Kim, Jin-Seog;Lee, Byung-Hoi;Kim, So-Hee;Min, Suk-Ki;Choi, Jung-Sup
Journal of Plant Biotechnology
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v.33
no.1
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pp.57-62
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2006
Several methods for determining the response of corn to glyphosate were investigated to provide a fast and reliable method for identifying glyphosate-resistant corn in vivo. Two bioassays were developed. One assay is named 'whole plant / leaf growth assay', in which the herbicide glyphosate is applied on the upper part of 3rd leaf and the growth of herbicide-untreated 4th leaf is measured at 3 day after treatment. in this assay, the leaf growth of conventional corn was inhibited in a dose dependent from 50 to $1600{\mu}g/mL$ of glyphosate and growth inhibition at $1600{\mu}g/mL$ was 55% of untreated control. The assay has the potential to be used especially in the case that the primary cause of glyphosate resistance is related with a reduction of the herbicide translocation. Another assay is named 'leaf segment / shikimate accumulation assay', in which the four excised leaf segments ($4{\times}4mm$) are placed in each well of a 48-well microtiter plate containing $200{\mu}L$ test solution and the amount of shikimate is determined after incubation for 24 h in continuous light at $25^{\circ}C$. In this assay, 0.33% sucrose added to basic test solution enhanced a shikimate accumulation by 3 to 4 times and the shikimate accumulation was linearly occurred from 2 to $8{\mu}g/mL$ of glyphosate, showing an improved response to the method described by Shaner et al. (2005). The leaf segment / shikimate accumulation assay is simple and robust and has the potential to be used as a high throughput assay in the case that the primary cause of glyphosate resistance is related with EPSPS, target site of the herbicide. Taken together, these two assays would be highly useful to initially select the lines obtained after transformation, to investigate the migration of glyphosate-resistant gene into other weeds and to detect a weedy glyphosate-resistant corn in field.
Taxonomic and genetic analysis of Phytophthora species belonging to six different morphological groups (GI, GII, GIII, GIV, GV, GVI) was conducted using RAPD method. Amplified fragments ranged $0.3{\sim}3.2$ kb in their molecular weights. Among total of 145 bands, there were 109 polymorphic bands. Seven isolates of P. infestans showed high similarities of $0.92{\sim}0.99$, and P. infestans isolate 3 from potato showed similarities of $0.93{\sim}0.95$ compared with other P. infestans. Among isolates of P. capsici, similarities of $0.77{\sim}0.86$ were observed and they were grouped in 80% level. P. cinnamomi and P. cryptogea isolates which belonging to group GVI showed very similar RAPD fingerprinting pattern. Primers OPA-04, OPA-17, OPA-18, OPA-19, and OPB-12 showed high level of differences among the tested isolates in major bands and molecular weights. The similarity between the isolates was 0.67. P. megasperma and P. sojae in group GV showed similarity of 0.65. These two isolates showed big differences in single major band in reactions with primers OPA-08, OPA-17, and OPA-19. Phytophthora-specific and P. infestans-specific molecular markers were also selected with one of the random primers tested. In reaction with primer OPA-20, all the genus Phytophthora showed common band at 600 bp, and all the P. infestans isolates showed specific band at 680 bp. These markers can be useful for identification of Phytophthora speices or P. infestans. As a result, P. infestans isolated from tomato and/or potato can easily be differentiated from other Phytophthora species with this primer.
Oyster mushrooms are widely cultivated and consumed in Korea. P. ostreatus 'Suhan(ASI 2504)' is an ideal cultivar for mushroom farmers due to its dark pileus and thick stipe; however, as it is very sensitive to environmental conditions, an alternative cultivar is required. To develop a new cultivar, parental strains 'Suhan(ASI 2504)' and 'ASI 0665 (Heuktari)' were selected from various collected strains according to morphological characteristics. P. ostreatus 'Soltari' was developed by DiMon crossing between the dikaryotic strain 'Suhan' and the monokaryotic strain derived from 'Heuktari'. Thirty-eight of the 100 crossed strains were selected following analysis of mitochondrial genetic characteristics, and 'Soltari' was ultimately selected by continuous cultivation tests. The mitochondrial DNA profile of 'Soltari' was found to be the same as that of 'Heuktari, and a nuclear DNA profile of 'Soltari' was similar as those of the parental strains, 'Suhan' and 'Heuktari.' 'Soltari' mycelium grows adequately in moderate to high temperatures of $12-20^{\circ}C$, although its optimum temperature was found to be $30^{\circ}C$. Fruiting body production per 1.1-L cultivation bottle was approximately 158.8 g. Its stipe length and thickness were comparable to those of diameter and thickness were somewhat lower (42.72 vs. 51.33 mm and 18.18 vs. 22.46 mm, respectively). 'Soltari' was found to be more resistant to high $CO_2$ atmosphere than 'Suhan', and the color of the pileus of 'Soltari' was dark gray at high temperature. Therefore, it is suggested that this new cultivar 'Soltari' is a good alternative cultivar and will contribute to energy saving in oyster mushroom farms.
Choi, Jin Hyoung;Choe, Young June;Hong, Ki Bae;Lee, Jina;Yoo, Won Joon;Kim, Han Soo;Park, Moon Seok;Cho, Tae Joon;Chung, Chin Youb;Lee, Hoan Jong;Choi, In Ho;Choi, Eun Hwa
Pediatric Infection and Vaccine
/
v.18
no.1
/
pp.31-39
/
2011
Purpose : This study was performed for the purpose of finding causative organisms and clinical features of septic arthritis or acute osteomyelitis in children. Methods : The study involved a retrospective review of the medical records of 63 microbiologically confirmed cases of acute pyogenic arthritis and osteomyelitis. All of the cases were brought about by community-associated infections and managed at the Seoul National University Children's Hospital or Seoul National University Bundang Hospital from June 2003 to July 2009. Results : The median age of all cases was 60 months and there were 35 males and 28 females. Major involved joints included the hip joint (15 cases), knee joint (7 cases), shoulder joint (4 cases), and elbow joint (4 cases). Also, major involved bones included the femur (20 cases), tibia (13 cases), humerus (7 cases), and radius (7 cases). Staphylococcus aureus was the most commonly identified causative organism, accounting for 49 cases (77.8%). Of the 49 isolates of S. aureus, methicillin-resistant S. aureus (MRSA) accounted for 8 cases (16.3%). Group B streptococcus spp. (GBS) and Salmonella spp. accounted for 3 cases, respectively. Nafcillin or cefazolin was often prescribed as an initial empirical antibiotic. There were 9 cases that were managed by a regimen that included vancomycin as the first choice. Fifty four cases (85.7%) recovered without any complications. Methicillin-sensitive S. aureus (MSSA), responsible for 41 cases, caused chronic complications in 3 cases. Of 8 cases caused by MRSA, 1 case showed chronic complication. There were no fatal cases. Conclusion : S. aureus remains the most common organism causing acute pyogenic arthritis and osteomyelitis of childhood acquired in the community. The high prevalence of methicillin resistance among S. aureus should be considered carefully in the selection of initial empirical antibiotics.
To know how the ribosomes involved in secretory protein synthesis were attached to the cytoplasmic membrane in Bacillus amyloliquefaciens, the cells were treated with puromycin combinated with magnesium at the logarithmic phase, and the variation of cell-bound and extracellular $\alpha$-amylase activity was assayed for determining the $\alpha$-amylase translocation blocking through the cytoplasmic membrane. In the abnormal $\alpha$-amylase producing mutant in which the C-terminal of the $\alpha$-amylase structure was deleted, B. umytotiquefaciens CH10-2, the $\alpha$-amylase was translocated normally through the cytoplasmic membranes, and the translocation blocking by puromycin was revealed to have a similar pattern as that in the wild type. This means that the C-terminal part of the enzyme structure may not have a signal for secretion. The cell death of the logarithmic phase cells in both strains was not affected much under 20$\mu\textrm{g}$/$m\ell$ of puromycin, however, the $\alpha$-amylase translocation was blocked markedly under less than 10$\mu\textrm{g}$/$m\ell$ of the puromycin concentration. The blocking of the enzyme secretion by puromycin may be due to the detachment of the ribosomes from cytoplasmic membranes by disturbing the nascent polypeptide synthesis. Further evidence for confirming this was that the detachment was increased in 50 mM of magnesium ion because the extracellular $\alpha$-amylase activity was decreased more under this condition. If the cells were treated with trypsin combinated with Iysozyme, the extracellular $\alpha$-amylase activity from the cultured medium was reduced markedly, however, the activity from the cells treated with trypsin only was not reduced. This means that the nascent polypeptides protruding from the cytoplasmic membrane were sensitive to the trypsin digestion, whereas the matured ones were not. Therefore, the protruding polypeptides from the cytoplasmic membranes may be truncated by trypsin before forming their final tertiary structures by folding in the cell wall layer.
During the Brewing of Soy Sauce on an industrial scale, Saccharomyces rouxii $T_9$ was cultured and added to the soy sauce mash. The comparative experiments of soy sauce mash in non-addition and addition group of yeast were examined in this report. The yeast flora and chemical composition of Soy sauce mash through out the brewing were observed and the results obtained were as follows. (1) The number of Osmophilic yeast in one ml of soy sauce mash showed $185{\times}10^3$ 1 month after mashing and $750{\times}10^3$ 4 months after mashing in case of yeast group, while presented as $98{\times}10^3$ 1 month after mashing and $394{\times}10^3$ 4 months after mashing in case of non-yeast group. And the number of Osmphilic yeast in yeast group was twice of that in non-yeast group. (2) The number of ordinary yeast of TTC red group was shown as $2132{\times}10^3\;to\;3252 ×10^3$ 5 to 6 months after mashing in case of yeast group, while presented $752{\times}10^3\;to\;1251{\times}10^3$ in case of non-yeast group. And the yeast group was shown more than non-yeast group in ordinary, red pink and pink yeast number. (3) TTC red yeast were strongly appeared in both addition and non-addition group of yeast from 1 month after mashing to 6 months after mashing. (4) Though total nitrogen, pure extract, pH and buffer action contents of soy sauce showed similiar tendency in yeast and non-yeast group, alcohol and color density contents were highly appeared in yeast group and reducing sugar content was in non-yeast group respectively. (5) By the results of Organic function test of soy sauce mash, the difference of taste quality in yeast and non-yeast group were not evidently appeared, however, the appearance and flavour of the soy sauce were better in yeast group than in non-yeast group.
This study was conducted to investigate the inhibitory activity of Lactobacillus spp., Bacillus ssp., and calf fecal isolates against pathogenic Salmonella typhimurium, E. coli, Listeria monocytogenes, and Staphylococcus aureus. Among thirteen strains of Lactobacillus ssp. tested, Lactobacillus helveticus CU631 showed the highest inhibition against three pathogens, whereas Bacillus spp. showed a weak inhibitory activity. Four calf fecal isolates were identified as Lactobacillus pentosus CU13, CU05, Pediococcus pentosaceus CUR02, and Lactobacillus lactis ssp. lactis CUM14. The whole cell and cell wall components of L. rhamnosus CU02 and L. pentosus CU13 were active in the inhibition of L. monocytogenes. The medium components and levels, which affect on the inhibitory activity, were revealed as Tween 80 1.0%, peptone 3.0%, yeast extract 3.0%, glucose 3.0%, beef extract 3.0%, and NaCl 1.0~3.0%, respectively. Inhibitory activity of the supernatant culture medium was not affected by catalase and proteinase K treatment but affected by heat treatment at 80℃ and netralization, which implies that the inhibitory activity is due to the production of organic acids during the growth. L. pentosus CU13 and L. rhamnosus CU02 exhibited broad inhibition spectrum against 16 out of 21 strains including some pathogens. Oral administration of L. rhamnosus CU02 to the mice infected with E. coli O157:H7 was proven to be effective to recover their body weight during the experimental period.
The Journal of the Korean Society for Microbiology
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v.17
no.1
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pp.21-34
/
1982
One hundred and twenty-one strains each of Escherichia coli isolated from stools of 60 patients who received various antimicrobial drugs in hospital for more than one week and apparently healthy 60 students who have no history of taking antimicrobial drugs during recent one month, were tested for their resistance to 13 antimicrobial drugs. The frequency of resistance strains was highest to tetracycline with 69.2%, and followed by streptomycin(Sm), sulfisomidine(Su), chloramphenicol(Cm), ampicillin(Ap), and carbenicillin(Cb) in the decreasing order, ranging from 61.2% to 39.3%. Strains resistant to kanamycin(Km), cephaloridine(Cr), and trimethoprim(Tp) occupied about one-fourth of strains, and only four strains were resistant either one or more of nalidixic acid, gentamicin and amikacin, and no strain was resistant to rifampicin. The frequency of resistant strains to Cm, Ap, Km, Cr, and Cb was much higher among patient isolates than student strains, but strains resistant to the other drugs showed almost the same frequencies between patient and student isolates. There was a marked difference in average minimum inhibitory concentrations of between resistant and susceptible strains, suggesting that the resistance to drugs is the plasmid origin. Seventy-six percent of strains were resistant to one to 10 drugs tested, and no much difference was observed between strains from patients and students. However, strains resistant to four or more drugs were much more frequently found among patient isolates than student strains, with the increasing tendency of multiply resistant strains among patient isolates following the increase in the number of resistant drugs. The transfer of drug resistance by conjugation was tested and 98 strains(67.5%) among 145 which were resistant to two or more drugs were found to transfer their drug resistance to E. coli. Among 74 strains resistant to 7 or more drugs, all except one transferred the resistance, and the number of strains with transferable resistance decreased, as the number of resistant drugs decrease. A R plasmid from randomly selected p13 strain was tested for the incompatibility group, and the plasmid was classified into Inc F II. R plasmM DNA bands were identified by polyacrylamide gel electrophoresis.
Journal of Korean Society of Environmental Engineers
/
v.30
no.8
/
pp.789-797
/
2008
This study was conducted to investigate removal characteristics of gaseous trimethylamine(TMA) through biofilter packed with waste tire-chips. The sludge in this experiment was collected from an activated sludge operated in a wastewater treatment facility treating malodorous pollutants. The nominal amount of collected sludge was inoculated through packing materials in the filter. The removal efficiencies for varying concentrations and SVs(Space velocity) were assessed based on TMA, COD$_{Cr}$, NO$_3{^-}$-N, NO$_2{^-}$-N, NH$_4{^+}$-N and EPS(Extracellular Polymeric Substances) in leachate, since biofilter had been steady-stately operated. The influent concentration of 10 ppm of TMA was removed to approximately 95% regardless of changing SV at 120 and 180 hr$^{-1}$, but it was lowered to 80 to 90% at SV 240 hr$^{-1}$. As influent concentration was gradually increased from 5 to 55 ppm, the removal efficiencies of TMA were initially high for 95% in the range of 5 to 10 ppm, but lowered to 80% for 10 to 30 ppm. As a part of kinetic study for TMA decomposition, V$_m$(maximum substrate removal rate) and $K_s$(substrate infinity coefficient) were 14.3 g$\cdot$m$^{-3}$$\cdot$h$^{-1}$ and 0.043 g$\cdot$m$^{-3}$, respectively while adapted period was shown in the range of 100 to 150 hr. Also, the EPS concentration was consistently observed from the leachate showing 100 to 200 ppm, which indicates that biofilm has been continuously formed and sustained throughout tire-chips packed reactor.
Lim, Hyun Taek;Lee, So Hee;Lee, Jung Hwa;Kim, Jeong Eun;Kim, Kyo Sun;Jeong, Eun Ju;Lee, Seung Hyun;Kang, Chang Gyu;Hong, Seong Jin
Clinical and Experimental Pediatrics
/
v.48
no.10
/
pp.1107-1115
/
2005
Purpose : An outbreak of ESBL-producing Shigella sonnei enteritis was unprecedented not only in Korea but throughout the world in the past. We intended to devise a management guideline for ESBL-producing shigellosis based on analysis of clinical manifestations and response to therapy. Methods : We analyzed 103 patients who were admitted to the hospital with acute GI symptoms and were shown positive result for S. sonnei on stool culture. We performed sensitivity test to the antibiotics and DNA sequencing of ESBL gene in the isolated S. sonnei colonies. In addition, we retrospectively analyzed their clinical characteristics, laboratory results, and clinical and microbiological responses to the antibiotics. Results : Among the clinical manifestations, fever was the most frequent(96.1%), followed by diarrhea(93.2%), abdominal pain(76.7%), headache(71.8%), vomiting(65.0%), and nausea(41.7%). The fever was sustained for average of 2.0 days and diarrhea for 3.9 days. Watery diarrhea was the most common(69%) followed by mucoid(26%), and bloody stool(5%). On peripheral blood smear, leukocytosis was noted in 53.4% of patients, and 78.6% of patients tested positive for serum CRP response. On stool direct smear, 11.7% of patients showed more than 50 WBCs/HPF, and 9.7% of patients between 5 to 20 WBCs/HPF. Stool occult blood was positive in 71% of patients. Production of CTX-M-14 type ESBL was reported for all S. sonnei strains isolated from this outbreak. Microbiological eradication rates to various antibiotics were as follows : 100%(9/9) to ciprofloxacin, 100% 5/5) to azithromycin, 6.9%(5/72) to cefdinir, 0%(0/8) to ceftriaxone, 12.5%(1/8) to ceftizoxime, 0%(0/ 8) to TMP/SMX, 42.9%(3/7) to ampicillin/sulbactam, 20%(1/5) to amoxicillin/clavulanic acid, and 68.8 %(11/16) to imipenem/cilastatin. Conclusion : It is presumed that azithromycin can be an attractive option for the treatment of ESBL-producing S. sonnei enteritis in pediatric population, given its cost-effectiveness and safety. Although ciprofloxacin is another cost-effective agent, its use in pediatric population may be a bit too premature.
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