Generation of a Constitutive Green Fluorescent Protein Expression Construct to Mark Biocontrol Bacteria Using P43 Promoter from Bacillus subtilis

Kong, Hyun-Gi,Choi, Ki-Hyuck,Heo, Kwang-Ryool,Lee, Kwang-Youll,Lee, Hyoung-Ju,Moon, Byung-Ju,Lee, Seon-Woo The Korean Society of Plant Pathology 2009 Plant Pathology Journal Vol.25 No.2

Marking biocontrol bacteria is an essential step to monitor bacterial behavior in natural environments before application in agricultural ecosystem. In this study, we presented the simple green fluorescent protein (GFP) reporter system driven by the promoter active in Bacillus species for tagging of the biocontrol bacteria. A constitutive promoter P43 from Bacillus subtilis was fused to an enhanced promoterless gfp gene by overlap extension PCR. The GFP expression was demonstrated by the high fluorescence intensity detected in B. subtilis and Escherichia coli transformed with the P43-gfp fusion construct, respectively. The GFP reporter system was further investigated in two bacterial biocontrol strains B. licheniformis and Pseudomonas fluorescens. When the reconstructed plasmid pWH34G was introduced into B. licheniformis, GFP level measured with the fluorescence intensity in B. licheniformis was almost equivalent to that in B. subtilis. However, GFP expression level was extremely low in other biocontrol bacteria P. fluorescens by transposon based stable insertion of the P43-gfp construct into the bacterial chromosome. This study provides information regarding to the efficient biomarker P43-gfp fusion construct for bio-control Bacillus species.

Spatial and Temporal Distribution of a Biocontrol Bacterium Bacillus licheniformis N1 on the Strawberry Plants

Kong, Hyun-Gi,Lee, Hyoung-Ju,Bae, Ju-Young,Kim, Nam-Hee,Moon, Byung-Ju,Lee, Seon-Woo The Korean Society of Plant Pathology 2010 Plant Pathology Journal Vol.26 No.3

Spatial and temporal distribution of Bacillus licheniformis N1 was investigated over time on the leaves, petioles and crowns of the strawberry plants. Bacterial population on the strawberry plants was quantified over time by selective plating. Bacterial population of N1 containing a plasmid pWH43G carrying green fluorescent protein (GFP) declined relatively faster on the plant surface as compared to the Strain N1 itself. However, this result was found to be enough to utilize the strain to visualize bacterial colonization on the plant surface. When B. licheniformis N1 was treated together with Silwet L-77 at 0.03%, the bacterial population on plant surface persisted for up to 7 days. B. licheniformis N1 (pWH43G) containing Silwet L-77 was applied on the strawberry plants and the GFP expressing bacteria were visualized by confocal laser scanning microscopy. Bacterial persistence was also investigated in a growth chamber and in a plastic house after N1 bioformulation treatment on the strawberry plant. The Strain N1 colonized three different tissues well and persisted over 3 to 5 days on the strawberry plants. They formed bacterial aggregates on plant surfaces for at least 3 days, resulting in a biofilm to resist fluctuating plant surface environment. However, the bacterial persistence dramatically declined after 7 days in all tested tissues in a plastic house. This study suggest that B. licheniformis N1 colonizes the strawberry plant surface and persists for a long time in a controlled growth chamber, while it can not persist over 7 days on the plant surface in a plastic house.

Development of transgenic rice lines expressing the human lactoferrin gene

Jin-Hyoung Lee,Hyo-Sung Kim,Kong-Sik Shin,Seok-Cheol Suh,Soon-Jong Kweon,임성렬,Il-Gi Kim 한국식물생명공학회 2010 식물생명공학회지 Vol.37 No.4

Lactoferrin is an 80-kDa iron-binding glycoprotein that is found in high concentrations in human milk. Human lactoferrin (hLF) has several beneficial biological activities including immune system modulation and antimicrobial activity. In the present study, we devolved a method of hLF expression through introducing the hLF gene construct into Oriza sativa cv. Nakdong using the Agrobacterium-mediated transformation system. The expression of the hLF gene under the control of the rice glutelin promoter was detected in the seeds of transgenic rice plants. Transformed rice plants were selected on media containing herbicide(DL-phosphinothricin) and the integration of hLF cDNA was confirmed by Southern blot analysis. The expression of the full length hLF protein from the grains of transgenic rice plants was verified by Western blot analysis. The lactoferrin expression levels in the transformed rice grains determined by enzyme-linked immunosorbant assay accounted for approximately 1.5% of total soluble protein. Taken together, these data indicate that rice grains expressing hLF can be directly incorporated into infant formula and baby food.

Development of transgenic rice lines expressing the human lactoferrin gene

Lee, Jin-Hyoung,Kim, Il-Gi,Kim, Hyo-Sung,Shin, Kong-Sik,Suh, Seok-Cheol,Kweon, Soon-Jong,Rhim, Seong-Lyul The Korean Society of Plant Biotechnology 2010 식물생명공학회지 Vol.37 No.4

Lactoferrin is an 80-kDa iron-binding glycoprotein that is found in high concentrations in human milk. Human lactoferrin (hLF) has several beneficial biological activities including immune system modulation and antimicrobial activity. In the present study, we devolved a method of hLF expression through introducing the hLF gene construct into Oriza sativa cv. Nakdong using the Agrobacterium-mediated transformation system. The expression of the hLF gene under the control of the rice glutelin promoter was detected in the seeds of transgenic rice plants. Transformed rice plants were selected on media containing herbicide(DL-phosphinothricin) and the integration of hLF cDNA was confirmed by Southern blot analysis. The expression of the full length hLF protein from the grains of transgenic rice plants was verified by Western blot analysis. The lactoferrin expression levels in the transformed rice grains determined by enzyme-linked immunosorbant assay accounted for approximately 1.5% of total soluble protein. Taken together, these data indicate that rice grains expressing hLF can be directly incorporated into infant formula and baby food.

Rhizosphere microbiome structure alters to enable wilt resistance in tomato

Kwak, Min-Jung,Kong, Hyun Gi,Choi, Kihyuck,Kwon, Soon-Kyeong,Song, Ju Yeon,Lee, Jidam,Lee, Pyeong An,Choi, Soo Yeon,Seo, Minseok,Lee, Hyoung Ju,Jung, Eun Joo,Park, Hyein,Roy, Nazish,Kim, Heebal,Lee, M Nature Pub. Co 2018 Nature biotechnology Vol.36 No.11

<P> Tomato variety Hawaii 7996 is resistant to the soil-borne pathogen Ralstonia solanacearum, whereas the Moneymaker variety is susceptible to the pathogen. To evaluate whether plant-associated microorganisms have a role in disease resistance, we analyzed the rhizosphere microbiomes of both varieties in a mesocosm experiment. Microbiome structures differed between the two cultivars. Transplantation of rhizosphere microbiota from resistant plants suppressed disease symptoms in susceptible plants. Comparative analyses of rhizosphere metagenomes from resistant and susceptible plants enabled the identification and assembly of a flavobacterial genome that was far more abundant in the resistant plant rhizosphere microbiome than in that of the susceptible plant. We cultivated this flavobacterium, named TRM1, and found that it could suppress R. solanacearum-disease development in a susceptible plant in pot experiments. Our findings reveal a role for native microbiota in protecting plants from microbial pathogens, and our approach charts a path toward the development of probiotics to ameliorate plant diseases. </P>

브랜드경험과 브랜드애착의 관계에서 즐거움의 매개효과

공란란,김동국,김형길,Kong,Lan-Lan,Jin,Dong-guo,Kim,Hyoung-Gi 한국소비문화학회 2015 소비문화연구 Vol.18 No.2

단순한 제품이나 서비스의 기능, 품질 등 물리적 측면을 넘어 소비 자체의 경험을 추구하는 소비자들의 욕구 변화에 따라 기업의 마케터들 뿐만 아니라 마케팅 분야의 학자들은 경험의 중요성에 주목하고 있다. 그러나 브랜 드로부터 유발되는 경험은 브랜드에 대한 소비자의 감정 형성, 소비자-브랜드 간의 관계 구축에 있어 아주 중요 함에도 불구하고 관련 연구는 미흡한 실정이다. 따라서 본 연구는 다차원 브랜드경험(감각적, 감정적, 인지적, 행 동적)이 소비자의 감정(정서적 즐거움, 인지적 즐거움)에 어떤 영향을 미치는지, 브랜드경험에 의한 형성된 소비자의 즐거움이 소비자-브랜드 간의 유대관계를 의미하는 브랜드애착을 증가시키는지, 그리고 브랜드경험과 브랜드애착의 관계에서 다차원 즐거움(정서적, 인지적)이 매개효과가 있는지에 대해서 검증할 것이다. 문헌고찰을 통 해 연구모형 및 가설을 도출하였으며, 실증분석은 스마트폰 브랜드를 조사대상으로 선정하여, 20대-30대 스마트 폰 사용자 357명으로부터 수집된 자료를 활용하였다. 분석결과, 첫째, 브랜드경험(감각적, 감정적, 인지적)은 각각 정서적 즐거움과 인지적 즐거움에 정(+)의 영향 을 미치는 것으로 확인되었으나, 행동적 경험은 가설과 일치하지 않아 기각되었다. 둘째, 즐거움(정서적, 인지적) 은 브랜드애착을 증가시키는 유의한 변수로 확인되었다. 셋째, 다차원 브랜드경험(감각적, 감정적, 인지적)과 즐거움(정서적, 인지적)의 상대적 영향력 비교 결과, 브랜드경험(감각적, 감정적, 인지적)의 정서적 즐거움에 대한 상대적 영향력은 인지적, 감정적, 감각적 경험의 순으로 판명되었다. 또한 브랜드경험(감각적, 감정적, 인지적)의 인지적 즐거움에 대한 상대적 영향력은 인지적, 감정적, 감각적 경험 순으로 나타났다. 넷째, 브랜드경험(감각적, 감정적, 행동적, 인지적)과 브랜드애착의 관계에서 즐거움(정서적, 인지적)의 매개효과를 살펴보면, 감각적 및 인 지적 경험과 브랜드애착의 관계는 정서적 및 인지적 즐거움에 의해 완전 매개되고, 감정적 경험과 브랜드애착은 정서적 및 인지적 즐거움에 의해 부분 매개되는 것으로 확인되었다. 이러한 결과는 스마트폰 시장의 브랜드 마케 팅전략 수립과 소비자-브랜드 간의 관계 구축에 중요한 시사점을 제공한다. According to the changes of consumers' needs for the pursuit of consumption experiences beyond the physical aspects(e.g., function, quality) of products or service, marketers as well as marketing scholars are focused on the importance of experience. However, research has ignored brand experience despite the important influence of brand experience on affective outcomes and the formation of relationship between consumers and brand. So, the present study is aimed to demonstrate the causal relationships of muli-dimensional brand experience(sensory, affective, behavioral, intellectual) to pleasure(emotional and cognitive), the effect of pleasure induced by brand experience on enhancing brand attachment and in addition, to investigate mediating effects of pleasure (emotional, cognitive) in the links between multi-dimensional brand experiences and brand attachment respectively. In order to effectively achieve the research objective, we built the research model based on the literature reviews and formulated the hypotheses. We selected three smartphone brands; GalaxyS, iPhone, and Optimus. We conducted the survey from 20-30's smartphone users. 357 responses were applied for analysis. The results indicated as follows. First, Brand experience(sensory, affective, intellectual) were positively associated with emotional pleasure and cognitive pleasure respectively, whereas the effect of behavioral experience to emotional and cognitive pleasure was not supported. Second, it was demonstrated that pleasure(emotional, cognitive) was a significant factor on enhancing brand attachment. Third, comparing the relative effects of construct of brand experiences to pleasure(emotional and cognitive), intellectual experience had the strongest impacts to pleasure (emotional, cognitive) better than sensory and affective, and had the same results in case of cognitive pleasure. Finally, with regard to the results of the mediating effect by pleasure(emotional and cognitive) in the relationships between brand experiences and brand attachment, emotional pleasure and cognitive pleasure had partially mediated in the relationship between sensory experience-brand attachment, and intellectual experience-brand attachment respectively, whereas, both pleasures were partially mediated the relationship of affective experience-brand attachment. The findings will help marketers establish more efficient brand strategies and the formation of relationship between consumers and brand for smartphone market.

Altered Gene Expression and Intracellular Changes of the Viable But Nonculturable State in Ralstonia solanacearum by Copper Treatment

Um, Hae Young,Kong, Hyun Gi,Lee, Hyoung Ju,Choi, Hye Kyung,Park, Eun Jin,Kim, Sun Tae,Murugiyan, Senthilkumar,Chung, Eunsook,Kang, Kyu Young,Lee, Seon-Woo The Korean Society of Plant Pathology 2013 Plant Pathology Journal Vol.29 No.4

Environmental stresses induce several plant pathogenic bacteria into a viable but nonculturable (VBNC) state, but the basis for VBNC is largely uncharacterized. We investigated the physiology and morphology of the copper-induced VBNC state in the plant pathogen Ralstonia solanacearum in liquid microcosm. Supplementation of $200{\mu}M$ copper sulfate to the liquid microcosm completely suppressed bacterial colony formation on culture media; however, LIVE/DEAD BacLight bacterial viability staining showed that the bacterial cells maintained viability, and that the viable cells contain higher level of DNA. Based on electron microscopic observations, the bacterial cells in the VBNC state were unchanged in size, but heavily aggregated and surrounded by an unknown extracellular material. Cellular ribosome contents, however, were less, resulting in a reduction of the total RNA in VBNC cells. Proteome comparison and reverse transcription PCR analysis showed that the Dps protein production was up-regulated at the transcriptional level and that 2 catalases/peroxidases were present at lower level in VBNC cells. Cell aggregation and elevated levels of Dps protein are typical oxidative stress responses. $H_2O_2$ levels also increased in VBNC cells, which could result if catalase/peroxidase levels are reduced. Some of phenotypic changes in VBNC cells of R. solanacearum could be an oxidative stress response due to $H_2O_2$ accumulation. This report is the first of the distinct phenotypic changes in cells of R. solanacearum in the VBNC state.

Formation of Biological Agents using Stenotrophomonas maltophilia BW - 13 stain to Bottom Rot caused by Rhizoctonia solani PY-1 on Chisphead Lettuce and Its Control Effects

Ok Ju Chun,Hyun Ju Kim,Han Woo Kim,Jung Woo Baek,Kwang Youll Lee,Hyoung Gi Kong,Jin Woo Lee,Soon Je Jung,Byung Ju Moon 한국생명과학회 2004 한국생명과학회 심포지움 Vol.43 No.-

Selection of effective nutrient sources for mass culture of Stenotrophomonas maltophilia BW - 13 against Rhizoctonia solani PY-1 causing Bottom Rot on Chisphead Lettuce

Ok Ju Chun,Hyun Ju Kim,Han Woo Kim,Jung Woo Baek,Kwang Youll Lee,Hyoung Gi Kong,Jin Woo Lee,Soon Je Jung,Byung Ju Moon 한국생명과학회 2004 한국생명과학회 심포지움 Vol.43 No.-