Journal
PLANT CELL TISSUE AND ORGAN CULTURE
Volume 101, Issue 1, Pages 31-39Publisher
SPRINGER
DOI: 10.1007/s11240-009-9659-y
Keywords
RAPD; Flow cytometry; Conservation; Germplasm
Funding
- Xunta de Galicia [PGIDT/PGIDIT 07MDS009200PR]
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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 N-6-(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 mu M 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 mu M 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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