Abstract
A high rate of plastome-encoded mutations was induced in Nicotiana by exposing seeds to N-nitroso-N-methylurea. Seeds then were subjected to nutritional and in vitro selection procedures for systematic isolation of plastome-dependent antibiotic-resistant plants. Multiple flowering lines resistant to streptomycin, spectinomycin, lincomycin, or chloramphenicol were obtained. A detailed analysis of the streptomycin-resistant lines is presented. Sexual hybridization, cybrid formation following protoplast fusion, and in organello protein synthesis were used to rigorously assign the mutations to the chloroplast genome. The efficient rates of mutagenesis combined with the in vitro mass-screening procedures described here should facilitate investigation of fundamental aspects of chloroplast genetics in higher plants.
Keywords: N-nitroso-N-methylurea, plant cell culture, protoplast fusion, in organello protein synthesis, cytoplasmic inheritance
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- Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
- Epp M. D. Nuclear Gene-Induced Plastome Mutations in OENOTHERA HOOKERI. I. Genetic Analysis. Genetics. 1973 Nov;75(3):465–483. doi: 10.1093/genetics/75.3.465. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fluhr R., Edelman M. Conservation of sequence arrangement among higher plant chloroplast DNAs: molecular cross hybridization among the Solanaceae and between Nicotiana and Spinacia. Nucleic Acids Res. 1981 Dec 21;9(24):6841–6853. doi: 10.1093/nar/9.24.6841. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Galun E., Arzee-Gonen P., Fluhr R., Edelman M., Aviv D. Cytoplasmic hybridization in Nicotiana: mitochondrial DNA analysis in progenies resulting from fusion between protoplasts having different organelle constitutions. Mol Gen Genet. 1982;186(1):50–56. doi: 10.1007/BF00422911. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Maliga P., Sz-Breznovits A., Marton L Joo F. Non-Mendelian streptomycin-resistant tobacco mutant with altered chlorplasts and mitochondria. Nature. 1975 May 29;255(5507):401–402. doi: 10.1038/255401a0. [DOI] [PubMed] [Google Scholar]