Plant Cell Reports

, Volume 14, Issue 5, pp 285–289

Transformation of sunflower (Helianthus annuus L.) following wounding with glass beads

  • W. Scott Grayburn
  • Brady A. Vick
Article

Abstract

A procedure was developed for transformation of Helianthus annuus (sunflower) using Agrobacterium tumefaciens. Cotyledons were removed from young seedlings, and the remaining tissue was uniformly wounded by shaking with glass beads. The wounded tissue was then co-cultivated with a hypervirulent strain of Agrobacterium tumefaciens harboring the binary plasmid pCNL56. Minimal use of defined medium was required, and no callus was observed. The polymerase chain reaction (PCR) followed by DNA hybridization demonstrated the presence of gusA DNA from pCNL56 in total leaf DNA of 6 primary transformants and 2 progeny plants. No Agrobacterium DNA was detected in total DNA from transformed sunflower leaves that was amplified with primers specific to the miaA chromosomal gene of Agrobacterium. Foreign DNA was also detected in the next generation. β-Glucuronidase (GUS) activity was demonstrated for 5 of the T2 transgenic plants. Grafting was used to increase the number of seeds present on plants that had undergone tissue culture manipulations.

Abbreviations

PCR

polymerase chain reaction

EMBL

European Molecular Biology Laboratory

M U

7-hydroxy-4-methylumbelliferone

GUS

β-Glucuronidase

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References

  1. Bidney D, Scelonge C, Martich J, Burrus M, Sims L, Huffman G (1992) Plant Mol Biol 18:301–313Google Scholar
  2. Chraibi KM, Castelle J-C, Latche A, Roustan J-P, Fallot J (1992) Plant Cell Rep 10:617–620Google Scholar
  3. Ditta G, Stanfield S, Corbin D, Helinski DR (1980) Proc Natl Acad Sci USA 77:7347–7351Google Scholar
  4. Espinasse A, Lay C (1989) Crop Sci 29:201–205Google Scholar
  5. Everett NP, Robinson KEP, Mascarenhas D (1987) Bio/technology 5:1201–1204Google Scholar
  6. Feinberg AP, Vogelstein B (1983) Anal Biochem 132:6–13PubMedGoogle Scholar
  7. Feinberg AP, Vogelstein B (1984) Addendum. Anal Biochem 137:266–267Google Scholar
  8. Gamborg OL, Miller RA, Ojima K (1968) Exp Cell Res 50:151–158PubMedGoogle Scholar
  9. Gray J, Wang J, Gelvin SB (1992) J Bacteriol 174:1086–1098Google Scholar
  10. Grayburn WS (1993) In: Proc 15th Sunflower Research Workshop, pp 93–97Google Scholar
  11. Harada WS (1981) PhD Dissertation. North Dakota State University, Fargo, NDGoogle Scholar
  12. Hood EE, Helmer GL, Fraley RT, Chilton M-D (1986) J Bacteriol 168:1291–1301PubMedGoogle Scholar
  13. Jefferson RA, Burgess SM, Hirsh D (1986) Proc Natl Acad Sci USA 83:8447–8451PubMedGoogle Scholar
  14. Lee S, Rasheed S (1990) BioTechniques 9:676–679Google Scholar
  15. Li X-Q, Liu C-N, Ritchie SW, Peng J-Y, Gelvin SB, Hodges TK (1992) Plant Mol Biol 20:1037–1048PubMedGoogle Scholar
  16. McCabe DE, Swain WF, Martinell BJ, Christou P (1988) Bio/technology 6:923–926Google Scholar
  17. Rogers SO, Bendich AJ (1985) Plant Mol Biol 5:69–76Google Scholar
  18. Schrammeijer B, Sijmons PC, van den Elzen PJM, Hoekema A (1990) Plant Cell Rep 9:55–60Google Scholar
  19. Seiler GJ (1992) Field Crops Res 30:191–194Google Scholar
  20. Tartof K, Hobbs CA (1987) Bethesda Res Lab Focus 9:12Google Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • W. Scott Grayburn
    • 1
  • Brady A. Vick
    • 1
  1. 1.United States Department of AgricultureAgricultural Research Service, Northern Crop Science LaboratoryFargoUSA

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