• Journal of Plant Biotechnology
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• v.5 no.3
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• pp.143-148
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• 2003

Bax, a mammalian pro-apoptotic member of the Bcl-2 family induces cell death when expressed in yeast. To investigate whether Bax expression can induce cell death in plant, we produced transgenic Arabidopsis plants that contained murine Bax cDNA under control of a glucocorticoid-inducible promoter. Transgenic plants treated with dexamethasone, a strong synthetic glucocorticoid, induced Bax accumulation and cell death, suggesting that some elements of cell death mechanism by Bax may be conserved among various organisms. Therefore, we developed novel yeast genetic system, and cloned several Plant Bax Inhibitors (PBIs). Here, we report the function of two PBIs in detail. PBI1 is ascorbate peroxidase (sAPX). Fluorescence method of dihydrorhodamine123 oxidation revealed that expression of Bax in yeast cells generated reactive oxygen species (ROS), and which was greatly reduced by co-expression with sAPX. These results suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Baxinduced cell death in yeast. PBI2 encodes nucleoside diphosphate kinase (NDPK). ROS stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Transgenic plants overexpressing AtNDPK2 have lower levels of ROS than wildtype plants. Mutants lacking AtNDPK2 had higher levels of ROS than wildtype. $H_2O_2$ treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6. In the absence of $H_2O_2$ treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased in the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the MSP phosphorylation activity of AtMPK3 in vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation in situ. Thus, AtNDPK2 appears to playa novel regulatory role in $H_2O_2$-mediated MAPK signaling in plants.

• Molecules and Cells
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• v.39 no.5
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• pp.426-438
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• 2016

Plant disease resistance occurs as a hypersensitive response (HR) at the site of attempted pathogen invasion. This specific event is initiated in response to recognition of pathogen-associated molecular pattern (PAMP) and subsequent PAMP-triggered immunity (PTI) and effector-triggered immunity (ETI). Both PTI and ETI mechanisms are tightly connected with reactive oxygen species (ROS) production and disease resistance that involves distinct biphasic ROS production as one of its pivotal plant immune responses. This unique oxidative burst is strongly dependent on the resistant cultivars because a monophasic ROS burst is a hallmark of the susceptible cultivars. However, the cause of the differential ROS burst remains unknown. In the study here, we revealed the plausible underlying mechanism of the differential ROS burst through functional understanding of the Magnaporthe oryzae (M. oryzae) AVR effector, AVR-Pii. We performed yeast two-hybrid (Y2H) screening using AVR-Pii as bait and isolated rice NADP-malic enzyme2 (Os-NADP-ME2) as the rice target protein. To our surprise, deletion of the rice Os-NADP-ME2 gene in a resistant rice cultivar disrupted innate immunity against the rice blast fungus. Malic enzyme activity and inhibition studies demonstrated that AVR-Pii proteins specifically inhibit in vitro NADP-ME activity. Overall, we demonstrate that rice blast fungus, M. oryzae attenuates the host ROS burst via AVR-Pii-mediated inhibition of Os-NADP-ME2, which is indispensable in ROS metabolism for the innate immunity of rice. This characterization of the regulation of the host oxidative burst will help to elucidate how the products of AVR genes function associated with virulence of the pathogen.

• Journal of Plant Biotechnology
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• v.44 no.1
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• pp.61-68
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• 2017

Gibberellic Acid-Stimulated Arabidopsis (GASA) genes are involved in plant hormone signaling, cell division and elongation, as well as in responses to stress conditions in plants. In this study, we isolated a GASA gene from hybrid poplar (Populus alba ${\times}$ P. glandulosa) and analyzed its physiological phenotype and molecular functions in poplar. PagGASA cDNA encodes a putative protein composed of 95 amino acids containing an N-terminal signal peptide and a conservative cysteine-rich C-terminal domain. Southern blot analysis revealed that one or two copies of the PagGASA are present in the poplar genome. The PagGASA transcripts were highly detected in flowers and roots. Moreover, the expression of PagGASA was induced by growth hormone (gibberellic acid) and stress hormones (abscisic acid, jasmonic acid, and salicylic acid). By using transgenic analysis, we showed that the upregulation of PagGASA in poplar provides high tolerance to drought stress. Therefore, our results suggest that PagGASA plays an important role in drought stress tolerance via stress-related plant hormone signaling in poplar.

• Biomedical Science Letters
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• v.12 no.2
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• pp.73-79
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• 2006

Nebulin is a giant ($600{\sim}900$ kDa), modular sarcomeric protein proposed to regulate the assembly, and to specify the precise lengths of actin filamints in vertebrate skeletal muscles. Recently, There is an evidence that the nebulin also expressed in non muscle tissue, brain and liver. We identified a new isoform of nebulin from adult brain library by PCR screening. It contains two simple-repeats exon 165, 166 and linker-repeats exon $154{\sim}161$ except exon 159. The nebulin modules M160 to M170 (exon 150 to exon 161) has been shown to bind desmin. In mature striated muscle, desmin intermediate filaments surround Z-discs and link individual myofibrils laterally at their Z-discs and to other intracellular structures, including the costameres and the intercalated discs of the sarcolemma, sarcoplasmic reticulum, mitochondria, T-tubules, and nuclei. Therefore, it is an interesting possibility that the differential splice pathways within the linker region of nebulin modify the affinity of nebulin's interaction with desmin. The specific interactions of nebulin and desmin were confirmed in vivo by yeast two hybrid experiments. To verify in the cellular level the interaction between nebulin isoform and desmin, we transfected COS-7 cell with EGFP-tagged nebulin and DsRed-tagged desmin. Based on evidence showing that despite exon 159 was deleted, the new isoform of nebulin was interact with desmin. This suggest that nebulin in brain may interact with another intermediate filament. The conservation of these ligand-binding capacity in brain and skeletal nebulins suggest that nebulins may have conserved roles in brain and skeletal muscle.

• Journal of The Korean Society of Grassland and Forage Science
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• v.35 no.2
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• pp.137-144
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• 2015

This study was carried out to investigate the growth characteristics, yield, and chemical compositions of corn hybrids for silage according to the drainage depths in a lowland paddy field. The experimental design was arranged in a randomized block design with four treatments and three replications. The drainage depths of the four treatments were 0 cm, 20 cm, 30 cm and 50 cm. Tip-filling degree, ear length, and ear circle were higher in the 30 cm and 50 cm treatments compared with the other treatments (p < 0.05); but the plant length, leaf length, leaf width, ear height, stem diameter, and stem hardness were not significantly different (p < 0.05). The brix was higher in the order of 20 cm > 0 cm > 50 cm > 30 cm treatment. The fresh yield, dry matter yield, and TDN yield increased as the drainage depth increased (p < 0.05). The crude-protein and crude-ash content were highest in the 50 cm treatment (p < 0.05), but the crude-fat, NDF, ADF, and crude-fiber levels were not significantly different (p < 0.05). The total mineral content was higher in the order of 0 cm > 30 cm > 20 cm > 50 cm treatment (p < 0.05). Fructose, glucose, and sucrose were highest in the 20 cm treatment. The total amino-acid (EAA + NEAA) content was the highest in the 50 cm treatment. Based on these results, greater drainage depths increased the yield and feed value of silage corn.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2003.04a
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• pp.65-71
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• 2003

Bax, a mammalian pro-apoptotic member of the Bcl-2 family, induces cell death when expressed in yeast. To investigate whether Bax expression can induce cell death in plant, we produced transgenic Arabidopsis plants that contained murine Bax cDNA under control of a glucocorticoid-inducible promoter. Transgenic plants treated with dexamethasone, a strong synthetic glucocorticoid, induced Bax accumulation and cell death, suggesting that some elements of cell death mechanism by Bax may be conserved among various organisms. Therefore, we developed novel yeast genetic system, and cloned several Plant Bax Inhibitors (PBIs). Here, we report the function of two PBIs in detail. PBI1 is ascorbate peroxidase (sAPX). Fluorescence method of dihydrorho-damine 123 oxidation revealed that expression of Bax in yeast cells generated reactive oxygen species (ROS), and which was greatly reduced by co-expression with sAPX. These results suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Baxinduced cell death in yeast. PBI2 encodes nucleoside diphosphate kinase (NDPK). ROS stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Transgenic plants overexpressing AtNDPK2 have lower levels of ROS than wildtype plants. Mutants lacking AtNDPK2 had higher levels of ROS than wildtype. $H_2O_2$ treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6. In the absence of $H_2O_2$ treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased in the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the MBP phosphorylation activity of AtMPK3 in vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation in situ. Thus, AtNDPK2 appears to play a novel regulatory role in $H_2O_2$-mediated MAPK signaling in plants.

• Journal of Ginseng Research
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• v.28 no.4
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• pp.191-195
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• 2004

An investigation was conducted to ascertain the basic information on characteristics of embryonic tissue development among Korean Ginseng, American ginseng, $F_{1}$ hybrids and $BC_{1}F_{1}(F_{1}$ pollen back cross to $BC_{1}=Korean\;Ginseng)seeds$. The specific activities of lipoxygenase (LOX) and the protein assays were made during the above embry­onic growth period at 5 or 25 degree C. The results are summarized as follows: The fresh weights(mg) of germinating seeds were significantly different among species. Ginseng seeds were stratified at $5^{\circ}C\;for\;60\;days$ the ratios of embryo/endosperms in $P.g.,\;P.q\;or\;F_{1}\;were\;about\;90{\%}$. The ratio was in $BC_{1}F_{1},\;79.2{\%}$ during the same period. The ratios of embryo/endosperms of seeds of P.g. or P.q. germinated at $5^{\circ}C$ showed rather higher values as compared to that at $25^{\circ}C$ LOX specific activity the highest in the $F_{1}$ and decreased in the order of P.g., P.q., and $BC_{1}F_{1}$. The highest LOX specific activity was shown at 80 days after sowing, followed by 70 days sowing, the least LOX activity was shown at 60 days.

• Proceedings of the Korean Information Science Society Conference
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• 2007.06b
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• pp.273-278
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• 2007

검사점 및 복구 도구(Checkpointing & Recovery Facility)를 이용하여 임베디드 시스템에서 결함 허용(Fault Tolerance) 기법을 적용할 경우 쓰기 작업의 오버헤드로 인해 실용성이 크게 떨어지게 된다. 실시간 운영체제와 함께 어떠한 한계 상황에서 결함 허용 및 복구 도구가 오히려 시스템의 성능을 저하시키는 요인으로 작용하게 되면 이는 결국 쓸모없는 도구가 되어 사용되지 않을 것이다. 따라서 프로세스의 복구를 위해 저장하는 프로세스 이미지의 기록에 소요되는 시간을 크게 낮추어야만 비로소 검사점 도구가 그 진가를 발휘하게 될 수 있다. 본 논문에서는 NVSRAM(Non Volatile SRAM)을 검사점 및 복구 도구의 저장 장치로 활용함으로써 기존의 검사점 도구에서 성능을 저하시키는 주원인이었던 검사점 기록의 오버헤드를 개선하기 위한 연구를 수행하였다. 검사점 기록 시간을 줄이기 위한 방법으로 주 메모리에 저장된 프로세스의 복구와 관련된 데이터를 SRAM 특성을 갖는 비휘발성 저장 장치인 NVSRAM에 저장하여 디스크 접근에 소요되는 시간을 최소화시킴으로써 임베디드 시스템에서 실용적으로 사용 가능한 검사점 도구를 구현하였고, 이러한 연구의 결과를 검증하기 위해 기존 시스템에서 저장 장치로 사용되던 플래시 메모리, 주 메모리, 원격 메모리를 사용하는 경우의 성능과 NVSRAM을 활용할 때의 성능을 비교해 보았다. 본 연구에서 제안하는 결함 허용 도구는 실제 시스템에 적용하여 효과적인 성능을 발휘할 수 있을 것이며, 차세대 메모리를 이용한 결함 허용 도구의 연구에 기여를 할 수 있을 것으로 기대된다.ate첨가배지(添加培地)에서 가장 저조(低調)하였다. vitamin중(中)에서는 niacin과 thiamine첨가배지(添加培地)에서 근소(僅少)한 증가(增加)를 나타내었다.소시켜 항이뇨 및 Na 배설 감소를 초래하는 작용과, 둘째는 신경 경로를 통하지 않고, 아마도 humoral factor를 통하여 신세뇨관에서 Na 재흡수를 억제하는 작용이 복합적으로 나타내는 것을 알 수 있었다.으로 초래되는 복합적인 기전으로 추정되었다., 소형과와 기형과는 S-3에서 많이 나왔다. 이상 연구결과에서 입도분포가 1.2-5mm인 것이 바람직한 것으로 나타났다.omopolysaccharides로 확인되었다. EPS 생성량이 가장 좋은 Leu. kimchii GJ2의 평균 분자량은 360,606 Da이었으며, 나머지 두 균주에 대해서는 생성 EPS 형태와 점도의 차이로 미루어 보아 생성 EPS의 분자구조와 분자량이 서로 다른 것으로 판단하였다.TEX>개로 통계학적으로 유의한 차이가 없었다. Heat shock protein-70 (HSP70)과 neuronal nitric oxide synthase (nNOS)에 대한 면역조직화학검사에서 실험군 Cs2군의 신경세포가 대조군 12군에 비해 HSP70과 nNOS의 과발현을 보였으며, 이는 통계학적으로 유의한 차이를 보였다(p<0.05). nNOS와 HSP70의 발현은 강한 연관성을 보였고(상관계수 0.91, p=0.000), nNOS를 발현하는 세포가 동시에 HSP70도 발현함을 확인할 수 있었다. 결론: 우리는 cyclosporin A가 토끼의 25분간의 척수허혈에 대해 척수보호 효과가 있었으며 이는 HSP70의

• Proceedings of the Microbiological Society of Korea Conference
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• 2002.10a
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• pp.162-172
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• 2002

Schizosaccharomyces pombe is suited for the study of cytokinesis as it divides by forming a septum in the middle of the cell at the end of mitosis. To enhance our understanding of the cytokinesis, we have carried out a genetic screen for temperature-sensitive S. pombe mutants that show defects in septum formation and cell division. Here we present the isolation and characterization of a new temperature-sensitive mutant, sun1(septum uncontrolled), which undergoes uncontrolled septation during cell division cycle at restrictive temperature $(37^{\circ}C)$. In sun1 mutant, actin ring and septum are positioned at random locations and angles, and nuclear division cycle continues. These observations suggest that the sun] gene product is required for the proper placement of the actin ring as well as precise septation. The sun] mutant is monogenic recessive mutation unlinked to previously known various cdc genes of S. pombe. In a screen for $sunl^+$ gene to complement the sun] mutant, we have cloned a gene, $susl^+$(suppressor of sun1 mutant), that encodes a protein of 689 amino acids. The predicted amino acid sequence of $susl^+$ gene is similar to the human hMadlp and Saccharomyces cerevisiae Mad1p, a component of the spindle checkpoint in eukaryotic cells. The null mutant of $susl^+$ gene grows normally at various temperatures and has the increased sensitivity to anti-microtubule drug, while $susl^+$ mutant shows no sensitivity to microtubule destabilizing drugs. The putative S. pombe Sus1p directly interacts with S. pombe Mad2p in yeast two-hybrid assays. These data suggest that the newly isolated susr gene encodes S. pombe Mad1p and suppresses sun] mutant defective in controlled septation in a cell division cycle.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.32 no.2
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• pp.144-150
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• 1987

To increase the feed value of corn stover after grain harvesting, these studies were conducted to evaluate the methods and effects of intercropping with corn and legume crops, soybean and cowpea. Basic studies were designed to detect the light receiving efficiency of intercrops with change of row-space and interrowspace under plant density of the level of 5,555 plants/l0a. Another study was conducted to select the favorable varieties for intercropping. The plant space of 90 x 20 cm was more effective to increase the light receiving effeciency of intercrop without decrease the yield of corn than that of 60 x 30 cm. Among several corn hybrid/varieties, erect leaf type was desirable for intercropping with high light penetration. Silage yield of intercropping with corn and legume crops increased above 20% without decrease of grain yield of corn compared with monocropping. The silage of intercropping appeared as a good quality roughage bulky feed with high content of crude protein and fat.