Plant Cell, Tissue and Organ Culture

, Volume 47, Issue 2, pp 135–144 | Cite as

A risk assessment study of plant genetic transformation usingAgrobacterium and implications for analysis of transgenic plants

  • Carol Barrette
  • Emily Cobb
  • Ronald McNicol
  • Gary Lyon
Original Research Papers

Abstract

Agrobacterium transformation systems forBrassica, Solanum andRubus, using carbenicillin, cefotaxime and ticaracillin respectively to eliminate contamination, were examined for the presence of residualAgrobacterium. The results indicated that none of the antibiotics in question, succeeded in eliminatingAgrobacterium and the contamination levels increased in explants from 12 to 16 weeks to such an extent thatSolanum cultures senesced and died. This may be due to the fact that four times the Minimum bactericidal concentration values (concentration to be used for elimination of contaminants in culture), for the three antibiotics, were higher than the concentrations employed in the culture medium. Contamination in shoot material decreased over 16 to 24 weeks possibly due to bacteriostatis and the use only of the apical node for further culture. The presence of the binary vector was also noted under non-selective conditions, even up to 6 months after transformation, where approx. 50% of contaminated material still harboured bacterial cells with the binary vector at levels of approx. 107 Colony forming units per gram.

Key words

binary vector minimum bactericidal concentration residual Agrobacterium 

Abbreviations

CFU

colony forming units

GUS

β-glucoronidase

PCR

polymerase chain reaction

MBC

minimum bactericidal concentration

NPTII

neomycin phosphotransferase

RAPD's

random amplified polymorphic DNA

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Copyright information

© Kluwer Academic Publishers 1997

Authors and Affiliations

  • Carol Barrette
    • 1
  • Emily Cobb
    • 2
  • Ronald McNicol
    • 2
  • Gary Lyon
    • 2
  1. 1.Teagasc, Kinsealy Research and Development CentreDublinIreland
  2. 2.Scottish Crop Research InstituteInvergowrieScotland

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