• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.121-121
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• 2003

The standard protocol for the production of transgenic mouse from ES-injected embryo has to process via chimera producing and several times breeding steps, In contrast, tetraploid-ES cell complementation method allows the immediate generation of targeted murine mutants from genetically modified ES cell clones. The advantage of this advanced technique is a simple and efficient without chimeric intermediates. Recently, this method has been significantly improved through the discovery that ES cells derived from hybrid strains support the development of viable ES mice more efficiently than inbred ES cells do. Therefore, the objective of this study was to generate transgenic mice overexpressing human resistin gene by using tetrapioid-ES cell complementation method. Human resistin gene was amplified from human fetal liver cDNA library by PCR and cloned into pCR 2.1 TOPO T-vector and constructed in pCMV-Tag4C vector. Human resistin mammalian expression plasmid was transfected into D3-GL ES cells by lipofectamine 2000, and then after 8~10 days of transfection, the human resistin-expressing cells were selected with G418. In order to produce tetraploid embryos, blastomeres of diploid embryos at the two-cell stage were fused with two times of electric pulse using 60 V 30 $\mu$sec. (fusion rate : 93.5%) and cultured upto the blastocyst stage (development rate : 94.6%). The 15~20 previously G418-selected ES cells were injected into tetraploid blastocysts, and then transferred into the uterus of E2.5d pseudopregnant recipient mice. To investigate the gestation progress, two El9.5d fetus were recovered by Casarean section and one fetus was confirmed to contain human resistin gene by genomic DNA-PCR. Therefore, this finding demonstrates that tetraploid-ES mouse technology can be considered as a useful tool to produce transgenic mouse for the rapid analysis of gene function in vivo.

• The Korean Journal of Mycology
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• v.17 no.3
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• pp.154-160
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• 1989

The nuclear transfer technique was employed to obtain intraspecific hybrids in Aspergillus nidulans. Nuclei isolated from either a wild type or an auxotrophic mutant strain (FGSC 475) were transferred into the protoplasts of a recipient strain (FGSC 514). The frequency of hybrid formation (4.8% and 10.1 %, respectively) by nuclear transfer was higher than the frequency (0.6%) by protoplast fusion. Furthermore, most of the hybrids formed showed increased activity of some components of cellulase system, xylanase system, and mannanase. The hybrids were analyzed to be either diploid or aneuploid. These results suggest that nuclear transfer technique is more efficient the formation of intraspecific hybrids than protoplast fusion method and is useful for the improvement of Aspergillus strains.

• Korean Journal of Ichthyology
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• v.24 no.1
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• pp.1-10
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• 2012

Reciprocal hybrids between the mud loach ($Misgurnus$ $mizolepis$) and cyprinid loach ($M.$ $anguillicaudatus$) were produced by artificial fertilization. The chromosome number of mud loach was 2n=48, consisting of 12M+4SM+32A chromosomes. The cyprinid loach has 2n=50, consisting of 10M+4SM+36A chromosomes. The chromosome numbers of the diploid reciprocal hybrids were 2n=49, consisting of 11M+4SM+34A chromosomes. All the karyotypes documented in this study had the same arm number of 64. There was no evidence of chromosomal polymorphisms or sex-related heteromorphism. The cytogenetic traits of the hybrid genotypes were intermediate between those of the parent species. In all genotypes, the chromosomal NORs localized to the terminal short arms of the same metacentric chromosome pair. These results suggest that Robertsonian translocation occurred between metacentric chromosome 1 of mud loach and acrocentric chromosome of cyprinid loach.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.27 no.1
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• pp.83-88
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• 1994

For the purpose of genetic stock indentification of three species of salmonid fishs and their hybrid, lactate dehydrogenase(LDH), malate dehydrogenase(MDH), isocitrate dehydrogenase(IDH), a-gylycerophosphate dehydrogenase(a-GPDH), malic enzyme(ME), 6-phospho-gluconate dehydrogenase(6-PGD), phosphoglucose isomerase(PGI) and phospho-glucomutase(PGM) from skeletal muscle, liver, heart and gill tissues in all three species were analyzed. Chum and masu salmon showed no polymorphic patterns in all isozyme loci, however rainbow trout were found to have polymorphic patterns at MDH-B, LDH and IDH loci. Especially, significant differences were found at MDH-B loci between the three species and the IDH patterns of rainbow trout were also different from the other two species. These loci therefore can be utilized as efficient genetic markers for the identification of hybrids and improve the efficiency of fish breeding. There was no difference except PGI between diploid and triploid isozyme patterns but PGI showed some potential as a marker for triploid in masu salmon.

• Korean Journal of Plant Tissue Culture
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• v.24 no.3
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• pp.175-181
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• 1997

Plants were regenerated from hypocotyl protoplast culture of cabbage (F$_1$hybrid Green Challenger) and were transplanted on fields. The ploidy level of regenerated and seed-grown plants was measured by flow cytometry. In total 125 regenerated plant, diploid (2n), tetraploid(4n), and mixoploid (2n+4n) were 72.8%, 25.6%, and 1.6%, respectively. Most of the regenerated plants had normal phenotype, whereas several plane showed abnormal phenotype such as heavy leaf incision, savoy, and wave. The regenerated plants with abnormal phenotype and different ploidy level were analysed by isozyme and RAPD, but no significant difference was found.

• Horticultural Science & Technology
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• v.19 no.1
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• pp.54-59
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• 2001

The objective of this research was to develop a screening system for selection of grape downy mildew (Plasmopara viticola)-resistant grape cultivars or seedlings under greenhouse condition. Inoculum concentrations ranging from $10^4$ to $10^5spores/mL$ for screening of resistance were evaluated and $5{\times}10^4spores/mL$ was optimum. Of the tested 14 grape cultivars, Campbell Early and Niagara were resistant to grape downy mildew, Sheridan and Cheongsoo were moderate resistant, while Kaiji, Red Queen and Ruby Okuyama were susceptible under both greenhouse and field conditions. Vitis vinifera grape cultivars were more susceptible to grape downy mildew than V. vinifera-labrusca hybrids and V. vinifera-labrusca-aestivalis hybrids. In V. vinifera-labrusca hybrids, tetraploid grape cultivars were more susceptible than diploid culivars. The evaluated results of grape downy mildew resistance under greenhouse condition were in accordance with those of field condition. Results of this study indicated that both greenhouse and field procedures could be used to screen grape cultivars and seedlings for downy mildew resistance.

• Journal of Life Science
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• v.22 no.7
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• pp.871-879
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• 2012

Polyploids are a potentially important germplasm source in seedless citrus breeding program. Seedlessness is one of the most promising traits of commercial mandarin breeds that mandarin triploid hybrids possess permanently. The formation of new constant triploid hybrids can be recovered through diploid species hybridization from the fusion of divalent gametes at low frequencyor intra-and inter-ploidy crosses. However, extensive breeding work based on small $F_1$ hybrid seeds developed is impossible without a very effective aseptic methodology and ploidy event. In this study, in vitro embryo culture was employed to recover natural hybrids from monoembryonic diploid, open-pollinated mandarin. Flow cytometry was used to determine ploidy level. A total of 10,289 seeds were extracted from 792 fruits having approximately 13 seeds per fruit. Average frequency of small seeds developed was 7.1%, while the average frequency of small seeds per fruit were: 8.9% for 'Clementine' 10.2% for 'Harehime' 2.6% for 'Kamja' 3.1% for 'Pyunkyool' 2.8% for 'Sadookam' and 7.0% for 'Wilking' mandarin. Average size of a perfect seed was $49.52{\pm}0.07mm^2$ ('Clementine') while the small seed measured $7.95{\pm}0.04mm^2$ ('Clementine'), which was about 1/6 smaller than the perfect seed. In total, 731 small seeds were obtained and all of them contained only one embryo per seed. The efficiency of 'Clementine' was 14 times higher than 'Wilking' and more than 109 times higher than 'Pyunkyool'. The basic information on spontaneous polyploidy provides for the hybridization of constant triploids and increases the efficiency of conventional cross.

• Korean Journal of Ichthyology
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• v.5 no.1
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• pp.113-121
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• 1993

Cytogenetic studies and hybridization experiment using two populations of Oryzias latipes from Korea were conducted in order to examine their systematic positions. In the chromosome number of specimens examined. O. sinensis had 46 with a pair of large metacentrics and O. latipes had 48 chromosomes without a pair of large metacentrics. However, the diploid chromosome number of the hybrid between O. latipes and O. sinensis was 47 and they had a large metarcentric chromosome in their karyotype. In the $F_2$ hybrids, distribution of chromosome number was variable among individuals. Hatching rates showed little difference between control and hybrids. However, hatchiabilties of $F_2$ hybrids were decreased by 83.3% as compared with $F_1$ offspring. Incidence of abnormal larvae was increased by 15.3% as compared with their parents. These results indicate that the two karyotype populations formerly regarded as two subspecies would be two distinct allopatric sibling species O. sinensis, and O. latipes.

• Korean Journal of Weed Science
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• v.30 no.4
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• pp.330-339
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• 2010

In order to suggest correct direction of researches on Miscanthus spp. which are promising bioenergy crop, authors had reviewed and summarized various literature about botanical taxonomy, morphology and present condition of breeding, cultivation and utilization of miscanthus. Among the genus of Miscanthus which are known 17 species, the most important species are M. sinensis and M. sacchariflorus which origin are East Asia including Korea, and M. x giganteus which is inter-specific hybrid of tetraploid M. sacchariflorus and diploid M. sinensis. Miscanthus is superior to other energy crops in resistance to poor environments including cold, saline and damp soil, nitrogen utilization efficiency, budget of input energy and carbon which are required for producing biomass and output which are stored in biomass. The major species for production of energy and industrial products including construction material in Europe, USA and Canada is M. x giganteus which was introduced from Japan in 1930s. In present, many breeding programs are conducted to supplement demerits of present varieties and to develop "Miscanes" which is hybrid of miscanthus and sugar cane. In Korea, the researches on breeding and cultivation of miscanthus were initiated in 2007 by collecting germplasms, and developed "Goedae-Uksae 1" which is high biomass yield and "mass propagation method of miscanthus" which can improve propagation efficiency in 2009. In order to develop "Korean miscanthus industry" in future, the superior varieties available not only domestic but also foreign market should be developed by new breeding method including molecular markers. Researches on production process of cellulosic bio-ethanol including pre-treatment and saccharification of miscanthus biomass also should be strengthen.

• Korean Journal of Plant Taxonomy
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• v.39 no.1
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• pp.12-23
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• 2009

To verify hybridity of Asplenium castaneo-viride, external morphology, spore morphology, anatomy and chromosomes of the species and of the two presumed parental species, A. incisum and A. ruprechtii, were examined. A. castaneo-viride usually had 1-pinnately divided frond. However, some individuals had almost simple fronds with pinnatisect basal parts similar to A. ruprechtii, while others had fronds similar to A. incisum in having oblanceolate blades and basal pinnae with triangular, 2-3 lobed apices. On the surface of the spores, sculpturing consisted of folds that were usually prominent; forming long wings, and irregular or incomplete reticulation. However, reticulation patterns varied among species. A. castaneo-viride showed a wide range of variation from sparse to dense patterns, whereas A. incisum showed only from sparse to intermediate patterns. A. ruprechtii showed from intermediate to dense patterns. The spore size of A. castaneo-viride was $54.63{\mu}m$, larger than other two species ($47.81{\mu}m$ in A. incisum and $44.22{\mu}m$ in A. ruprechtii). The level of undulation of epidermal cell wall was also different. A. incisum had the most shallowly undulated wall, and A. castaneo-viride had a pattern intermediate between the two presumed parental species. This same patterns was recognized in the density of stomata. The density of $45.91/mm^2$ in A. castaneo-viride was intermediate between the two presumed parental species ($67.00/mm^2$ in A. incisum, and $37.86/mm^2$ in A. ruprechtii). Chromosome number was constant (2x =2n = 72) as in A. incisum and A. ruprechtii. However, A. castaneo-viride showed a different ploidy level. The populations of Mt. Mai (Jeonbuk province) and Mt. Duryun (Jeonnam province) were diploid (2n = 72) which is a new record for this taxon, whereas the population of Mt. Buram (Seoul) was tetraploid (2n = 144). Conclusively, A. castaneo-viride was revealed to be a hybrid of A. ruprechtii and A. incisum based on evidence involving leaves, spores, epidermal cells, stomata and chromosome number.