Protoplast

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

"Arabidobrassica": A novel plant obtained by protoplast fusion.

Abstract: Using somatic hybrid cell lines Arabidopsis thaliana+Brassica campestris, obtained by cloning individual protoplast-fusion products as starting material, shoots and flowering plants have been regenerated. Cytological, biochemical, and morphological analyses indicate that genetic material of both species is present in the resultant plants. Shoots and plants obtained from different lines and different regeneration events differed morphologically and genetically. Most regenerants show morphological abnormalities and unusual organizational patterns. Flowering forms have so far been sterile. "Asymmetric" hybrids (i.e., hybrids bearing most genetic material of one of the parent species and only few chromosomes of the other) were more regular in morphology. The results represent the first case of intergeneric-intertribal hybridization of flowering plants.

Genetic Recombination in Streptomyces fradiae by Protoplast Fusion and Cell Regeneration

Abstract: SUMMARY: Conditions for highly efficient genetic recombination in Streptomyces by protoplast fusion are described. Protoplasts of S. fradiae and S. griseofuscus were formed by a modification of the glycine-lysozyme-lytic enzyme method (Okanishi, Suzuki & Umezawa, 1974). Regeneration of cells from protoplasts was monitored throughout the growth cycle and was most efficient when cells of either S. fradiae or S. griseofuscus were taken from the transition phase between the exponential and stationary growth phases. Fusion of protoplasts carrying different auxotrophic or chromosomal drug-resistance markers was achieved by treatment with polyethylene glycol, and high frequencies of stable genetic recombinants were obtained.

Expression of a foreign gene linked to either a plant-virus or a Drosophila promoter, after electroporation of protoplasts of rice, wheat, and sorghum

Abstract: The bacterial chloramphenicol acetyltransferase (CAT) gene was expressed in protoplasts of three important graminaceous plant species after introduction of the gene by electroporation. Gene transfer occurred when high-voltage electric pulses were applied either directly or indirectly (without anode contact) to a solution containing plasmid DNA and protoplasts of rice, wheat, or sorghum. The indirect method was more rapid, resulted in higher protoplast viability, and was less subject to contamination than the direct-contact method. Gene expression of approximately equal magnitude resulted when the CAT gene was fused to either the 35S promoter of cauliflower mosaic virus or the copia long terminal repeat promoter of Drosophila. Together with recent advances in regeneration of callus and whole plants from protoplasts, this system makes it possible to study inheritance and expression of genes introduced into graminaceous monocotyledonous plants.

Analysis of pre-mRNA processing in transfected plant protoplasts.

Abstract: Publisher Summary The analysis of RNAs transiently expressed in transfected plant protoplasts appears to be at present the most suitable approach for addressing the problems of plant mRNA processing. Transient expression experiments have been reported with protoplasts originating from a number of different tissues or suspension cultures from both dicotyledonous and monocotyledonous plants. In most instances the expression of introduced plasmids has been determined by assaying the activity of enzymes encoded by reporter genes. When protoplasts are to be transfected by electroporation, the filtrate is distributed into 12-ml polystyrene round-bottomed Falcon tubes (10 ml/tube), and the protoplast suspension in each tube is overlayed with 1 ml of EP solution. After centrifugation at 100 g for 10 min the floating protoplasts are collected, washed 2 times with EP, resuspended in EP, and used immediately for electroporation. Two alternative procedures can be used for transfecting protoplasts to study transient gene expression. Electroporation involves the use of an electric field to stimulate the uptake of exogenous DNA by the protoplasts.

Large-Scale Isolation of Intact Vacuoles and Isolation of Chloroplasts from Protoplasts of Mature Plant Tissues

Abstract: Intact plant vacuoles were prepared in large numbers (10/sup 6/) from protoplasts of mature leaves, flower petals, stems, pedicels, filaments, styles, and young fruits. Treatment of protoplasts with 0.2 molar K/sub 2/HPO/sub 4/-HCl, pH 8, with slow stirring resulted in gentle osmotic rupture of the protoplasts and release of intact vacuoles. Particulate components of the protoplast, less the vacuole, were largely shed as an aggregate, which was removed by filtration. Vacuoles were recovered from the filtrate by low-speed centrifugation. The general procedure was also used to isolate chloroplasts with a high degree of integrity and excellent photochemical activity. (auth)