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In vitro propagation of the endangered plant Centaurea ultreiae: assessment of genetic stability by cytological studies, flow cytometry and RAPD analysis

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Abstract

The effect of different cytokinins on multiple shoot regeneration from shoots of Centaurea ultreiae was studied. The culture system consisted of solid basal half-strength Murashige and Skoog medium supplemented with one of four cytokinins [6-benzyladenine (BA), zeatin, kinetin, or N6-(2-isopentyl) adenine (2-iP)] at each of five different concentrations. The highest multiplication rate (5.52 shoots per explant) was obtained in the medium supplemented with 4.44 μM BA. Shoots were successfully rooted (91% success) by dipping the basal end into a solution containing 10 M 1-naphthaleneacetic acid for 30 s. Genetic stability of the regenerated plants was assessed by random amplified polymorphic DNA (RAPD) analysis and flow cytometry. In the initial randomly selected plant material (control) and 20 of its regenerants, 2,688 bands were generated by RAPD with 12 different primers, and the same banding profiles were exhibited. Molecular and cytological analyses did not reveal genomic alterations in any of the regenerated plants obtained on medium containing 4.44 μM BA. The success of acclimatization to environmental conditions—100% of plants were successfully acclimatized—suggests that the micropropagation system described is a reliable method for propagation of C. ultreiae.

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Acknowledgments

R. M. was financially supported by a predoctoral fellowship from the Xunta de Galicia. The authors thank Patricia Corral for help with the flow cytometry analysis. The study was partially financed by the Xunta de Galicia, project PGIDT/PGIDIT 07MDS009200PR.

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  1. Rubén Mallón

    Present address: Instituto de Investigaciones Agrobiológicas de Galicia, CSIC, Avenida de Vigo s/n. Apartado 122, 15780, Santiago de Compostela, Spain

Authors and Affiliations

  1. Departamento de Fisiología Vegetal, Universidad de Santiago de Compostela, 15782, Santiago de Compostela, Spain

    Rubén Mallón & María Luz González

  2. Departamento de Botánica, Universidad de Santiago de Compostela, 15782, Santiago de Compostela, Spain

    Juan Rodríguez-Oubiña

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  1. Rubén Mallón
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Correspondence to María Luz González.

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Mallón, R., Rodríguez-Oubiña, J. & González, M.L. In vitro propagation of the endangered plant Centaurea ultreiae: assessment of genetic stability by cytological studies, flow cytometry and RAPD analysis. Plant Cell Tiss Organ Cult 101, 31–39 (2010). https://doi.org/10.1007/s11240-009-9659-y

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