• Journal of Plant Biotechnology
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• v.38 no.1
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• pp.30-41
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• 2011

The influences of the agitation as well as the addition of medium during culture on the production of embryos were invested in isolated microspore culture of hot pepper (Capsicum annuum L.). When the culture medium was added during initial liquid culture step of liquid-double layer culture, the embryo yield and quality greatly increased. The most effective time point for medium addition was 5 days after the culture commenced. On the other hand, the effect of medium addition at later double layer culture step in liquid-double layer culture on the embryo production was less compared to that of medium addition during the initial liquid culture step. Agitating the culture for 1 week during later double layer culture step in liquid-double layer culture effectively increased the production of normal cotyledonary embryos. In the case of liquid culture, agitating the culture for 1 week from 7 days after the culture commenced was also effective for embryo development. However, when the total agitation time was longer (2 to 3 weeks) during liquid-double layer culture or liquid culture, the embryos developed abnormally in both cases. The normal cotyledonary embryos obtained in this study successfully developed to plants when transferred to regeneration media. These regenerated plants were either diploid or haploid, and there was a difference in the number of chloroplasts between guard cells of diploid and haploid. These results can be used as an important data for developing an efficient microspore culture system with high quality embryo production in hot pepper.

• Journal of Life Science
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• v.20 no.11
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• pp.1634-1639
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• 2010

This study was conducted to increase the efficiency of farming practice in rainbow trout, Oncorhynchus mykiss, by sex reversal and chromosome-set manipulation techniques. To obtain phenotypic males, hormonal sex reversal was carried out using an exogenous hormone treatment method. 5 mg of 17 alpha-methyltestosterone per kg diet was supplied for 82 days after first feeding at $10^{\circ}C$ and $13^{\circ}C$. More than 93% of the male population was produced by this method and growth of hormone-treated fish at $13^{\circ}C$ was faster than that of untreated bi-sexual groups. Induced diploid gynogenesis was carried out using artificial insemination of UV-irradiated sperm into haploid eggs. Based on the appearance of the rate of haploid syndrome and survival of embryo, a UV ray dose of at least $3,600\;erg/cm^2$ was required to inactivate rainbow trout sperm genetically. Haploid embryos were restored to diploid by blocking the extrusion of the second polar body using heat shock treatment at $28^{\circ}C$ for 20 min, 10 min post insemination. Gynogenetic diploid sex ratios were confirmed after maturation of the fish erythrocyte measurements and chromosome counts.

• Korean Journal of Microbiology
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• v.33 no.3
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• pp.199-202
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• 1997

The effect of electric shock (AC 38 Vll.3 cm) on the lethal effect and induction rate of mutation of chemical mutagen N-methyl-N'-nitro-N-nitrosoguanidine (NTG) was examined by using the spores of Streptomyces and the cells of haploid and polyploid yeast strains of Saccharomyccrs. Spores of Sireptomyces were all alive after 180 min of electric shock, hut all dead after 960 min-treatment. When the spores of Streptomyces or the cells of haploid and polyploid yeast were treated with electric shock and NTG, the electric shock increased the lethal effect of NTG; the survival rate of Streptomyces dropped from 72 to 48% after 180 min-treatment and those of haploidand polyploid-yeast decreased from 8 to 3% and 25 to lo%, respectively, after 40 min-treatment. The electric shock also increased mutation rates of Streptomyces and haploid yeast. from 1.8 to 13.6%' after 120 min-treatment and from 2.4 to 4.8% after 40 min-treatment, respectively.

• Journal of Embryo Transfer
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• v.16 no.3
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• pp.223-231
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• 2001

The success of nuclear transplantation with mammalian oocytes depends critically on the potential of oocytes activation, which mainly caused to prevent the re-accumulation of maturation promoting factor (MPF). This study was conducted to compare the effect of combined treatment of lonomycin with a Hl-histone kinase inhibitor (dimethylaminopurine, DMAP) or cdc2 kinase inhibitor (sodium pyrophosphate, SPP) on activation of bovine oocytes. In vitro matured bovine oocytes with the first polar body (PB) and dense cytoplasm were assigned to 3 experimental groups. For activation treatment, oocytcs were exposed to 5 $\mu$M lonomycin for 5 min (Group 1), and followed by 1.9 mM dimethylaminopurine (DMAP) for 3 h (Group 2) or followed by 2 mM sodium pyrophosphate (SPP) for 3 h (Group 3). The activation effects in the three treatments and the control group (untreated) were judged by the extrusion of the second PB and formation of a pronucleus (PN). Differences among groups were analysed using one-way ANOVA after arc-sine transformation of proportional data. All three treatments led to high activation rates (90% to 95%), with significant difference from the control. However, the extrusion of the second PB and the rate of PN formation differed remarkably among treatments. In Group I and 3, about 95% of the oocytes had extruded the second polar body, but one PN had formed in a higher proportion of oocytes in Group 3 than in Group 1 (90% vs. 5%). In experiment 2, the rates of cleavage and development into blastocysts in Group 1 were significantly lower than those of Group 2 and 3 (8.7% and 0% vs. 50.5% and 11.6%, and 44.6% and 7.2%, respectively, P<0.05). In experiment 3, ~80% of parthenotes in Group 1 were developed with haploid chromosomal sets. However, when ionomycin was followed immediately by DMAP (Group 2). only 20% of parthenotes were haploid. In Group 3, combined treatment with ionomycin and SPP, the appearance of abnormal chromosomal tracts was significantly (P〈0.05) reduced and the proportion of haploid parthenotes was increased to 85% (17/20) than in Group 2. These results demonstrate that SPP acted as a cdc2 kinase inhibitor and formed the haploidy in oocyte activation. Thus, the present study suggests that cdc2 kinase inhibitor, such as sodium pyrophosphate, may have an effective role in oocyte activation for the production of cloned embryos/animals by nuclear transplantation.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 2005.10a
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• pp.78-80
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• 2005

• Korean Journal of Microbiology
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• v.22 no.3
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• pp.197-205
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• 1984

After the previous paper, this chromosomal studies on the fungi were dealt with 17 species in genus of Rhizopus. The results are sumarized as the followings; The haploid chromosome number of 17 species were confirmed as of 6(Rh. oligosporus), 8(Rh. homothallicus, Rh. liquefaciens, Rh. shanghaiensis, Rh, acetorinus), 12(Rh. microsporus, Rh. pseudochinensis, Rh, rhizopodiformis, Rh, thermosus, and Rh. kazanensis), 14(Rh. stolonifer), and 16(Rh. suinus), respectively. Referring to the above fact and the previous paper, it is strongly presumed that the basic chromosome number of Rhizopus are 4.

• Proceedings of the Korean Society of Crop Science Conference
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• 1999.05a
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• pp.126-127
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• 1999

• Proceedings of the Korean Society of Crop Science Conference
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• 2007.11a
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• pp.216.2-216.2
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• 2007

• Korean Journal of Plant Resources
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• v.3 no.1
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• pp.1-30
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• 1990

The traditional plant imprownent methods consisted of pure line selection, cross breeding, heterosis breeding, polyploid breeding, mutati-onbreeding, ect.Biotechmoiogy is divided into gene spliclng , monocle-nal antibodies , protein engineering , agricultural research, and microbiological engineering. Of these , high plants deal with agricultural research, and the importent part of which is tissue culture and celLculture , Tissue .culture and cell culture are again divided into embryoculture, test tube fertilization, anther and pollen culture, somatichybridization , transformation, recombination, recombinant DNA moleculehybrid plasmid, ect For these haploid production, protoplast culture,protoplast fusion, selection and propagation, ect. , the technical sett-lement is needed.

• 대한생식의학회:학술대회논문집
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• 2002.11a
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• pp.96-96
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• 2002