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Genomic Analysis of Satellite RNA of Cucumber mosaic virus-Paf Related with Mild Symptoms (Cucumber mosaic virus Paf 계통의 약독 병징과 관련된 satellite RNA의 유전자 해석)

  • Sung, Mi-Young;Jung, Min-Young;Lee, Sang-Yong;Ryu, Ki-Hyun;Choi, Jang-Kyung
    • Research in Plant Disease
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    • v.10 no.4
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    • pp.241-247
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    • 2004
  • Recently, we reported a satellite RNA (Paf-satRNA) which is encapsidated in a pepper isolate of Cucumber mosaic virus (CMV-Paf) regulated symptom attenuation of the helper virus. To characterize mild symptom domain of Paf-satRNA, a series of chimeric cDNAs of satRNAs were created by using full-length cDNA clones of Paf-satRNA and a Pep-satRNA, chlorosis-inducing satRNA in pepper plants, and analyzed for determinants of symptom attenuation. When compared the nucleotide sequences, the 3' and 5' terminal sequences of the two wild-type (wt) satRNAs contained relatively conserved sequences which are the typical to CMV satRNA. Ten bases insertions were found in PepY-satRNA, and two variable regions, 81st to 113th and 183rd to 265th from the 5'-end, were located in the middle parts of the satRNAs. To delineate the attenuated symptom-related domain for the Paf-satRNA, in vitro transcripts RNAs transcribed from the wt cDNAs and constructed chimeric cDNAs were combined with genomic RNAs, RNA1, RNA2 and RNA3, of CMV-Fny and inoculated onto Nicotiana benthamiana plants. These transcripts were fully infectious onto the N. benthamiana and infectivity was confirmed by the RT-PCR. Chimeric Paf(H/N)-satRNA and PepY(N/A)-satRNA as well as Paf-satRNA induced very mild mosaic or symptomless infection on N. benthamiana. By contrast, typical mosaic symptom and stunting of infected plants were induced when PepY-satRNA, PepY(H/N)-satRNA and Paf(N/A)-satRNA were infected to N. benthamiana. Paf-satRNA coinfected with CMV-Fny RNAs induced very mild to sympomless on pepper plants whereas PepY-satRNA-infected pepper expressed typical chlorosis mosaic symptom. Two kinds of chimeric mutants, Paf(H/N)-satRNA and PepY(N/A)-satRNA, induced mild mosaic or symptomless infection onto pepper plants, while PepY(H/N)-satRNA and Paf(N/A)-satRNA showed typical chlorosis and mosaic symptom with stunting. This results suggest that mild symptom-related domain for the Paf-satRNA was located on HpaI-NarI region.

Genetic Variations of Chicken TYR Gene and Associations with Feather Color of Korean Native Chicken (KNC) (한국 토종닭 모색 변이와 TYR 유전자형 간의 상관관계 분석)

  • Choi, Jin Ae;Lee, Jun-Heon;Jang, Hyun-Jun;Lee, Kyung-Tai;Kim, Tae-Hun;Lee, Hyun-Jeong;Heo, Kang-Nyeong;Kim, Chong-Dae;Han, Jae-Yong;Park, Mi Na
    • Korean Journal of Poultry Science
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    • v.41 no.1
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    • pp.7-14
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    • 2014
  • Tyrosinase (TYR) gene is located on chromosome 1 in chicken and it is composed of five exons and four introns. TYR gene is described as a key enzyme in melanin biosynthesis. Most examples of complete albinism in chicken have been due to defects in the tyrosinase gene. The association of feather color and sequence polymorphism in the Tyrosinase (TYR) gene was investigated using Korean Native chicken H breed (H_PL), Korean Native chicken L/W breed(L/W_PL) and 'Woorimatdag' commercial chickens (Woorimatdag_CC). From L_PL and W_PL breed analyses, 4 synonymous SNPs (locus G33A, G116A, C217T and C247T) and 2 SNPs (G838A and G958A) were detected in 4th exon and 4th intron of TYR gene respectively. The genotype frequencies for 6 SNPs were compared between L_PL and W_PL and W_PL represented homozygous SNP types in all the analyzed SNP positions while L_PL displayed various SNP types.

Genetic Diversity and Relationship by SSR Markers of Korean Soybean Cultivars (한국 콩 육성품종의 SSR마커에 의한 유전적 다양성과 유연관계)

  • Kim Seong-Hun;Jung Jong-Wook;Moon Jung-Kyung;Woo Sun-Hee;Cho Yong-Gu;Jong Seung-Keun;Kim Hong-Sig
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.3
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    • pp.248-258
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    • 2006
  • Genetic diversity of 91 Korean soybean cultivars was assessed with 20 simple sequence repeat (SSR). Twenty SSR loci generated a total of 149 alleles. The number of alleles for each SSR locus ranged from 3 to 15 with a mean of 7.5 alleles. Genetic diversity estimated by PIC value of 91 cultivars was ranged from 0.424 to 0.905 with an average of 0.711. Cluster analysis based on Nei's genetic distances classified 91 soybean cultivars except Geomjeongkong 4 into 7 groups. The majority groups were I, IV, and VI which included 26, 24, and 18 cultivars, respectively. Obvious differences in genetic diversity appeared to be related with the released periods of cultivars and utilization type of cultivars, but not with breeding sites. Cultivars released in 1970's and in 1990's showed the lowest and the highest genetic diversities with 0.576 and 0.706, respectively. Soybean cultivars for vegetable and early maturity showed the lowest genetic diversity with 0.514, while those for soy sauce and tofu showed the highest genetic diversity with 0.691. Genetic distance between soybean cultivar groups developed before 1969 and during 1970's was the nearest, while genetic distance between those developed in 1970's and 1990's was the furthest. Cultivar group for vegetable and early maturity showed the furthest genetic distance with cultivar group for soy sauce and tofu, while it showed the nearest genetic distance with cultivar group for cooking with rice. Genetic distance was greater between soybean cultivar groups developed in Suwon and Iksan than between those developed in Milyang and Iksan.

Current status, mechanism and control of herbicide resistant weeds in rice fields of Korea (한국 논에서 제초제 저항성잡초의 발생 현황, 메카니즘 및 방제)

  • Park, Tae Seon;Seong, Ki Yeong;Cho, Hyun Suk;Seo, Myung Chul;Kang, Hang Won;Park, Kee Woong
    • Korean Journal of Agricultural Science
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    • v.41 no.2
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    • pp.85-99
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    • 2014
  • Sulfonylurea (SU)-resistant weeds of eight annual weeds, Monochoria vaginalis, Scirpus juncoides and Cyperus difformis, etc., and four perennial weeds, Scirpus planiculmis, Sagittaria pigmaea, Eleocharis acicularis and Sagittaria trifolia as of 2013 since identification Monochoria korsakowii in the reclaimed rice field in 1998. And the resistant Echinochloa oryzoides to ACCase and ALS inhibitors has been confirmed in rice fields of the southern province, Korea in 2009. In the beginning, the M. vaginalis, S. juncoides and C. difformis of these SU-resistant weeds were rapidly and individually spreaded in different fields, however, these resistant weeds have been occurring simultaneously in the same filed recently. The resistant biotype by weed species demonstrated about 10-to 1,000-fold resistance, based on $GR_{50}$ values of the SU herbicides tested. And the resistant biotype of E. oryzoides to cyhalofop-butyl, pyriminobac-methyl, and penoxsulam was about 14, 8, and 11 times more resistant than the susceptible biotype base on $GR_{50}$ values. The products and applied area of SU-included herbicides have been increased rapidly, and have accounted for about 69% and 96% in Korea, respectively. In Korea, the main cause of SU-resistant weed is extensive use of these herbicides. The top ten herbicides by applied area were composed of all SU-included herbicides by 2003. The concentrated and successive treatment of ACCase and ALS inhibitors for control of barnyardgrass in rice led up to the resistance of E. oryzoides. Also, SU-herbicides like pyrazosulfuron-ethyl and imazosulfuron which effective to barnyardgrass can be bound up with the resistance of E. oryzoides. The ALS activity isolated from the resistant biotype of M. korsakowii to SU-herbicides tested was less sensitive than that of susceptible biotype. The concentration of herbicide required for 50% inhibition of ALS activity ($I_{50}$) of the SU-resistant M. korsakowii was 14-to 76-fold higher as compared to the susceptible biotype. No differences were observed in the rates of [$^{14}C$]bensulfuron uptake and translocation. Acetolactate synthase (ALS) genes from M. vaginalis resistant and susceptible biotypes against SU-herbicides revealed a single amino acid substitution of proline (CCT), at 197th position based on the M. korsakowii ALS sequence numbering, to serin (TCT) in conserved domain A of the gene. Carfentrazone-ethyl and pyrazolate were used mainly to control SU-resistant M. vaginalis by 2006 in Korea. However, the alternative herbicides such as benzobicyclone, to be possible to control simultaneously the several resistant weeds, have been developing and using broadly, because the several resistant weeds have been occurring simultaneously in the same fieled. The top ten herbicides by applied area in Korea have been occupied by products of 3-way mixture type including herbicides with alternative mode of action for the herbicide resistant weeds. Mefenacet, fentrazamide and cafenstrole had excellent controlling effects on the ACCase and ALS inhibitors resistant when they were applied within 2nd leaf stage.

Preliminary Study on the MR Temperature Mapping using Center Array-Sequencing Phase Unwrapping Algorithm (Center Array-Sequencing 위상펼침 기법의 MR 온도영상 적용에 관한 기초연구)

  • Tan, Kee Chin;Kim, Tae-Hyung;Chun, Song-I;Han, Yong-Hee;Choi, Ki-Seung;Lee, Kwang-Sig;Jun, Jae-Ryang;Eun, Choong-Ki;Mun, Chi-Woong
    • Investigative Magnetic Resonance Imaging
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    • v.12 no.2
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    • pp.131-141
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    • 2008
  • Purpose : To investigate the feasibility and accuracy of Proton Resonance Frequency (PRF) shift based magnetic resonance (MR) temperature mapping utilizing the self-developed center array-sequencing phase unwrapping (PU) method for non-invasive temperature monitoring. Materials and Methods : The computer simulation was done on the PU algorithm for performance evaluation before further application to MR thermometry. The MR experiments were conducted in two approaches namely PU experiment, and temperature mapping experiment based on the PU technique with all the image postprocessing implemented in MATLAB. A 1.5T MR scanner employing a knee coil with $T2^*$ GRE (Gradient Recalled Echo) pulse sequence were used throughout the experiments. Various subjects such as water phantom, orange, and agarose gel phantom were used for the assessment of the self-developed PU algorithm. The MR temperature mapping experiment was initially attempted on the agarose gel phantom only with the application of a custom-made thermoregulating water pump as the heating source. Heat was generated to the phantom via hot water circulation whilst temperature variation was observed with T-type thermocouple. The PU program was implemented on the reconstructed wrapped phase images prior to map the temperature distribution of subjects. As the temperature change is directly proportional to the phase difference map, the absolute temperature could be estimated from the summation of the computed temperature difference with the measured ambient temperature of subjects. Results : The PU technique successfully recovered and removed the phase wrapping artifacts on MR phase images with various subjects by producing a smooth and continuous phase map thus producing a more reliable temperature map. Conclusion : This work presented a rapid, and robust self-developed center array-sequencing PU algorithm feasible for the application of MR temperature mapping according to the PRF phase shift property.

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Cloning of the β-Lactamase Gene from Bacillus sp. J105 and Analysis of Its Expression in E. colis Cells (Bacillus sp. J105 유래 β-lactamase 유전자의 cloning 및 E. coli 내에서의 발현 분석)

  • Kang, Won-Dae;Lim, Hak-Seo;Seo, Min-Jeong;Kim, Min-Jeong;Lee, Hye-Hyeon;Cho, Kyeong-Soon;Kang, Byoung-Won;Seo, Kwon-Il;Choi, Yung-Hyun;Jeong, Yong-Kee
    • Journal of Life Science
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    • v.18 no.11
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    • pp.1592-1599
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    • 2008
  • The $\beta$-lactamase gene was cloned into E. coli DH5$\alpha$ from Bacillus sp. J105 with strong resistance against $\beta$-lactam antibiotics. The chromosomal DNA was partially digested with Sau3AI and ligated to BamHI digested pLAFR3. $\beta$-Lactamase positive clones were obtained by using in vitro packaging kit. The pKL11-${\Delta}4.6$ with $\beta$-lactamase activity was obtained by subcloning of the recombinant plasmid ($\beta$-lac +). The 6.5 kb fragment in the subcloned plasmid was sequenced. The DNA fragment that contains the $\beta$-lactamase gene encodes 309 amino acids. The 0.17 kb upstream region was similar to those of B. thuringinesis and B. cereus with 97% identity. The deduced amino acids sequence was also similar to those of $\beta$-lactamase from B. thuringinesis and B. cereus with 97% and 94% identity, respectively. The phylogenetic tree also showed the relationships of the $\beta$-lactamase gene of Bacillus sp. J105 to genetically related that of other Bacillus strains. Analysis of expression pattern of the pKL11-${\Delta}4.6$ in E. coli, revealed that the secretion efficiency of $\beta$-lactamase was $4{\sim}5%$ and the molecular weight was as same as that of original $\beta$-lactamase (31 kDa) from Bacillus sp. J105.

Susceptibility-Weighted MR Imaging for the Detection of Developmental Venous Anomaly: Comparison with T2 and FLAIR Imaging (자화율강조 MR영상을 이용한 뇌정맥 기형의 진단: T2강조영상과 FLAIR영상과의 비교)

  • Cho, Soo Bueum;Choi, Dae Seob;Ryu, Hyeon Gyu;Shin, Hwa Seon;Kim, Ji-Eun;Choi, Hye Young;Park, Mi Jung;Choi, Ho Cheol;Son, Seungnam
    • Investigative Magnetic Resonance Imaging
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    • v.18 no.3
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    • pp.200-207
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    • 2014
  • Purpose : We evaluated the diagnostic value of susceptibility-weighted imaging (SWI) for the detection of developmental venous anomaly (DVA). Materials and Methods: Retrospective review of 1068 brain MR examinations found 28 DVAs in 28 patients (2.6%) on contrast-enhanced T1-weighted images. SWI, T2, and FLAIR images of 28 patients with DVA and 28 sex- and age-matched control patients without DVA were analyzed by blinded readers on each type of sequences. All images were independently reviewed by two radiologists who were blinded to other MR imaging finding. In cases of discrepancy, two reviewers reached a consensus later. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of each MR sequence for the detection of DVA were determined. Statistical analysis was performed by using the Mcnemar test. The significance level was p < 0.05. Results: The sensitivity, specificity, PPV, and NPV of SWI for the detection of DVA were 85.7%, 92.9%, 92.3%, and 86.7%, respectively. T2 and FLAIR images showed sensitivity of 35.7% and 35.7%, specificity of 92.9% and 96.4%, PPV of 83.3% and 90.9%, and NPV of 59.1% and 60.0%, respectively. On SWI, the sensitivity and NPV for the detection of DVAs were significantly higher than those of T2 and FLAIR images (p < 0.05). Conclusion: SWI was sensitive and specific for the detection of DVA.

Improvement of Selection Efficiency for Bacterial Blight Resistance Using SNP Marker in Rice (SNP 마커를 이용한 벼 흰잎마름병 저항성 선발 효율 증진)

  • Shin, Woon-Chul;Baek, So-Hyeon;Seo, Chun-Sun;Kang, Hyeon-Jung;Kim, Chung-Kon;Shin, Mun-Sik;Lee, Gang-Seob;Hahn, Jang-Ho;Kim, Hyun-Soon
    • Journal of Plant Biotechnology
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    • v.33 no.4
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    • pp.309-313
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    • 2006
  • Discovery of single nucleotide polymorphisms (SNPs), including small insertions and deletions, is one of the hot topics in genetic research. The most common type of sequence variant consists of single base differences or small insertions and deletions at specific nucleotide positions. Significance of SNPs in rice is increasing for genetic research, positional cloning and molecular breeding. $F_2$ 170 lines and $F_3$ 194 lines derived from Sangjuchalbyeo/HR13721-53-3-1-3-3-2-2 Were used for Searching SNP markers related to bacterial blight resistance. Sangjuchalbyeo is susceptible to bacterial blight, but HR13721-53-3-1-3-3-2-2 has Xa1 gene resistant to bacterial blight. Individual lines were inoculated with $K_1$ race of bacterial blight and resistant or susceptible was evaluated after 3 weeks from inoculation. The genotypes of population were analysed by PCR-RFLP for SNP marker developing. The segregation of $F_2\;and\;F_3$ population showed almost 3:1, 1:1 ratio, respectively. Analysis of genotype using SNP marker is capable of confirming resistance for $K_1$ race and genotype through amplifying the gene using 16PFXal primer and digested the PCR product with Eco RV. There were close relation between resistance test for $K_1$ race and SNP marker genotype. Especially, DNA analysis using SNP marker is capable of judging homozygote/heterozygote in $F_2$ population compared with resistant test for Kl race. So, it seems to improve the selection efficiency in disease resistant breeding.

U-Pb(SHRIMP) and K-Ar Age Dating of Intrusive Rocks and Skarn Minerals at the W-Skarn in Weondong Deposit (원동 중석 스카른대에서의 관입암류와 스카른광물에 대한 U-Pb(SHRIMP) 및 K-Ar 연대)

  • Park, Changyun;Song, Yungoo;Chi, Se Jung;Kang, Il-Mo;Yi, Keewook;Chung, Donghoon
    • Journal of the Mineralogical Society of Korea
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    • v.26 no.3
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    • pp.161-174
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    • 2013
  • The geology of the weondong deposit area consists mainly of Cambro-Ordovician and Carboniferous-Triassic formations, and intruded quartz porphyry and dyke. The skarn mineralized zone in the weondong deposit is the most prospective region for the useful W-mineral deposits. To determine the skarn-mineralization age, U-Pb SHRIMP and K-Ar age dating methods were employed. The U-Pb zircon ages of quartz porphyry intrusion (WD-A) and feldspar porphyry dyke (WD-B) are 79.37 Ma and 50.64 Ma. The K-Ar ages of coarse-grained crystalline phlogopite (WD-1), massive phlogopite (WDR-1), phlogopite coexisted with skarn minerals (WD-M), and vein type illite (WD-2) were determined as $49.1{\pm}1.1$ Ma, $49.2{\pm}1.2$ Ma, $49.9{\pm}3.6$ Ma, and $48.3{\pm}1.1$ Ma, respectively. And the ages of the high uranium zircon of hydrothermally altered quartz porphyry (WD-C) range from 59.7 to 38.7 Ma, which dependson zircon's textures affected by hydrothermal fluids. It is regarded as the effect of some hydrothermal events, which may precipitate and overgrow the high-U zircons, and happen the zircon's metamictization and dissolution-reprecipitation reactions. Based on the K-Ar age datings for the skarn minerals and field evidences, we suggest that the timing of W-skarn mineralization in weondong deposit may be about 50 Ma. However, for the accurate timing of skarn mineralization in this area, the additional researches about the sequence of superposition at the skarn minerals and geological relationship between skarn deposits and dyke should be needed in the future.

Paenibacillus kimchicus sp. nov., an antimicrobial bacterium isolated from Kimchi (김치로부터 분리된 항균 활성 세균 Paenibacillus kimchicus sp. nov.)

  • Park, A-rum;Oh, Ji-Sung;Roh, Dong-Hyun
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.319-326
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    • 2016
  • An antimicrobial bacterium to pathogenic microorganisms, strain $W5-1^T$ was isolated from Korean fermented-food Kimchi. The isolate was Gram-staining-variable, strictly aerobic, rod-shaped, endospore-forming, and motile with peritrichous flagella. It grew at $15-40^{\circ}C$, at pH 6.0-10.0, and in the presence of 0-4% NaCl. Strain $W5-1^T$ could hydrolyze esculin and xylan, and assimilate $\small{D}$-mannose, but not $\small{D}$-mannitol. Strain $W5-1^T$ showed antimicrobial activity against Listeria monocytogens, Pseudomonas aeruginosa, Staphylococcus aureus, and Salmonella typhi. The G+C content of the DNA of strains $W5-1^T$ was 52.6 mol%. The predominant respiratory quinone was menaquinone-7 (MK-7) and the major cellular fatty acids were $C_{16:0}$, antieiso-$C_{15:0}$, $C_{18:0}$, and $C_{12:0}$. The strain contained meso-diaminopimelic acid in cell-wall peptidoglycan. On the basis of 16S rRNA gene sequence and phylogenetic analysis, the strain W5-1 was shown to belong to the family Paenibacillaceae and was most closely related to Paenibacillus pinihumi $S23^T$ (98.4% similarity) and Paenibacillus tarimensis $SA-7-6^T$ (96.4%). The DNA-DNA relatedness between the isolate and Paenibacillus pinihumi $S23^T$ was 8.5%, indicating that strain $W5-1^T$ represented a species in the genus Paenibacillus. On the basis of the evidence from this polyphasic study, it is proposed that strain $W5-1^T$ is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus kimchicus sp. nov. is proposed. The type strain is $W5-1^T$ (=KACC $15046^T$ = $LMG 25970^T$).