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BSA-Seq Technologies Identify a Major QTL for Clubroot Resistance in Chinese Cabbage (Brassica rapa ssp. pekinesis)

  • Yuan, Yu-Xiang;Wei, Xiao-Chun;Zhang, Qiang;Zhao, Yan-Yan;Jiang, Wu-Sheng;Yao, Qiu-Ju;Wang, Zhi-Yong;Zhang, Ying;Tan, Yafei;Li, Yang;Xu, Qian;Zhang, Xiao-Wei
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.41-41
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    • 2015
  • BSA-seq technologies, combined Bulked Segregant Analysis (BSA) and Next-Generation Sequencing (NGS), are making it faster and more efficient to establish the association of agronomic traits with molecular markers or candidate genes, which is the requirement for marker-assisted selection in molecular breeding. Clubroot disease, caused by Plasmodiophora brassicae, is a serious threat to Brassica crops. Even we have breed new clubroot resistant varieties of Chinese cabbage (B. rapa ssp. pekinesis), the underlying genetic mechanism is unclear. In this study, an $F_2$ population of 340 plants were inoculated with P. brassicae from Xinye (Pathotype 2 on the differentials of Williams). Resistance phenotype segregation ratio for the populations fit a 3:1 (R:S) segregation model, consistent with a single dominant gene model. Super-BSA, using re-sequencing the parents, extremely R and S DNA pools with each 50 plants, revealed 3 potential candidate regions on the chromosome A03, with the most significant region falling between 24.30 Mb and 24.75 Mb. A linkage map with 31 markers in this region was constructed with several closely linked markers identified. A Major QTL for clubroot resistance, CRq, which was identified with the peak LOD score at 169.3, explaining 89.9% of the phenotypic variation. And we developed a new co-segregated InDel marker BrQ-2. Joint BSA-seq and traditional QTL analysis delimited CRq to an 250 kb genomic region, where four TIR-NBS-LRR genes (Bra019409, Bra019410, Bra019412 and Bra019413) clustered. The CR gene CRq and closely linked markers will be highly useful for breeding new resistant Chinese cabbage cultivars.

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The Photoreactivity of 2-Halobenzyl Phenyl Ether (2-할로벤질 페닐 에델의 광반응성)

  • Park, Yong Tae;Kim, Young Hee;Shin, Hyun Il
    • Journal of the Korean Chemical Society
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    • v.42 no.2
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    • pp.203-208
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    • 1998
  • The photochemical reactivities of 2-halobenzyl phenyl ether, in which 2-halobenzyl moiety are tethered to phenyl moiety by the etheral alkyl linkage, has been studied. In the presence of nitrogen, the photochemical reaction of 2-chlorobenzyl phenyl ether (1) produces mainly phenol and photo-Fries type reaction products, while the corresponding bromo analog 2 produces photocyclization and photoreduced products, along with phenol and photo-Fries type products. The former result implies that since chlorine is bound to the benzyl ring firmly, the rather weaker $CH_{2}-O$ bond of 1 is cleavaged to produce the photo-Fries type product. The latter implies that the photoinduced fission of phenyl-bromine bond of 2 can compete with the fission of $CH_{2}-O$ bond, since the bond energy of phenyl-bromine is lower than that of phenyl-chlorine. Since by the presence of oxygen the formation of phenol is not affected much, the formation of photo-Fries type products is changed a little, and the formations of photocyclization and photoreduced products are affected effectively, a singlet state is involved in the formation of phenol, and both singlet and triplet state may be involved in the formation of photo-Fries type reaction, while a triplet state is involved in the formation of photocyclization and photoreduction products.

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Source Apportionment Study and Chemical Composition of PM10 and PM2.5 in the Industrial Complex of Busan City, Korea (SEM-EDX 분석법에 의한 부산 S공업단지의 PM10과 PM2.5의 화학적 조성 및 발생원 추정)

  • Kim, Yong-Seog;Choi, Kum-Chan;Suh, Jeong-Min
    • Journal of Environmental Science International
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    • v.26 no.11
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    • pp.1297-1306
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    • 2017
  • This study identified physical characteristics and aerosol particle sources of $PM_{10}$ and $PM_{2.5}$ in the industrial complex of Busan Metropolitan City, Korea. Samples of $PM_{10}$, $PM_{2.5}$ and also soil, were collected in several areas during the year of 2012 to investigate elemental composition. A URG cyclone sampler was used for collection. The samples were collected according to each experimental condition, and the analysis method of SEM-EDX was used to determine the concentration of each metallic element. The comparative analysis indicated that their mass concentration ranged from 1% to 3%. The elements in the industrial region that were above 10% were Si, Al, Fe, and Ca. Those below 5% were Na, Mg, and S. The remaining elements (1% of total mass) consisted of elements such as Ni, Co, Br and Pb. Finally, a statistical tool was applied to the elemental results to identify each source for the industrial region. From a principal components analysis (SPSS, Ver 20.0) performed to analyze the possible sources of $PM_{10}$ in the industrial region, five main factors were determined. Factor 1 (Si, Al), which accounted for 15.8% of the total variance, was mostly affected by soil and dust from manufacturing facilities nearby, Factors 2 (Cu, Ni), 3 (Zn, Pb), and 4 (Mn, Fe), which also accounted for some of variance, were mainly related to iron, non-ferrous metals, and other industrial manufacturing sources. Also, five factors determined to access possible sources of $PM_{2.5}$, Factor 1 (Na, S), accounted for 13.5% of the total variance and was affected by sea-salt particles and fuel incineration sources, and Factors 2 (Ti, Mn), 3 (Pb, Cl), 4 (K, Al) also explained significant proportions of the variance. Theses factors mean that the $PM_{2.5}$ emission sources may be considered as sources of incineration, and metals, and non-ferrous manufacturing industries.

Effects of physico-chemical treatment on 'Nunkeunhukchal' (black sticky rice with giant embryo) for the enhancement of GABA (γ- aminobutyric acid) contents ('눈큰흑찰'의 가바(GABA) 함량 증진을 위한 이화학적 처리 효과)

  • Han, Sang-Ik;Ra, Ji-Eun;Seo, Kyung-Hye;Park, Ji-Young;Seo, Woo Duck;Park, Dong-Soo;Cho, Jun-Hyun;Lee, Jong-Hee;Sim, Eun-Yeong;Nam, Min-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.59 no.4
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    • pp.398-405
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    • 2014
  • We assessed the GABA accumulation and other components after the 'Nunkeunhukchal (BGE)', 'Josanghukchal (BR)', and 'Ilmibyeo (IB)' grain was soaked in water for 24, 36, 48, 60, 72 and 96 hr. The results showed a continuous accumulation of GABA in soaking treated brown rice of BGE and IB. Among the treated hours, 72 hours of soaking had the maximal accumulation of GABA (51.4 mg/100 g), amino acid, polyphenol and other components. The activities of glutamate decarboxylase (GAD) in brown rice and rice-bran were the same in BGE rice. However, the formation of GABA treated with L-glutamate as substrate showed dramatic increase of 354.6 (fourteen times higher than normal extraction) and 726.4 mg/100 g in BGE rice and rice-bran, respectively. These results suggested that the soaking and extraction with L-glutamate buffer could be better methods for the harvest of increased GABA.

The Effect of Red Cabbage (Brassica oleracea L. var. capitata f. rubra) Extract on the Apoptosis in Human Breast Cancer MDA-MB-231 Cells (적양배추 추출물이 인체 유방암 세포 MDA-MB-231의 세포사멸에 미치는 영향)

  • Nam, Mi Kyung;Kang, Keum Jee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.1
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    • pp.8-16
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    • 2013
  • We investigated the effect of red cabbage extract (RCE) on cell death in MDA-MB-231 human breast cancer cells. Cells were cultured in the presence 1.0, 1.5, and 2.0 mg/mL concentrations of RCE for 24 hours. MTT assays demonstrated that mitochondrial dehydrogenase activities decreased in a dose-dependent manner in cells (p<0.05). In contrast, the proportion of dual staining with Hoechst 33342/ethidium bromide (EtBr) for cell death increased in a dose-dependent manner in cells (p<0.05). Flow cytometry assays revealed that cell death caused by an apoptotic program increased in a dose-dependent (p<0.05). Also, increased ROS accumulation in cells, as revealed by DCF-DA staining, was observed in a dose-dependent fashion (p<0.05). The apoptosis suppressor gene Bcl-2 decreased significantly at the mRNA level. Pro-apoptotic genes Bax and caspase-3, genes that are related to the last stage of apoptosis significantly increased. The Bcl-2/Bax ratio which is an important indicator of apoptosis, was found to have significantly decreased dose dependence. These results taken together indicate that the effect of red cabbage extract induces cell death in MDA-MB-231 human breast cancer cells.

Response Mechanism of 5, 10, 15, 20-tetraphenyl(porphyrinato) Manganase(III) chloride-Based Ion-Selective Membranes (망간포르피린을 함유한 고분자형 이온선택성 막전극의 감응 메카니즘)

  • Hong, Young Ki;Kang, You Ra;Shin, Dae Ho;Shin, Doo Soon;Cha, Geun Sig;Nam, Hakhyun
    • Analytical Science and Technology
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    • v.9 no.3
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    • pp.270-278
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    • 1996
  • Response mechanism for the chloride-selective membrane doped with 5, 10, 15, 20-tetraphenyl(porphyrinato)manganase(III) chloride(Mn(TPP)Cl) in PVC/DOS matrix is proposed by examining the visible spectra of the corresponding optode membrane. The visible spectra of Mn(TPP)Cl-doped membrane placed in aqueous solution show that the chloride ligand is easily replaced with water molecule. When other halogen ions, such as $F^-$, $Br^-$ and $I^-$, are added to the sample solution, they replace the water ligand, exhibiting distinctive change in the Soret band of Mn(TPP). On the other hand, bulky anions, such as SCN and salicylate, do not form a bond with the central metal. These results suggests that the potentiometric response of Mn(TPP)-based membrane results either from the ligand exchange (water with halides) at the central metal or from the counter ion exchange (chlorides with bulky lipophilic anions) around the positively charged porphyrin molecule in membrane phase. It was also noted that both hydration enthalpies of anions and their binding constants to Mn(TPP) play critical role in determining the potentiometric selectivity pattern of the membrane.

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Comparison of Meat Quality and Sensory Characteristics of Different Native Chickens in Korean Market (시중 유통 토종닭의 품종별 품질 및 관능 특성 비교)

  • Cha, Ju-Su;Kim, Sun Hyo;Jung, Samuel;Kang, Ho Jin;Jo, Cheorun;Nam, Ki-Chang
    • Korean Journal of Poultry Science
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    • v.41 no.1
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    • pp.53-59
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    • 2014
  • To evaluate the meat quality differences of Korean Native Chickens (KNC) available in Korean market, the physicochemical and sensory characteristics of a broiler and two KNC (HH and WD breed) were analyzed. The fat content of KNC WD breast meat was higher than that of KNC HH. The breast and thigh meats of KNC HH had higher $L^*$ and $a^*$ values than the broiler or the KNC WD. WD meats showed greater DPPH radical scavenging activity and higher pH values than the HH. For the fatty acid composition, there was no significant difference in total content of the saturated fatty acids among chicken breeds (p>0.05). KNC had greater content of polyunsaturated fatty acids, especially arachidonic acid (20:4) and DHA (22:6), compared with the broiler. KNC WD meats had higher composition of linolenic acid (18:3) than the KNC HH. For sensory evaluation, the WD breast meat showed higher taste and tenderness scores than the HH. On the other hand, the soup made by broiler had higher taste scores than the KNC. The breast and thigh meat from two KNC breeds showed different quality parameters that may come from the differences of genetic attributes, feed, and production strategy between two breeds.

Analysis of Aliphatic Carboxylic Acids Using Ion-Exchange Chromatography: Application to Groundwater Affected by Landfill Leachates (이온-교환 크로마토그래피를 활용한 유기산 분석: 매립지 침출수의 영향을 받은 지하수에 대한 적용)

  • Cheon, Su-Hyun;Koh, Dong-Chan;Ko, Kyung-Seok
    • Journal of Soil and Groundwater Environment
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    • v.12 no.2
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    • pp.55-64
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    • 2007
  • An analytical method using ion-exchange chromatography was developed for simultaneous quantification of low-molecularweight organic acids ($C_1-C_6$ aliphatic carboxylic acids) and inorganic anions, and then applied to the assessment of ground water contaminated by leachates from a municipal solid waste landfill. Peak interferences of halide ions to organic acids were removed by pretreatment of water samples with Ag-containing cartridges. This method allowed accurate detection of low-molecular weight organic acids (i.e., formate, acetate, propionate, pyruvate, succinate, and oxalateas) low as 0.5 mg/L with a linear dynamic range up to 20 mg/L within 11 min run time along with typical inorganic anions. High level of pyruvate and low level of formate and acetate were detected in groundwater and landfill leachates using the analytical method. Pyruvate concentration in groundwater showed a significant correlation with concentrations of $Cl^-$ and $HCO_3^-$, and pyruvate levels decreased along the downgradient from the landfill, indicating the sources of pyruvate are landfill leachate.

젖산균의 Plasmid DNA 분리방법 및 Electroporation에 의한 젖산균의 형질전환에 관한 연구

  • Kim, Seon-Gi
    • 한국유가공학회:학술대회논문집
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    • 1997.05a
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    • pp.41-61
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    • 1997
  • 젖산균의 유전자 연구를 촉진하기 위해 간단하고 신속한 plasmid DNA의 분리방법과 electro-poration을 이용하여 vector plasmid의 간단하고 신속한 전이방법을 얻기 위해 젖산균의 형질전환에 영향하는 요인에 대하여 연구하였으며 연구결과는 다음과 같다. 1. O'Sullivan과 Klaenhammer의 방법을 개선하여 젖산균 plasmid DNA의 분리에 좋은 결과를 얻을 수 있는 신속하고 쉬운 방법을 고안하였으며, genomic DNA 분리에 이용되는 guanidium thio-cyanate 처리방법을 plasmid의 분리에 적용할 수 있었다. 2. L. casei, L. acidophilus. L. delbruekii var. bulgaricus. L. brevis와 L. plantarum 균주에서 plasmid를 확인하였으며, 돼지 분에서 분리된 L. lactis ssp. lactis. L. fermentum과 L. plantarum에서도 plasmid를 분리 확인하였다. 3. Lactococci의 plasmid분리는 lactobacilli와는 달리 mutanolysin의 처리없이도 잘 되었으며, L. lactis ssp. lactis와 Ent. faecalis에서 plasmid를 확인하였다. 4. E. coli plasimd 분리에 이용되는 MPS membrane filter 방법으로 젖산균 plasmid pLZ12의 분리가 가능하였으나, 세포파편이 filter를 막아 사용에 어려움이 있는 것으로 확인되었다. 5. Plasmid 분리없이 electroporation을 이용한 세포 대 세포 전이법으로 간편하고 빠르게 E. coli DH5${\alpha}$에 E. coli Jm109의 plasmid pBX19, pBR322를 전이시켰다. 6. L. lactis ssp. lactis 균주에 lysozyme 처리시 30${\sim}$80%의 생존율을 보였으며, 대부분의 L. acidophilus 균주의 경우 약 70%의 생존율을 보였다. L. casei 102S의 경우는 45분간 처리 시에도 100%의 생존율을 보였다. 8. L. lactis ssp. lactis 균주에 pLZ12를 6.0kV에서 전이시킨 결과 12.5kV에서보다 형질전환 효율이 훨씬 높았으며 lysozyme 처리에 의해 형질전환 효율이 증가되었다. 9. L. acidophilus 균주에 pLZ12를 전이시 6.0kV에서는 전이가 모두 이루어졌으나, 12.5kV에서는 L. acidophilus WIESBY와 NCFM에서 전이가 이루어지지 않았으며, lysozyme 처리 후 pLZ12를 전이시켰을 때 12kV보다 6.0kV에서 형질전환 효율이 증가되었다. 10. Gene Pulser와 Progenitor II를 사용하여 pLZ12를 L. lactis ssp. lactis 균주에 전이하였을 때 Gene Pulser에 비해 Progenitor II의 형질전환 효율이 현저히 떨어졌다. L. acidophilus HY7008과 HY7001은 두 기기 모두 형질전환이 이루어졌으나, L. acidophilus WEISBY와 NCFM은 Progeni-tor II에서 전이가 일어나지 않았으며, Gene Pulser에서 전이균주를 얻어 두 electroporator간에 형질전환 효율의 차이를 보였다. 11. L. casei 102S에 pLZ12를 electroporation시 낮은 전압에서 형질전환 효율이 비교적 좋았으며, 배양 시기를 달리하여 전이시켰을 때 대수생장 말기의 세포가 형질전환 효율이 좋았다. 12. L. casei 102S세포를 각각 10% glycerol, EB, 2차 증류수 등에 녹여 electroporation을 실시하였을 때 각각 $3.8{\times}10^3$, $5.0{\times}10^2$,1.5${\times}10^2$cfu의 형질전환 효율을 보였으며, 1.0mM HEPES, TE buffer를 사용하였을 때에는 전이가 이루어지지 않았다. 13. Plasmid pLZ12의 농도를 달리하여 electroporation을 하였을 때 형질전환 효율이 농도에 비례하여 증가하였다. 14. L. casei 102S에 대수생장 말기의 세포를 채취하여 10% glycerol, 200 Ohms, 25 ${\mu}$FD, 10kV/cm로 plasmid pLZ12를 electroporation할 때 최대 형질전환 효율인 3.8${\times}$10$^{3}$cfu를 얻었으며, lysozyme 처리가 다른 젖산균과는 달리 형질전환 효율을 증가시키지 못하였다. 15. L. casei 102S 세포를 10% glycerol과 EB에 녹여 -20$^{\circ}C$에서 냉동시킨 다음 1일과 7일 후의 세포를 electroporation한 결과 냉동시 세포에 손상을 주는 것으로 인식되었다.

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Effect of Solar Salt on the Quality Characteristics of Doenjang (천일염이 된장의 품질특성에 미치는 영향)

  • Chang, Mi;Kim, In-Cheol;Chang, Hae-Choon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.1
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    • pp.116-124
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    • 2010
  • Bacterial-koji was prepared by using B. subtilis DJI and Doenjang was prepared by using the bacterial-koji and purified salt or solar salt at 12% (w/v) concentration. Characteristics of the Doenjangs were examined after 2 months of aging. Moisture contents of the purified salt-Doenjang was decreased from 51.55% to 45.38% whereas that of the solar salt-Doenjang was decreased from 51.88% to 47.90%. Acidities of the Doenjangs were gradually increased. Hunter's L (lightness) color values of the Doenjangs were decreased. Browning reaction rate of purified salt-Doenjang was faster than solar salt-Doenjang. The content of free amino acids in solar salt-Doenjang was higher than those in purified salt-Doenjang. The major free amino acids of the Doenjang were glutamic acid (252.34 mg%~284.07 mg%), phenylalanine (184.72 mg%~224.71 mg%), leucine, tyrosine and valine.