Plastid transformation in Lesquerella fendleri, an oilseed Brassicacea.
A plastid transformation protocol was developed for Lesquerella fendleri, a species with a high capacity for plant regeneration in tissue culture. Transformation vector pZS391B carried an aadA16gfp marker gene conferring streptomycin-spectinomycin resistance and green fluorescence under UV light. Biolistic transformation of 51 Lesquerella leaf samples, followed by spectinomycin selection, yielded two transplastomic clones. The AAD-GFP fusion protein, the marker gene product, was localized to chloroplasts by confocal laser microscopy. Fertile plants and seed progeny were obtained in line Lf-pZS391B-1. In the 51 samples a large number (108) of spontaneous mutants were identified. In five of the lines spectinomycin resistance was localized to a conserved stem structure by sequencing 16S rRNA genes. Success in L. fendleri, a wild oilseed species, extends plastid transformation beyond Arabidopsis thaliana in the Brassicaceae family.
Skarjinskaia
Marina
M
Waksman Institute, Rutgers, The State University of New Jersey, 190 Frelinghuysen Road, Piscataway, NJ 08854-8020, USA.
Svab
Zora
Z
Maliga
Pal
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D000900:Anti-Bacterial Agents / Q000494:pharmacology; D015139:Blotting, Southern; D019607:Brassicaceae / Q000235:genetics*; D002478:Cells, Cultured; D002736:Chloroplasts / Q000235:genetics*; D004352:Drug Resistance, Microbial; D005819:Genetic Markers; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000302:isolation & purification; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009154:Mutation; D018087:Plastids / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification; D012639:Seeds / Q000235:genetics*; D000198:Spectinomycin / Q000494:pharmacology; D013307:Streptomycin / Q000494:pharmacology; D014026:Nicotiana / Q000235:genetics; D014170:Transformation, Genetic; D014466:Ultraviolet Rays
D000900:Anti-Bacterial Agents; D005819:Genetic Markers; D008164:Luminescent Proteins; D012336:RNA, Ribosomal, 16S; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D000198:Spectinomycin; D013307:Streptomycin