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Spectinomycin resistance mutations in the rrn16 gene are new plastid markers in Medicago sativa

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Abstract

We report here the isolation of spectinomycin-resistant mutants in cultured cells of Medicago sativa line RegenSY-T2. Spectinomycin induces bleaching of cultured alfalfa cells due to inhibition of protein synthesis on the prokaryotic type 70S plastid ribosomes. Spontaneous mutants resistant to spectinomycin bleaching were identified by their ability to form green shoots on plant regeneration medium containing selective spectinomycin concentrations in the range of 25–50 mg/l. Sequencing of the plastid rrn16 gene revealed that spectinomycin resistance is due to mutations in a conserved stem structure of the 16S rRNA. Resistant plants transferred to the greenhouse developed normally and produced spectinomycin-resistant seed progeny. In light of their absence in soybean, a related leguminous plant, the isolation of spectinomycin-resistant mutants in M. sativa was unexpected. The new mutations are useful for the study of plastid inheritance, as demonstrated by detection of predominantly paternal plastid inheritance in the RegenSY-T2 × Szapko57 cross, and can be used as selective markers in plastid transformation vectors to obtain cisgenic plants.

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Acknowledgments

This research was supported by the HSRF Grant K-82037 and the Research and Development Fund of the Ministry of Agriculture, Hungary. We thank Ms. Ágnes Mihály and Ms. Magdolna Péli for skillful technical assistance. Research on alfalfa plastid inheritance in PM’s laboratory at Rutgers University is supported by the USDA National Institute of Food and Agriculture Biotechnology Risk Assessment Research Grant Program Award No. 2010-2716.

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Authors and Affiliations

  1. Agricultural Biotechnology Center, Szent-Györgyi Albert u. 4, 2100, Gödöllő, Hungary

    Brigitta Dudas, Barnabas Jenes & Gyorgy Botond Kiss

  2. Waksman Institute of Microbiology, Rutgers The State University of NJ, 190 Frelinghuysen Road, Piscataway, NJ, 08854-8020, USA

    Pal Maliga

  3. Department of Plant Biology, Rutgers University, 59 Dudley Road, New Brunswick, NJ, 08901, USA

    Pal Maliga

Authors
  1. Brigitta Dudas
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  2. Barnabas Jenes
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  3. Gyorgy Botond Kiss
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  4. Pal Maliga
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Correspondence to Pal Maliga.

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Communicated by M. Havey.

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Dudas, B., Jenes, B., Kiss, G.B. et al. Spectinomycin resistance mutations in the rrn16 gene are new plastid markers in Medicago sativa . Theor Appl Genet 125, 1517–1523 (2012). https://doi.org/10.1007/s00122-012-1930-7

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  • DOI: https://doi.org/10.1007/s00122-012-1930-7

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