• Title/Summary/Keyword: wild type

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Pathogen Associated Molecular Pattern (PAMP)-Triggered Immunity Is Compromised under C-Limited Growth

  • Park, Hyeong Cheol;Lee, Shinyoung;Park, Bokyung;Choi, Wonkyun;Kim, Chanmin;Lee, Sanghun;Chung, Woo Sik;Lee, Sang Yeol;Sabir, Jamal;Bressan, Ray A.;Bohnert, Hans J.;Mengiste, Tesfaye;Yun, Dae-Jin
    • Molecules and Cells
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    • v.38 no.1
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    • pp.40-50
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    • 2015
  • In the interaction between plants and pathogens, carbon (C) resources provide energy and C skeletons to maintain, among many functions, the plant immune system. However, variations in C availability on pathogen associated molecular pattern (PAMP) triggered immunity (PTI) have not been systematically examined. Here, three types of starch mutants with enhanced susceptibility to Pseudomonas syringae pv. tomato DC3000 hrcC were examined for PTI. In a dark period-dependent manner, the mutants showed compromised induction of a PTI marker, and callose accumulation in response to the bacterial PAMP flagellin, flg22. In combination with weakened PTI responses in wild type by inhibition of the TCA cycle, the experiments determined the necessity of C-derived energy in establishing PTI. Global gene expression analyses identified flg22 responsive genes displaying C supply-dependent patterns. Nutrient recycling-related genes were regulated similarly by C-limitation and flg22, indicating re-arrangements of expression programs to redirect resources that establish or strengthen PTI. Ethylene and NAC transcription factors appear to play roles in these processes. Under C-limitation, PTI appears compromised based on suppression of genes required for continued biosynthetic capacity and defenses through flg22. Our results provide a foundation for the intuitive perception of the interplay between plant nutrition status and pathogen defense.

Biological Effect of Metronidazole Resistance in Helicobacter Pylori (Helicobacter pylori에서 metronidazole 내성이 미치는 생물학적 영향)

  • Lee Gin Hyug;Lee Sun-Mi;Nam Won-Hee;Kim Eun Sil;Kim Jin Ho;Kang Ho Young;Jeong Jin-Yong
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.955-960
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    • 2005
  • Resistance to metronidazole, a key component of therapies against Helicobacter pylori, is common in clinical isolates. Resistance generally requires inactivation of rdxA (HP0954), and sometimes also frxA (HP0642), two related nitroreductase genes. Here we studied the effect of resistance to metronidazole on fitness of the gastric pathogen H. pylori. The effect of metronidazole resistance for H. pylori in culture was assessed first by looking at colonies formed by freshly constructed mutant derivatives of H. pylori strain 26695. Mutations resulting in metronidazole resistance caused premature death of H.pylori in stationary phase, but had no significant effect on early exponential growth. The effect of nitroreductase deficiencies on fitness in vivo was tested by infecting C57BL/6 mice with 1:1 mixtures of SS1 wild type and its isogenic metronidazole resistant derivatives. Inactivation of rdxA caused an inability to colonize mice in SS1 H. pylori strain. Derivatives of a metronidazole resistant strain that survived better in stationary phase, although remaining metronidazole resistant, could again colonize mice. In conclusion, metronidazole resistance diminishes H. pylori's fitness, but their costs can be suppressed by additional mutation.

Characterization of a Drought-Tolerance Gene, BrDSR, in Chinese Cabbage (배추의 건조 저항성 유전자, BrDSR의 기능 검정)

  • Yu, Jae-Gyeong;Lee, Gi-Ho;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.34 no.1
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    • pp.102-111
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    • 2016
  • The goal of this study was to characterize the BrDSR (Drought Stress Resistance in B. rapa) gene and to identify the expression network of drought-inducible genes in Chinese cabbage under drought stress. Agrobacterium-mediated transformation was conducted using a B. rapa inbred line ('CT001') and the pSL100 vector containing the BrDSR full length CDS (438 bp open reading frame). Four transgenic plants were selected by PCR and the expression level of BrDSR was approximately 1.9-3.4-fold greater than that in the wild-type control under drought stress. Phenotypic characteristics showed that BrDSR over-expressing plants were resistant to drought stress and showed normal growth habit. To construct a co-expression network of drought-responsive genes, B. rapa 135K cDNA microarray data was analyzed to identify genes associated with BrDSR. BrDSR was directly linked to DARK INDUCIBLE 2 (DIN2, AT3G60140) and AUTOPHAGY 8H (ATG8H, AT3G06420) previously reported to be leaf senescence and autophagy-related genes in plants. Taken together, the results of this study indicated that BrDSR plays a significant role in enhancement of tolerance to drought conditions.

Selection of Mutant Lines with Useful Traits by Irradiation of Proton Ion Beams and Gamma Ray in Rapeseed (Brassica napus L.) (양성자 및 감마선처리에 의한 유채 유용형질의 변이계통 선발)

  • Kim, Jun-Su;Eun, Jong-Seon
    • Korean Journal of Breeding Science
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    • v.43 no.2
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    • pp.145-153
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    • 2011
  • To develop new varieties of rapeseed (Brassica napus L.), the seeds of three varieties 'Naehan', 'Tammi', and 'Halla' were treated with proton ion beams and gamma rays (0~2,000 Gy), and then the characteristics of the mutants induced were examined up to $M_5$ generation to select the lines with fixed useful traits. In $M_5$ generation, we had selected several lines that were highly fixed for some useful traits such as plant height, maturity and flower size; one line with both earlier maturity and shorter stem than wild type, one line with only earlier maturity, two lines with shorter stem, one line with large flower, and one line with chlorophyll mutation. Among them, NP600-1-1-198-2 (induced from variety 'Naehan' was treated with proton ion beams 600 Gy) was superior for its distinction from the original variety, uniformity and stability. The unique characteristics of NP600-1-1-198-2 were dark green leaves, green stem, yellow flower, and black seed coat. Its flowering date was April 14, eight days earlier than its original variety, while seed maturity date was June 16 (five days earlier) and plant height 105 cm (shorter by 10 cm). NP600-1-1-198-2 has 52 silique per panicle, 6.2 cm silique length, 23 seeds per silique, 4.2 g per 1000 seeds and 45.9% oil content. The seed oil contained 67.8% of oleic acid, 16.7% of linoleic acid and 7.3% of linolenic acid but no erucic acid.

Induction of Tumor Suppressor Gene p53-dependent Apoptosis by Sanguinarine in HCT116 Human Colorectal Cancer Cells (결장암세포에서 sanguinarine에 의한 종양억제 유전자 p53 의존적 apoptosis 유도)

  • Choi, Yung Hyun
    • Journal of Life Science
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    • v.31 no.4
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    • pp.400-409
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    • 2021
  • Sanguinarine, a natural benzophenanthridine alkaloid, has been considered a potential therapeutic target for the treatment of cancer because it can induce apoptosis in human cancer cells; however, the underlying mechanisms of action still remain unclear. Tumor suppressor p53 deletion or mutation is an important reason for the resistance of colorectal cancer cells to anticancer agents. Therefore, in the present study, the role of p53 during apoptosis induced by sanguinarine was investigated in p53wild type (WT, p53+/+) and p53null (p53+/+) HCT116 colon carcinoma cells. Sanguinarine significantly caused greater reductions in cell viability in HCT116 (p53+/+) cells than in HCT116 (p53-/-) cells. Consistently, sanguinarine promoted more DNA damage and apoptosis in HCT116 (p53+/+) cells than in HCT116 (p53-/-) cells while increasing the expression of p53 and cyclin-dependent kinase inhibitor p21WAF1/CIP1. Sanguinarine increased the activity of caspase-8 and caspase-9, which are involved in the initiation of extrinsic and intrinsic apoptosis pathways, respectively, and it activated caspase-3, a typical effect caspase, in HCT116 (p53+/+) cells. Sanguinarine also increased the generation of reactive oxygen species (ROS), and the Bax/Bcl-2 ratio, while destroying the integrity of mitochondria in HCT116 (p53+/+) cells, but not in HCT116 (p53-/-) cells. Overall, the results indicate that sanguinarine induced p53-dependent apoptosis through ROS-mediated activation of extrinsic and intrinsic apoptotic pathways in HCT116 colorectal cancer cells.

CRISPR/Cas9-mediated knockout of CD47 causes hemolytic anemia with splenomegaly in C57BL/6 mice

  • Kim, Joo-Il;Park, Jin-Sung;Kwak, Jina;Lim, Hyun-Jin;Ryu, Soo-Kyung;Kwon, Euna;Han, Kang-Min;Nam, Ki-Taek;Lee, Han-Woong;Kang, Byeong-Cheol
    • Laboraroty Animal Research
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    • v.34 no.4
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    • pp.302-310
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    • 2018
  • CD47 (integrin-associated protein), a multi-spanning transmembrane protein expressed in all cells including red blood cells (RBCs) and leukocytes, interacts with signal regulatory protein ${\alpha}$ ($SIRP{\alpha}$) on macrophages and thereby inhibits phagocytosis of RBCs. Recently, we generated a novel C57BL/6J CD47 knockout ($CD47^{-/-}$ hereafter) mouse line by employing a CRISPR/Cas9 system at Center for Mouse Models of Human Disease, and here report their hematological phenotypes. On monitoring their birth and development, $CD47^{-/-}$ mice were born viable with a natural male-to-female sex ratio and normally developed from birth through puberty to adulthood without noticeable changes in growth, food/water intake compared to their age and sex-matched wild-type littermates up to 26 weeks. Hematological analysis revealed a mild but significant reduction of RBC counts and hemoglobin in 16 week-old male $CD47^{-/-}$ mice which were aggravated at the age of 26 weeks with increased reticulocyte counts and mean corpuscular volume (MCV), suggesting hemolytic anemia. Interestingly, anemia in female $CD47^{-/-}$ mice became evident at 26 weeks, but splenomegaly was identified in both genders of $CD47^{-/-}$ mice from the age of 16 weeks, consistent with development of hemolytic anemia. Additionally, helper and cytotoxic T cell populations were considerably reduced in the spleen, but not in thymus, of $CD47^{-/-}$ mice, suggesting a crucial role of CD47 in proliferation of T cells. Collectively, these findings indicate that our $CD47^{-/-}$ mice have progressive hemolytic anemia and splenic depletion of mature T cell populations and therefore may be useful as an in vivo model to study the function of CD47.

Generation of Bacterial Blight Resistance Rice with Transcription Factor OsNAC69-overexpressing (전사인자 OsNAC69-과발현을 통한 흰잎마름병 저항성 벼 제작)

  • Park, Sang Ryeol;Cha, Eun-Mi;Moon, Seok Jun;Shin, Dongjin;Hwang, Duk-Ju;Ahn, Il-Pyung;Bae, Shin-Chul
    • Korean Journal of Breeding Science
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    • v.43 no.5
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    • pp.457-463
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    • 2011
  • Plant specific gene family, NAC (NAM, ATAF, and CUC) transcription factors have been characterized for their roles in plant growth, development, and stress tolerance. In this study, we isolated OsNAC69 gene and analyzed expression level by inoculation of bacterial leaf blight pathogen, Xanthomonas oryzae pv. oryzae (Xoo). NAC transcription factor family can be divided into five groups (I-V). On the basis of phylogenetic analysis, OsNAC69 was fall into group II. OsNAC69 was strongly induced 1 hr after infected with Xoo. To investigate its biological function in the rice, we constructed vector for overexpression in rice, and then generated transgenic rice lines. Gene expression of OsNAC69-overexpressed transgenic rice lines were analyzed by northern blot. Analysis of disease resistance to pathogen Xoo, nine OsNAC69-overexpressed transgenic rice lines showing high expression level of OsNAC69 were shown more resistant than wild type. These results suggest that OsNAC69 gene may play regulatory role during pathogen infection.

Characteristics of agronomy to vitamin A strengthening rice at large scale GMO field (대규모 GMO 포장에서 비타민 A 강화 벼의 농업특성 검정)

  • Lee, Hyun-Suk;Ryu, Tae-Hun;Jung, Hee-Young;Park, Soon-Ki;Park, Gyu-Hwan;Sohn, Jae-Keun;Kim, Kyung-Min
    • Korean Journal of Breeding Science
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    • v.42 no.1
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    • pp.56-60
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    • 2010
  • This study was carried out to develop of macro-protocol and the biosafety guide Golden Rice, modified vitamin A at large scale GMO field. Commercialization of genetically modified (GM) plants will be required the assessment of risks associated with the release of GM plants that should include a detailed risk assessment of their impacts in the environment and human health. Prior to GM plant release, applicants should provide the information on GM crops for approval. A total of $4,700m^2$ GM field, each of Golden Rice and 'Nakdong' were used for further analysis on agronomic traits and pathogenesis. Generally plant growth and panicle number in the transgenic lines were lower than in the wild type under field condition. The major agronomic traits such as plant growth, grain quality in Golden Rice were similar than in 'Nakdong'. The yield difference was due to reduced ripened rate and the 1,000 grain weight. The grain shape was similar donor plant, however, color of brown rice was different from segregation. Also the difference of pathogenesis reaction was not significant in the leaf blast and sheath blight between Golden Rice and 'Nakdong'. These results provided the agronomic data and pathogenesis for risk assessment analysis of Golden Rice and suggested that the macro-protocol could be useful to detect GM plants.

Seroprevalence of Dengue Virus Antibody in Korea (한국인에서 뎅기바이러스 항체의 혈청 유병률 연구)

  • Lee, Ji Hyen;Kim, Han Wool;Kim, Kyung-Hyo
    • Pediatric Infection and Vaccine
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    • v.25 no.3
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    • pp.132-140
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    • 2018
  • Purpose: The number of dengue fever cases is rising due to increasing overseas travel. Vaccination makes severe dengue fever in seronegative individuals after vaccination when they exposure to wild-type dengue virus. We investigated the seroepidemiology of the dengue virus for monitoring of Korean dengue virus immunity and establishing the prevention of dengue infection. Methods: The study was based on 446 residual sera collected from 98 infants (2 months to 1 year old), 152 adolescents (13 to 19 years old), 90 adults (20 to 50 years old), and 106 elderly participants (more than 65 years old) for other studies. Antibody levels for dengue virus immunoglobulin G (IgG) in each age group were measured using an enzyme-linked immunosorbent assay (ELISA). For each dengue virus IgG positive or equivocal result, an IgG ELISA was performed for Japanese encephalitis virus. Results: Of the 446 serum samples, only 1 (0.2%) adolescent had a positive result from the dengue IgG antibody test. In the dengue virus IgG antibody test, 14 (3.1%) samples showed equivocal results (10 adolescents and 4 elderly). In the 1 positive case of dengue virus IgG, the Japanese encephalitis IgG test was also positive. In the 14 equivocal cases of dengue virus IgG, there were 6 positive, 3 equivocal, and 5 negative of Japanese encephalitis IgG. Conclusions: The seroprevalence rate of dengue virus was very low in Koreans. This study provides important data for establishing the policy for preventive measures of dengue fever. It will be necessary to continuously monitor for dengue virus immunity.

Changes in Miscanthus sacchariflorus Growth and Heading Rate Influenced by Water Stress Treatment at Reproductive Growth Stage (생식생장기 수분스트레스 처리가 억새의 출수율 및 생육 변화에 미치는 영향)

  • Lee, Ji-Eun;Cha, Young-Lok;Moon, Youn-Ho;Kim, Kwang-Soo;Kwon, Da-Eun;Kang, Yong-Ku
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.63 no.4
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    • pp.390-398
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    • 2018
  • Miscanthus is a perennial energy plant that reproduces via rhizomes and has C4 metabolism. The flowering pattern of a wild type M. sacchariflorus (WTM) is affected by environmental conditions such as photoperiod and soil water status. Geodae-Uksae 1 (Geodea), which is a new M. sacchariflorus cultivar, has a lower heading rate than WMS; however the mechanism with why this happens is unknown. To confirm the effects of drought or waterlogging stresses on the growth of WTM and Geodea at the reproductive stage, we investigated variations in morphological characteristics and nutrient contents of the two Miscanthus species after four months of three water treatments. Morphological traits of the two Miscanthus species under the drought condition were similar to those under the control condition. But, the height of Geodea increased by 30% in response to the waterlogging stress. In WTM, the heading rate under the drought condition was lowest, while there was no significant difference between the waterlogging and control conditions. In the two Miscanthus species, nutrient contents, such as sucrose, total N, $P_2O_5$, K and Mg, were the highest under the drought condition, wherea Ca and Mg contents under the waterlogging condition were more than three times lower than those under the drought condition. The current study results showed that drought stress accelerated senescence and then inhibited nutrient mobilization in WTM, while waterlogging stress promoted the growth. This study is the first report to confirm that waterlogging stress promotes flowering of M. sacchariflorus.