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Genetics, Vol. 152, 1111-1122, July 1999, Copyright © 1999

Homeologous Plastid DNA Transformation in Tobacco Is Mediated by Multiple Recombination Events

Tony A. Kavanagha, Nguyen Duc Thanhb, Nga Thi Laoa, Noreen McGrathb,c, Stefan O. Peterb, Eva M. Horváthb, Philip J. Dixc, and Peter Medgyesyb
a Department of Genetics, Trinity College, University of Dublin, Dublin 2, Ireland,
b Biological Research Center, Hungarian Academy of Sciences, H-6701 Szeged, Hungary
c Department of Biology, National University of Ireland, Maynooth, County Kildare, Ireland

Corresponding author: Peter Medgyesy, Biological Research Center, Hungarian Academy of Sciences, P.O. Box 521, H-6701 Szeged, Hungary., pmedgyesy{at}nucleus.szbk.u-szeged.hu (E-mail)

Communicating editor: K. J. NEWTON

Efficient plastid transformation has been achieved in Nicotiana tabacum using cloned plastid DNA of Solanum nigrum carrying mutations conferring spectinomycin and streptomycin resistance. The use of the incompletely homologous (homeologous) Solanum plastid DNA as donor resulted in a Nicotiana plastid transformation frequency comparable with that of other experiments where completely homologous plastid DNA was introduced. Physical mapping and nucleotide sequence analysis of the targeted plastid DNA region in the transformants demonstrated efficient site-specific integration of the 7.8-kb Solanum plastid DNA and the exclusion of the vector DNA. The integration of the cloned Solanum plastid DNA into the Nicotiana plastid genome involved multiple recombination events as revealed by the presence of discontinuous tracts of Solanum-specific sequences that were interspersed between Nicotiana-specific markers. Marked position effects resulted in very frequent cointegration of the nonselected peripheral donor markers located adjacent to the vector DNA. Data presented here on the efficiency and features of homeologous plastid DNA recombination are consistent with the existence of an active RecA-mediated, but a diminished mismatch, recombination/repair system in higher-plant plastids.




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