Skip to main content
SpringerLink
Log in

In vitro morphogenesis in zygotic embryo cultures of neem (Azadirachta indica A. Juss.)

  • Cell Biology and Morphogenesis
  • Published:
Plant Cell Reports Aims and scope Submit manuscript

Abstract

Immature zygotic embryo cultures of neem yielded highly regenerative cultures, with the response varying with the embryo stage at culture. Early dicotyledonous stage embryos were the most responsive followed by torpedo stage embryos. The embryo cultures differentiated three types of regenerants: somatic embryos (SEs), shoot buds and neomorphs. SEs exhibited morphological abnormalities such as pluricotyledony, fusion of cotyledons and absence of cotyledons. Although these SEs showed secondary embryogenesis, the occurrence of normal dicotyledonous embryos was extremely rare. On MS basal medium 3% of SEs developed a long tap root but a plumular shoot did not appear. However, it was possible to regenerate plantlets from immature zygotic embryo cultures of neem via neomorph formation and adventitious shoot bud formation. The transplantation survival of these plants was more than 80%.

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
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

BAP :

6-Benzylamino purine

CH :

Casein hydrolysate

2,4-D :

2,4-Dichlorophenoxyacetic acid

GA 3 :

Gibberellic acid

IAA :

Indole-3-acetic acid

IBA :

Indole-3-butyric acid

NAA :

α-Naphthaleneacetic acid

SE :

Somatic embryo

References

  • Abubacker MN, Alagumanian S (1999) In vitro organogenesis of various explants of Azadirachta indica A. Juss. (Neem). Plant Tissue Cult 9:177–180

    Google Scholar 

  • Akhtar N, Kumari N, Pandey S, Ara H, Singh M, Jaiswal U, Jaiswal VS, Jain SM (2000) Somatic embryogenesis in tropical fruit trees. In: Jain SM, Gupta PK, Newton RJ (eds) Somatic embryogenesis in woody plants, vol 6. Kluwer, Dordrecht, pp 93–140

  • Bhojwani SS, Razdan MK (1996) Plant tissue culture: theory and practice. Elsevier, Amsterdam

    Google Scholar 

  • Chaturvedi R, Razdan MK, Bhojwani SS (2003a) Production of haploids of neem (Azadirachta indica A. Juss.) by anther culture. Plant Cell Rep 21:531–537

    CAS  PubMed  Google Scholar 

  • Chaturvedi R, Razdan MK, Bhojwani SS (2003b) An efficient protocol for the production of triploid plants from endosperm callus of neem, Azadirachta indica A.Juss. J Plant Physiol 160:557–564

    CAS  PubMed  Google Scholar 

  • Cruz GS, Canhato JM, Abreu MAV (1990) Somatic embryogenesis and plant regeneration from zygotic embryos of Feijoa sellowiana Berg. Plant Sci 66:263–270

    Article  CAS  Google Scholar 

  • Custers JBM, Bergervoet JHW (1990) In vitro culture of embryos of Cucumis spp.: heart-stage embryos have a higher ability of direct plant formation than advanced stage embryos. Sex Plant Reprod 3:152–159

    Google Scholar 

  • Custers JBM, den Nijs APM (1986) Effects of aminoethoxyvinylglycine (AVG), environment, and genotype in overcoming hybridization barriers between Cucumis species. Euphytica 35:639–647

    CAS  Google Scholar 

  • Ezumah BS (1986) Germination and storage of neem (Azadirachta indica A. Juss.) seed. Seed Sci Technol 14:593–600

    Google Scholar 

  • Gamene CS, Kraak HL, Van Piljen JG, de Vos CHR (1996) Storage behavior of neem (Azadirachta indica A. Juss.) seeds from Burkina Faso. Seed Sci Technol 24:441–448

    Google Scholar 

  • Garin E, Grenier E, Grenier-De-March G (1997) Somatic embryogenesis in wild cherry (Prunus avium). Plant Cell Tissue Organ Cult 48:83–92

    Article  CAS  Google Scholar 

  • Islam R, Joarder N, Joarder OI, Hossain M, Rahman SM, Naderuzzaman (1993) Somatic embryogenesis and organogenesis in neem. In: Proceedings of World Neem Conference, Bangalore, pp 955–960

  • Liu W, Xu Z, Chua N (1993) Auxin polar transport is essential for the establishment of bilateral symmetry during early plant embryogenesis. Plant Cell 5:621–630

    CAS  PubMed  Google Scholar 

  • Maithani GP, Bahuguna V, Rawat MMS, Sood OP (1989) Fruit maturity and interrelated effects of temperature and container on longevity of neem (Azadirachta indica) seed. Indian For 115:89–97

    Google Scholar 

  • Merkle SA, Parrott WA, Williams EG (1990) Applications of somatic embryogenesis and embryo cloning. In: Bhojwani SS (ed) Plant tissue culture: applications and limitations. Elsevier, Amsterdam, pp 67–101

  • Monnier M (1990) Zygotic embryo culture. In: Bhojwani SS (ed) Plant tissue culture: applications and limitations. Elsevier, Amsterdam, pp 366–393

  • Muralidharan EM, Mascarenhas AF (1989) In vitro morphogenesis in Azadirachta indica A. Juss. and Eucalyptus citriodora Hook. f. Tissue culture and biotechnology of medicinal and aromatic plants, CIMAP, Lucknow, pp 49–55

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

    CAS  Google Scholar 

  • Nirmalakumari A, Ramaswamy NM, Sree Rangaswamy SR (1993) Tissue culture studies in neem (Azadirachta indica A. Juss.). In: Proceedings of the World Neem Conference, Bangalore, pp 981–992

  • Rodriguez APM, Wetzstein HY (1994) The effect of auxin type and concentration on pecan (Carya illinoinensis) somatic embryo morphology and subsequent conversion into plants. Plant Cell Rep 13:607–611

    CAS  Google Scholar 

  • Sacande M, Golovina EA, Van Aelst AC, Hoekstra FA (2001) Viability loss of neem (Azadirachta indica) seeds associated with membrane phase behaviour. J Exp Bot 52:919–931

    Article  CAS  PubMed  Google Scholar 

  • Salvi ND, Singh H, Tivarekar S, Eapen S (2001) Plant regeneration from different explants of neem. Plant Cell Tissue Organ Cult 65:159–162

    Article  CAS  Google Scholar 

  • Sarker RH, Islam MR, Hoque MI (1997) In vitro propagation of Neem (Azadirachta indica A. Juss.) plants from seedling explants. Plant Tissue Cult 7:125–133

    Google Scholar 

  • Sharma AK, Sharma M, Chaturvedi HC (1999) Conservation of phytodiversity of Azadirachta indica A. Juss. through in vitro strategies. In: National symposium on role of plant tissue culture in biodiversity conservation and economic development. G.B. Pant Institute of Himalayan Environment and Development, Almora, pp 1–13

  • Shrikhande M, Thengane SR, Mascarenhas AF (1993) Somatic embryogenesis and plant regeneration in Azadirachta indica A. Juss. In vitro Cell Dev Biol 29P:38–42

    CAS  Google Scholar 

  • Su WW, Hwang WI, Kim SY, Sagawa Y (1997) Induction of somatic embryogenesis in Azadirachta indica. Plant Cell Tissue Organ Cult 50:91–95

    Article  Google Scholar 

  • Thorpe TA, Stasolla C (2001) Somatic embryogenesis. In: Bhojwani SS, Soh WY (eds) Current trends in the embryology of angiosperms. Kluwer, Dordrecht, pp 219–236

Download references

Acknowledgements

Rakhi Chaturvedi thanks Professor S.P. Bhatnagar for his critical analysis of the manuscript and the Council for Scientific and Industrial Research (CSIR), India, for financial assistance.

Author information

Author notes

  1. R. Chaturvedi

    Present address: 310, School of Life Sciences, Jawaharlal Nehru University, New Delhi 110 067, India

  2. M. K. Razdan

    Present address: Department of Botany, Ramjas College, University of Delhi, Delhi 110 007, India

  3. S. S. Bhojwani

    Present address: Dayalbagh Educational Institute (Deemed University), Dayalbagh, Agra 282 005, India

Authors and Affiliations

  1. Department of Botany, University of Delhi, Delhi-7, India

    R. Chaturvedi, M. K. Razdan & S. S. Bhojwani

Authors
  1. R. Chaturvedi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. K. Razdan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S. S. Bhojwani
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to R. Chaturvedi.

Additional information

Communicated by W. Harwood

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chaturvedi, R., Razdan, M.K. & Bhojwani, S.S. In vitro morphogenesis in zygotic embryo cultures of neem (Azadirachta indica A. Juss.). Plant Cell Rep 22, 801–809 (2004). https://doi.org/10.1007/s00299-004-0768-0

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-004-0768-0

Keywords

Search

Navigation