• Title/Summary/Keyword: Resistant screening

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Probiotic Properties of Lactobacillus strains Isolated from Kimchi (김치로부터 분리된 Lactobacillus strains의 probiotic 특성)

  • Choi, Hye Jung;Lim, Bo Ram;Kim, Dong Wan;Kwon, Gi-Seok;Joo, Woo Hong
    • Journal of Life Science
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    • v.24 no.11
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    • pp.1231-1237
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    • 2014
  • The objective of this study was to evaluate the safety and functional properties of four potential probiotic strains isolated from Kimchi, traditional Korean fermented vegetables. Based on being higher tolerance to bile salts and showing higher acid resistance or hydrophobic properties, one Lactobacillus arizonensis strain (BCNU 9032) and three L. brevis strains (BCNU 9037, BCNU 9098 and BCNU 9101) were selected in the screening experiment. All strains can survived up to 99% after 3h culture in pH 2.5 and resistant to 1% bile salts. These strains also showed good antimicrobial activities against a number of food borne pathogens, especially against Escherichia coli and Shigella sonnei. The ability to lower cholesterol levels of L. arizonensis BCNU 9032 and L. brevis 9037 were demonstrated by bile salt hydrolytic activity and cholesterol assimilation tests. Moreover, L. brevis BCNU 9098 and BCNU 9101 showed higher adherence to Caco-2 cells (12.76 and 11.86%, respectively) than Lactobacillus rhamnosus GG, a commercial probiotic strain used worldwide. The results suggest that these strains could be used as probiotics.

Growth Characteristics of Barley Seedlings in Hydroponic Culture Conditioned Artificial Wet Injury (양액재배를 이용한 인위적 습해유발조건에서 보리 유묘의 생육특성)

  • 윤성중;박미은;최혜란;최재성;김정곤;서세정;강현중;김정곤;최경구
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.48 no.3
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    • pp.160-168
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    • 2003
  • This study was carried out to investigate the effects of induced wet-injury conditions on barley seedling growth. Barley seedlings at the three leaf stage were grown in culture solutions with various combinations of dissolved oxygen (DO), nutrients and citric acid (CA) concentrations. Seedling growth was reduced by hypoxia. Root length and root fresh weight were most severely reduced under 1 ppm DO. Shoot growth was slightly reduced but not root growth under low nutrient condition. Seedling growth was reduced by CA in a concentration-dependant manner, Response of seedling growth was efficiently differentiated in a solution containing 1 ppm DO, one tenth of the normal P, K and Mg concentration and 3 mM CA. Under this induced wet-injury condition, overall seedling growth of 6 cultivars was reduced by 11 to 19% compared to the control condition. The results suggest that the induced wet-injury condition can be used for the screening of resistant genotypes at the seedling stage.

Isolation of Mutants Susceptible to Rice Blast from DEB-treated Rice Population (DEB 처리에 의해 유도된 벼 돌연변이 집단으로부터 도열병 감수성 돌연변이 분리)

  • Kim, Hye-Kyung;Lee, Sang-Kyu;Han, Mu-Ho;Jeon, Yong-Hee;Lee, Gi-Hwan;Lee, Youn-Hyung;Bhoo, Seong-Hee;Hahn, Tae-Ryong;Jeon, Jong-Seong
    • Applied Biological Chemistry
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    • v.48 no.4
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    • pp.339-344
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    • 2005
  • Rice blast, which is caused by the fungus Magnaporthe grisea, is one of the most destructive diseases of rice. To identify genes involving in the signal transduction pathways that mediate rice blast resistance, we screened over 2,000 mutant lines of a highly resistant variety RIL260 that were generated by using a DEB (1, 3-Butadiene diepoxide) treatment method. In the mutant population, the frequency of albino plants was 6.7%, indicating that this population has a high frequency of mutations in the genome. The primary screening identified 29 mutant plants that exhibit a complete or partial loss of the resistance to rice blast. Among them, M5465, the most susceptible line, was subsequently examined by DNA gel-blot experiments using DNA molecular markers of Pi5(t) that has been previously identified as a durable resistance locus in RIL260. The result revealed that a large deletion and rearrangement of genomic DNA occurred in the Pi5(t) locus. The results suggest that DEB can be used as an efficient mutagen to induce large scale mutations in the rice genome. The isolated mutants should be useful for elucidating the Pi5(t)-mediated signaling pathways of rice blast resistance.

Efficient Gene Targeting using Nuclear Localization Signal (NLS) and Negative Selection Marker Gene in Porcine Somatic Cells

  • Kim, Hye Min;Lee, Sang Mi;Park, Hyo Young;Kang, Man-Jong
    • Reproductive and Developmental Biology
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    • v.38 no.2
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    • pp.71-77
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    • 2014
  • The specific genetic modification in porcine somatic cells by gene targeting has been very difficult because of low efficiency of homologous recombination. To improve gene targeting, we designed three kinds of knock-out vectors with ${\alpha}1,3$-galactosyltransferase gene (${\alpha}1,3$-GT gene), DT-A/pGT5'/neo/pGT3', DT-A/NLS/pGT5'/neo/pGT3' and pGT5'/neo/ pGT3'/NLS. The knock-out vectors consisted of a 4.8-kb fragment as the 5' recombination arm (pGT5') and a 1.9-kb fragment as the 3' recombination arm (pGT3'). We used the neomycin resistance gene (neo) as a positive selectable marker and the diphtheria toxin A (DT-A) gene as a negative selectable marker. These vectors have a neo gene insertion in exon 9 for inactivation of ${\alpha}1,3$-GT locus. DT-A/pGT5'/neo/pGT3' vector contain only positive-negative selection marker with conventional targeting vector. DT-A/NLS/pGT5'/neo/pGT3' vector contain positive-negative selection marker and NLS sequences in upstream of 5' recombination arm which enhances nuclear transport of foreign DNA into bovine somatic cells. pGT5'/neo/pGT3'/NLS vector contain only positive selection marker and NLS sequence in downstream of 3' recombination arm, not contain negative selectable marker. For transfection, linearzed vectors were introduced into porcine ear fibroblasts by electroporation. After 48 hours, the transfected cells were selected with $300{\mu}g/ml$ G418 during 12 day. The G418-resistant colonies were picked, of which 5 colonies were positive for ${\alpha}1,3$-GT gene disruption in 3' PCR and southern blot screening. Three knock-out somatic cells were obtained from DT-A/NLS/ pGT5'/neo/pGT3' knock-out vector. Thus, these data indicate that gene targeting vector using nuclear localization signal and negative selection marker improve targeting efficiency in porcine somatic cells.

Development of Ethanol Producing Saccharomyces cerevisiae Strain Using High Concentration Galactose (고농도 Galactose로부터 에탄올을 생산하는 Saccharomyces cerevisiae 균주의 육성)

  • Kim, Ju-Hye;Yoon, Min-Ho
    • Journal of Applied Biological Chemistry
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    • v.54 no.1
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    • pp.41-46
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    • 2011
  • A galactose-fermenting yeasts, Saccharomyces cerevisiae No. 9, was selected by screening their abilities to produce carbon dioxide gas when grown on galactose. The selected strain, No. 9 and the reference strains NRRL Y-1528 which was exceptionally resistant to high concentration of substrate, were acclimated on sugars such as glucose, mannose, and galactose, and then their ethanol productivities were investigated during fermentation on these three carbon sources. Ethanol productivity of the strain No. 9 reached to the maximum levels after 18 h of fermentation and the ethanol yield was from 36 to 38% when presented as $[EtOH]_{max}/[Sugar]_{ini}(g/g)$, regardless of the conditions of acclimation. From the results obtained by acclimation and fermentation, it was concluded that the ethanol yields from galactose were not affected by the sugars acclimated. Improvements of the strain S. cerevisiae No. 9 were attempted to increase the fermentation efficiency and/or ethanol yields on high concentration of substrate by the conventional mutation methods employing methanesulfonic acid, ethyl ester (EMS). Mutants, Mut-5 (SJ1-40), -17 (LK4-25) and -24 (LK2-48) fermented galactose at the concentration of 20% in the levels of higher 39.9~51.6% than the mother strain, No. 9, however, their ethanol yields never exceeded those of the reference strain.

Isolation of Biogenic Amine Non-producing Lactobacillus brevis SBB07 and Its Potential Probiotic Properties (바이오제닉 아민 비생성 Lactobacillus brevis SBB07의 분리 및 잠재적 프로바이오틱스 특성 분석)

  • Yang, Hee-Jong;Jeong, Su-Ji;Jeong, Seong-Yeop;Ryu, Myeong Seon;Jeong, Do-Youn
    • Journal of Life Science
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    • v.28 no.1
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    • pp.68-77
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    • 2018
  • The purpose of this study was to isolate the probiotic lactic acid bacteria, and verify the possibility of the final selection strain as probiotic material. For screening of biogenic amines non-producing microorganisms, 42 lactic acid bacteria were isolated from various berries, extract and vinegar grown in Sunchang. Isolates were investigated for various physiological activities such as extracellular enzyme, antimicrobial and antioxidant activities, and 5 isolates were firstly screened. SBB07 was finally selected by analyzing the biogenic amine, and named Lactobacillus brevis SBB07 by 16S rRNA sequencing analysis. Next, SBB07 was assayed for their survival ability when exposed to acidic and bile conditions as well as heat and antibiotic resistance. As a result, SBB07 showed more than 86% and 54% higher survival rate in acidic condition at pH 2.0 and bile resistance with 0.5% oxgall. In addition, SBB07 showed a survival rate of more than 113% in $60^{\circ}C$, and also confirmed that it has resistant to various antibiotics. As a result of confirming the possibility of prebiotics, SBB07 showed the best utilization of GOS as a prebiotic substrate, and utilization of FOS and inulin were also high. These results suggest that SBB07 have good potential for application as probiotic lactic acid bacteria.

Screening of Tomato Spotted Wilt Virus Resistance in Tomato Accessions (토마토반점위조바이러스(TSWV) 저항성 토마토 유전자원 탐색)

  • Han, Jung-Heon;Choi, Hak-Soon;Lee, Jun-Dae;Kim, Jae-Deok;Lee, Won-Phil;Choi, Hong-Soo;Kim, Jung-Soo;Yoon, Jae-Bok
    • Horticultural Science & Technology
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    • v.30 no.2
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    • pp.171-177
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    • 2012
  • A total of 94 tomato accessions were evaluated for the resistance to $Tomato$ $spotted$ $wilt$ $virus$ (TSWV) using a Sw5-2 SCAR marker and bioassay. PCR products of the marker were approximately 574 bp, 500 bp, and 462 bp, among which the longest was linked to TSWV resistance allele of Sw5-b. This allele was only found in three accessions (09-438, 10-318, and 10-321) in which some individuals showed apparent recovery or stem necrosis symptom to a tomato isolate of TSWV-pb1. Thirty-five individuals (one per each accession) which were non-infected by ELISA were selected for further observation. Among these, 26 individuals that did not show any symptom at 5 months after inoculation were confirmed for viral infection by RT-PCR. TSWV-specific PCR amplicon was weakly detected in all 26 individuals including 'Eureta', a commercial F1 possessing the resistance allele of Sw5-b. The resistant genes in the selected individuals may play an important role for reducing the viral concentration in tissues of inoculated tomato plants and seems to be quantitatively controlled by several factors including Sw5-b gene.

The Effect of Newly Synthesized Compounds on the Photosynthetic Electron Transport of Cyanobacteria (Anacystis nidulans $R_2$) (신규(新規) 합성화합물들이 cyanobacteria의 광합성전자전달계에 미치는 영향)

  • Hwang, I.T.;Kim, J.S.;Cho, K.Y.;Yoneyama, K.;Yoshida, S.
    • Korean Journal of Weed Science
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    • v.13 no.2
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    • pp.89-95
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    • 1993
  • The Inhibiting activity of newly synthesized phenol (E-series) and triazine (T-series) derivatives was evaluated by using thylakoid membranes extracted from cyanobacteria (Anacystis nidulans $R_2$). There were no significant differences between phenol derivatives and dinoseb to the thylakoid membrane extracted from wild type in the Hill reaction. However, a phenol derivative, E-24 which has no -Cl at phenyl ring, did not show any activity. The longer the length of R substituents was in phenol derivatives, the lower inhibiting activity was in the Hill reaction. Triazine derivatives, T-27, T-28, T-40, T-41, T-47 and T-48 were also compared with diuron and atrazine. Among triazine compounds, T-27 and T-28 showed 10 and 30 times activity as high as atrazine to wild type, respectively. Other triazine derivatives, T-40, T-41, T-47 and T-48 showed low inhibiting activity to wild and mutant type. A structural difference of T-27 and T-28 from T-40, T-41, T-47 and T-48 was the presented of -C-NH-. Both T-27 and T-28 were very closely associated with serine, an amino acid located at the 264th position of D1 protein because of the resistant ratio(R/S) to mutant G-264 were higher than that of atrazine.

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An Evaluation Method for Sesame (Sesamum indicum L.) Resistance to Phytophthora nicotianae var. parasitica (참깨발병 품종저항성 검정방법 연구)

  • Seong Ho, Choi;Eui Kyoo, Cho;Young Am, Chae
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.32 no.2
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    • pp.173-180
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    • 1987
  • Symptom development and disease severity of Phytophthora blight in the sesame plants varied depending upon age of the plants tested, inoculation method, watering method, and inoculum density in both susceptible Suweon 9 and Suweon 26 and moderately resistant B-67 and IS 103 sesame lines to Phytophthora nicotianae var. parasitica when inoculated. However, successful differentiation of the sesame lines for varietal resistance was possible using 20-day old seedling, inoculation by soil infestation, saturated soil water condition by half immersion of pots in water tank, and 200 sporangia per one ml of inoculum. Spraying or soil inoculation to 70-day old plants also was effective in differentiating the varietal resistance. By the screening method Suwon 26 showed 100% diseased plants and symptom severity index 9.0, while B-67 showed 20% diseased plants and symptom severity index 1.7. The rating scale given was from 0 through 9. For example, the scale 0 signified no symptom development, 5 signified discoloration of basal part of stem, and 9 signified discoloration of stem more than 10 cm high above the soil surface with blighted leaves. Differentiation in symptom severity also was made by percentage of the lesion area. Results evaluated using both parameters were well corresponded in varietal reaction of sesame to Phytophthora blight.

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Identification and Annual Change of Races of Pyricularia oryzae in Korea (한국에 있어 벼도열병균레이스의 분화 및 년차적변동)

  • Lee Eun Jong;Joo Won Joon;Chung Bong Jo
    • Korean journal of applied entomology
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    • v.14 no.4 s.25
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    • pp.199-204
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    • 1975
  • One thousand forty-four isolates of Pyricularia oryzae obtained during the period 1962-1974 front blast-infected rice samples from all rice-growing areas of Korea were tested on the Japanese differential set of twelve rice varieties. Twenty-seven races of the fungus were differentiated. The number of races falling in groups T,C and N were 4, 15 and 8, respetively. Since only Japonica type varieties had been cultivated widely before 1966, N-race groups were found to be most Prevalent with a frequency of about $85\%$ of the total isolates. After 1966, T-and C-race groups gradually increased to $21\%$ and $42\%$ respectively, as a result of the widespread cultivation of Tongil bred from a cross between IR8 x(Yukara x T(N)1. Before 1966, a few T, and C-race groups occurred only in the Suweon region; since 1966, most of the races differentiated have been uniformly distributed throughout the country regardless of region. The most prevalent races in Korea were N-2 and C-8. The variety Tetep was resistant to all races previously identified in Korea, but a new race (T-d) to which Tetep is susceptible has been found each year since 1973. It appears likely that races capable of attacking Indica types of rice will become increasingly prevalent and a programme of continuous screening is needed to detect new races as soon as they appear.

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