Regeneration protoplast

Collen J, Roeder V, Rousvoal S, Collin O, Kloareg B, Boyen C (2006a) An expressed sequence tag analysis of thallus and regenerating protoplasts of Chondrus crispus (Gigartinales, Rhodophyceae). J Phycol 42 104-112... 

Collen J, Davison IR (1999c) Stress tolerance and reactive oxygen metabolism in the intertidal red seaweeds Mastocarpus stellatus and Chondrus crispus. Plant Cell Environ 22 1143-1151 Collen J, Pedersen M, Bornman CH (1994) A stress-induced oxidative burst in Eucheuma platycladum (Rhodophyta). Physiol Plant 92 417 122 Collen J, Roeder V, Rousvoal S, Collin O, Kloareg B, Boyen C (2006a) An expressed sequence tag analysis of thallus and regenerating protoplasts of Chondrus crispus (Gigartinales, Rhodophyceae). J Phycol 42 104-112... 

While we did not test sufficient numbers of regenerated protoplasts to accurately determine the transformation frequency to aromatic hydrocarbon resistance, these frequencies were certainly high. Chang and Cohen (37) reported transformation frequencies of 10-80% using the same lysozyme-PEG method and similar quantities of plasmid DNA. The naphthalene transformants retained their degradative ability through at least six successive transfers. 

Pardo, M., Monteoliva, L., Pla, J., Sanchez, M., Gil, C., and Nombela, C. 1999. Two-dimensional analysis of proteins secreted by Saccharomyces cerevisiae regenerating protoplasts A novel approach to study the cell wall, feast 15, 459-472. 

Schumann, B., Maiei W. and Grogeg D. (1987) Characterization of some Claviceps strains derived from regenerated protoplasts. Z. Naturforsch., 42c, 381-386. 

J. Spiral and co-workers, "Protoplast Culture and Regeneration in Coffea Species," in Ref. 40. 

Abdullah, R., Cocking, E.C. Thompson, J.A. (1986). Efficient plant regeneration from rice protoplasts through somatic embryogenesis. Biol Technology, 4, 1087-90. 

Our two network model of the primary wall receives support from a variety of indirect observations. For example it has been shown Aat when a cell wall is regenerated by a carrot protoplast a homogalacturonan/ rhamnogalacturonan shell is laid down first, through which the cellulose/ hemicellulose network is later intercalated (8). Further evidence that pectin may form an independent network is seen in the fact that walls from suspension-cultured cells of tofnato Lycopersicon esculentum VF 36),... 

Shea, E.M., Gibeaut, D.M., and Carpita, N.C. (1989) Structural analysis of the cell walls regenerated by carrot protoplasts. Planta, 179 293-308. 

Bacillus subtilis transformants were identified by their resistance to naphthalene, parachlorobiphenyl, or dibenzofuran. Controls subtilis protoplasts regenerated without the addition of B. megaterium plasmid DNA) exhibited no increased reistance to these three compounds. That each transformant was B. subtilis and not a contaminant was verified by testing for the B. subtilis chromosomal markers. Only streptomycin resistant strains that showed arginine and leucine dependent growth were used in subsequent experiments. 

Agrobacterium-mQdi iQd DNA transfer technique and direct gene transfer into protoplasts on the basis of the electroporation technique have frequently been used for the gene transfer methods in plants. However, these methods were hardly adapted to monocotyledonous plants or to varieties in which regeneration system was not established. These deficiencies found in gene transfer techniques will be largely overcome if pollen could be used as a DNA vector. 

Protoplast fusion has also been successfiilly used to produce novel mold isoprenoids, such as citreoanthrasteroids and citreohybridones (Nakada 2000). Long chain aldehydes, valuable as a bio-flavor, have also been secured along similar procedures fiom tMli cultures of the green seaweed Ulva pertusa, regenerated fiom protoplasts (Fujimura 1990). 

Finally, we tested the particle bombardment method, a biolistic approach that was shown before to work when other approaches have failed. In particular, particle bombardment has allowed the generation of transgenic plants such as crop species, which are not susceptible to Agrobacterium tumefaciens or cannot be regenerated from protoplasts. Furthermore, particle bombardment has facilitated organelle transformation in intact cells and the genetic modification of cultured cells that were not accessible to other transfection techniques. 

Ethylene production was restored to some extent when the protoplasts were cultured for 3 or more days (Fig. 10) (58). Restoration of ethylene producing ability by culturing was correlated with regeneration of some cell-wall material, as shown by incorporation of myoinositol in the ethanol-insoluble fraction of the protoplasts and by increased fluorescence with calcafluor-white (58). Regeneration of cell-wall material was correlated with ethylene production in response to methionine added to the cultured protoplasts. Production of ethylene by these cultured protoplasts was not only dependent on addition of... 

If the DNA is maintained stably within the cell and if the protoplasts can be induced to regenerate a whole plant, transgenic plants can be obtained. 

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