Plant regeneration from protoplasts: a literature review

The results are reported for the first time on successful plant regeneration from mesophyll-derived protoplasts of 'rapid cycling' B. oleracea. Comparative data were also presented on plant regeneration from mesophyll-derived protoplasts of two other varieties namely var. botrytis and var. gemmifera. It was found that a modified Pelletier (Pelletier et al. 1983) protocol is highly beneficial for protoplast culture and plant regeneration from mesophyll-derived protoplasts. The plating efficiency of B. oleracea 'rapid cycling' protoplasts was, in the best combination of isolation method, culture technique and culture media 4.5% + 0.4% and the plant regeneration frequency approximately 15%. Plant regeneration was further improved by transferring the calli from the D medium of Pelletier to a callus growth medium (MSll) and subsequently to the K 3 regeneration medium of Glimelius (Glimelius 1984). Various factors influencing plating efficiency and plant regeneration from mesophyll protoplasts of B. oleraeea are discussed.

“…oleracea was attributed to the genotypic response, a common phenomenon observed in many other species (Roest & Gilissen 1989). In our experiments, the protoplast culture response of var.…”

Section: Discussionmentioning

confidence: 72%

Protoplast culture and regeneration from Brassica oleracea ?rapid cycling? and other varieties

Kik

Zaal

1993

“…Plants have been regenerated from protoplasts for about ten species of the Gramineae (Roest & Gilissen 1989) including rice, maize and wheat. In these studies embryogenic cell suspensions were selected for fast growing capacity to obtain dividing protoplasts.…”

mentioning

confidence: 99%

Efficient plant regeneration through somatic embryogenesis from callus induced on mature rice embryos (Oryza sativa L.)

Rueb

Leneman

Schilperoort

et al. 1994

103

To obtain a reproducible efficient procedure for regeneration of rice plants through somatic embryogenesis from callus four published methods of callus induction and regeneration were compared. Callus was initiated from mature embryos of the Japonica cultivar Taipei 309 of rice (Oryza sativa L.). The number, mass and morphology of the callus formed on the scutellum were dependent on the medium used. A limited humidity and an optimal aeration of the culture vessels enhanced the frequency of embryogenesis and plant regeneration. A method described by Poonsapaya et al. (1989) was found to be the most efficient and was slightly modified. As a result 98% of the T309 embryos formed callus, of which 63% regenerated into plants. Each callus yielded an average of 6 plants. Plant morphology, fertility and seed set of the regenerants were found to be normal.

“…Since the regeneration of green plants from protoplasts of grasses and sugarbeet is difficult (Roest & Gilissen 1989;Krens et al 1990), it may prove to be important to use protoplasts from cell lines that are capable of regeneration both as the cytoplasm acceptor and as the cytoplasm donor in fusion experiments. This may 231 increase the likelihood of green plant regeneration from the cybrid calli obtained.…”

Section: Discussionmentioning

confidence: 99%

High yields of cytoplasts from protoplasts of Lolium perenne and Beta vulgaris using gradient centrifugation

Ark¹,

Hall²,

Creemers-Molenaar³

et al. 1992

Methods for the isolation of cytoplasts from suspension culture-derived protoplasts of the monocot Lolium perenne (perennial ryegrass) and the dicot Beta vulgaris (sugarbeet) have been determined. After comparing a range of gradients it was found that a discontinuous sucrose/mannitol gradient gave the highest cytoplast yields for both species tested: of the protoplasts loaded onto the gradient, for Beta >30% and for Lolium up to 45% could be recovered as cytoplasts. Sufficient protoplasts could be loaded onto the gradient to produce suitable numbers of cytoplasts for use in asymmetric somatic hybridisation experiments. Cytoplasts could be isolated from several suspension cultures of different ages. The cytoplast fraction was recovered from the upper part of the gradient in all cases and was only slightly contaminated (2-8%) with protoplasts. Lolium cytoplasts were small, evacuolate cells with granular cytoplasm. In contrast, Beta cytoplasts were larger and predominantly vacuolate. Both contained mitochondria as determined using fluorescence staining.