Engineering the plastid genome of <i>Nicotiana sylvestris</i>, a diploid model species for plastid genetics

Maliga, P, Svab Z.  2011.  

Journal:

Methods in Molecular Biology

Volume Number:

701

Pages:

37-50

Abstract:

The plastids of higher plants have their own approximately 120-160-kb genome that is present in 1,000-10,000 copies per cell. Engineering of the plastid genome (ptDNA) is based on homologous recombination between the plastid genome and cloned ptDNA sequences in the vector. A uniform population of engineered ptDNA is obtained by selection for marker genes encoded in the vectors. Manipulations of ptDNA include (1) insertion of transgenes in intergenic regions; (2) posttransformation excision of marker genes to obtain marker-free plants; (3) gene knockouts and gene knockdowns, and (4) cotransformation with multiple plasmids to introduce nonselected genes without physical linkage to marker genes. Most experiments on plastome engineering have been carried out in the allotetraploid Nicotiana tabacum. We report here for the first time plastid transformation in Nicotiana sylvestris, a diploid ornamental species. We demonstrate that the protocols and vectors developed for plastid transformation in N. tabacum are directly applicable to N. sylvestris with the advantage that the N. sylvestris transplastomic lines are suitable for mutant screens.

Notes:

Maliga, PalSvab, ZoraClifton, N.J.Methods Mol Biol. 2011;701:37-50.

Related External URL:

http://www.ncbi.nlm.nih.gov/pubmed/21181523
Citation:
Maliga, P, Svab Z.  2011.  Engineering the plastid genome of Nicotiana sylvestris, a diploid model species for plastid genetics. Methods in Molecular Biology. 701:37-50.