Protoplast

About: Protoplast is a research topic. Over the lifetime, 5474 publications have been published within this topic receiving 122468 citations.

Stable Transformation of Soybean Callus by DNA-Coated Gold

Abstract: Immature soybean (Glycine max L.) embryos from commercially important cultivars were the targets of rapidly accelerated, DNA-coated, gold particles. Protoplasts were prepared from these tissues and propagated in culture under selection conditions for the introduced neomycin phosphotransferase II gene. Kanamycin-resistant calli were obtained at a rate of approximately 10-. Enzyme assays and Southern blot hybridization confirmed the expression of the foreign gene and its stable integration into the soybean genome. Our results show that particle acceleration can be used for the introduction of foreign DNA into the soybean genome and indicate the technique may be useful in the recovery of engineered plants by transformation of regenerable tissues. Successful application of genetic engineering procedures to soybean has been limited by the inability to regenerate plants from transformed tissues. Successful use of nononcogenic Agrobacterium Ti vectors has not been reported, although oncogenic transformation of soybean by virulent Agrobacterium strains and the axenic culture of excised tumors on hormone-free media has been achieved (7, 15, 16, 22, 23). Various assays and markers have been used to confirm transformation events, but caution should be exercised in drawing conclusions regarding putative transformation events (3). We have recently described efficient transformation of soybean protoplasts by electroporation and the recovery of stably transformed calli and roots (4). This technique, however, will not be useful in the recovery of genetically engineered soybean plants until regeneration from protoplasts is accomplished. Klein et al. (13) used DNA-coated tungsten microprojectiles to obtain transient expression of foreign DNA in intact epidermal cells of Allium cepa (onion). However, these authors did not demonstrate stable transformation of the plant cells. Activity through transient expression does not necessarily indicate that stable transformants will be obtained. We have used a microparticle technique to deliver foreign DNA into intact soybean embryo cells and have recovered stable transformants using a protoplast selection regimen (4). Enzyme assays and Southern blot hybridization confirm the integration of the foreign gene and its expression in the soybean genome. Results from these experiments indicate that particle acceleration can lead to stable transformation at rates comparable to electroporation. The properties of the transformants obtained by these two

Method of transforming monocotyledon.

Abstract: A method of transforming monocotyledon which necessitates only a short period from the transformation to the generation of a plant body as compared with the conventional methods, thus reducing the frequency of occurence of mutants, and can be generally applied to the plant for which any system of regenerating the plant body from the protoplast has not been established, and in which the material to be used can be readily prepared. The method comprises transforming cultured tissues of a monocotyledon under or after dedifferentiation with a bacterium of the genus (Agrobacterium) containing desired genes.

Comparison of single cell culture derived Solanum tuberosum L. plants and a model for their application in breeding programs.

Abstract: The techniques of microspore and protoplast regeneration starting from dihaploid Solanum tuberosum plants has been improved to such an extent that the production of more than 2000 microspore derived A1 plant lines and of several hundred protoplast derived plantlets has become possible. Further, from the dihaploid Solanum species S. phureja the regeneration of microspores to plants, and from the species S. infundibuliforme, S. sparsipilum and S. tarijense the regeneration of protoplasts to calluses, has been achieved. The plants descending from the two single cell culture systems are compared with reference to phenotypic markers and economic qualities. Some principles characteristic for either microspore or protoplast derived plants are examined and their significance is discussed. The results are compiled into an extended analytical synthetic breeding scheme based on a stepwise reduction of the autotetraploid to the monohaploid level and a subsequent controlled combination to a new synthetic completely heterozygous tetraploid potato.

Plant Regeneration from Protoplasts Isolated from Embryogenic Maize Cell Cultures

Abstract: Genetic engineering of maize via protoplast technology has been limited due to lack of plant regeneration from maize protoplasts. We have developed a system of protoplast culture that results in high plating efficiency from embryogenic protoplasts and can be followed by plant regeneration. Maize protoplasts were grown on filters directly over a feeder layer of nurse cells in liquid medium. Initial condition and subsequent growth of feeder cells were critical in obtaining a plating efficiency of 10% from protoplasts. Protoplasts were digested from embryogenic cell suspension cultures, and recovered callus retained the morphogenic potential of initial donor cultures. The system has been successfully used with protoplasts of two maize inbreds, one of which is an important commercial line.

Introduction to plant biotechnology

Abstract: Plant tissue culture: laboratory organization nutrition medium sterilization techniques types of culture micropropagation cell suspension and secondary metabolites in vitro production of haploids protoplast isolation and fusion somaclonal variation germplasm storage and cryopreservation. Genetic material and its organization: genetic material organization of DNA and gene expression. Recombinant DNA technology: gene cloning gene transfer methods polymerase chain reaction molecular markers and marker-assisted selection genome mapping transgenics in crop improvement intellectual property rights.