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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45 citations
TL;DR: In this paper, the authors describe the successful use of protoplast fusion in Escherichia coli and obtain prototrophic strains by recombination in fused protoplasts at frequencies of 0.05-0.7% based on the number of proteins subjected to fusion.
45 citations
TL;DR: The method described represents a highly efficient and reliable technique for protoplast production from Aspergillus nidulans and corresponded to both the point of maximum autolysis and maximum lytic enzyme activity.
Abstract: The rate and degree of autolysis of Aspergillus nidulans was investigated during a 16 day incubation period. Changes in the activity of chitinase, B -glucanase, α-glucanase, invertase, esterase, acid and alkaline phosphatase and protease liberated into the culture fluid were monitored during growth and autolysis. Protoplasts were released from A. nidulans mycelium by lytic enzymes present in autolytic-phase culture filtrates of the same organism. Maximum protoplast-liberating activity corresponded to both the point of maximum autolysis and maximum lytic enzyme activity. The method described represents a highly efficient and reliable technique for protoplast production from Aspergillus nidulans.
45 citations
TL;DR: The PEG-mediated banana protoplast transformation system can serve as a rapid and effective tool for transient expression assays and sgRNA validation in banana and enables the generation of banana plants engineered by DNA-free gene editing.
Abstract: To date, CRISPR/Cas9 RNP editing tools have not been applied to the genetic modification of banana. Here, the establishment of a PEG-mediated banana protoplast transformation system makes it possible to build an efficient DNA-free method for a site-directed mutagenesis system. Protoplasts constitute a versatile platform for transient expression in plant science. In this study, we established a PEG-mediated banana protoplast transformation system. This system was further optimized for successfully delivering CRISPR/Cas9 and CRISPR/Cas12a plasmids and CRISPR/Cas9 ribonucleoproteins (RNPs) for targeted delivery of the PDS gene into banana protoplasts. Specific bands were observed in PCR-Restriction Enzyme Digestion (PCR-RE) assays, and Sanger sequencing of single clones further confirmed the occurrence of indels at target sites. Deep amplicon sequencing results showed that the editing efficiency of the CRISPR/Cas9 system was higher than that of the other two systems. The PEG-mediated banana protoplast transformation system can serve as a rapid and effective tool for transient expression assays and sgRNA validation in banana. The application of the CRISPR/Cas9 RNP system enables the generation of banana plants engineered by DNA-free gene editing.
45 citations
TL;DR: Carrot cells cultured in vitro as well as protoplasts isolated from these cells formed embryos and grew into mature plants that exhibited a higher frequency of tetraploids than those grown from untreated protoplast and from the original cells.
Abstract: Carrot cells cultured in vitro as well as protoplasts isolated from these cells formed embryos and grew into mature plants. Treatment of the protoplasts with polyethylene glycol (PEG) did not interfere with the capacity of this material to multiply and differentiate. Plants grown from PEG-treated protoplasts exhibited a higher frequency of tetraploids than those grown from untreated protoplasts and from the original cells.
45 citations