Skip to main content

Transformation of Nuclear DNA in Meristematic and Embryogenic Tissues

  • Protocol
  • First Online:
Biolistic DNA Delivery

Part of the book series: Methods in Molecular Biology ((MIMB,volume 940))

Abstract

Particle bombardment/biolistic delivery is a very popular method of genetic transformation of diverse targets including cells and intact tissues. Delivery of DNA through particle bombardment is genotype and species independent, nevertheless, an efficient protocol for large-scale generation of transgenic plants through embryogenic tissues with a high (≥80%) shoot regeneration efficiency is a prerequisite. Young embryogenic tissues or multiple shoot buds in early stages of induction are the most suited target tissues for recovery of transgenic plants. We describe the protocol for delivery of foreign genes using particle delivery system (Biorad gene gun, PDS-1000/He) in to the meristematic tissues of embryonic axes derived from mature seeds of castor. With the optimized physical and biological parameters, putative transformants were obtained at a frequency of 1.4% through particle gun bombardment of castor embryo axes. Also, transformation of embryogenic calli of sorghum using particle inflow gun (PIG) is described.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Sanford JC et al (1987) Delivery of substances into cells and tissues using a particle bombardment process. Particul Sci Technol 5:27–37

    Article  CAS  Google Scholar 

  2. Birch RG, Bower R (1994) Principles of gene transfer using particle bombardment. In: Yang N, Christou P (eds) Particle bombardment technology for gene transfer. Oxford University Press, New York, pp 3–38

    Google Scholar 

  3. Vasil V, Vasil IK (1981) Somatic embryogenesis and plant regeneration from tissue cultures of Pennisetum americanum and P. americanum x P. purpureum hybrid. Am J Bot 68:864–872

    Article  Google Scholar 

  4. McCabe DE et al (1988) Stable transformation of soybean (Glycine max L.) by particle acceleration. Bio/Technol 6:923–926

    Article  Google Scholar 

  5. Brar GS et al (1994) Recovery of transgenic peanut (Arachis hypogaea L.) plants from elite cultivars utilizing ACCELL technology. Plant J 5:745–753

    Article  Google Scholar 

  6. Sailaja M, Tarakeswari M, Sujatha M (2008) Stable genetic transformation of castor (Ricinus communis L.) via particle gun-mediated gene transfer using embryo axes from mature seeds. Plant Cell Rep 27:1509–1519

    Article  PubMed  CAS  Google Scholar 

  7. Sujatha M et al (2009) Expression of the cry1EC gene in castor (Ricinus communis L.) confers field resistance to tobacco caterpillar (Spodoptera litura Fabr) and castor semilooper (Achoea janata L.). Plant Cell Rep 28:935–946

    Article  PubMed  CAS  Google Scholar 

  8. 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 

  9. Walter C et al (1994) A biolistic approach for the transfer and expression of a gusA reporter gene in embryogenic cultures of Pinus radiata. Plant Cell Rep 14:69–74

    Article  CAS  Google Scholar 

  10. Lis JT (1980) Fractionation of DNA fragments by polyethylene glycol induced precipitation. Methods Enzymol 65:347–353

    Article  PubMed  CAS  Google Scholar 

  11. Medford J (1992) Vegetative apical meristems. Plant Cell 4:1029–1039

    PubMed  Google Scholar 

  12. Vain P, McMullen MD, Finer JJ (1993) Osmotic treatment enhances particle bombardment-mediated transient and stable transformation of maize. Plant Cell Rep 12:84–88

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Mulpuri Sujatha .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2013 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Sujatha, M., Visarada, K.B.R.S. (2013). Transformation of Nuclear DNA in Meristematic and Embryogenic Tissues. In: Sudowe, S., Reske-Kunz, A. (eds) Biolistic DNA Delivery. Methods in Molecular Biology, vol 940. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-110-3_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-110-3_3

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-109-7

  • Online ISBN: 978-1-62703-110-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics