• Journal of Aquaculture
• /
• v.13 no.4
• /
• pp.359-362
• /
• 2000

Mitotic gynogenesis was induced in the far eastern catfish, Silurus asotus using UV-irradiated heterospecific sperm and cold shock treatment. Eggs were activated with the sperm of mud loach, Misgurnus mizolepis which has been irradiated with UV at dose of 9,000 ergs/$mm^2$. To determine the optimum duration required to prevent the first cleavage, a cold shock at 4$^{\circ}C$ with duration of 20, 30 or 40 min was applied to the eggs 50 min after activation. To induce diploidization of mitogenesis, the most effective protocol was to apply cold shock to 50-min old (after activation) eggs at 4$^{\Circ}C$ for 30min.

• Journal of Aquaculture
• /
• v.12 no.3
• /
• pp.167-173
• /
• 1999

Blocking the 1st mitotic cleavage was performed in mud loach (Misgurmus mizolepis) using UV-irradiated cyprinid loach (M. anguillicaudatus) sperm and ternal shocks Optimum UV range for inactivation of cyprinid loach sperm and thermal shocks. Optimum UV range for inactivation of cyprinid loach sperm was between 3,150 to 4,050 ergs/m$m^2$. Heat shock treatment ($41^{\circ}C$ for 3mins) with various treatment initiation times ranged from 22 to 50 min post insemination resulted wide range of success for induced gynogenesis. Best result was obtained when haploid egges were shocked at 28 min after insemination (corresponding to metaphase division of the 1st cleavage); 26% of total eggs inseminated were viable diploid gynogens. The hatching success and early survival of the both meiotic and mitotic gynogenetic groups were significantly lower than those of control crosses (P<0.05). Maternal origin of induced gynogenetic mud loach was verified by multi-locus DNA fingerprinting.

• Proceedings of the Korean Aquaculture Society Conference
• /
• 2003.10a
• /
• pp.38-38
• /
• 2003

Blocking the first mitotic cleavage of the zygote is a key tool for chromosome-set manipulations in fish. We developed an improved method for inducing tetraploidy by blocking the mitosis with a combination of heat shock at 40.5$^{\circ}C$ for 1, 2 or 3 min followed by cold shock at $1.5^{\circ}C$ for 30, 45 or 60 min. When applied during the first cleavage metaphase of mud loach (Misgurnus mizolepis) zygotes, the optimal combination was heat for 2 min followed by cold for 45 min. At 1 month, the frequency of 4N survivors and the yield from total eggs fertilized was 55.7% and 14.4%, respectively, compared to heat shock alone with 20.0% efficiency and 3.6% yield. The effectiveness of the procedure was confirmed by diploid mitotic gynogenesis using transgenic markers. The overall yield of homozygous diploids, 34.0%, was better than that for single heat shock, 17.3%. The tetraploids and homozygous diploids had higher early mortality than normal diploid controls. However at 1 month, the viability of the tetraploids was the same as normal diploids. For gynogenetic diploids, the survival was similar to normal diploids after 3 months. The high efficiency of this new protocol extends the opportunity to study polyploidy in basic and applied research.

• Development and Reproduction
• /
• v.4 no.1
• /
• pp.79-85
• /
• 2000

This study was made to optimize the conditions needed to produce two types of gynogenetic diploids in the sweet fish, Plecoglossus altivelis. Firstly, ultraviolet (UV) ray doses between 3,000 erg to 14,000 erg/$\textrm{mm}^2$ were tested to inactivate sperm genetically. Based on the appearance of the haploid syndromes in the embryo, a dose of UV ray 6000~7000 erg was required to inactivate sperm genetically. Then, cold shock treatment at 1~2$^{\circ}C$ for 15~30 min were conducted to retain the 2nd polar body in inseminated egg. The best elapsed time before the start of the cold shock was examined between 5~8 min. The experiments in which began 5 min after insemination at 1~2$^{\circ}C$ during 17.5 min gave 21.2％ survival rate and 89.7% normal eyed embryo rate. The gynogenetic diploid produced by suppression of the first cleavage, a considerably high number of heteroploids appeared and high mortality was observed at the metamorphosis stage, so further investigation is needed. The production of gynogenetic diploids were confirmed by GPI isozyme marker. The heterozygous type in Gpi-1 locus was observed in the meiotic-G2N as a result of gene-centromere recombination during meiosis. The heterozygous type was never observed in mitotic-G2N and showed segregation into two homozygous types at Gpi-1 locus.

• Development and Reproduction
• /
• v.4 no.2
• /
• pp.153-160
• /
• 2000

In this study, genetic variation in the 5 strains of 2N-cont, meiotic-G2N, mitotic-G2N and two types of clones with different genetic backgrounds was investigated by developing their tolerance to water temperature and salinity, which is a physiological trait, into a quantitative trait. The temperature was set at 19$^{\circ}C$, 22.5$^{\circ}C$, 25$^{\circ}C$ and 30$^{\circ}C$, each of which was combined with 0$\textperthousand$, 15$\textperthousand$ and 30$\textperthousand$ of salinity respectively, making 12 groups in all. In the mean survival time (MST), samples with 15$\textperthousand$ of salinity showed the longest survival time at all temperatures. The 2N-cont had the longest 126.16 h followed by clone-11 and clone-15 surviving for 113.22 h and 91.05 h respectively. Gynogenetic diploids showed the shortest 87,32 h and 36.56 h. At 22.5 and 25$^{\circ}C$, MST of each strain was significantly short, showing similar results to those of the groups at 19$^{\circ}C$. The 2N-cont had the longest MST while clones had a longer MST than gynogenetic diploids. This could be due to gynogenesis which causes homozygosis among malignant harmful genes, leading to its appearance in populations and resulting in early death in individuals with such genes. On the other hand, MST of clones was longer than that of gynogenetic fish. This could be because the 1st gynogenetic generation, which is a parental population, has already had its malignant genes removed, while the clones of the 2nd gynogenetic generation have had their superior genes fixed as well as their tolerance and survival improved. When temperature was raised to 22.5$^{\circ}C$ and 25$^{\circ}C$, increase in variation was observed in gynogenetic diploids and decrease in clones in 15$\textperthousand$ of salinity. This shows that such a trait is genetic to a certain extent. Consequently, if this character is developed into a quantitative trait and applied to selective breeding, it could be a useful character to secure superior strains and individuals, and also it would be possible to improve populations genetically through selection.