Regeneration of Mesophyll Protoplasts Isolated from Dihaploid Clones of Solanum tuberosum

Abstract

Protoplasts have been isolated from leaves of shoot cultures of six dihaploid clones of Solanum tuberosum L. (2n = 2x = 24). In the KM medium (Kao and Michayluk 1975), sustained cell divisions were obtained in up to 50% of the plated protoplasts of four clones, whereas only a few divisions occurred in the other two clones. The first mitosis appeared 2–8 days after plating, dependent on the clones. In the clones showing sustained cell divisions, a protoplast titre of about 5 × 103 per ml turned out to be optimal. The culture conditions for protoplasts of one of the poorly growing clones, clone H2 140, have been improved using modified KM media, plating at a concentration of as high as 5 × 104 cells per ml, and subsequent diluting at intervals 5 days. The dilutions were carried out with media containing 0.25% agar. Up to 60% of the plated protoplasts underwent divisions within 10 days under these conditions. After about 15 days, the regenerants were transferred onto media inducing organogenesis. Shoots and roots were formed on modified media MS (Murashige and Skoog 1962) and B5 (Gamborg et al. 1968). Plants have been regenerated in four of the investigated clones. Countings of chromosomes revealed a satisfactory stability of the karyotype in shoot culture and protoplast regeneration.

Plant regeneration from callus culture in potato

“…Plant regeneration in potato (Sohum tuberosum L.) has been reported from callus culture (SKIRVIN et al, 1975;WANG & HUANG, 1975;ROCA et al, 1978) and from single cell and protoplast culture derived from callus (BEHNKE, 1976;LAM, 1977;SHEPARD & TOTTEN, 1977;BINDING et al, 1978;WENZEL et al, 1979) and protoplast fusion (MELCHERS et al, 1978) This paper describes a relatively simple procedure for callus induction, plant regeneration and multiplication in potato.…”

Plant regeneration from mesophyll protoplasts of tomato cv. Ponderosa

“…The 8p medium was originally devised for the culture of Vicia ha jastana Grossh. protoplasts but has also been found beneficial for several other Solanaceous crops (3,12).…”

Regeneration of Plants from Potato Protoplasts

“…This may be part of the source of success of the method developed by Binding and collaborators (Binding et al 1978). Axenic cultures have been used in producing source material in several laboratories (Butenko et al 1977a;Thomas 1981;Helgeson et al 1982;Bokelmann and Roest 1983;Pellow and Towill 1984).…”

“…No one method has been designed, as yet, that functions uniformly well for all Solanum section Tuberarium species or even for all dihaploid or tetraploid clones of commercial potatoes (Binding et al 1978;Gunn and Shepard 1981;Shepard 1982;Radke 1983). Therefore, a protocol is presented here of methods that have been found to be successful when used on some genotypes, but these state-of-the-art techniques are still subject to modifications.…”