Abstract
A number of Solanum nigrum mutants resistant to the antibiotics spectinomycin, streptomycin and lincomycin have been isolated from regenerating leaf strips after mutagenesis with nitroso-methylurea. Selection of streptomycin- and spectinomycin-resistant mutants has been described earlier. Lincomycin-resistant mutants show resistance to higher levels of the antibiotic than used in the initial selection, and in the most resistant mutant (Ll7A1) maternal inheritance of the trait was demonstrated. The lincomycin-resistant mutant L17A1 and a streptomycin plus spectinomycin resistant double mutant (StSpl) were chosen for detailed molecular characterisation. Regions of the plastid DNA, within the genes encoding 16S and 23S rRNA and rps12 (3′) were sequenced. For spectinomycin and lincomycin resistance, base changes identical to those in similar Nicotiana mutants were identified. Streptomycin resistance is associated with an A → C change at codon 87 of rps 12 (converting a lysine into a glutamine), three codons upstream from a mutation earlier reported for Nicotiana. This site has not previously been implicated in streptomycin resistance mutations of higher plants, but has been found in Escherichia coli. The value of these mutants for studies on plastid genetics is discussed.
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Bookjans G, Stumman BM, Henningsen KW (1984) Preparation of chloroplast DNA from pea plastids isolated in a medium of high ionic strength. Anal Biochem 141:244–247
Cseplo A, Maliga P (1982) Lincomycin resistance, a new type of maternally inherited mutation in Nicotiana plumbaginifolia. Curr Genet 6:105–107
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 PH (1988) Point mutations in 23S rRNA genes of four lincomycin resistant Nicotiana plumbaginifolia mutants could provide selectable markes for chloroplast transformation. Mol Gen Genet 214:295–299
D'Arcy WG (1979) The classification of the Solanaceae. In: Hawkes JG, Lester RN, Skelding AD (eds) The biology and taxonomy of the Solanaceae. Academic Press, London, pp 3–47
Dix PJ, McKinley CP, McCabe PF (1990) Antibiotic resistant mutants of Solanum nigrum. In: Nijkamp HJJ, Vander Plas LHW, Van Aartrijk J (eds) Progress in plant cellular and molecular biology. Kluwer Academic Publisher, Dordrecht, Boston, London, pp 169–174
Etayebi M, Prasad SM, Morgan EA (1985) Chloramphenicol-erythromycin resistance mutations in a 23S rRNA gene of Escherichia coli. J Bacteriol 162:551–557
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
Fromm H, Edelman M, Aviv D, Galun E (1987) The molecular basis for rRNA-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 12:494–505
Funatsu G, Wittman HG (1972) Ribosomal proteins XXXIII. Location of amino acid replacements in protein S12 isolated from E. coli mutants resistant to streptomycin. J Mol Biol 68:547–550
Galili S, Fromm H, Aviv D, Edelman M, Galun G (1989) Ribosomal protein S12 as a site for streptomycin resistance in Nicotiana chloroplasts. Mol Gen Genet 218:289–292
Gasquez J, Darmency H, Compoint C-P (1981) Étude de la transmission de la résistance chloroplastique aux triazines chez Solanum nigrum L. C R Acad Sci 3:847–849
Glas C, Kamp JC, Jongsma C, Nijkamp HJJ, Hille J (1990) The induction of lincomycin resistance in Lycopersicon peruvianum and Lycopersicon esculentum. Plant Sci 70:231–247
Goloubinoff P, Edelman M, Hallick RB (1984) Chloroplast-coded atrazine resistance in Solanum nigrum: psbA loci from susceptible and resistant biotypes are isogenic except for a single codon change. Nucleic Acids Res 12:5489–5496
Guri A, Levi A, Sink KC (1988) Morphological and molecular characterization of somatic hybrid plants between Lycopersicon esculentum and Solanum nigrum. Mol Gen Genet 212:191–198
Hagemann R (1982) Induction of plastome mutations by nitrosourea compounds. In: Edelman M, Hallick RB, Chua N-H (eds) Methods in chloroplast molecular biology. Elsevier Biomedical Press, Amsterdam, pp 119–127
Hamill J, Ahuja PS, Davey MR, Cocking EC (1986) Protoplast-derived streptomycin-resistant plants of the forage legume Onobrychis viciifolia Scop (sainfoin). Plant Cell Rep 5:439–441
Jansen CE, Snel EAM, Akerboom MJE, Nijkamp HJJ, Hille J (1990) Induction of streptomycin resistance in the wild tomato Lycopersicon peruvianum. Mol Gen Genet 220:261–268
Maliga P (1984) Cell culture procedures for mutant selection and characterization in Nicotiana plumbaginifolia. In: Vasil IK (ed) Cell culture and somatic cell genetics of plants, vol 1. Academic Press, New York, pp 552–562
Maliga P, Moll B, Svab Z (1990) Toward manipulation of plastid genes in higher plants. In: Selitch I (ed) Perspectives in genetic and biochemical regulation of photosynthesis. Alan R Liss, New York, pp 133–137
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Marsh JL, Erfle M, Wykes EJ (1984) The pIC plasmid and phage vectors with versatile cloning sites for recombinant selection by insertional inactivation. Gene 32:481–485
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
McCabe PF, Cseplo A, Timmons AM, Dix PJ (1990) Selection of chloroplast mutants. In: Pollard JW, Walker JM (eds) Methods in molecular biology, vol 6. Plant cell and tissue culture. Humana Press, pp 467–475
Medgyesy P (1990) Selection and analysis of cytoplasmic hybrids. In: Dix PJ (ed) Plant cell line selection. VCH, Weinheim, pp 287–316
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Murphy G, Kavanagh T (1988) Speeding-up the sequencing of double-stranded DNA. Nucleic Acids Res 16:5198
O'Neill CM, Horvath GV, Horvath E, Dix PJ, Medgyesy P (1993) Chloroplast transformation in plants: polyethylene glycol (PEG) treatment of protoplasts is an alternative to biolistic delivery systems. Plant J 3:729–738
Shinozaki K, Ohme M, Tanaka M, Wakasugi T, Hayashida N, Matsubayashi T, Zaita N, Chunwongse J, Obokata J, Shinozaki Svab Z, Hajdukiewitz P, Maliga P (1990) Stable transformation of plastids in higher plants. Proc Natl Acad Sci USA 87:8526–8530
Thanh ND, Medgyesy P (1989) Limited chloroplast gene transfer via recombination overcomes plastome-genome incompatibility between Nicotiana tabacum and Solanum tuberosum. Plant Mol Biol 12:87–93
Thanh ND, Pay A, Smith MA, Medgyesy P, Marton L (1988) Intertribal chloroplast transfer by protoplast fusion between Nicotiana tabacum and Salpiglossis sinuata. Mol Gen Genet 213:186–190
KY, 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. Plant Mot Biol Rep 4:110–147
Staub JM, Maliga P (1992) Long regions of homologous DNA are incorporated into the tobacco plastid genome by transformation. Plant Cell 4:39–45
Svab Z, Maliga P (1991) Mutation proximal to the rRNA binding region of the Nicotiana plastid 16SrRNA confers resistance to spectinomycin. Mol Gen Genet 228:316–319
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Kavanagh, T.A., O'Driscoll, K.M., McCabe, P.F. et al. Mutations conferring lincomycin, spectinomycin, and streptomycin resistance in Solanum nigrum are located in three different chloroplast genes. Molec. Gen. Genet. 242, 675–680 (1994). https://doi.org/10.1007/BF00283422
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DOI: https://doi.org/10.1007/BF00283422