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Applied and Environmental Microbiology, January 2003, p. 673-678, Vol. 69, No. 1
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.1.673-678.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Degradation and Transformability of DNA from Transgenic Leaves

MariaTeresa Ceccherini,1,2 John Poté,1,3 Elisabeth Kay,1 Van Tran Van,1,4 Joëlle Maréchal,1 Giacomo Pietramellara,2 Paolo Nannipieri,2 Timothy M. Vogel,1 and Pascal Simonet1*

Écologie Microbienne, UMR CNRS 5557, Université Claude Bernard, Lyon 1,1 Libragen, 69622 Villeurbanne Cedex, France,4 Dipartimento di Scienza del Suolo e Nutrizione della Pianta, Università degli Studi di Firenze, 50144 Florence, Italy,2 Institut F. A. Forel—Université de Genève, CH-1290 Versoix, Switzerland3

Received 17 June 2002/ Accepted 6 October 2002

The fate of transplastomic (chloroplast genome contains the transgene) tobacco plant DNA in planta was studied when the plant leaves were subjected to decay conditions simulating those encountered naturally, including grinding, incubation with cellulase or enzymes produced by Erwinia chrysanthemi, and attack by the plant pathogen Ralstonia solanacearum. Direct visualization of DNA on agarose gels, gene extraction yield (the number of amplifiable aadA sequences in extracted plant DNA), and the frequency that recipient bacteria can be transformed by plant DNA were used to evaluate the quality and quantity of plant DNA and the transgene. These measurements were used to monitor the physical and biological degradation of DNA inside decaying plant tissues. Our results indicate that while most of the DNA will be degraded inside plant cells, sufficient DNA persists to be released into the soil.


* Corresponding author. Mailing address: Ecologie Microbienne, UMR 5557, Université Lyon I, 43 bd du 11 Novembre 1918, 69622 Villeurbanne, France. Phone: 33 472 44 82 89. Fax: 33 472 43 12 23. E-mail: simonet{at}biomserv.univ-lyon1.fr.


Applied and Environmental Microbiology, January 2003, p. 673-678, Vol. 69, No. 1
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.1.673-678.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.




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