Brown leaf spot, caused by necrotrophic Cochliobolus miyabeanus (imperfect; Bipolaris oryzae), is one of the devastating disease in rice (Oryza sativa). Especially, recommended agricultural system such as diminishing fertilizer and environmental alteration like temperature increment result in the favorable conditions for the outbreak of this disease. Lack of water supply also requires drought-tolerant rice cultivar. We hypothesized that regulation of programmed cell death (PCD) should be a common solution conferring resistance and tolerance against above biotic and abiotic stresses at the same time. Among 17 CYCLIC NUCLEOTIDE-GATED ION CHANNEL in rice (OsCNGCs), over expression of a CNGC resulted in lesion mimic phenotypes in Dongjin background. Further, knock out of a CNGC resulted in enhanced resistance against rice brown spot in the field. These results indicate that selected OsCNGC should be involved in the PCD regulation and fungal infection-specific regulation of OsCNGC expression might induce resistance against rice brown spot because of pathogen’s necrotrophic nature. Vitamin E, tocopherol, is involved in the accumulation inhibition of reactive oxygen species involving superoxide and hydrogen peroxide. Tocopherol cyclase in Nicotiana benthamiana (NtTC), which is included in tocopherol systhesis, conferred tolerance against drought stress to rice. We already have settled down the recombination system effectively removing selection markers in vector. Based on these systems, we will define fungal secretome and genome, confirmation of PAMPs/effectors, identification of rice pattern recognition receptor, and functional characterization of these rice genes in the respect of PCD and disease resistance. We will also develop marker-free transgenic rice tolerant to drought and/or salinity stresses. These works should give us a bundle of rice genes conferring resistance and tolerance against biotic and abiotic stresses and amount of information useful for the analyses of common cross talk points between disease resistance and stress-tolerance.

In higher plants, formate dehydrogenase (FDH; EC 1.2.1.2) is a NAD-dependent enzyme that catalyzes the oxidation of formate into carbon dioxide. Although FDHs have been reported to be induced by various abiotic stresses, the function of FDHs in biotic stress is rarely known. In this work, we describe the identification of pepper mitochondrial FORMATE DEHYDROGENASE1 (CaFDH1) as a positive regulator of cell death and disease resistance. Transient expression of CaFDH1 in Nicotiana benthamiana leaves caused hypersensitive response (HR)-like cell death, where D-isomer specific 2-hydroxyacid dehydrogenase signature was crucial for the induction of the cell death. Localization analysis using green fluorescence protein showed that CaFDH1 targeted to the mitochondria with the mitochondria targeting sequence at N-terminal region. However, the mitochondrial localization of CaFDH1 is not essential for the induction of HR-like cell death, because the cytosolic CaFDH1 could elicit cell death response. Silencing of CaFDH1 in pepper significantly compromised the cell death response and salicylic acid (SA) levels, but enhanced growth of Xanthomonas campestris pv. vesicatoria. In contrast, transgenic Arabidopsis overexpressing FDH1 exhibited enhanced resistance to Pseudomonas syringae pv. tomato. Taken together, these data suggest that CaFDH1 has a role in HR cell death and defense response to microbial pathogens.

Environmentally inflicted stress (abiotic stress) such as high drought stress could be limiting the plant productivity. The mechanism of drought stress signaling in plant related with anti-apoptosis has not yet been full described. Understanding drought stress signaling is key to producing drought-tolerant plant. In this study we recently have identified Oryza sativa genes related abiotic stress water deficit. Abiotic stress related genes were screened from Oryza sativa cDNA library and identified gene by yeast functional screening. The yeast expression showed that they east cell grow well on SD-galactose-Leu-Ura-. The screening of over than 7000 clones from Oryza sativa cDNA libraries has been identified. 28 clones that survived following BAX-expression on inducible galactose medium. R12H780 clones confirmed protein prediction like putative senescence-associated-protein. This gene contains an open reading frame (ORF) of 108 amino acids. Transcription of R12H780 was induced in response to drought stresses, RT-PCR analysis showed transcript level in plant strongly detected in earliest time of drought stress treatment. Yeast transformed with R12H780 gene displayed markedly improved tolerance to PEG treatment, and high salinity in comparison to the control yeast (vector only). The results indicate R12H780 expression represents a new type of drought stress related gene with anti-apoptotic in Oryza sativa and endows tolerance to several types abiotic stress.

선행연구에서 본 연구자는 IEX-1 단백질이 난소 암세포에서 세포사멸(apoptosis)을 유도하는 기능을 수행함을 확인하였으나, 세포사멸과 세포생존의 여러 단계에서 어떠한 신호전달 체계로 IEX-1이 작용하는지는 정확히 알지 못하고 있다. 따라서 IEX-1 단백질과 결합하는 새로운 단백질을 찾기 위해 yeast two-hybrid system을 이용하였다. 그 결과 IEX-1이 여러 다양한 인간 암 세포에서 세포사멸을 유도하는 CATHEPSIN B 단

Ethanol treatment during the brain growth spurt period has been known to induce the death of Purkinje cells. The underlying molecular mechanisms and the role of reactive oxygen species (ROS) in triggering ethanol-induced Purkinje cell death are, however, largely unresolved. We undertook TUNEL staining, western blotting assay and immunohistochemistry for the cleaved forms of caspase-3 and -9, with calbindin D28K double immunostaining to identify apoptotic Purkinje cells. The possibility of ROS-induced Purkinje cell death was immunohistochemically determined by using anti-8-hydroxy-2'deoxyguanosine (8-OHdG), a specific cellular marker for oxidative damage. The results show that Purkinje cell death of PD 5 rat cerebellum following ethanol administration is mediated by the activation of caspase-3 and -9. However, unexpectedly, TUNEL staining did not reveal any positive Purkinje cells while there were some TUNEL-positive cells in the internal and external granular layer. 8-OHdG was detected in the Purkinje cell layers at 8 h, peaked at 12-24 h, but not at 30 h post-ethanol treatment. No 8-0HdG immunoreactive cells were detected in the internal and external granular layer. The lobule specific 8-OHdG staining patterns following ethanol exposure are consistent with that of ethanol-induced Purkinje cell loss. Thus, we suggest that ethanol-induced Purkinje cell death may not occur by the classical apoptotic pathway and oxidative damage is involved in ethanol-induced Purkinje cell death in the developing cerebellum.

Cerebral ischemia results from a transient or permanent reduction in cerebral blood flow that decreases oxygen and glucose supply. When the cellular oxygen supply is reduced to critical level, damage to cells and induction of cell death are occurred by excitotoxicity, oxidative stress and inflammation. Ischemia remains one of the leading causes of death, but there is no effective treatment that might protect neurons gainst ischemia by interrupting the cascade of cell death. In this study, human neuroblastoma SH-SY5Y cells are exposed to oxygen and glucose deprivation (OGD) followed by reoxgenation. OGD can mimic the acute restriction of metabolite and oxygen supply caused by ischemia and is widely used as a model of ischemic conditions. SH-SY5Y cells are treated samples at the commencement of OGD to achieve different final concentrations, and cell viabilities were quantified using the measurement of flow cytometry analysis. Of those tested, the extracts of Polygala tenuifolia (roots), Dictamnus dasycarpus (barks), Polygala tenuifolia (roots), Eucommia ulmoides (branches), Eucommia ulmoides (barks), Poria cocos (whole), Sophora flavescens (roots) showed neuroprotective effects, with EC50 values of 4.5±0.6, 7.9±1.5, 10.5±0.7, 18.4±1.9, 19.6±0.3, 21.6±1.9, and 30.7±3.9μg/ml, respectively.

This study investigated the protective roles and mechanism of magnolol, from the stem bark of Magnolia officinalis against potential neurotoxin 3-hydroxykynurenine (3-HK)-induced neuronal cell death. For the evaluation of protective role of magnolol, we examined cell viability, apoptotic nuclei, change of mitochondrial membrane potential and caspase activity in human neuroblastoma SH-SY5Y cells. It was found that 3-HK induces neuronal cell death in the human neuroblastoma SH-SY5Y cell line. The reduced cell viability produced characteristic features such as cell shrinkages, plasma membrane blebbing, chromatin condensation, and nuclear fragmentation. The cells treated with 3-HK showed an increase in the concentration of reactive oxygen species (ROS) as well as in caspase activity. In addition, both are involved in the 3-HK-induced apoptosis. Magnolol attenuated the cell viability reduction by 3-HK in both a dose- and time-dependent manner. Optical microscopy showed that magnolol inhibited the cell morphological features in the 3-HK-treated cells. Furthermore, the increase in the ROS concentration and the caspase activities by 3-HK were also attenuated by magnolol. These results showed that magnolol has a protective effect on the 3-HK induced cell death by inhibiting ROS production and caspase activity.

바둑에 있어 사활문제는 컴퓨터 바둑을 구현하기 위해 반드시 극복해야 하는 기본적인 문제이다. 사활문제와 같은 국부적인 바둑 문제를 해결하기 위하여 고려해야 될 중요한 사항은 게임 트리의 엄청난 분기수와 그 깊이를 어떻게 처리하느냐이다. 본 논문에서 수행된 실험의 기본 착상은 둘러싸인 돌들을 죽이기 위해 인식된 첫 수들을 찾아내는 인간의 습성을 모방한 것이다. 바둑에 있어, 유사한 사활문제(패턴)들은 자주 유사한 해들을 갖는다. 유사한 패턴을 분류 하기 위하여 코호넨 신경망(KNN)을 기반으로 한 군집화를 수행하였으며, 실험 결과는 고무적이며 사활문제를 풀기 위해 신경망으로 통제 학습을 사용하는 패턴 일치와 경쟁할 수 있음을 알아냈다.

Iris nertschinsk has been used generally as a decorative plant. However, it has been almost used as a medicine for therapy on various human diseases. In this study, we demonstrate the anti-tumor effect of Iris nertschinsk on human breast cancer cells. Firstly, we found that Iris nertschinsk dose-dependently induced cell death in human breast cancer cell lines, MCF7 and MDA-MB231. Moreover, phosphorylation of p53 was induced after Iris nertschinsk treatment in MCF7 cells, which has a functional p53, but not in MDA-MB231 cells, which has a dysfunctional p53. We next examined whether Iris nertschinsk induces caspase-dependent cell death. Caspase-7 was cleaved after Iris nertschinsk treatment in MCF7 cells. Interestingly, either caspase-3 or caspase-7 was cleaved in MDA-MB231 cells that p53 had been phosphorylated by Iris nertschinsk treatment, indicating that Iris nertschinsk induces apoptosis through the cleavage of caspase-3, -7 in human breast cancer cell lines, MCF7 and MDA-MB231, but related to the status of p53. Therefore, these results suggest that Iris nertschinsk could be used as a treatment for human breast cancer. This research is supported by National Institute of Agricultural Biotechnology research grant.

배아줄기세포는 다양한 분화 유도 방법을 통해 신경계세포로 분화시킬 수 있을 뿐만 아니라, 보다 더 엄격한 선발조건을 적용함으로써 특정 종류의 신경세포만을 확보할 수도 있게 되었다. 세포사멸연구를 포함한 신경생리학적 연구의 대상으로써 중요한 요건은 이렇게 확보한 배아줄기세포 유래의 신경계세포들이 정상적인 신경생리학적 특성을 갖고 있어야 하며, 동시에 그런 신경생리학적 특성이 체외에서 일정기간 동안 이상 자연적인 세포사멸없이 유지되어야 한다는 것이다. 생쥐

Amyloid 에 의해 유도되는 세포사멸을 보호하는 물질을 검색하기 위하여 250여 식물 재료 및 식품성분으로부터 스크리닝한 결과 가장 효과가 있는 시금치 추출물을 이용하여 뇌신경세포사멸(neuronal cell death)을 어느 정도 보호할 수 있는지를 알아보았다. 시금치 추출물이 항산화 활성과 acetylcholinesterase 활성에 대한 저해효과는 시금치 추출물 처리농도가 높을수록 유의적으로 높게 나타났다. 과산화수소와 amyloid

퇴행성 뇌질환의 하나인 알츠하이머는 가벼운 기억력의 장애에서부터 전반적인 인지기능의 장애를 나타내는 질환으로 해마 (hippocampus)를 포함한 신경세포에서 세포사와 관련이 있는 것으로 보고 되어왔다. AP의 자체 독성과 산화 스트레스, plaque에서 나오는 free radical은 세포내 를 높이고 이로 인해 calpain이 활성화되어 신경 세포사가 촉진되며 microtubule과 같은 cytoskeleton을 파괴시킨다. 이러한 ROS

From the soils of soybean fields in Cotton Branch Station (CBS) and Pine Tree Station (PTS), Arkansas, USA, various single spore isloates of sudden death syndrome (SDS) pathogen were obtained on modified Nash ＆ Snyder's medium (MNSM) with dilution plating technique and transferred to potato dextrose agar (PDA) medium to identify the cultural colony shape. The colony shapes of these isolates resembled F. solani isolate 171 which was white and chalky shaped on MNSM and most of them had unique form of morphology which produced white margin and blue center colony on PDA. Although, some of these isolates had more dark blue or showed slightly different color, all isolates that were selected randomly for green-house inoculation assay produced typical foliar symptoms on leaves of soybean, Hartz 6686. To determine the genetic differences among the isolates, mitochondrial DNA restriction fragment length polymorphism (RFLP) was conducted with fourty isolates from both fields, using mtDNA probes, 2U18 and 4U40, derived from Colletotrichum orbiculare. We obtained distinctive RFLPs in each treatment of restriction enzyme, EcoRI and HaeⅢ. Isolates, 11-2-5 and 14-3-1-1, from CBS and isolates, 104-3-1-2 and 701-1-5-1, from PTS showed different band patterns from 171 in both or in either treatment of restriction enzymes. Even if some of these isolates showed heterogeneous, they were more closer to 171 than PN603. And, also, rest of the thirty-six isolates had exactly same polymorphisms as 171 in each treatment of restriction enzyme. Although, some of the isolates showed the different morphological shape on PDA and slightly different band patterns on RFLPs, all of the isolates selected on MNSM due to their distinctive colony shape from other fungi produced the typical foliar symptoms on soybean leaves in greenhouse inoculation assay. It might be suggested that these isolates were not genetically different from check isolate 171 and they were unique strain of F. solani

In order to identify the responses of Korean soybean cultivars to sudden death syndrome (SDS), forty-two Korean cultivars and three check cultivars (Hartwig and PI 520733 are resistant; Hartz 6686 is susceptible) were tested with sorghum seed inoculum infested with Fusarium solani f. sp. glycines isolate 171 in the greenhouse. This isolate has blue pigment cultural shape on potato dextrose agar (PDA) medium. All Korean cultivars inoculated with F. solani isolate 171 showed the typical SDS symptoms and disease severity on soybean leaves in each cultivar varied at 4 weeks after inoculation. Nine cultivars were included in the most SDS susceptible group and six cultivars were included in the most susceptible group based on Duncan＇s multiple range tests (P~leq 0.05). In results of the LSD analysis for SDS the resistant group, a total of twenty-five Korean cultivars were included in the same SDS resistant group as PI 520733 or Hartwig and fourteen Korean cultivars were included in the same SDS susceptible group as Hartz 6686. In the second experiment, ten Korean cultivars, ten U.S. cultivars, and one introduced line were compared in the same way as the first experiment Disease severity ranking of check cultivars, Hartwig, PI 520733, and Hartz 6686, were the same as in the first experiment. Within Korean cultivars, seven cultivars showed the consistent severity proportions of leaf symptoms. Disease rankings of these cultivars in this experiment were the same as those in the first experiment. Three US cultivars: Hartwig, Hartz 5454, and Forrest, three Korean cultivars： Keunolkong, Myeongjunamulkong, and Jinpumkong 2, and one introduced line, PI 520733, were included in the highest SDS resistant group. Shinphaldalkong 2, Milyang 87, and Samnamkong consistently showed the highest SDS susceptibility in both experiments. Average disease severity in the first and the second experiment were 49.56% and 45.39%, respectively.

Six soybean cultivars having different SDS susceptibility were planted with sorghum seedinoculum infested with F. solani isolate 171 in the greenhouse. First leaf symptoms appeared on unifoliar leaves at 9 days after inoculation and all cultivars showed the typical leaf symptoms at 13 days after inoculation, when trifoliar leaves emerged. Leaf symptoms development in susceptible cultivars was faster than in resistant cultivars. Leaf symptom severities during the period of 25 to 29 days after inoculation showed a significant difference between cultivars which had SDS resistance and sus ceptibility. In this period, area under the diseaseprogress curve (AUDPC) of Hartz 6686 was the highest and that of PI 520733 was the lowest. SDS caused serious damage to the growth of soybean in all cultivars. Average reductions of growth rate of root fresh weight and dry weight were greater than those of plant tops. Duyu-kong showed less severe leaf symptoms than that of SDS suscetible cultivars; however, average growth rate of plants top and roots of this cultivar was less but not significantly different than those of SDS susceptible cultivars. In all cultivars, as severity of leaf symptoms increased, plant top weight decreased. Root rot symptoms were observed in all cultivars before leaf symptoms appeared. Average proportions of tap root reddish-brown discoloration of all cultivars was up to 75 % at 15 days after inoculati on; however there was no significant differenc between cultivars at each rating date. Appearances of leaf symptoms on leaves varied in each cultivar. SDS resistant cultivars had a significantly higher level of crinkling than susceptible cultivars and SDS susceptible cultivars had a significantly higher level of necrosis than resistant cultivars. Further study will be needed to identify the relationships between the physiological growth rate and SDS severities in soybeans.

본 연구에서는 자연세포사 (apoptosis)를 유발시키는 것으로 알려진 ceramide를 배양중인 생쥐 과립세포에 처리한 뒤 형광염색, in-situ 3'-end labeling(ISEL), 그리고 flow cytometry 기법을 이용하여, 자연세포사 및 세포주기에 미치는 ceramide의 영향을 조사하였다. Ceramide를 처리하지 않은 대조군에 비하여, ceramide를 처리한 실험군에서 세로의 생존율은 농도에 비레하여 유의하게 감소하였다. 또