Skip to main content
SpringerLink
Log in

Role of phytohormones and nitrogen in somatic embryogenesis induction in cell culture derived from leaflets of Azadirachta indica

  • Brief Communication
  • Published:
Biologia Plantarum

Abstract

A protocol for somatic embryogenesis in Azadirachta indica A Juss. has been standardized using in vivo leaflets. Experiments were carried out to examine the effect of various auxins, cytokinins, sucrose, inorganic and organic salts on subsequent somatic embryo induction and maturation. Embryogenic calli were induced on Murashige and Skoog (MS) medium supplemented with 1.5 mg dm−3 kinetin and 1.5 mg dm−3 indole-3-acetic acid and subsequently all the stages of somatic embryo development (globular, cordate, torpedo and cotyledonary) were observed. Maturation of these embryos was accomplished with the same growth regulators after three subcultures. The histological study of somatic embryos showed resembles to zygotic embryos. The matured somatic embryos were transferred onto half strength MS-medium devoid of growth regulators for their germination (82 %). Plantlets were acclimatized in the field with a survival rate of 80–83.5 %.

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

Abbreviations

2,4-D:

2,4-dichlorophenoxyacetic acid

BAP:

N 6-benzylaminopurine

IAA:

indole-3-acetic acid

IBA:

indole-3-butyric acid

KIN:

kinetin

MS:

Murashige and Skoog medium

NAA:

naphthaleneacetic acid

References

  • Akula, C., Akula, A., Drew, R.: Somatic embryogenesis in clonal neem, Azadirachta indica A juss and analysis for in vitro azadirachtin production. — In Vitro cell. dev. Biol. Plant 39: 304–310, 2003.

    CAS  Google Scholar 

  • Arora, D.K., Suri, S.S., Ramawat, K.G., Merillan, J.M.: Factors affecting somatic embryogenesis in long term callus cultures of ’safed musli’ (Chlorophytum orivilianum), an endangered wonder herb. — Indian J. exp. Biol. 37: 75–82, 1999.

    Google Scholar 

  • Chengalrayan, K., Sathaya, S.S., Hazra, S.: Somatic embryogenesis from mature embryo-derived leaflets of peanut (Arachis hypogaea L.). — Plant Cell Rep. 13: 578, 1994.

    Article  CAS  Google Scholar 

  • Cuenca, B., San-Jose, M.C., Martinez, M.T., Ballester, A., Vieitez, A.M.: Somatic embryogenesis from stem and leaf explant of Quercus robur L. — Plant Cell Rep. 18: 538–543, 1999.

    Article  CAS  Google Scholar 

  • Eapen, S., George, L.: Somatic embryogenesis in peanut: influence of growth regulators and sugars. — Plant Cell Tissue Organ Cult. 35: 151–156, 1993.

    Article  CAS  Google Scholar 

  • Emons, A.M.C., Droppers, A.S., Toorn, C.V.D.: The influence of sucrose, mannitol, L-proline, abscisic acid and gibbrellic acid on the maturation of somatic embryos of Zea mays L. from suspension culture. — J. Plant Physiol. 142: 597–604, 1993.

    CAS  Google Scholar 

  • Ganesh Kumari, K., Ganesan, M., Jayabalan, N.: Somatic embryogenesis and plant regeneration in Ricinus communis. — Biol. Plant. 52: 17–25, 2008.

    Article  Google Scholar 

  • Garin, E., Grenier, E., March, G.G.: Direct somatic embryogenesis in wild cherry (Prunus avium). — Plant Cell Tissue Organ Cult. 48: 83–91, 1997.

    Article  CAS  Google Scholar 

  • Martin, G.G., González-Benito, M.E., Manzanera, J.A.: Quercus suber L. somatic embryo germination and plant conversion: pretreatments and germination conditions. — In vitro cell. dev. Biol. Plant 37: 190–198, 2001.

    Article  Google Scholar 

  • Mathur, S., Shekhawat, G.S., Batra, A.: Somatic embryogenesis and plantlet regeneration from cotyledon explants of Salvadora persica L. — Phytomorphology 58: 57–63, 2008.

    Google Scholar 

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

    Article  CAS  Google Scholar 

  • Murthy, B.N.S., Saxena, P.K.: Somatic embryogenesis and plant regeneration of neem (Azadirachta indica A. Juss). — Plant Cell Rep. 17: 469–475, 1998.

    Article  CAS  Google Scholar 

  • Rout, G.R., Debata, B.K., Das, P.: Somatic embryogenesis in callus culture of Rosa hybrida L. cv. ‘Landora’ — Plant Cell Tissue Organ Cult. 27: 65–69, 1991.

    Article  CAS  Google Scholar 

  • Satdive, R.K., Fulzele, D.P., Eapen, S.: Biotechnology of neem: Azadirachta indica A Juss. — In: Khan, I.A., Khanum, A. (ed.). Role of Biotechnology in Medicinal and Aromatic Plants. Vol. IV. Pp. 168–181. Ukaaz Publication, Hyderabad 2001.

    Google Scholar 

  • Shekhawat, G.S., Batra, A., Mathur S.: A reliable in vitro protocol for rapid mass propagation of Azadirachta indica A. Juss. — J. Plant Biol. 29: 109–112, 2002.

    Google Scholar 

  • Srivastava, P., Singh, M., Mathur, R., Chaturvedi, R.: In vitro organogenesis and plant regeneration from unpollinated ovary cultures of Azadirachta indica. — Biol. Plant. 53: 360–364, 2009.

    Article  CAS  Google Scholar 

  • Su, W.W., Hwang, W.L., Kim, S.Y., Sangawa, Y.: Induction of somatic embryogenesis in Azadirachta indica. A. Juss. — Plant Cell Tissue Organ Cult. 50: 91–95, 1997.

    Article  Google Scholar 

  • Terzi, M., Loschiavo, F.: Somatic embryogenesis. — In: Bhojwani, S.S.: Plant Tissue Culture, Applications and Limitations. Pp. 54–66. Elsevier, Amsterdam 1990.

    Google Scholar 

  • Thengane, S., Joshi, M., Mascarenhas, A.F.: Somatic embryogenesis in neem (Azadirachta indica. A Juss). — In: Jain, S. M., Gupta, P.K., Newton, R.J. (ed.): Somatic Embryogenesis in Woody Plants. Vol. 2. Angiosperms. Pp. 357–374. Kluwer Academic Publishers, Dordrecht 1995.

    Google Scholar 

  • Tiwari, D.W.: Monograph on Neem (Azadirachta indica. A.Juss.). — International Book Distributors, Deharadun 1992.

    Google Scholar 

  • Vijaya Kumar, J., Ranjitha Kumari, B.D., Castaño, E.: Cyclic somatic embryogenesis and efficient plant regeneration from callus of safflower. — Biol. Plant. 52: 429–436, 2008.

    Article  Google Scholar 

  • Wilhelm, E.: Somatic embryogenesis in oak Quercus sps. — In Vitro cell. dev. Biol. Plant 36: 349–357, 2000.

    Article  CAS  Google Scholar 

  • Yang, X.M., An, L.Z., Xiong, Y.C., Zhang, J.P., Li, Y., Xu, S.J.: Somatic embryogenesis from immature zygotic embryos and monitoring genetic fidelity of regenerated plants in grapevine. — Biol. Plant. 52: 209–214, 2008.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Bioscience and Biotechnology, Banasthali University, Banasthali, 304022, Rajasthan, India

    G. S. Shekhawat

  2. Department of Botany, Government Post Graduate College, Kot Putli, 303108, Jaipur, India

    S. Mathur

  3. Biotechnology Laboratory, Department of Botany, University of Rajasthan, Jaipur, 302004, India

    A. Batra

Authors
  1. G. S. Shekhawat
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S. Mathur
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A. Batra
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to G. S. Shekhawat.

Additional information

Acknowledgement: G.S. Shekhawat greatly acknowledges the grant received from CSIR, New Delhi, as Senior Research fellow.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Shekhawat, G.S., Mathur, S. & Batra, A. Role of phytohormones and nitrogen in somatic embryogenesis induction in cell culture derived from leaflets of Azadirachta indica . Biol Plant 53, 707–710 (2009). https://doi.org/10.1007/s10535-009-0127-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10535-009-0127-7

Additional key words

Search

Navigation