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

• Published in: Plant cell, tissue and organ culture Vol. 101; no. 1; pp. 31 - 39
• Main Authors: Mallon, Ruben, Rodriguez-Oubina, Juan, Gonzalez, Maria Luz
• Format: Journal Article
• Language: English
• Published: Dordrecht Dordrecht : Springer Netherlands 01.04.2010; Springer Netherlands; Springer Nature B.V
• Subjects: acclimation; Acclimatization; Adenine; application rate; Benzyladenine; Biomedical and Life Sciences; Centaurea; Centaurea ultreiae; culture media; Cytokinins; Deoxyribonucleic acid; DNA; Endangered plants; endangered species; Environmental conditions; explants; Flow cytometry; Flow stability; genetic variation; genome; in vitro regeneration; isopentenyladenine; Kinetin; Life Sciences; Micropropagation; Multiplication; Naphthaleneacetic acid; Original Paper; Plant Genetics and Genomics; Plant Pathology; Plant Physiology; Plant propagation; Plant Sciences; Polymerase chain reaction; Primers; Propagation; Random amplified polymorphic DNA; random amplified polymorphic DNA technique; Regeneration; rooting; Shoots; Stability analysis; Zeatin; Flow cytometry; Germplasm; RAPD; Conservation
• ISSN: 0167-6857; 1573-5044
• DOI: 10.1007/s11240-009-9659-y

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.