Plant regeneration from protoplasts of dessert banana cv. Grande Naine (Musa spp., Cavendish sub-group AAA) via somatic embryogenesis

Regeneration of flowering plants from difficile lily protoplasts by means of a nurse culture

“…In rice, the regeneration of plants from protoplasts was first reported by Fujimura et al (1985), and the most efficient protocols were developed from novel nurse-culture methods (Kyozuka et al 1987). Subsequently, it was reported that the initial division of monocotyledonous protoplasts was dependent on nurse cells (Hahne et al 1990;Funatsuki et al 1992;Stoldt et al 1996;Moura et al 1997;Assani et al 2001). Despite this, there have been no reports on using nurseculture methods for the division of lily protoplasts.…”

“…However, we were not able to obtain divided cells from our preliminary culturing of protoplasts isolated from various kinds of Oriental hybrid cultivars according to their protocol. The accumulation of results showing the positive effect of nurse cells on the division of protoplasts derived from monocotyledonous species such as rice (Kyozuka et al 1987;Moura et al 1997), oats (Hahne et al 1990), barley (Funatsuki et al 1992;Stoldt et al 1996), and bananas (Assani et al 2001) has led us to introduce the nurse-culture method to our lily protoplast culture systems.…”

Production and characterization of somatic hybrids between upland cotton (Gossypium hirsutum) and wild cotton (G. klotzschianum Anderss) via electrofusion

“…Following fusion, the protoplasts were cultured in the dark in KM 8 P medium supplemented with 0.5 mM mannitol, 3% (w/v) glucose, 2.685 μM NAA (α-naphthaleneacetic acid), and 0.465 μM kinetin through a modified Feeder-layer culture (Assani et al 2001) with a final density of 2.5×10 5 ml −1 ; the embryogenic calluses of Coker 201 were used as nurse cells. Fresh KM 8 P medium supplemented with 1.343 μM NAA and 0.233 μM kinetin was added to the culture medium at weekly intervals.…”

Protoplast Fusion in Banana (Musa spp.): Comparison of Chemical (PEG: Polyethylene Glycol) and Electrical Procedure

“…Protoplast-derived microcalluses were individually picked from the feeder layer and gently transferred onto A 0.4 B 0.5 regeneration medium containing MS salts, Morel vitamins, 88 mM sucrose, 2.3 lM IAA, 2.2 lM BA and 7.5 g l )1 agarose sea plaque (pH 5.7) (Assani et al, 2001). The cultures were maintained at 27°C in the dark.…”

“…A goal of banana improvement is the release of disease-resistant triploid hybrids , protoplast fusion is considered as useful complementary tool for the production of somatic hybrid tetraploid parents that could be used in interploid crosses with other diploid lines or for direct release of triploid somatic hybrids by haplo/ diploid protoplast fusion (Assani et al, 2003). Since a protoplast regeneration system in banana has already been established (Megia et al, 1993;Panis et al, 1993;Matsumoto and Oka, 1998;Assani et al, 2001;Assani et al, 2002); the use of cell fusion techniques in banana breeding had become a realizable objective.…”