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

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

Tony A. Kavanagh^a, Nguyen Duc Thanh^b, Nga Thi Lao^a, Noreen McGrath^b,c, Stefan O. Peter^b, Eva M. Horváth^b, Philip J. Dix^c, and Peter Medgyesy^b
^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 Nicotianatabacum using cloned plastid DNA of Solanum nigrum carryingmutations conferring spectinomycin and streptomycin resistance.The use of the incompletely homologous (homeologous) Solanumplastid DNA as donor resulted in a Nicotiana plastid transformationfrequency comparable with that of other experiments where completelyhomologous plastid DNA was introduced. Physical mapping andnucleotide sequence analysis of the targeted plastid DNA regionin the transformants demonstrated efficient site-specific integrationof the 7.8-kb Solanum plastid DNA and the exclusion of the vectorDNA. The integration of the cloned Solanum plastid DNA intothe Nicotiana plastid genome involved multiple recombinationevents as revealed by the presence of discontinuous tracts ofSolanum-specific sequences that were interspersed between Nicotiana-specificmarkers. Marked position effects resulted in very frequent cointegrationof the nonselected peripheral donor markers located adjacentto the vector DNA. Data presented here on the efficiency andfeatures of homeologous plastid DNA recombination are consistentwith the existence of an active RecA-mediated, but a diminishedmismatch, recombination/repair system in higher-plant plastids.

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