• Title/Summary/Keyword: genetic resistance

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Development of an Efficient In Vitro Screening Method for Selection of Resistant Lily Cultivars Against Fusarium oxysporum f. sp. lilii (백합 구근부패병 저항성 검정을 위한 기내 간편 검정법 개발)

  • Jang, Ji-Young;Moon, Ki-Beom;Ha, Jang-Ho;Park, Ji-Sun;Kim, Mi-Jin;Jeon, Jae-Heung;Lee, Geung-Joo;Kim, Hyun-Soon
    • Horticultural Science & Technology
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    • v.33 no.6
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    • pp.883-890
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    • 2015
  • The soilborne fungus Fusarium oxysporum f. sp. lilii (Fol) is a serious threat to all lily cultivars, especially infecting bulbs and flowers. It has become increasingly important to develop varieties resistant against the bulb rot disease. Genetic diversity of cultivars and reliable screening methods are required for this purpose. Here, an efficient in vitro screening system for evaluating resistance to Fol in 38 in vitro-grown lily plants was investigated. Various factors including culture conditions of Fol, inoculum density, appropriate plant materials, inoculation method and duration, and incubation period of plant materials after inoculation were combined to optimize the screening method. As a result, we suggest optimal conditions for an in vitro screening system for the selection of Fol-resistant lily cultivars as follows. Fol was grown on potato dextrose agar (PDA) medium for 6 days at $25^{\circ}C$ in darkness and used as working inoculation. Spore suspensions were prepared (inoculum density: $1.0{\times}10^4$ $spores{\cdot}mL^{-1}$), and then leaf segments $1.5{\times}2.0cm^2$ were inoculated by dipping for 22 hours at $25^{\circ}C$ in dark. Later, leaves were cultured on 0.6% agar plates at $25^{\circ}C$ and 50% humidity with a photoperiod of 16 hours light/8 hours dark (fluence rate of $40{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$) to examine the progress of bulb rot. After 7 days, disease levels were classified into indices 1 (no symptom) to 6 (serious bulb rot). Soil inoculation of Fol carried out with resistant or susceptible lily cultivars that had been selected through in vitro screening confirmed the reproducibility of results. Therefore, the in vitro screening method established in this study is efficient and reliable for selection of lily cultivars resistant against bulb rot disease.

Characteristics of Vibrio vulnificus Isolated in Incheon (인천지역에서 분리된 비브리오 패혈증균의 특성)

  • Oh, Bo-Young;Kim, Jung-Hee;Gong, Young-Woo;JeGal, Seung;Kim, Hye-Yeung;Lee, Mi-Yeon;Hwang, Kyoung-Wha;Koh, Yeon-Ja;Lee, Jae-Mann;Go, Jong-Myoung;Kim, Yong-Hee
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.256-263
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    • 2007
  • We performed the biochemical characteristics, molecular epidemiologocal analysis, and drug susceptibility test on V. vulnificus isolated from environmental sources in Incheon. For this study, 233 strains were isolated from seawater, sediment, shellfish. V. vulnificus isolates were divided into 15 biochemical groups, which were positive for ONPG and Amygdalin test. Among the 209 strains, 206 (98.6%) strains and 110 (52.6%) strains revealed positive for vvhA and viuB gene, and the viuB gene detection rates of V. vulnificus from seawater, shellfish and sediment were 48%, 48.5% and 61.6%, respectively. From disc diffusion test on 175 isolates, most of strains were sensitive to Imipenem (100.0%), Sulfamethoxazole/trimethoprim (98.9%), Tetracycline, Ciprofloxacin (98.3%), Ampicillin/sulbactam (97.1%), Ohloramphenicol (96.6%), Cefepime (94.9%) and Ceftriaxone (94.8%), multi-drug resistance rates was 31.5% of seawater, 34.4% of sediment and 29.2% of shellfish. PFGE was performed on 233 V. vulnificus isolates with the objective of investigating the extent of genetic diversity of these isolates in our region. We could find that at least 126 different PFGE patterns were generated according by 90% of similarity and 13 clusters by 58% of similarity. The major cluster was type I (44.6%) during the most of the year, and type J was frequent pattern in June and October. There were 9 distinct PFGE types in July, 8 types in August, 7 types in June, 6 types in September, 5 types in October 3 types in May and 1 type in March.

Genetic Variance and Combining Ability of Shattering and Growth Characters by Diallel Crosses of Sesame (이면교잡에 의한 참깨의 탈립성 및 생육형질의 유전변이와 조합능력)

  • Kim, Dong-Hwi;Kang, Chul-Whan;Shim, Kang-Bo;Park, Chang-Hwan;Lee, Sung-Woo;Seong, Nak-Sul
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.7
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    • pp.652-657
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    • 2006
  • Combining ability and heterosis for $F_{1}$ population obtained $5{\times}5$ half diallel cross in sesame were estimated for six quantitative characters. The parental varieties used for diallel cross were Yangbaek and Ansan of normal type, Suwon 177 and 195 of placenta adhesion type (PA) and SIG960320-5-1-1 of indehiscent type (ID). PA and ID type are shattering-resistant sesames and Yangbaek and Ansan are shattering sesames. The high heterosis of $F_{1}$ hybrids was expressed in the number of capsules per plant (NC), grain yield per plant (GY) and number of branches per plant (NB). Shattering.ate (SR) showed high positive heterosis toward increase of seeds shattering in all the combination of $F_{1}$ generation which was presumed because the characters connected with shattering resistance of seeds were governed by recessive gene. At the analysis of combining ability, highly significant general combining ability (GCA) effects were observed on all the characters in $F_{1}$ generation, significant specific combining ability (SCA) effects were observed on the NC, GY and SR. GCA variances were greater than SCA variance, which suggested that additive gene effects were greater than other nonadditive gene effects. Considering the estimated GCA effects, Suwon 195 (PA type) was found to be good parent for SR and GY, SIG960320-5-1-1 (ID type) was poor for GY and plant height. SCA effect for NC was great in Yangbaek/Suwon 195, SCA for GY was great in Ansan/SIG960320-5-1-1, and SCA for decrease of SR was great in Ansan/Suwon177.

Genetic Polymorphisms of the $\beta_2$-Adrenergic Receptor in the Severity of Bronchial Asthma (기관지 천식 환자에서 천식 증상의 정도에 따른 $\beta_2$ 교감신경 수용체의 유전자 다형성)

  • Shim, Jae-Jeong;Kim, Jei-Hyung;Lee, Seung-Yong;Kwan, Young-Hwan;Lee, So-Ra;Lee, Sang-Youb;Kang, Se-Yong;Kang, Yong-Koo;Cho, Jae-Youn;In, Kwang-Ho;Won, Nam-Hee;Yoo, Se-Hwa;Kang, Kyung-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.1
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    • pp.77-89
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    • 1998
  • Background: Genetic and environmental factors are known to affect the incidence and severity of asthma. Stimulation of $\beta_2$-Adrenergic Receptor ($\beta_2$AR) results in smooth muscle relaxation, leading to decrease in resistance of airflow. The gene encoding the $\beta_2$AR has recently been seguenced. The $\beta_2$AR genotype at the polymorphic loci of codons 16, 27, 34, and 164 was known to cause changes in the amino acids. The relationships between the structure of the $\beta_2$AR and its functions are being elucidated. Purpose : The gene encoding the $\beta_2$AR was carried out to assess the frequency of polymorphisms in bronchial asthma, to determine wheather these polymorphisms have any relation to the severity, or nocturnal symptoms in bronchial asthma. Methods: The subjects studied were 103 patients with bronchial asthma, which consisted of 30 mild episodic, 32 mild persistent, 17 moderate, and 24 severe asthma patients. The polymorphisms of the $\beta_2$AR gene were detected by mutated allele specific amplification (MASA) method at the codons 16,27,34, and 164. Results: The most frequent polymorphism was arginine 16 to glycine. The other two polymorphisms, valine 34 to methionine and glutamine 27 to glutamic acid occured in 11 and 6 patients respectively. The polymorphism of threonine 164 to isoleucine was not found in our enrolled patients. The homozygous polymorphism of $\beta_2$AR gene was found in only arginine 16 to glycine (12.6%). The heterozygous polymorphisms of $\beta_2$AR gene were in arginine 16 to glycine, valine 34 to methionine, and glutamine 27 to glutamic acid, as 65.1 %,10.7%, and 5.8% respectively in asthma patients. The presence of agrginine 16 to glycine heterozygous or/and homozygous polymorphism was associated in severe asthma (p=0.015), but there was no association between the other three polymorphisms and the severity of asthma. The frequency of the $\beta_2$AR gene polymorphisms was no relation in nocturnal asthma as compared with non-nocturnal asthma. Conclusion: The arginine 16 to glycine polymorphism of the $\beta_2$AR gene is the most frequently found in asthma patients and association with severe asthma. But there was no association between the polymorphism of the $\beta_2$AR gene and nocturnal asthma.

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Studies of Soybean Improvement by ${\gamma}$-ray and Chemical Mutagens -I. Comparison of the effects of EMS, Ei, and ${\gamma}$-rays in soybean(preliminary report) (돌연변이 유기에 의한 대두 품종개량연구 -I. 대두에 대한 EMS, EI 및 ${\gamma}$선처리효과비교(예보))

  • Shin-Han Kwon;Kun-Hyuk Im;Cheong-Yeol Sohn;Jong-Sun Eun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.7 no.1
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    • pp.139-144
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    • 1969
  • 1. In order to obtain useful mutants such as early maturity, resistance to lodging, high protein and oil content, and capability of high yield, dormant seeds of two soybean varieties, Jang Dan Baik Mok and Clark, were treated with ${\gamma}$-ray, Ethyl Methane Sulfonate(EMS), Ethylene Imine(EI)and combinations of ${\gamma}$-ray and EMS or EI. 2. The germination rate and survival rate in a variety Jang Dan Baik Mok were significantly decreased with ${\gamma}$-ray treatment while it was not the same in the Clark variety. A significant decrease for seedling height measured at 14 and 21 days after sowing was found with the increase of ${\gamma}$-ray dose in both varieties. 3. Germination rates in both varieties were significantly decreased as EI concentration increases, particularly severe damage in germination was observed at 0.008 Mo. concentration. Germination rate damages were found with EMS concentration increases in the variety Jang Dan Baik Mok while no regular responses in seedling height were observed in the variety Clark. 4. Germination rate was significantly lowered with the combined treatment of EMS and ${\gamma}$-ray 24KR than that of EMS alone. In the treatments of ${\gamma}$-ray with three levels of EI concentration, the combined treatments except 24KR+EI 0.002 Mol. resulted in better germination than of EI alone. In both varieties, significant reduction in seedling height was observed in the combined treatments of ${\gamma}$-ray with various concentrations of EMS, whereas stimulation effect on seedling height was found with treatment of ${\gamma}$-ray EI +0.08Mol. 5. Germination rate, survival rate, and plant height as criteria of radio sensitivity, variety Jang Dan Baik Mok is moze sensitive to ${\gamma}$-ray, EMS, and EI than the variety Clark, and the varietal difference in responsibility to mutagen may be due to the genetic-constitution of the varieties.

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Characterization of Entomopathogenic Fungus from Trialeurodes vaporariorum and Evaluation as Insecticide (온실가루이 병원성 곰팡이의 특성 및 살충제 개발을 위한 평가)

  • Yoon, Hwi Gun;Shin, Tae Young;Yu, Mi Ra;Lee, Won Woo;Ko, Seung Hyun;Bae, Sung Min;Choi, Jae Bang;Woo, Soo Dong
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.64-70
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    • 2013
  • The greenhouse whitefly, Trialeurodes vaporariorum, is an economically important pest for greenhouse crops because they cause direct damage by feeding on plant nutrients and indirect damage as transmits many virus vectors. It has recently become a serious problem because of the continuous use of insecticide resulting in resistance among greenhouse whitefly population. To overcome these problems, in this study, the biological characteristics and virulence of an entomopathogenic fungus isolated from the cadaver of nymph greenhouse whitefly were investigated. Isolated fungus was identified as Isaria fumosorosea by morphological examinations and genetic identification using sequences of the ITS, ${\beta}$-tubulin, and EF1-${\alpha}$ regions. This fungus was named as I. fumosorosea SDTv and tested for the virulence against nymphs T. vaporariorum and the cold activity, the thermotolerance and the stability of UV-B irradiation on conidia. Mortality rate of greenhouse whitefly showed from 84 to 100% and the virulence increased with increasing conidial concentrations, $1{\times}10^5$ to $10^8$ conidia/ml. Conidia were stable at $35^{\circ}C$, 0.1 $J/cm^2$ of UV irradiation and germinated after 8 days at $4^{\circ}C$. Additionally, the activities of chitinases and proteases produced by I. fumosorosea SDTv were varied according to the medium. In conclusion, I. fumosorosea SDTv which showed high mortality rate against greenhouse whitefly will be used effectively in the integrated pest management programs against the greenhouse whitefly.

Studies on Antimicrobial Susceptibility and Characteristics of R-plasmids and Antigens of High-level Gentamicin Resistant Enterococcus faecalis (Gentamicin 고도내성 Enterococcus faecalis균주의 항균제감수성, R-플라스미드 및 항원의 특성연구)

  • Kang, Hyun
    • Biomedical Science Letters
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    • v.1 no.1
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    • pp.55-72
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    • 1995
  • Forty gentamicin-resistant isolates of Enterococcus faecalis were selected from various clinical materials, determined their antimicrobial susceptibility, and studied there R-plasmid characteristics and polypeptide patterns. All of the isolates were susceptible to vancomycin. The MICs($\mu$/ml) of antimicrobial agents to the isolates were as follows; the MIC of gentamicin was 128 and $\geq$2040, ampicillin 1 and 1, chlorarmphenicol 2 and 8, erythromycin 32 and 256, and vancomycin 1 and 2. E. faecalis HL-1 strain had 8 plasmid DNA elements, HL-2 and HL-3 strains had 6, HL-4 had 7, HL-5 had 4, and HL-6 had 5. The 51.7 Kb of gentamicin resistance plasmid DNA was conjugally transferred from two strains of E. faecalis HL-1 and HL-6 to S. aureus SK 982. The plasmid transfer frequency between S. aureus SK 982 and E. faecalis HL-1 or E. faecalis HL-6 was 6.3$\times10^{-4} and 3.7$\times10^{-5}$, respectively. Plasmid curing ratio after the treatment of ethidium bromide(10$\mu$/ml) to E. faecalis tarnsconjugants R-1 and R-6 were about 51% and 67%, respectively. The tetracycline gene was located in 2.15 Kb plasmid of E. faecalis HL-1, but it was not found in the E. faecalis HL-6 by Southern blot analyses. The antigenic components of E. faecalis HL-1, HL-6, R-1 and R-6 strains were analyzed by SDS-PAGE and immunoblotting. The E. faecalis strains had 7 to 16 polypeptide bands, however their major proteins were 97.8 and 26.8 Kd. At the Immunoblotting, 97.8, 95.8, 74.8, 63.5, 33.7 and 26.8 Kd polypeptides of the strains showed major antigenic activities with patient's sera infected intra-abdominally with an E. faecalis strain.

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Cell Death Study in Embryonic Stem Cell-derived Neurons and Its Applications (배아줄기세포 유래 신경계세포에서의 세포사멸 연구와 그 응용)

  • Lee, Chul-Sang
    • Development and Reproduction
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    • v.12 no.1
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    • pp.1-8
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    • 2008
  • Specific protocols to increase the differentiation of neuronal cells from embryonic stem (ES) cells have been well established, such as retinoic acid induction and lineage selection of neuronal cells. For the neuropathological studies, ES-derived neurons (ES neurons) must show normal physiological characteristics related to cell death and survival and should be maintained in vitro for a sufficient time to show insults-specific cell death without spontaneous death. When mouse ES cells were plated onto astrocytes monolayer after retinoic acid induction, most ES cells differentiated into neuronal cells, which were confirmed by the presence of specific neuronal markers, and the cultures were viable for at least four weeks. When these cultures were examined for vulnerability to glutamate excitotoxicity, ES neurons were vulnerable to excitotoxic insults mediated by agonist-specific receptors. The vulnerability to excitotoxic death increased with developmental age of ES neurons in vitro. Specific receptors for Neurotrophin and GDNF family ligands were present in ES neurons. GDNF and NT-3 could modulate the survival and excitotoxic vulnerability of ES neurons. The vulnerability and resistance to toxic insults, which are essential requirements of model culture systems for neuropathological studies, make ES neurons to a useful model culture system. Especially ES cell are highly amenable to genetic modification unlikely to primary neuronal cells, which will give us a chance to answer more complicated neurophysiological questions. Recently there was an outstanding attempt to explore the cellular toxicity using human ES cells (Schrattenholz & Klemm, 2007) and it suggested that ES cells could be a new model system for neurophysiological studies soon and go further a large-scale screening system for pharmacological compounds in the future.

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Structural Analysis of Class I Integron Gene Cassette and Assessment of Genetic Relationships by PFGE of Salmonella enterica Serovar Typhimurium Isolated in Gyeongbuk Area (경북지역에서 분리된 Salmonella enterica Serovar Typhimurium의 Class I Integron Gene Cassette 특성과 PEGE 유형분석)

  • Sohn, Chang-Kyu;Lee, Jung-A;Lee, Do-Young;Hun, Wan;Jung, Jung-Kyo
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.12-18
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    • 2006
  • Thirty five Salmonella enterica serovar Typhimurium strains were isolated from diarrheic patients and pigs in Gyeongbuk area from 2003 to 2004. All 35 strains (17 strains from diarrheic patients and 18 from pigs) were resistant to more than one drug and most of strains isolated from pigs were resistant to ampicillin, ohloram-phenicol, streptomycin, sulfamethoxazole-trimethoprime, tetracyclin and nalidixic acid. Each isolate was also screened or the presence of class I, II and III integron gene cassettes. Among 35 strains,3 out of 17 strains isolated from diarrheic patients, carried dhfrX-orfF-aadA2 integron gene cassette and among 18 strains isolated from diseased pigs, 11 strains carried dhfrX-orfF-aadA2 integron gene cassette and 1 strain carried aadA2 integron only. But any class II and class II integron gene cassette were not detected in 35 strains. Thirty five strains were divided by five pulsotypes. Thirty one strains out of thirty five were pulsotype A. Among the remaining 4 strains, one each strain belonged to pulsotype B, C, D and pulsotype E. This data of pulsotypes showed that the widespread of pulsotype A, Salmonella enterica serovar Typhimurium in human and pigs in Gyeongbuk area may have been caused by the dissemination of a few epidemic strains in this area. Thirteen strains contain dhfrX-orfF-aadA2 integron gene cassette showed pulsotype A and one strain contains dhfrX-orfF-aadA2 integron gene cassette showed pulsotype B. One strain contains aadA2 integron showed pulsotype E. But fifteen strains do not contain any integron showed pulsotype A.

Construction of a Transgenic Tobacco Expressing a Polydnaviral Cystatin (폴리드나바이러스 유래 시스타틴 유전자 발현 형질전환 담배 제작)

  • Kim, Yeongtae;Kim, Eunsung;Park, Youngjin;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.54 no.1
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    • pp.7-15
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    • 2015
  • CpBV (Cotesia plutellae bracovirus) is a polydnavirus and encodes a cystatin (CpBV-CST1) gene. Its overexpression suppresses insect immunity and alters insect developmental processes. This study aimed to construct a genetically modified (GM) tobacco to further explore the physiological function of the viral cystatin and to apply to control insect pests. To this end, the transgenic tobacco lines were screened in expression of the target gene and assessed in insecticidal activity. A recombinant vector (pBI121-CST) was prepared and used to transform a bacterium, Agrobacterium tumefasciens. The transformed bacteria were used to generate transgenic tobacco lines, which were induced to grow callus and resulted in about 92% of shoot regeneration. The regenerated plants were screened by PCR analysis to confirm the insertion of the target gene in the plant genome. In addition, the expression of the target gene was assessed in the regenerated plants by quantitative real-time PCR (qRT-PCR). The qRT-PCR analysis showed that the transgenic line plant expressed the target gene about 17 times more than the control tobacco, indicating a stable insertion and expression of the target gene in the transgenic tobacco line. The insecticidal activity was then analyzed using the screened transgenic tobacco lines against the teneral 1st instar larvae of the oriental tobacco budworm, Helicoverpa assulta. Though there was a variation in the insecticidal efficacy among transgenic lines, T9 and T12 lines exhibited more than 95% mortality at 7 days after feeding treatment. These results suggest that CpBV-CST1 is a useful genetic resource to be used to generate GM crop against insect pests.