The various activated chemical particles RF APGD can alter DNA sequences by widely broken down the C-N bond between base and ribose, amino groups on the base, as well as the P-O bonds in phosphodiester. When working, the plasma gas (helium with purity of 99.99% or better) flowing through the discharge region between the two electrodes is ionized by the externally applied RF electric field, and thus, a non-thermal plasma jet consisting of various activated chemical particles is formed at the downstream of the plasma torch nozzle exit. RF APGD can be produced between two water-cooled bare-metallic electrodes driven by a radio-frequency power supply.
#MEGATUNE GENERATE AFR TABLE GENERATOR#
Moreover, atmosphere and room temperature plasma (ARTP) is a newly developed mutation system for microbes that uses a helium radio-frequency atmospheric-pressure glow discharge (RF APGD) plasma generator as its core component. In medaka, a specific locus test system for environmental mutagenesis was established using 137Cs γ-ray irradiation. In addition to chemical mutagens, physical mutagens such as γ-ray, X-ray, UV, or particle radiation also play important roles in mutation induction. While consider mutation rate of specific loci based on phenotype, the mutation rate was 0.15% in zebrafish and 0.1%~ 0.195% in medaka. When consider the mutation rate that based on specific gene, the mutation rate was 1 per 297 kb (0.0003%) in fugu, 1 per 345 kb in medaka (0.0003%), 1 per 235 kb in zebrafish (0.0004%) and 0.41% in grass carp ( Ctenopharyngodonidellus). The ENU has shown relatively high mutation frequency. One of the widely used chemical mutagens, N-ethyl- N-nitrosourea (ENU) acts as an alkylating agent, transferring its ethyl group to nucleophilic nitrogen or oxygen sites on deoxyribonucleotides, leading to base mismatching during DNA replication, and thus, it mainly induces single-base substitutions. Artificially induced mutation in fish usually employs physical radiation or chemical mutagens. In fish, the natural mutation rate at specific loci is generally lower than 1.0 × 10 − 6. Mutations are the basis of genetic variation, and naturally occurring mutations play important roles in evolution. No matter which kind of breeding method is used, the successful breeding of improved strains basically depends on the mutations present in the base population. The purpose of breeding in aquaculture fish is to discover and generate economically important traits such as fast growth, disease resistance or meat quality. ConclusionsĪRTP mutagenesis is a useful method for breeding of fish species to accelerate the selection of economically important traits that would benefit the aquaculture industry, given the variety of mutations detected. Gene ontology clustering indicated that genes associated with cell components, binding function, catalytic activity, cellular process, metabolic process and biological regulation processes had higher mutation rates. The average mutation rate reached 0.064% at the genome level. In total, 69.25Gb clean data were obtained from three controls and eight randomly selected ARTP-treated individuals, revealing 240,722 to 322,978 SNPs and 82,149 to 86,798 InDels located in 17,394~18,457 and 12,907~13,333 genes, respectively. Whole-genome sequencing was used to reveal the mutation features of ARTP-treated individuals collected at day 120 after hatching. The ARTP-treated group presented differences in morphological traits such as body height, total length among individuals at day 90 after hatching. Under an ARTP radio-frequency power input of 200 W, the optimal treatment time for sperm diluted with Ringer’s solution by 1:40 v/v was 10 min. Eggs fertilized for 60 min were treated by ARTP with a radio-frequency power input of 120 W, and the ARTP treatment time was 25 min. ARTP treatment time was optimized for the fertilized eggs and sperm, respectively. We reported the first successful mutagenesis in a marine aquaculture fish species, Japanese flounder, Paralichthys olivaceus, using a novel atmosphere and room temperature plasma (ARTP) mutagenesis tool. Hence, development of a simple, safe and effective mutagenesis method with a high mutation rate and applicability to multiple fish species, is needed. However, the traditional mutation methods for fish breeding are not convenient or safe for daily use. Artificial induction of mutagenesis is effective for genetic resource innovation and breeding.
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