Protoplasts as tools for plant genome modifications

Abstract

We present here the application of protoplast technology in the selection and recovery of rare, spontaneous plant genome alterations. Using protoplasts as a cell cloning system allowed the detection and molecular characterization of intrachromosomal recombination events between genomic repeats. The mechanism, frequencies and the induction of intrachromosomal recombination are discussed as well as its application for genome mutagenesis.

Gene replacement

“…Treat-ments that induce damage to DNA have been shown to increase rates of intrachromosomal recombination in plant cells [34,53,75]. Such treatments may also increase targeting frequencies but they will have an unwanted side-effect of introducing other mutations into the plant material.…”

Identification and Characterization of an Antifungal Gene Mt1 from Bacillus subtilis by Affecting Amino Acid Metabolism in Fusarium graminearum

“…The library preparation for whole-genome sequencing (WGS) using next-generation sequencing (NGS) begins with DNA fragmentation, and sonication is the most commonly used approach due to its ease and reliability [15,16]. In recent years, PEG-mediated protoplast transformation and homologous recombination methods for knocking out and knocking in target genes have become widely popular, and researchers using both can study functional genes, metabolic regulatory networks, and genetic engineering in plants, animals, and microorganisms [17][18][19][20][21]. Gene knock-in allows researchers to study the function of specific genes in fungi as well as engineer fungi for various biotechnological applications [22][23][24].…”