Summary
Nicotiana tabacum lines carrying maternally inherited resistance to spectinomycin were obtained by selection for green callus in cultures bleached by spectinomycin. Two levels of resistance was found. SPC1 and SPC2 seedlings are resistant to high levels (500 μg/ml), SPC23 seedlings are resistant to low levels (50 μg/ml) of spectinomycin. Lines SPC2 and SPC23 are derivatives of the SR1 streptomycin-resistant plastome mutant. Spectinomycin resistance is due to mutations in the plastid 16S ribosomal RNA: SPC1, an A to C change at position 1138; SPC2, a C to U change at position 1139; SPC23, a G to A change at position 1333. Mutations similar to those in the SPC1 and SPC2 lines have been previously described, and disrupt a conserved 16S ribosomal RNA stem structure. The mutation in the SPC23 line is the first reported case of a mutation close to the region of the 16S rRNA involved in the formation of the initiation complex. The new mutants provide markers for selecting plastid transformants.
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References
Cseplo A, Maliga P (1984) Large scale isolation of maternally inherited lincomycin resistance mutations in diploid Nicotiana plumbaginifolia protoplast cultures. Mol Gen Genet 196:407–412
Cseplo A, Etzold T, Schell J, Schreier P (1988) Point mutations in the 23S rRNA genes of four lincomycin-resistant Nicotiana plumbaginifolia mutants could provide new selectiable markers for chloroplast transformation. Mol Gen Genet 214:295–299
Dahlberg AE (1989) The functional role of ribosomal RNA in protein synthesis. Cell 57:525–529
Etzold T, Fritz CC, Schell J, Schreier PH (1987) A point mutation in the chloroplast 16S rRNA gene of a streptomycin-resistant Nicotiana tabacum. FEBS Lett 219:343–346
Fejes E, Engler D, Maliga P (1990) Extensive homologous chloroplast DNA recombination in the pt14 Nicotiana somatic hybrid. Theor Appl Genet 79:28–32
Fromm H, Edelman M, Aviv D, Galun E (1987) The molecular basis for rDNA-dependent spectinomycin resistance in Nicotiana chloroplasts. EMBO J 6:3233–3237
Fromm H, Galun E, Edelman M (1989) A novel site for streptomycin resistance in the ‘530 loop’ of chloroplast 16S ribosomal RNA. Plant Mol Biol 12:499–505
Harris EH, Burkhart BD, Gillham NW, Boynton JE (1989) Antibiotic resistance mutations in the chloroplast 16S and 23S rRNA genes of Chlamydomonas reinhardtii: Correlation of genetic and physical maps of the chloroplast genome. Genetics 123:281–292
Jansen CE, Snel EAM, Akerboom HJJ, Nijkamp HJJ, Hille J (1990) Induction of streptomycin resistance in the wild tomato Lycopersicon peruvianum. Mol Gen Genet 220:261–268
Kolodner R, Tewari KK (1975) The molecular size and conformation of the chloroplast DNA from higher plants. Biochim Biophys Acta 402:372–390
Maliga P (1984) Cell culture procedures in mutant selection and characterization in Nicotiana plumbaginifolia. In: Vasil IK (ed) Cell culture and somatic cell genetics of plants. Vol 1. Academic Press, Orlando, pp 552–562
Maliga P, Breznovits A, Marton L (1973) Streptomycin-resistant plants from callus culture of haploid tobacco. Nature New Biol 224:29–30
Maliga P, Breznovits A, Marton L, Joo F (1975) Non-Mendelian streptomycin-resistant tobacco with altered chloroplasts and mitochondria. Nature 255:401–402
Maliga P, Svab Z, Harper EC, Jones JDG (1988) Improved expression of streptomycin resistance in plants due to a deletion in the streptomycin phosphotransferase coding sequence. Mol Gen Genet 214:456–459
Maliga P, Moll B, Svab Z (1990) Toward manipulation of plastid genes in higher plants. In: Zelitch I (ed) Perspectives in genetic and biochemical regulation of photosynthesis, Alan R. Liss, New York, pp 133–143
McCabe PF, Timmons AM, Dix PJ (1989) A simple procedure for the isolation of streptomycin resistant plants in Solanaceae. Mol Gen Genet 216:132–137
Moazed D, Noller HF (1987) Interaction of antibiotics with functional sites in 168 ribosomal RNA. Nature 327:389–394
Newman SM, Boynton JE, Gillham NW, Randolph-Anderson BL, Johnson AM, Harris AH (1990) Transformation of chloroplast ribosomal RNA genes in Chlamydomonas: Molecular and genetic characterization of integration events. Genetics 126:875–888
Shinozaki K, Ohme M, Tanaka M, Wakasugi T, Hayashida N, Matsubayashi T, Zaita N, Chunwongse J, Obokata J, Yamaguchi-Shinozaki K, Ohto C, Torazawa K, Meng BY, Sugita M, Deno H, Kamogashira T, Yamada K, Kusuda J, Takaiwa F, Kato A, Tohdoh N, Shimada H, Sugiura M (1986) The complete nucleotide sequence of the tobacco chloroplast genome: its organization and expression. EMBO J 5:2043–2049
Svab Z, Hajdukiewitz P, Maliga P (1990) Stable transformation of plastids in higher plants. Proc Natl Acad Sci USA 87:8526–8530
To KY, Chen CC, Lai YK (1989) Isolation and characterization of streptomycin resistant mutants in Nicotiana plumbaginifolia. Theor Appl Genet 78:81–86
Vieira J, Messing J (1987) Production of single-stranded plasmid DNA. Methods Enzymol. 153:3–11
Wallace BJ, Tai PC, Davis BD (1974) Selective inhibition of initiating ribosomes by spectinomycin. Proc Natl Acad Sci USA 71:1634–1638
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Svab, Z., Maliga, P. Mutation proximal to the tRNA binding region of the Nicotiana plastid 16S rRNA confers resistance to spectinomycin. Molec. Gen. Genet. 228, 316–319 (1991). https://doi.org/10.1007/BF00282483
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DOI: https://doi.org/10.1007/BF00282483