Protoplast-Independent Production of Transgenic Plants
Problems encountered with protoplast-based methods for the generation of transgenic plants have prompted the development of alternative techniques for gene transfer in grasses. These problems relate mainly to plant regeneration from protoplasts and do not reflect specific barriers to the uptake of foreign DNA by isolated protoplasts. Examples of these difficulties are relatively low plating efficiencies and low plant regeneration frequencies from protoplasts, species and genotype dependence often observed in the regeneration process, and albinism and somaclonal variation revealed in protoplast-derived plants (Potrykus 1990). Plant regeneration from protoplasts is thus a delicate process depending upon parameters that are not under experimental control, such as wound response and genotype-dependent competence for regeneration (Vasil 1988; Potrykus 1990). Furthermore, transgenic plants recovered from protoplasts may show serious fertility constraints and undesired integration of multiple and rearranged transgene copies (Spangenberg et al. 1995a).
KeywordsTransgenic Plant Tall Fescue Perennial Ryegrass Immature Zygotic Embryo Italian Ryegrass
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