Isolation of antibiotic-resistant and antimetabolite-resistant mutants of Frankia strains EuI1c and Cc1.17

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Abstract:

Antibiotic-resistant and antimetabolite-resistant mutants of the nitrogen-fixing symbiotic bacterium Frankia were isolated to provide strains with genetic backgrounds amenable to genetic analysis. The lethal and mutagenic effects of ethyl methanesulfonate (EMS) and UV light on four Frankia strains were investigated. UV irradiation or EMS treatment of strain EuI1c cells resulted in the formation of two different colony types: rough and smooth. The smooth colonies were conditional sporulation mutants. In the case of EMS-induced cells of strain Cc1.17, resistance to lincomycin, ampicillin, and 5-fluorouracil occurred at a frequency of 1 × 10–5, 1 × 10–5, and 4 × 10–5, respectively. The lincomycin-resistant mutants produced a yellow–tan pigment that was released into the growth medium. Resistance to tetracycline and lincomycin with EMS-induced cells of strain EuI1c occurred at a frequency of 3.2 × 10–3 and 4.7 × 10–4, respectively. These strains will be useful for the development of genetic methods for Frankia.Key words: genetics, genetic markers, Frankia, actinorhizal symbiosis, nitrogen fixation, mutagenesis, EMS, UV light.

Des mutants résistants aux antibiotiques et aux anti-métabolites de la bactérie symbiotique fixatrice d'azote Frankia ont été isolés afin de fournir des souches ayant des fonds génétiques pouvant se prêter à des analyses génétiques. Les effets létaux et mutagènes du méthanesulfonate d'éthyle (MSE) et du rayonnement UV sur quatre souches de Frankia ont été étudiés. L'irradiation UV ou le traitement au MSE des cellules de la souche EiI1c a entraîné la formation de deux types distincts de colonies: des colonies lisses et rugueuses. Les colonies lisses étaient des mutants de sporulation conditionnelle. Chez les cellules de la souche Cc1.17 induites au MSE, la résistance à la linomycine, à l'ampicilline et au 5-fluorouracile est apparue à une fréquence de 1 × 10–5, 1 × 10–5 et 4 × 10–5, respectivement. Les mutants résistants à la linomycine ont libéré un pigment jaune–bronzé dans le milieu de culture. La résistance à la tétracycline et à la linomycine chez les cellules de la souche EuI1c induites au MSE est apparue à une fréquence de 3,2 × 10–3 et 4,7 × 10–4, respectivement. Ces souches seront d'une utilité pour le développement de méthodes d'analyse génétique chez Frankia.Mots clés : génétiques, marqueurs génétiques, Frankia, symbiose actinorhizienne, fixation de l'azote, mutagénèse, MSE, lumière UV.[Traduit par la Rédaction]

Document Type: Research Article

Publication date: April 1, 2004

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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