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

Abstract

The effect of different cytokinins on multiple shoot regeneration from shoots of Centaurea ultreiae was studied. The culture system consisted of solid basal halfstrength 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 lM 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 lM 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.

Efficient in vitro propagation of Artemisia nilagirica var. nilagirica (Indian wormwood) and assessment of genetic fidelity of micropropagated plants

“…4), showing genetic stability of the micropropagated plants. Similar reports showing the genetic stability of micropropagated plants was reported in Centaurea ultreiae and Nepenthes khasiana (Mallon et al 2010;Devi et al 2013).…”

ASSESSMENT OF GENETIC STABILITY OF MICROPROPAGATED Eucalyptus globulus Labill HYBRID CLONES BY MEANS OF FLOW CYTOMETRY AND MICROSATELLITES MARKERS

“…Flow cytometry has been widely used to detect variation in ploidy level of plants micropropagated because of relatively low cost, high yield and precision of analysis (Rahman and Rajora, 2001;Loureiro et al, 2007;Mallón et al, 2010). Brito et al (2010) found no genetic variation when two wild olives (Olea maderensis L. and O. europaea spp.…”

“…Somaclonal variation and guarantee completely genetic fidelity (Mallón et al, 2010) can´t be detected by any techique. Therefore, the adoption of different methodologies is necessary to assess genetic stability of micropropagated plants (Peredo et al, 2009).…”

“…Flow cytometry to determine accurately and quickly, the amount of DNA in large numbers of cells (Loureiro et al, 2005;Orbovic et al, 2008) provide an excellent alternative to assess the genetic stability (Jin et al, 2008). The change in ploidy level is the main genetic variation that occurs in plant tissue culture (Mallón et al, 2010) and flow cytometry has been effective in detecting this type of variation (Brito et al, 2010). Genetic stability several woody species was performed by flow cytometry (Conde et al, 2004;Loureiro et al, 2005Loureiro et al, , 2007Brito et al, 2010), including Eucalyptus (Pinto et al, 2004).…”

Mass propagation of Plectranthus bourneae Gamble through indirect organogenesis from leaf and internode explants

“…Consequently, special solicitude should be remunerate to protect and preserve the endemic flora that is normally found in limited areas (Fay 1992;Sarasan et al 2006). Conservation of endemic species by plant tissue culture has attracted the attention of many researchers (Mallon et al 2010;Piovan et al 2010). Lamiaceae herbs namely basil, oregano, sage, marjoram and rosemary are popular aromatic herbs.…”