Direct somatic embryogenesis and plant regeneration from leaf explants of Limonium sinensis (Girard) Kuntze

The potentialities of direct somatic embryogenesis and plant regeneration from leaf explants of Limonium sinensis var. Golden Diamond in vitro were investigated. Young whole leaf and cut leaf explants when cultured on MS basal medium supplemented with each of the growth regulators N-6-benzyladenine (BA) (0.44-2.2 mu M) or thidiazuron (TDZ) (4.54 mu M) alone or in combination with a fixed concentration of alpha-naphthalene acetic acid (NAA) (1.07 mu M) produced somatic embryos directly. More than 90% of the leaf explants produced white, globular somatic embryos on BA (2.2 mu M) and NAA (1.07 mu M) supplemented MS basal medium within 1 week of inoculation. Most of the embryos matured further and converted after 8 weeks of culture on the same medium. Histological observation showed that the somatic embryos originated front single cells of epidermal layer of leaf. Histological evidence of formation of shoot and root poles during conversion of the embryos confirmed that these structures were true somatic embryos. After conversion the plantlets were further placed on MS medium containing 0.44 mu M BA and 4.5 mu M IBA for better shoot and root growth. About 90% of the plantlets transferred to the mixture of soil:perlite:vermiculite (1:1:1) in small plastic pots acclimatized successfully. Of these 85.5% plants survived after transferring into earthen pots containing a mixture of soil, coarse sand and cattle manure (1:1:1) under greenhouse or shady open condition. (C) 2010 Elsevier B.V. All rights reserved.