• Journal of Embryo Transfer
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• v.24 no.1
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• pp.57-64
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• 2009

This study was carried out to investigate expression of apoptosis-related differentially expressed gene (DEG) in ovaries of Korean cattle with follicular and luteal cysts and to identify the relationship between cyst and apoptosis using microarray, real-time PCR, TUNEL staining, and Western blot analysis. Microarray data showed that PIK3R2 and AKT1 were significantly up-regulated in follicular cyst, and TNF-RAF2, PRLR, FOXL2, STK4, and COL4A3 were up-regulated whereas INHA, CIDEB, BCL10, and FASLG were down-regulated in luteal cyst. Real-time PCR was performed to validate DEGs altered in luteal cyst. Of nine DEGs, four DEGs down-regulated in luteal cyst showed a positive corelation between microarray data and real-time PCR data. In this study, we focused on INHA, among many DEGs, which was highly down-regulated in both follicular and luteal cysts. Real-time PCR and micro array data showed that INHA was down-regulated by 12.3-fold and by 1.4-fold, respectively, in the bovine follicular cyst. TUNEL assay and Western blot analysis for ERK, JNK, p38, PI3K, and Akt, which were used to detect whether apoptosis is occurred, showed no significant changes in cystic ovaries (p>0.05). In the expression and activity of caspase-3, Bax, Bel-2, and Bel-xL, there was no significant changes between follicular cystic ovary and normal ovary. Rather, the expression levels of PI3K and p-Akt were decreased in follicular cystic ovary. These results suggest that deficiency of apoptosis in cystic ovary is associated with decreased expression of apoptotic effectors.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.47 no.6
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• pp.395-401
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• 2002

The combination of early heading time, maturing time and short grain-filling period is very important to develop early varieties in winter barley. The 4 parental half diallel crosses (parents, $F_1$s, $F_2$s) were cultivated at the field. The heading date was from April 3 to 26, maturing date from May 15 to 27 and grain-filling period from 31 days to 42 days, showing that the varietal differences about the 3 traits were remarkable. According to half diallel cross analyses, Dongbori 1 for heading time (late heading) was dominant, but Oweolbori (early heading) was recessive, showing partial dominance with high additive component of genetic variance. Dongbori 1 for maturing time was dominant, but Oweolbori was recessive, showing partial dominance with high additive variance. Reno for grain-filling period (short grain-filling period) was dominant, but Oweolbori (long grain-filling period) was recessive with additive, and partial dominance. There were highly significant mean squares for both GCA and SCA effects on the heading and maturing times, and GCA/SCA ratios for all traits were high, showing the additive gene effects more important. Sacheon 6 and Oweolbori had greater GCA effects for early heading and maturing times, and Dongbori 1 and Reno had greater GCA effects for late times. GCA effects were highly significant in $F_1$ and $F_2$ generations, showing high GCA/SCA ratios (7.02). The heading and maturing times in field were positively correlated with antifreeze proteins concentrations, accumulation, resistance to photoinhibition and winter survival, respectively) but the grain-filling period did negatively correlated with the trails.

• Biomedical Science Letters
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• v.1 no.1
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• pp.55-72
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• 1995

Forty gentamicin-resistant isolates of Enterococcus faecalis were selected from various clinical materials, determined their antimicrobial susceptibility, and studied there R-plasmid characteristics and polypeptide patterns. All of the isolates were susceptible to vancomycin. The MICs($\mu$/ml) of antimicrobial agents to the isolates were as follows; the MIC of gentamicin was 128 and $\geq$2040, ampicillin 1 and 1, chlorarmphenicol 2 and 8, erythromycin 32 and 256, and vancomycin 1 and 2. E. faecalis HL-1 strain had 8 plasmid DNA elements, HL-2 and HL-3 strains had 6, HL-4 had 7, HL-5 had 4, and HL-6 had 5. The 51.7 Kb of gentamicin resistance plasmid DNA was conjugally transferred from two strains of E. faecalis HL-1 and HL-6 to S. aureus SK 982. The plasmid transfer frequency between S. aureus SK 982 and E. faecalis HL-1 or E. faecalis HL-6 was 6.3$\times10^{-4} and 3.7$\times10^{-5}$, respectively. Plasmid curing ratio after the treatment of ethidium bromide(10$\mu$/ml) to E. faecalis tarnsconjugants R-1 and R-6 were about 51% and 67%, respectively. The tetracycline gene was located in 2.15 Kb plasmid of E. faecalis HL-1, but it was not found in the E. faecalis HL-6 by Southern blot analyses. The antigenic components of E. faecalis HL-1, HL-6, R-1 and R-6 strains were analyzed by SDS-PAGE and immunoblotting. The E. faecalis strains had 7 to 16 polypeptide bands, however their major proteins were 97.8 and 26.8 Kd. At the Immunoblotting, 97.8, 95.8, 74.8, 63.5, 33.7 and 26.8 Kd polypeptides of the strains showed major antigenic activities with patient's sera infected intra-abdominally with an E. faecalis strain.

• Journal of Plant Biotechnology
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• v.45 no.3
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• pp.171-182
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• 2018

Papaya (Carica papaya L.) is one of the crops widely planted in tropical and subtropical areas. The papaya fruit has low calories and are plentiful in vitamins A and C and in minerals. A major problem in papaya production is a plant disease caused by the papaya ringspot virus (PRSV). The first PRSV-resistant GM papaya expressing a PRSV coat protein gene was developed by USA scientists in 1992. The first commercial GM papaya cultivars derived from the event was approved by the US government in 1997. Development of transgenic papayas has been focused on vaccine production and limited agricultural traits, including insect and pathogen resistance, long shelf life, and aluminum and herbicide tolerance. Approximately 17 countries, including the USA and China, produced transgenic papayas and/or commercialized them, which provoked studies on biosafety assessment and development of GM-detection technologies. For the biosafety assessment of potential effects on human health, effects of long-term feeding to model animals have been studied in terms of toxicity and allergenicity. Studies on environmental safety assessment include influence on soil-microbial biodiversity and transfer to soil bacteria of GM selection markers. Many countries, such as Korea, the European Union, and Japan, that have strict regulations for GM crops have serious concerns about unintended introduction of GM cultivars and food commodities using unauthorized GM crops. Transgene- and/or GM event-specific molecular markers and technologies for genomics-based detection of unauthorized GM papaya have been developed and have resulted in the robust detection of GM papayas.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.10-10
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• 2017

Rice consumption has been continuously decreasing as the eating habits of Koreans have become westernized and diversified. The per capita annual rice consumption in Korea has dropped sharply from 136.4 kg in 1970 to 61.9 kg in 2016. The Korean government, therefore, has been trying to promote rice consumption by invigorating the processed food industry using rice flour. To facilitate the market for processed rice foods, it is essential to develop proper milling technology in terms of flour particle size and damaged starch content to produce high quality rice flour at competitive cost. Dry milling and wet milling are the two major processes used to produce rice flour. Although the dry milling process is relatively simple with a lower production cost, damaged starch content increases because of the high grain hardness of rice. In wet milling, the quality of rice flour is improved by reducing flour particle size as well as damaged starch content through soaking procedures. However, the production costs are high because of the additional expenses associated with the disposal of waste water, sterilization and drying of the wet flour. Recently developed technologies such as jet milling and cryogenic milling also require expensive investment and production. Therefore, developing new rice cultivars with dry milling adaptability as well as good processing properties is an important goal of rice breeding in Korea. 'Suweon 542' is a floury endosperm mutant line derived from sodium azide treatment on a high-yield, early maturing, and non-glutinous japonica rice cultivar, 'Namil'. Compared with the wild type, after dry milling process, the grain hardness of 'Suweon 542' was significantly lower because of its round and loosely packed starch granules. Also, the flour of 'Suweon 542' had significantly smaller particles and less damaged starch than 'Namil' and other rice cultivars and its particle size distribution was similar to a commercial wheat cultivar. Recently, through collaborations with nine universities and food companies, a total of 21 kinds of processed prototypes, using the dry milling flour of 'Suweon 542', were evaluated. In the production of major rice processing products, there was no significant quality difference between the flours prepared by wet milling and dry milling. Although the amount of water added to the dough was slightly increased, it was confirmed that the recipe applying the wet flour could be used without significant change. To efficiently transfer the floury endosperm characteristics of 'Suweon 542' to other commercial rice cultivars, it is essential to develop DNA marker tightly linked to the target gene. Association analysis using 70 genome-wide SSR markers and 94 F2 plants derived from 'Suweon 542'/'Milyang 23' showed that markers on chromosome 5 explained a large portion of the variation in floury grains percentage (FGP). Further analysis with an increased number of SSR markers revealed that the floury endosperm of 'Suweon 542' was directed by a major recessive locus, flo7(t), located in the 19.33-19.86 Mbp region of chromosome 5, with RM18639 explaining 92.2% of FGP variation in the F2 population. Through further physical mapping, a co-segregate and co-dominant DNA marker with the locus, flo7(t) was successfully developed, by which, thereby, breeding efficiency of rice cultivars having proper dry milling adaptability with high yield potential or useful functional materials would be improved. 'Suweon 542' maintained the early maturity of the wild type, Namil, which can be used in rice-wheat double cropping systems in Korea not only for improved arable land but also for sharing flour production facilities. In addition to the high susceptibility against major rice diseases, nevertheless, another possible drawback of 'Suweon 542' is the high rate of viviparous under prolonged rainfall during the harvesting season. To overcome susceptibility and vivipary of 'Suweon 542', the progeny lines, derived from the crosses 'Suweon 542' and 'Jopyeong', an early maturing rice cultivar with multiple resistance against rice blast, bacterial blight, and rice strip virus, and 'Heugjinju', a anthocyanin pigment containing black rice cultivar, were intensively evaluated. As the outputs, three dry milling suitable rice elite lines, 'Jeonju614', 'Jeonju615', and 'Jeonju616' were developed.

• Korean Journal of Plant Tissue Culture
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• v.27 no.6
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• pp.423-428
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• 2000

In 1997 when cloned sheep Dolly and soon after Polly were born, it had become head-line news because in the former the nucleus that gave rise to the lamb came from cells of six-year-old adult sheep and in the latter case a foreign gene was inserted into the donor nucleus to make the cloned sheep produce human protein, factor IX, in e milk. In the last few years, once the realm of science fiction, cloned mammals especially in livestock have become almost commonplace. What the press accounts often fail to convey, however, is that behind every success lie hundreds of failures. Many of the nuclear-transferred egg cells fail to undergo normal cell divisions. Even when an embryo does successfully implant in the womb, pregnancy often ends in miscarriage. A significant fraction of the animals that are born die shortly after birth and some of those that survived have serious developmental abnormalities. Efficiency remains at less than one % out of some hundred attempts to clone an animal. These facts show that something is fundamentally wrong and enormous hurdles must be overcome before cloning becomes practical. Cloning researchers now tent to put aside their effort to create live animals in order to probe the fundamental questions on cell biology including stem cells, the questions of whether the hereditary material in the nucleus of each cell remains intact throughout development, and how transferred nucleus is reprogrammed exactly like the zygotic nucleus. Stem cells are defined as those cells which can divide to produce a daughter cell like themselves (self-renewal) as well as a daughter cell that will give rise to specific differentiated cells (cell-differentiation). Multicellular organisms are formed from a single totipotent stem cell commonly called fertilized egg or zygote. As this cell and its progeny undergo cell divisions the potency of the stem cells in each tissue and organ become gradually restricted in the order of totipotent, pluripotent, and multipotent. The differentiation potential of multipotent stem cells in each tissue has been thought to be limited to cell lineages present in the organ from which they were derived. Recent studies, however, revealed that multipotent stem cells derived from adult tissues have much wider differentiation potential than was previously thought. These cells can differentiate into developmentally unrelated cell types, such as nerve stem cell into blood cells or muscle stem cell into brain cells. Neural stem cells isolated from the adult forebrain were recently shown to be capable of repopulating the hematopoietic system and produce blood cells in irradiated condition. In plants although the term$\boxDr$ stem cell$\boxUl$is not used, some cells in the second layer of tunica at the apical meristem of shoot, some nucellar cells surrounding the embryo sac, and initial cells of adventive buds are considered to be equivalent to the totipotent stem cells of mammals. The telomere ends of linear eukaryotic chromosomes cannot be replicated because the RNA primer at the end of a completed lagging strand cannot be replaced with DNA, causing 5' end gap. A chromosome would be shortened by the length of RNA primer with every cycle of DNA replication and cell division. Essential genes located near the ends of chromosomes would inevitably be deleted by end-shortening, thereby killing the descendants of the original cells. Telomeric DNA has an unusual sequence consisting of up to 1,000 or more tandem repeat of a simple sequence. For example, chromosome of mammal including human has the repeating telomeric sequence of TTAGGG and that of higher plant is TTTAGGG. This non-genic tandem repeat prevents the death of cell despite the continued shortening of chromosome length. In contrast with the somatic cells germ line cells have the mechanism to fill-up the 5' end gap of telomere, thus maintaining the original length of chromosome. Cem line cells exhibit active enzyme telomerase which functions to maintain the stable length of telomere. Some of the cloned animals are reported prematurely getting old. It has to be ascertained whether the multipotent stem cells in the tissues of adult mammals have the original telomeres or shortened telomeres.

• Journal of Plant Biotechnology
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• v.38 no.1
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• pp.15-21
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• 2011

We developed 14 transgenic lines of Chinese cabbage (Brassica rapa) harboring the T-DNA border sequences and CryIAc1 transgene of the binary vector 416 using Agrobacterium tumefaciens-mediated DNA transfer. Six lines had single copy cryIAc1 gene and four of them contained no vector backbone DNA. Of the left border (LB) flanking sequences six nucleotides were deleted in transgenic lines 416-2 and 416-3, eleven nucleotides in line 416-9, and 65 nucleotides including the whole LB sequences in line 416-17, respectively. And we defined 499 bp of genomic DNA (gDNA) of transformed Chinese cabbage, and blast results showed 96% homology with Brassica oleracea sequences. PCR with specific primer for the right border (RB) franking sequence revealed 834 bp of PCR product sequence, and it was consisted of 3' end of cryIAc1, nosterminal region and 52 bp of Chinese cabbage genomic DNA near RB. RB sequences were not found and the 58 nucleotides including 21 bp of nos-terminator 3' end were deleted. Also, there were deletion of 10 bp of the known genomic sequences and insertion of 65 bp undefined genomic sequences of Chinese cabbage in the integration site. These results demonstrate that the integration of T-DNA can be accompanied by unusual deletions and insertions both in transgenic and genomic sequences.

• Journal of Mushroom
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• v.12 no.1
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• pp.41-51
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• 2014

Cauliflower mushroom widely known high concent of ${\beta}$-glucan for farm cultivation invigoration verified characteristics of mycelia growth, genetic diversity, resistance to Trichoderma by replacement culture with Trichoderma and growth characteristics of new variety crossbleeding strain. The result of replacement culture with Trichoderma for verification resistance about Trichoderma, 6951 (T. viride) strain did not show special change after formation of confrontation line and 6952 (T. spp.) strain was showed more formation of spore after formation of confrontation line. But 6426 (T. harzianum) strain found to encroach part of growth area of cauliflower mushroom mycelia. Among 10 kinds cauliflower mushroom strain, JF02-06 strain collected by Gurye, found did not spore of Trichoderma and thought to be resistant to Trichoderma. The result of crossbleeding after selected that mother strain good growth and formation of fruit body, verified good mycelia growth at JF02-47, 49 and 50 strain in Korean pine of wood-chip media. The result of gene sequence about ITS1, 5.8S and ITS4 for analysis of genetic diversity at crossbleeding strain, found high significance to other cauliflower mushroom in registered Genebank. The result of growth characteristic of spore and mycelia of cauliflower mushroom by observation microscope, size of spore showed water drop shape to major axis $6{\mu}m$ and minor axis $5{\mu}m$ and clamp showed 3 types in mycelia. The wide of mycelia was $3{\mu}m$. The characteristic of mycelia of cauliflower mushroom found to grow mycelia in clamp at approximately 50%. The growth speed of mycelia was $0.507{\mu}m/min$ and 2nd mycelia grown similar speed to mother mycelia at parallel with mother mycelia after growth speed at $0.082{\mu}m/min$. The formation of clamp made small clamp for 5 hours after shown transfer of electrolyte in mycelia inside. The septum formation started after 3 hours and then finally completed after 2 hours. In this study, strain of cauliflower mushroom verified resistance of Trichoderma, genetic diversity and characteristic of mycelia growth. Therefore, basic knowledge of cauliflower mushroom will improve and further contribute to development of mushroom industry.

• Journal of Life Science
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• v.20 no.3
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• pp.381-387
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• 2010

To find offspring of Jeju Black cattle (JBC) produced by embryo transfer (ET) and artificial insemination (AI), a molecular genetic study was carried out in candidate cattle populations collected from cattle farms in Jeju Island, Korea. The genetic marker set was composed of 11 ISAG microsatellite (MS) markers, 11 SAES MS markers selected by our preliminary analysis for population diversity of JBC and two major coat color related genes: MC1R and ASIP. The results showed a combined non-exclusion probability for first parent (NE-P1) that was higher than that recommended by ISAG (above 0.9995), and a combined non-exclusion probability for sib identity of $5.3{\times}10^{-10}$. Parentage analysis showed that the cases identified the candidate's father only (77.0%), mother only (54.0%), and both parents (40.5%) in the candidate offspring population. The ET and AI calves were identified as 14.7% in the in vitro fertilized eggs provided and 32.4% in total population, respectively. However, the result from ISAG marker analysis showed 3 identical allele-combinations in 7 calves, and that from ISAG/SAES MS marker combination also showed 1 identical allele-combination in 2 calves. Data from MS and coat-color gene analyses provided information for complete identification of all animals tested. Because the present JBC population was mostly bred using small nuclear founders through bioengineering techniques such as AI and ET, the genetic diversity levels obtained from MS analysis in the JBC population were relatively lower than those of other cattle populations, including Hanwoo. The results suggested that the more efficient marker combinations, including coat color related genotypes, should be studied and used for constructing a system for identification and molecular breeding of JBC as well.

• Current Research on Agriculture and Life Sciences
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• v.5
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• pp.119-126
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• 1987

This study was conducted to elucidate the most appropriate method to obtain auxotrophic mutants from Valsa ceratosperma, the causal fungus of apple canker, which may be used as a gene marker in detecting the transfer of the factors of avirulent strains to virulent strains. Among the 3 kinds of synthetic media tested, each have two formula for minimal and complete, the medium which has been used in study of Endothia parasitica (E. P medium) was turned out to be most appropriate for the growth of V. ceratosperma. A medium for single colony formation from pycnidiospore of this fungus was developed by adding 0.5% L - sorbose to the E. P minimal medium. The period of incubation in dark for preventing the photoreactivation after U. V irradiation was estimated as about 60hrs at which most of the spores become binucleate. Largest number of putative auxotrophs were obtained at about 50second of irradiation to the spores smeared on the medium for single colony formation, at which the survival rate of spores was 5 to 6 percent. With these method developed in this experiment, 161 isolates of putative auxotrophs were detected among which the nutrient requirement for 10 isolates were determined. Five out of 10 mutants were still virulent to apple tree and all but one could not sporulate.