Topic
Protoplast
About: Protoplast is a research topic. Over the lifetime, 5474 publications have been published within this topic receiving 122468 citations.
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TL;DR: Electron microscopy has been used to assess the efficiency of four different techniques as potential methods for fusing isolated plant protoplasts, and the methods are discussed in terms of their likely applicability as a practical fusion technique.
Abstract: Electron microscopy has been used to assess the efficiency of four different techniques as potential methods for fusing isolated plant protoplasts. Protoplast aggregation has been achieved by the use of an antiserum, high pH, polyethylene glycol 6000 or poly-L-lysine. The ultrastructural characteristics of each treatment are described. Fusion of adjacent surface membranes does not necessarily fellow aggregation, and has not been observed in protoplasts aggregated by antisera. The other three methods all result in some degree of fusion, depending upon the state of the protoplasts and the temperature of the experiment. Fusion is favoured by a high density of cytoplasm in the protoplasts and by higher temperatures. The methods are discussed in terms of their likely applicability as a practical fusion technique.
35 citations
TL;DR: Conditions that were found to be optimal for preparing and regenerating Schizosaccharomyces pombe protoplasts for cell fusion, and cell fusion is optimal when both fusion partners are fully protoplasted, although considerable fusion occurs between spheroplasted cells as well.
Abstract: Preparation and regeneration of protoplasts is essential for somatic hybridization and transformation of yeasts. We present conditions that were found to be optimal for preparing and regeneratingSchizosaccharomyces pombe protoplasts for cell fusion. In contrast to these conditions, genetic transformation ofS. pombe requires spheroplasts that are osmotically sensitive, but still have some wall material attached to the cell. The main finding were as follows: (a) For protoplast formation with Novozym SP234, 0.9M sorbitol was found to be the optimal osmotic milieu and β-mercaptoethanol is not necessary. (b) Embedding in soft agar yields considerably better regeneration frequencies than direct plating. (c) Cell fusion is optimal when both fusion partners are fully protoplasted, although considerable fusion occurs between spheroplasted cells as well. (d)Schizosaccharomyces pombe transformation frequencies are much higher with spheroplasts than with protoplasts. Inclusion of β-mercaptoethanol did not enhance transformation frequency.
35 citations
TL;DR: Embryogenic callus cultures were initiated from the mature caryopses of Zoysia japonica through precocious germination of somatic embryos and Numerous calli were recovered after transferring protocolonies onto an agar medium.
Abstract: Embryogenic callus cultures were initiated from the mature caryopses ofZoysia japonica. Plant regeneration was through precocious germination of somatic embryos. Protoplasts were isolated from the callus and cultured in a medium solidified with agarose. Numerous calli were recovered after transferring protocolonies onto an agar medium.
35 citations
TL;DR: The results show increased cellulase secretion by P. echinulatum strains and show that protoplast fusion techniques associated with an efficient selection process could be a useful tool in genetic improvement to generate better cellulose-producing strains aimed at economically hydrolyzing cellulose with a view to second-generation ethanol.
35 citations
TL;DR: From chromosomal and biochemical analysis, it can be concluded that the procedure of fusing microspore protoplasts of species A carrying a dominant selection marker with WT somatic cell protoplast of species B can be a convenient selection method for the synthesis of triploid plants.
Abstract: Microspore protoplasts (n) isolated at the tetrad stage from plants of Nicotiana tabacum Km+ (2n=4x=48) were fused with somatic cell protoplasts (2n) of WT N. rustica (2n=4x=48) to produce triploid plants. A total of 21.2×106 microspore protoplasts were fused with 11.2×106 somatic cell protoplasts using the high pH/Ca+ + method. Microspore protoplasts did not divide and WT N. rustica protoplasts stopped dividing when the protoplast-derived colonies were transferred to a selection medium containing kanamycin. A total of 104 actively growing green colonies were recovered on the selection medium. Ninety-six of these colonies were tested for their hybrid nature by PAGE of peroxidases and were found to contain bands characteristic of both parents. Hybrid nature of the plants regenerated from some of the selected colonies was confirmed by IEF of leaf esterases, by NPT II activity assay and by hybridizing total DNAs restricted with EcoR I to a cloned 18s rDNA fragment. Root tip squashes of six of the hybrid plants revealed chromosome numbers ranging from 58–72. From chromosomal and biochemical analysis, it can be concluded that the procedure of fusing microspore protoplasts (n) of species A carrying a dominant selection marker with WT somatic cell protoplasts (2n) of species B can be a convenient selection method for the synthesis of triploid plants. The significance of triploids lies in their subsequent use for transferring alien chromosomes and genes of species A to species B.
35 citations