• Korean Journal of Breeding Science
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• v.41 no.4
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• pp.456-462
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• 2009

Bacterial pustule (BP), caused by Xanthomonas axonopodis pv. glycines, is prevalent disease in major soybean production areas. BP can reduce seed yield as well as seed quality. To identify the genomic region associated with the resistance to BP, QTL analysis was conducted using $F_{10}$ RIL (recombinant inbred lines) population, Keunolkong${\times}$Shinpaldalkong. Four QTLs for BP disease were identified on the linkage group B2, D2, I and K in field accounts for 36.4% of the phenotypic variation. Especially, QTL at near of Satt135 on LG D2 was identified in green house experiment explaining 20.9% of the phenotypic variation was found to be a major QTL conferring BP. One of these QTLs, Satt135 on the LG D2, was also identified in green house experiment. In both field and green house condition, the position of major QTL for BP was detected between Satt135 and Satt397 on the LG D2. The major QTL for BP may be used for minimizing soybean BP through effective marker-assisted selection (MAS).

• Korean Journal of Breeding Science
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• v.43 no.6
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• pp.620-624
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• 2011

'Hwaweon 4' was developed from a cross between the African upland cultivar, 'Moroberekan' and 'Ilpumbyeo' based on marker-aided backcross selection. The recurrent parent 'Ilpumbyeo' is a high grain quality cultivar with medium to late maturity. 'Hwaweon 4' is nearly isogenic to 'Ilpumbyeo' except a small Moroberekan introgressed segment on chromosome 4 harboring the resistance gene for blast. The preliminary and replicated yield trial was conducted at Chungnam National University in 2006 and 2007. The local adaptability test was carried out by the National Seed Management Office (NSMO) from 2008 to 2009. This cultivar was registered to NSMO with a cultivar designated as 'Hwaweon 4'. This cultivar averaged 76 cm in culm length and has medium growth duration. Milled rice of 'Hwaweon 4' is translucent and the grain quality traits are comparable to those of the recurrent parent. It has low protein content. The yield potential of 'Hwaweon 4' in grain was about 6.31 MT/ha at the ordinary fertilizer level for two years. This variety showed highly resistance reaction at the blast nursery test at four locations and also at the sequential planting method. This resistance is due to the resistance gene designated as Pi45(t) on chromosome 4 introgressed from the donor parent, 'Moroberekan'. The Pi45(t) gene would be useful inenhancing resistance to blast in rice breeding program.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.52-52
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• 2001

Many nucleoside diphosphate (NDP) kinases are ubiquitous enzymes responsible for the exchange of ${\gamma}$-phosphates between tri- and diphosphonucleosides. The catalytic Many nucleoside diphosphate (NDP) kinases are ubiquitous enzymes responsible for the exchange of ${\gamma}$-phosphates between tri- and diphosphonucleosides. The catalytic reaction follows a ping-pong mechanism in which the enzyme is transiently phosphorylated on a histidine residue conserved in all nucleoside diphosphate kinases. Beside their role in nucleotide synthesis, these enzymes present additional functions, possibly independent of catalysis, in processes such as differentiation, cell growth, tumor progression, metastasis and development. To clone murine nm23-M5, several expressed sequence tags (ESTs) of the GenBank data base, selected according to their homology to nm23-H5 cDNA, reconstituted a complete open reading frame (GenBank AF222750). To test whether murine NDPKs (1, 2, 3, 4, 5, and 6) can inhibit Bax-mediated toxicity in yeast, co-transformation was performed respectively. The yeast S.cerevisiae was transformed with a copy expression plasmid containing the histidine selection marker and expressing murine Bax under the control of a galactose-inducible promoter. Several clones were selected and found to be growth inhibited when Bax expression was induced with galactose. A representative clone was transformed again with a copy expression plasmid containing the tryptophane selection marker and expressing either murine Bcl-xL or NDPK under the control of a galactose-inducible promoter. Several subclones of the double-transformants were selected and characterized. The ability of Bcl-xL and NDPKs to suppress Bax-mediated toxicity was determined by growing yeast cells overnight in galactose media and spot-testing on galactose plates starting with an equal number of yeast cells as determined by taking the OD$_{600}$. Ten-fold serial dilutions were used in the spot-test. Plates were grown at 3$0^{\circ}C$ for 2-3 days. All murine NDPKs suppressed Bax dependent apoptosis. Futher study will be peformed whether Bax-toxicity inhibition was caused by NDP kinase activity or additional function.n.

• Journal of Plant Biotechnology
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• v.50
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• pp.63-69
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• 2023

This study includes the transformation of genes such as ORE7, the increase of gene expression, and the use of the bar gene as a selectable marker that shows herbicide resistance with Agrobacterium tumefaciens using hypocotyls from the oilseed rape "Youngsan" cultivar. To establish an Agrobacterium transformation system for the production of oilseed rape with a high-yield trait, infection time and co-cultivation period with Agrobacterium were tested. Therefore, when hypocotyls from the oilseed rape "Youngsan" cultivar were infected with Agrobacterium for 20 min and co-cultivated for 3 days, approximately 32-36 putatively transformed hypocotyls with shoots including roots survived from 100 inoculated hypocotyls after 4 weeks of transformation on a selection medium containing 20 mg/L of phosphinothricin (PPT) as a selectable agent. Additionally, a PCR assay was performed to confirm the insertion of target genes and showed the presence of the ORE7 gene as a high-yielding trait and the bar gene as a selectable marker. Treatment with 0.5% (v/v) Basta solution as a selectable agent for 6 days with leaves from transformed oilseed rape expressed the bar gene. Therefore, this study can contribute to the development of special oilseed rapes containing agriculturally useful traits such as herbicide resistance, drought tolerance, high yielding traits, and high oleic acid content.

• Food Science and Preservation
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• v.24 no.7
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• pp.891-897
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• 2017

Apple (Malus domestica) is a climacteric fruit because of its high respiration and ethylene production. Ethylene affects the fruit by decreasing its quality and storability. Md-ACS1 and Md-ACO1 genes are involved in ethylene biosynthesis in apple; the Md-ACS1-2 and Md-ACO1-1 alleles are associated with low ethylene production. We conducted an analysis to study Md-ACS1 and Md-ACO1, and to examine ethylene production and softening rate of fruit at room temperature ($20^{\circ}C$) storage in 'Fuji (FJ)', 'Golden Supreme (GS)', and 5 cultivars of Korean apples ('RubyS (RS)', 'Hongro (HR)', 'Arisoo (AS)', 'Summer King (SK)', 'Greenball (GB)'). The result showed that an increase in the number of the alleles (ACS1-2, ACO1-1) decreased the ethylene production and softening rate. The presence of ACS1-1/1, ACO1-1/2 was confirmed in GS and the highest ethylene production and softening rate was observed. Ethylene production and softening rate of SK and GB expressing ACS1-1/2, ACO1-1/2 were higher than that of HR and AS, expressing ACS1-2/2, ACO1-1/2, but lower than GS. FJ with ACS1-2/2, ACO1-1/1 showed the lowest ethylene production and softening rate among all cultivars except RS. The Md-ACS1 and Md-ACO1 DNA markers could potentially be used to estimate storability and applied in marker assisted selection the improve the efficiency of apple breeding.

• Korean Journal of Medicinal Crop Science
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• v.10 no.1
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• pp.46-50
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• 2002

In DNA level, genetic study of Angelica species was firstly conducted to discriminate the bolting-resistant or low bolting variety, so called as Manchu, from other Korea collected lines and also this technuque was applied to identify the origin of commercial dried materials obtained from current oriental medicinal market. By RAPD analysis with 72 primers including sixty of 10-mers and twelve of 20-mers, respectively, three primers, which were related to the bolting resistant traits of Angelica gigas, were identified. Comparing the RAPD bands, URP04 primer showed the 1.7 kb specific band, which seemed to be related to delaying bolting traits, since it was observed only in Jinbu elite lines but not in others. On the other hand, since 1.2 kb band amplified by OPD11 was observed in other collected lines but not in Manchu var. and Jinbu line, this primer also could be considered as a selection marker for identifying bolting resistant or delaying bolting traits. In the same manner, since OPP09 did not show 1 kb major band but produced 0.8 kb and 1.2 kb bands in Manchu var., these three bands amplified by the primer could be considered one of the important key specifying Manchu var. related with the trait of Angelica gigas. OPC02 primer showed the same band patterns in all Korean collected lines, but not in other foreign introduced lines, such as A. sinensis from China, and A. acutiloba from Japan. Since these four RAPD primers, OPD11, OPP09, URP04, and OPC02 showed the specific polymorphisms in Angelica species, thus, these were useful to discriminate the three Angelica species, A. gigas, A. sinensis, and A. acutiloba.

• Journal of Embryo Transfer
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• v.16 no.3
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• pp.193-201
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• 2001

This study was designed to examine the factors affecting in fertilization and development of embryos in vitro, and to examine whether zone drilling by laser irradiation can improve the hatching rate of IVF embryos from DNA marker-proved Hanwoo. DNA markers related to marbling score were identified using DNA fingerprinting with Ml3 probe and restriction enzyme Hae III. Oocytes were aspirated from immature ovarian follicles using a combined method of rectal ovarian-palpation and transvaginal ultrasound-guidance(6.5MHz) under local anesthesia. The aspirated oocytes were washed twice with fresh D-PBS containing 5% FBS and were rewashed 4 to 5 times with TCM-199 containing 5% FBS. A morphological grade of I to IV was assigned to each oocyte. Data were analyzed using the GLM procedure of SAS. Sperm separation methods did not have any significant effect on cleavage or developmental abilities of IVF embryos. Significantly(P<0.05) higher cleavage rate was observed in embryos from GI(60.0%, 3/5), GII(69.2%, 18/26) and GIII(62.1%, 59/95) compared to embryos from GIV oocytes(36.2%, 25/69). And the developmental rate to blastocyst stage was higher(P<0.05) in embryos from GI(33.3%, 1/3) and GII oocytes(38.9%, 7/18) than those from GIII(16.9%,10/59) and GIV oocytes(4.0%, 1/25). There was no significant difference in development of IVF embryos to blastocyst by media for in vitro culture. Proportion of hatched blastocyst was significantly(P<0.05) higher in embryos received zona drilling by laser than those of non-drilled.

• Journal of Embryo Transfer
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• v.16 no.3
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• pp.183-191
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• 2001

This study was designed to establish the superior method for IVF embryos from DNA marker-proved Hanwoo cattle. DNA markers related to marbling score were identified using DNA fingerprinting with Ml3 probe and restriction enzyme Hae III. Oocytes were aspirated from unstimulated. immature ovarian follicles using a combined method of rectal ovarian-palpation and transvaginal ultrasound-guidance(6.5MHz) under local abesthesia. The aspirated oocytes were washed twice with fresh D-PBS containing 5% FBS and were rewashed 4 to 5 times with TCM-199 containing 5% FBS. A morphological grade of I to IV was assigned to each oocyte. Data were analysed using the GLM procedure of SAS. Mean number of follicles identified on ultrasound was 5.5 $\pm$2.9 in right and 4.3 $\pm$2.8 in left ovaries, respectively. The highest follicles(16.6$\pm$2.6) were found in 5101 cow compared to others. Recovery rate of follicular oocytes in individual cow was highest in 5101 cow with 89.3% in > 2mm and 94.0% in $\leq$ 2mm follicles. Total recovery rate was significantly(P<0.01) higher in $\leq$ 2mm(85.7%, 130/154) than > 2mm follicles(74.2%, 201/271). Significantly more oocytcs of Grade IV were recovered from > 2mm follicles. Mean number follicles recovered was 4.8$\pm$3.7. 3.0$\pm$3.4 and 0.3$\pm$0.6 in $\leq$2mm, 2~6mm and $\geq$6mm follicles. respectively. Our results imply that the more fertilizablc oocytes can be recovered from invisible-immature follicles by the combination of simultaneous rectal ovarian-palpation and ultrasound-guided approach in Hanwoo cattle.

• Journal of Life Science
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• v.22 no.9
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• pp.1152-1158
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• 2012

Development of transgenic plant increasing crop yield or disease resistance is good way to solve the world food shortage. However, the persistence of marker genes in crops leads to serious public concerns about the safety of transgenic crops. In the present paper, we developed marker-free transgenic rice inserted high molecular-weight glutenin subunit (HMW-GS) gene ($D{\times}5$) from the Korean wheat cultivar 'Jokyeong' using Agrobacterium-mediated co-transformation method. Two expression cassettes comprised of separate DNA fragments containing only the $D{\times}5$ and hygromycin resistance (HPTII) genes were introduced separately into Agrobacterium tumefaciens EHA105 strain for co-infection. Each EHA105 strain harboring $D{\times}5$ or HPTII was infected into rice calli at a 3: 1 ratio of EHA105 with $D{\times}5$ gene and EHA105 with HPTII gene expressing cassette. Then, among 66 hygromycin-resistant transformants, we obtained two transgenic lines inserted with both the $D{\times}5$ and HPTII genes into the rice genome. We reconfirmed integration of the $D{\times}5$ and HPTII genes into the rice genome by Southern blot analysis. Wheat $D{\times}5$ transcripts in $T_1$ rice seeds were examined with semi-quantitative RT-PCR. Finally, the marker-free plants containing only the $D{\times}5$ gene were successfully screened at the $T_1$ generation. These results show that a co-infection system with two expression cassettes could be an efficient strategy to generate marker-free transgenic rice plants.

• Korean Journal of Breeding Science
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• v.40 no.4
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• pp.408-413
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• 2008

Rice blast resistance is considered one of the most important traits in rice breeding and the disease, caused by Magnaporthe grisea Barr, has brought significant crop losses annually. Moreover, breakdown of resistance normally occurs in two to five years after cultivar release, thus a more durable resistance is needed for better control of this disease. We developed a new variety, Keumo3, which showed strong resistance to leaf blast. It was tested in 2003 to 2007 at fourteen blast nursery sites covering entire rice-growing regions of South Korea. It showed resistance reactions in 12 regions and moderate in 2 regions without showing susceptible reactions. Durability test by sequential planting method indicated that this variety had better resistance. Results showed that Keumo3 was incompatible against 19 blast isolates with the exception of KI101 by artificial inoculation. To understand the genetic control of blast resistance in rice cultivar Keumo3 and facilitate its utilization, recombinant inbred lines (RIL) consisting of 290 F5 lines derived from Akidagomachi/Keumo3 were analyzed and genotyped with Pizt InDel marker zt56591. The recombination value between the marker allele of zt56591 and bioassay data of blast nursery test was 1.1%. These results indicated that MAS can be applied in selecting breeding populations for blast resistance using zt56591 as DNA marker.