Abstract
Clonal fishes are useful tools in biology and aquaculture studies due to their isogenicity. In Japanese flounder (Paralichthys olivaceus), a group of homozygous clones was created by inducing meiogynogenesis in eggs from a mitogynogenetic homozygous diploid. As the clones reached sexual maturity, meiogynogenesis was again induced in order to produce a 2nd generation clonal group of Japanese flounder. After 3 months, there were 611 healthy, surviving individuals. Twenty-four microsatellite markers, that covered all the linkage groups of Japanese flounder, were used to identify the homozygosity of the 2nd generation clones; no heterozygous locus was detected. This indicates that the production of a 2nd generation clonal group of Japanese flounder was successful. Restriction-site DNA associated sequencing at the genomic level also confirmed the homozygosity and clonality of the 2nd generation clonal group. Furthermore, these 2nd generation clones had a small coefficient of variation for body shape indices at 210 days of age and showed a high degree of similarity in body characteristics among individuals. The successful production of 2nd generation clones has laid the foundation for the large-scale production of clonal Japanese flounder.
Highlights
Laboratory animals are commonly used for research purposes in several fields, including medicine, biology, and environmental toxicology etc
The use of eggs or sperm that are spawned by the double haploid (DH) to induce meiogynogenesis or androgenesis could establish a clonal line that has a coefficient of inbreeding of F = 1.00
The control group consisted of eggs from the 1st generation clones that were fertilized by sperm from wild-type males or double haploid males
Summary
Laboratory animals are commonly used for research purposes in several fields, including medicine, biology, and environmental toxicology etc. The matured eggs are spawned out of the body to be fertilized by sperm in the water when they are in the metaphase of second meiosis[5]. These reproductive traits make it possible to artificially induce polyploidy, androgenesis, and gynogenesis. The use of eggs or sperm that are spawned by the DHs to induce meiogynogenesis (eggs) or androgenesis (sperm) could establish a clonal line that has a coefficient of inbreeding of F = 1.00. Compared with full-sib mating, use of the chromosome manipulation method can establish a clonal line that has full homozygosity in short a period of time (2 generations). We describe the creation of a 2nd generation clonal group of Japanese flounder by induced meiogynogenesis using eggs spawned by 1st generation clones (Fig. 1)
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