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

• Fisheries and Aquatic Sciences
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• v.4 no.1
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• pp.39-46
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• 2001

To examine the effect of copy number-dependent transgenic genotype on the expression of foreign gene, stable hemizygous and homozygous transgenic breeding line was established using artificial parthenogenesis. For this purpose, induced diploid gynogenetic transgenesis was optimized in mud loach (Misgurnus mizolepis) using UV-irradiated cyprinid loach (M. anguillicaudatus) sperm and thermal shocks. Optimum UV range for inactivation of cyprinid loach sperm was between 3,150 to $4,050\;ergs/mm^2$ The UV-irradiated sperm were inseminated into eggs from recessive color strain (yellow) or heterozygous transgenic mud loach containing CAT gene. Cold shock at $2^{\circ}C$ for 60 min, 5 min post fertilization successfully restored the diploidy of eggs inseminated with UV-irradiated sperm. Restoration to diploidy was confirmed by flow cytometry and gynogenetic status was verified by examining maternal exclusive inheritance of multi-locus DNA fingerprints, body color and transgenic marker. Putative isogenic transgenic fish clearly showed homozygous status at trans gene locus based on Southern blot hybridization and progeny testing. Further, such homozygous gynogenetic diploids revealed the increased levels of transgene expression, when compared to those of heterozygous (hemizygous) transgenic fish.

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