Study of factors affecting the culture of Brassica napus L. and B. juncea Coss. mesophyll protoplasts

Abstract

Various factors affecting the culture of Brassica napus and B. juncea mesophyll protoplasts were examined in order to develop suitable culture media for these species. The basic components (salts and vitamins) of culture media K3 and Kao best supported cell division and colony development in protoplast culture of both species. The addition of casamino acids to Kao's medium resulted in colony browning in B. napus genotypes. B. napus protoplasts grew well with glucose as the osmotic stabilizer, whereas B. juncea protoplasts responded better to sucrose. High NAA and low 2,4-D combinations were effective in stimulating colony growth. Colony development was rapid for a range of genotypes cultured with these recommendations in these media and plant regeneration was obtained from protoplast-derived calli in both species.

Studies of cotyledon protoplast cultures from Brassica napus, B. campestris and B. oleracea. I: Cell wall regeneration and cell division

“…Kao & Seguin-Swartz (1987) also observed that the frequency of protoplast division was low and colony development was poor when mesophyll protoplasts of B. napus and B. juncea were cultured in this medium. All Brassica cultivars gave their lowest frequency of cell division when their protoplasts were cultured in a modified NN medium.…”

The blackleg fungus: phytotoxins and phytoalexins

“…The protoplast bioassays were conducted with mesophyll protoplasts of a number of cruciferous species (B. carinata Braun, B. juncea, B. napus, and Sinapsis alba L.). The protoplasts were isolated from plants grown in vitro and cultured in vitro according to Kao and Séguin-Swartz (1987). Protoplast viability was assessed visually or with fluorescein diacetate (Heslop-Harrison & HeslopHarrison 1970).…”

High plating efficiency and plant regeneration frequency in low density protoplast cultures derived from an embryogenic Brassica napus cell suspension

“…Plant regeneration has been reported from protoplasts derived from many different types of tissues, including leaves (Kartha et al 1974;Thomas et al 1976;Li & Kohlenbach 1982;Pelletier et al 1983;Glimelius et al 1984;Kao & Seguin-Swartz 1987), roots (Xu et al 1982), stem embryos , hypocotyls (Glimelius 1984;Chuong et al 1985; Barsby et al 1986), and stem cortical strips (Klimaszewska & Keller 1987). Generally, these systems required high plating densities of protoplasts to initiate division, and plating efficiencies were low.…”