Direct somatic embryogenesis through thin cell layer culture in Panax ginseng

Abstract

Direct somatic embryos were differentiated on cotyledon transverse Thin Cell Layers (tTCLs) of Panax ginseng after 9 weeks in the Murashige and Skoog basal (MS) medium containing 2,4-D (5#M). When MS medium containing 2,4-9 (5#M) was used for seedling pretreatment and for tTCLs culture, somatic embryos were observed 2 weeks earlier, i.e. after 7 weeks of culture. On the tTCLs from seedlings pretreated with 2,4-D (5#M) combined with benzyladenine and zeatin at 0.1/zM (BZ), somatic embryos were observed after 6 weeks of culture and the percentage of embryogenesis was higher (62%) than when 2,4-D was used alone for pretreatment (40%). Similar results were also obtained from pretreatment with combinations of 2,4-D (5/zM) and thidiazuron (TDZ) (0.01, 0. I#M). When a combination of 2,4-D (5#M) and BZ (0.1#M) was used both for seedling pretreatment and for tTCLs culture, both somatic embryos and shoots were observed after only 3 weeks. As the concentration of BZ increased, the percentage of somatic embryogenesis decreased but the percentage of organogenesis increased. Similar responses were obtained with a combination of 2,4-9 (5#M) and TDZ (0.01#M). On the medium containing both NAA (0.3#M) and BZ (I#M), globular-and heart-stage embryos developed after 4 weeks of culture into cotyledonary-staged embryos which remained dormant after a short elongation of the embryo axis. The importance of seedling pretreatment by growth substances in enhancing somatic embryogenesis is reported.

Somatic embryogenesis in Acrocomia aculeata Jacq. (Lodd.) ex Mart using the thin cell layer technique

“…The advantage of TCL is that cells have greater contact with the nutrients and growth regulators present in the culture medium than do whole explants (Ahn et al 1996). Furthermore, TCL provides a simple and efficient system of micropropagation that can be used in the propagation of elite clonal plants ( Monja-Mio & Robert 2013).…”

In vitro propagation of Psoralea corylifolia L. by somatic embryogenesis in cell suspension culture

“…Through tTCLs technology, micropropagation could be obtained in recalcitrant (Mulin and Bellio-Spataru 2000;Leguillon et al 2003) medicinal (Ahn et al 1996) and other crop plant species (Baskaran et al 2006;Ben Ghnaya et al 2008) in rapid multiplication. Moreover, tissue culture technology provides means of mass propagation of a species of interest with profound potential economic spin-offs (Teixeira da Silva 2003).…”

Thin cell layer culture system in Lilium: Regeneration and transformation perspectives

“…The TCL system, first described in tobacco (Tran Thanh Van 1973), is an efficient model for the study of the mechanisms of control of morphogenesis (Tran Thanh Van, 1981; Tran Thanh Van, 1991;Compton and Veilleux, 1992; Tran Thanh and transformation (Charest et al, 1988;Tran Thanh Van et al, 1990;Creemers-Molenaar et al, 1994;Bui et al, 1998aBui et al, , 1998bBui et al, , 1998c, and has shown to be very efficient in many and dicotyledonous species (Klimaszewska and Keller, 1985;Detrez et al, 1988;Pelissier et al, 1990;Ahn et al, 1996;Bui et al, 1998a) and monocotyledonous species (Golds et al, 1993;Goh et al, 1995;Lakshmanan et al, 1995;Gendy et al, 1996;Okole and Schultz, 1996;Bui et al, 1997, 1998c, 1999, Nhut et al, 2000a.…”