Plant regeneration via somatic embryogenesis from protoplasts of six plant species related to Citrus

Abstract

Protoplasts isolated from embryogenic callus of Fortunella polyandra (Ridl.), Atalantia bilocularis (Pieree ex Guill.), Hesperethusa crenulata (Roxb.), Glycosmis pentaphylla (Retz.) Corr., Triphasia trifolia (Burm. f.) P. Wils. and Murraya koenigii (L.) Spreng. were cultured in MT (Murashige and Tucker 1969) basal medium containing 5% sucrose supplemented with 0.0, 0.001, 0.01, 0.1 or 1.0 mg l(-1) BA and 0.6 M sorbitol. The highest plating efficiencies for all species were obtained on MT basal medium containing 5% sucrose supplemented with 0.001 mg l(-1) BA. F. polyandra produced higher percentages of globular somatic embryo development, while A. bilocularis consistently showed a lower percentage of globular somatic embryo development in all 5 concentrations of BA. MT basal medium containing 5% sucrose and supplemented with 0.001 mg l(-1) BA was found to be a suitable medium for development of globular somatic embryos derived from protoplasts to form heart-shaped somatic embryos with cotyledon-like structures. The highest percentages of shoot formation for all 6 species were obtained using 0.1 mg l(-1) GA3. A complete protoplast-to-plant system was developed for F. polyandra, A. bilocularis and T. trifolia, which could facilitate the transfer of nuclear and cytoplasmic genes from these species into cultivated Citrus through protoplast fusion.

The effect of several factors on somatic embryogenesis and plant regeneration in protoplast cultures of Gentiana kurroo (Royle)

“…Efficiency of isolation 31.5 · 10 5 d 38.8 · 10 5 c 44.1 · 10 5 bc 52.6 · 10 5 a 40.7 · 10 5 c 47.0 · 10 5 ab K/C, K/H = Cotyledon and hypocotyl derived cell suspension, respectively Until now two methods of plants regeneration using protoplast culture have been described: organogenesis and somatic embryogenesis. Next to indirect way (via callus), direct somatic embryogenesis is well documented but possible only for sparse species (Rybczyński 1997;Jumin and Nito 1996). Such way of regeneration was also observed in protoplasts culture of G. kurroo but only when protoplasts derived from young and freshly established cell suspensions with very high morphologic potential ( Fig.…”

An efficient regeneration system via direct and indirect somatic embryogenesis for the medicinal tree Murraya koenigii

“…There are only a few reports of in vitro studies of M. koenigii which are restricted to in vitro shoot multiplication from intact seedling (Bhuyan et al 1997) and nodal cuttings (Nirmal Babu et al 2000). Jumin and Nito (1996) reported plant regeneration through somatic embryogenesis; however, their work was not focused on this species only. In addition, their conversion rate of somatic embryos into plantlets was relatively low and the survival of the regenerants in the field were limited.…”

Plant regeneration from Gossypium davidsonii protoplasts via somatic embryogenesis

“…An effective protoplast culture system is necessary for obtaining somatic hybrids via cell fusion. It was reported that the combination of hormones and osmotic potential is the sticking points to many plants (Kao and Seguin-Swartz 1987, Kyozuka et al 1987, Jumin and Nito 1996, Hassanein and Soltan 2000. Until recently, plant regeneration from protoplasts was reported in upland cotton by several researchers (Firrozabady and DeBoer 1986, Chen et al 1989, Peeters et al 1994, Sun et al 2005, and the effect of nitrogen, the combination of hormones, the source of explants and the density of protoplasts on the protoplast regeneration in upland cotton has been evaluated (Bhojwani et al 1977, Saka et al 1987, Chen et al 1989, Sun et al 2005.…”