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Stability of pUC-Derived Plasmids with a Fluorescence Marker in Pectobacterium carotovorum subsp. carotovorum and subsp. betavasculorum

  • Hur, Woon-Yung;Roh, Eun-Jung;Oh, Chang-Sik;Han, Man-Wi;Lee, Seung-Don;Kim, Doo-Ho;Heu, Sung-Gi
    • The Plant Pathology Journal
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    • v.25 no.3
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    • pp.286-290
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    • 2009
  • The stability of three different kinds of pUC-derived plasmids, pDsRed, pZsYellow, and pGFPuv, was investigated in Pectobacterium strains to utilize those plasmids as tracers. All three plasmids pDsRed, pZsYellow and pGFPuv showed their specific colors in Pectobacterium strains. Especially, the plasmid pDsRed conferred bright pink colonies on the Pectobacterium strains. When the bacteria lost the plasmid pDsRed, the colonies turned white, suggesting that the plasmid could be a good marker system for Pectobacterium strains on different environmental conditions. The effect of the antibiotic pressure on the stability of the plasmid was different depending on the host bacteria. P. carotovorum subsp. betavasculorum was more sensitive to the antibiotic pressure than P. carotovorum subsp. carotovorum Pcc21. However, temperature change significantly affected plasmid stability on both Pectobacterium strains. Almost all strains lost the plasmids with the shift in temperature from $28^{\circ}C$ to $37^{\circ}C$. Presence of the plasmids did not affect bacterial pathogenicity on their own host plants. Among three plasmids, pZsYellow was not useful as a marker because the yellow fluorescent proteins from pZs Yellow were interfered with the yellow natural fluorescence of the plant tissues induced by the defense system. Since the red color of DsRed can be seen with naked eyes, plasmid pDsRed was applicable as a marker. However, the color change was slow so that additional manipulation to increase the expression speed was necessary. Plasmid pGFPuv could serve as a perfect marker without any problem, tracing the reproduction and spread of the plant pathogens perfectly.

Diversity of VC and incidence of hypovirulence-associated ds-RNAs in the chestnut blight fungus Cryphonectria parasitica in Korea

  • Byeongjin Cha;Jinyoung Lim;Ju, Young-Jik;Kim, Dae-Hyuk
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.23-23
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    • 2003
  • Chestnut blight, caused by Cryphonectria parasitica, is the most destructive disease of American and European chestnut trees. A total of 672 C prasitica was isolated from blight lesion on chestnut twigs, which were collected from major chestnut plantations all over Korea in 1999. Isolation rates were over 30% in Kyunggj-, Kyongnam-, and Chonnam-do. The highest isolation rate was 37.4% and recorded in Kyongnam-do. On the other hand, Chonbuk-do had the lowest isolation rate as 13.5%. In grouping of C parasitica by colony shape and color, yellow colony with irregular margin were the most dominant colony type with a frequency of 65.2%. When the 672 isolates were inoculated on the chestnut twigs, 380 isolates (56.5%) caused lesions larger than the standard virulent isolate EP155-2, while 158 isolates (23.4%) caused smaller lesions than the standard hypovirulent isolate UEP-1. In Bavendamm test that determines phenol oxidase activity, 97.1% of all the isolates resulted the same or darker discoloration than EP155-2, and only 12.2% resulted the same or lighter discoloration than UEP-1. In the vegetative compatibility (VC) tests, total 670 isolates were divided into 121 VC groups (VCGs). Kyongnam-, Chonnam-, and Chungnam-do, the three principal chestnut plantation area, had 49, 33, and 27 VCGs, respectively. Among the VCGs, the biggest VCG, KR-VC104, was composed of 164 isolates and the second biggest VCG had 62 isolates. But, 64 of 121 VCGs consisted of sole member. More than 65.8% of KR-VC104, was isolated from the three provinces, Kyongnam-, Kangwon-, and Chungbuk-do. In KR-VC104, 62.8%, 59.1%, and 85.9% of the isolates looked like virulent in colony type, pathogenicity test, and Bavendamm test. In ds-RNA detection tests using cellulose chromatography, 77 of total 650 isolates were ds-RNA positive and detected ds-RNA segments were approximately 12kb, 3kb, 2.7kb, 2kb, and 1.8kb in size. Among the 77 isolates, 46 isolates had 12kb and 25 isolates had 12kb and 2.7kb. Other 6 Isolates had small ds-RNA segments. Kyongnam-, Chonnam-, and Chungnam-do had 43, 16, and 5 ds-RNA positive isolates, respectively. Among the 121 VCGs, only 29 VCGs had ds-RNA positive isolates.(중략)

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Correlation Analysis of Binder Stiffness with Deformation Strength and Dynamic Creep Property of Asphalt Mixtures for Evaluation of Kim Test Applicability (바인더 Stiffness와 혼합물의 변형강도 및 동적크리프 특성과의 상관성 분석을 통한 김테스트 적용성 고찰)

  • Kim, Kwang-Woo;Doh, Young-Soo;Cho, Mun-Jin
    • International Journal of Highway Engineering
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    • v.7 no.4 s.26
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    • pp.91-102
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    • 2005
  • This study dealt with correlation analysis of binder stiffness with deformation strength and dynamic creep property of asphalt mixtures for evaluation of applicability of deformation strength $(S_D)$. Two aggregates, with maximum size of 13mm and eight binders were used to produce 16 different mixtures. The stiffness of binder $(G^*/sin\delta)$ was measured using DSR at $64^{\circ}C$. Final deformation(FD) and dynamic stability(DS) were measured by dynamic creep (DC) test, and SD was measured by Kim test for each mixture. Results of correlation analysis between $G^*/sin\delta$, and $S_D$, m and DS showed that correlation with binder stiffness and deformation strength was the highest $(R^2>0.88)$. There was good correlation between DS, FD with $S_D$. The results indicated that rut-resistance property of mixture is better reflected in $S_D$ test than FD or DS of dynamic creep test. Therefore, it is concluded that $S_D$ can be possibly used for evaluation of rut resistance of asphalt concretes with a good reliability if the procedure is standardized.

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Performance Analysis or MC / DS CDMA System with Hybrid SC / MRC Diversity over Multi-path Fading Channels (다중경로 페이딩 채널에서 Hybrid SC/MRC 기법이 적용된 MC / DS CDMA 시스템의 성능분석)

  • 김원섭;박진수
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.29 no.7A
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    • pp.748-755
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    • 2004
  • In this paper, the hybrid SC/MRC technique was applied to the MC/DS-CDMA system in the Nakagami-m multi-path fading environment. The applied scheme was compared with other cases when the MRC technique was applied and then performances were analyzed. The result of analysis showed that the performance of the applied system, whose number of branches (L) for the input diversity of each carrier wave was 3 and かme selected number of branches ( Lc) was 2, was better than that of the MRC technique when the Lc of the MRC technique was 2. The performance of the applied system was lower than that of the MRC technique, then the Lc of the MRC technique was 3. However, as the fading index was lowered, which means the telecommunication environment deteriorated, the performance of the hybrid SC/MRC-(2/3) diversity technique when Lc of MRC technique was 3, was almost the same as that of the MRC technique. The hybrid SC/MRC-(2/3) diversity technique shows a similar performance to that of the MRC technique, when the Lc of the MRC technique was 4. Therefore, it was confirmed that the MC/DS-CDMA system could reduce complexity and achieve similar performance to that of the MRC by applying the hybrid SC/MRC technique, which utilized the advantages of the SC technique and high performance of the MRC scheme, rather than the MRC diversity technique that requires bit synchronization, frequency of fading, and phase synchronization for every diversity branch.

Degradative Solidification/Stabilization of Liquid Waste Containing Chloroform and Methylene Chloride by Cement/Slag/Fe(II) Systems (Cement/Slag/Fe(II) 시스템에 의한 클로로포름과 메틸렌클로라이드 함유 액상폐기물의 분해성 고형화/안정화)

  • Seong, Joseph;An, Cheol-Hong;Choi, Won-Ho;Park, Joo-Yang
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.10
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    • pp.1034-1038
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    • 2008
  • Degradative Solidification/Stabilization(DS/S) is a modification of conventional Solidification/Stabilization(S/S) that incorporates degradative processes for organic contaminant destruction with the low cost of conventional S/S. Inorganic contaminants are immobilized and chlorinated organic contaminants are destroyed by DS/S treatment. In this study, a DS/S using cement/slag/Fe(II) systems as binder was investigated to assess its effectiveness in degrading chloroform(CF) and methylene chloride(MC) contained in hazardous liquid wastes. The initial concentration of CF was 0.26 mM, 1.0 mM, 8.4 mM, 25 mM and 42 mM and Fe(II) was 200 mM. The result showed that degradation of CF in various concentration was in one kind reaction as pseudo-first-order and 95% of 0.26 mM initial concentration of CF was removed in five days. 50 mg/L of heavy metal was added in order to accelerate the rate of degradation of MC and initial concentration of MC was 3.50 mM however, degradation did not occur in system. Thus additional studies needed for degradation of MC and more studies on other reaction pathways products will help elucidate reaction mechanisms and pathways for chlorinated methanes in cement/slag/Fe(II) systems.

Improvement of DS Simple Reception System for Advance of Duty Rate on Meteor Burst Transmission Path (유성 버스트 전송로의 도통을 향상을 위한 DS 간이 수신시스템의 개선)

  • Kwon Hyeog-Soong
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.30 no.11A
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    • pp.1073-1080
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    • 2005
  • In spite of many advantages of Meteor Burst Communications(MBC) on its transmission channel, the fact that its duty rate is less than 10 percents is a considerable deficiency of MBC. To overcome with this deficiency without paying large cost, we use a direct sequence(DS) simple reception system. This method doesn't need to add the components for acquisition or tracking the signal so that it keeps the cost efficiency of the MBC. But it has a disadvantage that its duty rate shows a large decline by noise. For this problem, we adopt the proposed system applying a positive feedback circuit. The improvement by this method is more significant when the received signal is weak. Simulation results show as tl is l00ms, then the burst length would be increased by $35\%$ when the initial value of the received signal power increases by 3 dB, t2 is 135ms. If the power increases by 6dB the burst length would be 170ms thus increased by $70\%$, which shows a great enhancement. Suppose now that tl is 50ms and the power is increased by 3dB, then t2 would be increased by $70\%$ to 85ms, and increasing the power by 6dB it would be increased by $130\%$ to 115ms.

Study of Viral Effects of the Mycovirus (LeV) and Virus-Free Commercial Line in the Edible Mushroom Lentinula edodes

  • Kim, Jung-Mi;Song, Ha-Yeon;Yun, Suk-Hyun;Lee, Hyun-Suk;Ko, Han-Kyu;Kim, Dae-Hyuk
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.37-37
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    • 2015
  • dsRNA was found in malformed cultures of Lentinula edodes strain FMRI0339, one of the three most popular sawdust cultivated commercial strains of shiitake, and was also found in healthy-looking fruiting bodies and actively growing mycelia. Cloning of the partial genome of the dsRNA revealed the presence of the RdRp sequence of a novel L. edodes mycovirus (LeV), and sequence comparison of the cloned amplicon showed an identical sequence to known RdRp genes of LeV found in strain HKA. The meiotic stability of dsRNA was examined by measuring the ratio of the presence of dsRNA among sexual monokaryotic progeny. More than 40% of the monokaryotic progeny still contained the dsRNA, indicating the persistence of dsRNA during sexual reproduction. Comparing the mycelia growth of monokaryotic progeny suggested that, although variations in the growth rate existed among progeny and virus infection was observed in highly actively growing progeny, there appeared to be a tendency toward a lower frequency of virus incidence in actively growing progeny. This study attempted to cure the edible mushroom L. edodes strain FMRI0339 of the L. edodes mycovirus (LeV) in order to obtain an isogenic virus-free fungal strain as well as a virus-infected strain for comparison. Mycelial fragmentation, followed by being spread on a plate with serial dilutions resulted in a virus-free colony. Viral absence was confirmed with gel electrophoresis after dsRNA-specific virus purification, Northern blot analysis, and PCR using reverse transcriptase (RT-PCR). Once cured, all of fungal cultures remained virus-free over the next two years. Interestingly, the viral titer of LeV varied depending on the culture condition. The titer from the plate culture showed at least a 20-fold higher concentration than that grown in the liquid culture. However, the reduced virus titer in the liquid culture was recovered by transferring the mycelia to a plate containing the same medium. In addition, oxygen-depleted culture conditions resulted in a significant decrease of viral concentration, but not to the extent seen in the submerged liquid culture. Although no $discernable phenotypic changes in colony morphology were observed, virus-cured strains showed significantly higher growth rates and mycelial mass than virus-infected strains. We were also explored effects of LeV on fruiting body formation and mushroom yield. The fruiting body formation yield of virus-free L. edodes was larger than virus-infected L. edodes. These results indicate that LeV infection has a deleterious effect on mycelial growth and fruiting body formation. In addition, we have been investigated host-parasite interaction between L. edodes and its mycovirus interaction to study viral mechanism by establishment of proteomics.

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A Study on the Dyadic Sorting method for the Regularization in DT-MRI (Dyadic Sorting 방법을 이용한 DT-MRI Regularization에 관한 연구)

  • Kim, Tae-Hwan;Woo, Jong-Hyung;Lee, Hoon;Kim, Dong-Youn
    • Journal of the Institute of Electronics Engineers of Korea SC
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    • v.47 no.4
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    • pp.30-39
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    • 2010
  • Since Diffusion tensor from Diffusion Tensor Magnetic Resonance Imaging(DT-MRI) is so sensitive to noise, the principle eigenvector(PEV) calculated from Diffusion tensor could be erroneous. Tractography obtained from PEV could be deviated from the real fiber tract. Therefore regularization process is needed to eliminate noise. In this paper, to reduce noise in DT-MRI measurements, the Dyadic Sorting(DS) method as regularization of the eigenvalue and the eigenvector is applied in the tractography. To resort the eigenvalues and the eignevectors, the DS method uses the intervoxel overlap function which can measure the overlap between eigenvalue-eigenvector pairs in the $3\times3$ pixel. In this paper, we applied the DS method to the three-dimensional volume. We discuss the error analysis and numerical study to the synthetic and the experimental data. As a result, we have shown that the DS method is more efficient than the median filtering methods as much as 79.97%~83.64%, 85.62%~87.76% in AAE, AFA respectively for the corticospinal tract of the experimental data.