Plant Cell Reports

, Volume 12, Issue 7–8, pp 468–473 | Cite as

Stable transformation of the food yam Dioscorea alata L. by particle bombardment

  • Mahmut Tör
  • Charles Ainsworth
  • Sinclair H. Mantell


A biolistic particle gun was used to deliver genetic material into intact yam cells. Cultured suspension cells of D. alata were bombarded with microprojectiles coated with pBI221.2 DNA and histochemical assays were carried out to show transient GUS expression in bombarded cells. Stably transformed D. alata cells were recovered from cultured cells after bombardment with microprojectiles coated with pRT99gus harbouring both the nptII and uidA genes. Bombarded cells were selected on a medium containing geneticin (G418). Two months after bombardment, calli resistant to G418 were assayed for GUS expression. There was a 100% correlation between resistance to G418 and GUS expression. From these calli, four cell lines were established and GUS activity in each line was determined fluorometrically. The use of a specific GUS inhibitor showed that the GUS activity was due to the introduced uidA gene rather than to any intrinsic GUS-like activity originating from the plant. Incorporation of the introduced DNA into the plant genomic DNA was confirmed by Southern analysis.

Key words

Dioscorea alata GUS transformation particle gun 











Sodium dodecyl sulphate


Tris-acetate-EDTA buffer




Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Ammirato PV (1984) In: Ammirato PV, Evans DA, Sharp WR and Yamada Y, (eds) Handbook of Plant Cell Culture. Macmillan, New York pp. 327–354.Google Scholar
  2. Coursey DG (1967) Yams. Longmans, London.Google Scholar
  3. Creissen G, Smith C, Francis R, Reynolds H and Mullineaux P (1990) Plant Cell Reports 8:680–683.Google Scholar
  4. Degras L (1986) In: Maisonneuve G-P et Larose, (ed) L'igname: Plante a Tubercule Tropicale. Techniques Agricoles et Productions Tropicales. Paris.Google Scholar
  5. Draper J and Scott R (1988) In: Draper RSJ, Armitage P and Walden R (eds) Plant Genetic Transformation and Gene Expression. Blackwell Scientific Publications, Oxford pp. 199–136.Google Scholar
  6. Finer JJ and McMullen MD (1990) Plant Cell Reports 8:586–589.Google Scholar
  7. Fromm ME, Morrish F, Armstrong C, Williams R, Thomas J and Klein TM (1990) Bio/Technology 8:833–839Google Scholar
  8. Gordon-Kamm WJ, Spencer TM, Mangano ML, Adams TR, Daines RJ, Start WG, O'Brien JV, Chambers SA, Adams WR, Willets NG, Rice TB, Mackey CJ, Krueger RW, Kausch AP and Lemaux PG (1990) The Plant Cell 2:603–618.Google Scholar
  9. Hahn SK, Osiru DSO, Akoroda AO and Otoo A (1987) Outlook on Agriculture 16:105–110.Google Scholar
  10. Jefferson RA (1987) Plant Molecular Biology Reporter 5:387–405.Google Scholar
  11. Jefferson RA, Kavanagh TA and Bevan MW (1987) EMBO J. 6:3901–3907.Google Scholar
  12. Klein TM, Wolf ED, Wu R and Sanford JC (1987) Nature 327:70–73.Google Scholar
  13. Klein TM, Gradziel T, Fromm ME and Sanford JC (1988) Bio/Technology 6:559–563.Google Scholar
  14. Lassner MW, Peterson P and Yoder JI (1989) Plant Molecular Biology Reporter 7:116–128.Google Scholar
  15. Mendel RR, Müller B, Schulze J, Kolesnikov V and Zelenin A (1989) Theoretical Applied Genetics 78:31–34.Google Scholar
  16. Mendel RR, Clauss E, Hellmund R, Schulze J, Steinbiss, HH and Tewes A (1990) In: Nijkamp HJJ, Van Der Plas LHW and Van Aartrijk J (eds) Progress in Plant Cellular and Molecular Biology. Kluwer Academic Publishers, Amsterdam, The Netherlands, pp. 73–78.Google Scholar
  17. Onwueme IC (1978) The Tropical Tuber Crops. Yams, Cassava, Sweet Potato and Cocoyams. John Wiley & Sons., Chichester, New York.Google Scholar
  18. Paskowski J and Saul MW (1986) Methods in Enzymology 118:668–684.Google Scholar
  19. Potrykus I (1990) Bio/Technology 8:535–542.Google Scholar
  20. Sadik S (1976) A review of sexual propagation for yam improvement. Fourth Symp. Int'l. Soc. Tropical Root Crops, CIAT, Call, Colombia, International Development Research Centre.Google Scholar
  21. Sambrook J, Fritsch EF and Maniatis T (dy1989) Molecular Cloning. A laboratory manual. Cold Spring Harbor Laboratory Press.Google Scholar
  22. Schäfer W, Görz A and Kahl G (1987) Nature 327:529–532.Google Scholar
  23. Spencer TM, Gordon-Kamm WJ, Daines RJ, Start WG and Lemaux PG (1990) Theoretical Applied Genetics 79:625–631.Google Scholar
  24. Töpfer R, Schell J and Steinbiss H-H (1988) Nucleic Acid Research 16:8725.Google Scholar
  25. Tör M (1991) Ph.D. Dissertation, University of London, England.Google Scholar
  26. Tör M, Mantell SH and Ainsworth C (1992) Plant Cell Reports 11:452–456.Google Scholar
  27. Twyford CT (1992) Ph.D. Dissertation (in preparation), University of London, England.Google Scholar
  28. Vasil V, Castillo AM, Fromm ME and Vasil IK (1992) Bio/Technology 10:667–674.Google Scholar
  29. Waldon R (1988) Genetic Transformation in Plants. Open University Press, Milton Keynes.Google Scholar
  30. Wilson LA (1986) In: Harlevy AA (ed), Handbook of Flowering. CRC Press Florida, pp 450–454.Google Scholar
  31. Xinhua F, Qiquan S and Xingcun J (1986) Genetic Manipulation Crops Newsletter 2:52–59.Google Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Mahmut Tör
    • 1
  • Charles Ainsworth
    • 1
  • Sinclair H. Mantell
    • 1
  1. 1.Wye College, University of LondonWye, AshfordUK

Personalised recommendations