex Fr. under the influence of ethylene action inhibitor-silver nitrate. Acta Physiol Plantarum 26:299–305

    Article  CAS  Google Scholar 

  • Hatanaka T, Choi YE, Kusano T, Sano H (1999) Transgenic plants of coffee Coffea canephora from embryogenic callus via Agrobacterium tumefaciens-mediated transformation. Plant Cell Rep 19:106–110

    Article  CAS  Google Scholar 

  • Kong L, Yeung EC (1994) Effects of ethylene and ethylene inhibitors on white spruce somatic embryo maturation. Plant Sci 104:71–80

    Article  CAS  Google Scholar 

  • Kumar V, Satyanarayana KV, Sarala Itty S, Indu EP, Giridhar P, Chandrasekar A, Ravishankar GA (2006) Stable transformation and direct regeneration in Coffea canephora P ex. Fr. by Agrobacterium rhizogenes mediated transformation without hairy-root phenotype. Plant Cell Rep 25:214–222

    Article  CAS  PubMed  Google Scholar 

  • Kumar V, Giridhar P, Ravishankar GA (2009) AgNO3—a potential regulator of ethylene activity and plant growth modulator. Electron J Biotechnol vol 12, no 2, Issue of April 15, 2009

  • Mishra MK, Sreenath HL (2004) High-efficiency Agrobacterium-mediated transformation of Coffee (Coffea canephora Pierre ex. Frohner) using hypocotyls explants. In: Proceedings of the 19th International Coffee Science Congress (ASIC Colloquium), Bangalore, India

  • 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 

  • Nissen P (1994) Stimulation of somatic embryogenesis in carrot by ethylene: effects of modulators of ethylene biosynthesis and action. Physiol Plant 92:397–403

    Article  CAS  Google Scholar 

  • Pestana MC, Lacorte C, de Freitas VG, de Oliveira DE, Mansur E (1999) In vitro regeneration of peanut (Arachis hypogaea L.) through organogenesis: effect of culture temperature and silver nitrate. In Vitro Cell Dev Biol Plant 35:214–216

    Article  Google Scholar 

  • Parvatam G, Mahesh V, Ravishankar GA, De Kochko A, Campa C (2006) Functional validation of Coffea PAL genes using genetic engineering. In: Proceedings of the 21st International Coffee Congress, ASIC_2006, Montpellier, France, 10–15 September, 2006, pp 702–705

  • Reddy BO, Giridhar P, Ravishankar GA (2001) In vitro rooting of Decalepis hamiltonii Wight and Arn., an endangered shrub by auxins and root-promoting agents. Curr Sci 81:1479–1482

    CAS  Google Scholar 

  • Ribas AF, Pereira LFP, Vierira LGE (2006) Genetic transformation of coffee. Braz J Plant Physiol 18:83–94

    Article  CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning—a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Santa RA, Sreenath HL, Srinivasan CS (1999) Morphogenetic responses of cultured root and hypocotyl tissues of Coffea arabica L. J Coffee Res 27:87–92

    Google Scholar 

  • Santana-Buzzy N, Rojas-Herrera R, Galaz-Ávalos MR, Ku-Cauich JR, Mijangos-Cortés J, Gutiérrez-Pacheco LC, Canto A, Quiroz-Figueroa F, Loyola-Vargas VM (2007) Advances in coffee tissue culture and its practical applications. In Vitro Cell Dev Biol Plant 43:507–520

    Article  CAS  Google Scholar 

  • Singh SK, Meghwal PR, Sharma HC, Singh SP (2002) Direct shoot organogenesis on hypocotyl explants from in vitro germinated seedlings of Psidium guajava L. cv. Allahabad Safeda. Scientia Horticult 95:213–221

    Article  CAS  Google Scholar 

  • Sondahl MR, Sharp WR (1977) High frequency induction of somatic embryos in cultured leaf explants of Coffea arabica L. Z Pflanzenphysiol 81:395–408

    Google Scholar 

  • Sonia PKJ, Gulati A, Dahiya S (1998) Direct organogenesis in hypocotyl cultures of Tamarindus indica. Biol Plant 41:331–337

    Article  Google Scholar 

  • Sridevi V, Giridhar P (2008) Recent trends in coffee biotechnology towards quality improvement—a review. Indian J Bot Res 4(1):5–12

    Google Scholar 

  • Staritsky G, Van Hasselt GAM (1980) The synchronized mass propagation of Coffea canephora in vitro. In: Proceedings of the 9th International Scientific Colloquium on Coffee, ASIC, London, 1980, pp 597–602

  • Uefuji H, Ogita S, Yamaguchi Y, Koizumi N, Sano H (2003) Molecular cloning and functional characterization of three distinct N-methyltransferases involved in the caffeine biosynthetic pathway in coffee plants. Plant Physiol 132:372–380

    Article  CAS  PubMed  Google Scholar 

  • Yasuda T, Fujii Y, Yamaguchi T (1985) Embryogenic callus induction from Coffea arabica leaf explants by benzyladenine. Plant Cell Physiol 26:595–597

    CAS  Google Scholar 

Download references

Acknowledgments

The authors are thankful to the Department of Biotechnology, Government of India, New Delhi, for the financial assistance.

Author information

Authors and Affiliations

  1. Plant Cell Biotechnology Department, Central Food Technological Research Institute, Council of Scientific and Industrial Research, Mysore, 570 020, India

    V. Sridevi, P. Giridhar, P. S. Simmi & G. A. Ravishankar

Authors
  1. V. Sridevi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. P. Giridhar
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. P. S. Simmi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. G. A. Ravishankar
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to P. Giridhar.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sridevi, V., Giridhar, P., Simmi, P.S. et al. Direct shoot organogenesis on hypocotyl explants with collar region from in vitro seedlings of Coffea canephora Pierre ex. Frohner cv. C × R and Agrobacterium tumefaciens-mediated transformation. Plant Cell Tiss Organ Cult 101, 339–347 (2010). https://doi.org/10.1007/s11240-010-9694-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-010-9694-8

Keywords

Search

Navigation