• Korean Journal of Soil Science and Fertilizer
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• v.25 no.4
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• pp.387-393
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• 1992

The fate of inoculum strain of Bradyrhizobium japonicum was studied by using genetically marked strain. RJB6 $str^rnal^rneo^r$. A spontaneous mutant of B. japonicum isolated from nodules was made to have antibiotic resistance against streptomycin and nalidixic acid. In order to make genetically marked strain, neomycine resistant gene(Tn5) was introduced into this spontaneous mutant by conjugation with E. coli containing pSUP2021. The southern hybridization was carried out to confirm the plasmid insertion. Hybridization of chromosome DNA using pSUP2021(Tn5) as a probe showed that Tn5 was located on the 4.9kb fragment of chromosome. Soybean seeds were planted into a soil previously cultivated with soybean and inoculated with different cell densities of marked strain. Fourty days after planting, the inoculation effects on nodule number, nodule fresh weight, plant height and nitrogen content in the plot inoculated with heavy cell suspension was a little better than those in the plot with low inoculation. The recovery percentage of the marked strains was about 12% in the plot inoculated with heavy density cell suspension, while 5% in the plot inoculated with low cell suspension.

• Asian Journal of Turfgrass Science
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• v.18 no.3
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• pp.141-148
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• 2004

In this report, several factors such as infection time and concentration of bacterial suspension, influencing on transient gene expression in Agrobacterium-mediated transformation were evaluated. An appropriate concentration (O.D 600nm = 1.0-1.2) of bateria and 30 min of infection time showed a higher level of GUS expression. To improve transformation efficiency (TE), friable embryogenic calli (FEC) were bombarded by tungsten particles without plasmid DNA, and then co-cultivated with A. tumefaciens LBA4404 which contains pTOK233 super binary vector, carrying neomycin phosphotransferase (NPTII), hygromycin phosphotransferase (hpt) and$\beta-glucuronidase$ (GUS) genes. Three days after co-cultivation with A. tumefaciens and particle bombardment, FEC cultures were transferred to the selection medium (SM: MS medium supplemented with BA 1mg/l, hygromycin 100mg/l, cefotaxime 250 mg/l and vancomycin 200mg/l). They were cultured for 2 weeks and then transferred to the second SM containing hygromycin 50mg/l, cefotaxime 200 mg/l and vancomycin 100mg/l. Later, stable GUS expression was detected 4 to 6 weeks after transfer to the SM. Further, TE from Agrobacterium-mediated transformation after particle bombardment increased to about 3-folds compared with Agrobacterium-mediated transformation without particle bombardment. In the present study, we established an efficient transformation protocol of zoysiagrass by using A. tumefaciens in the combination with particle bombardment for the first time.

• Journal of fish pathology
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• v.7 no.1
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• pp.37-46
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• 1994

A total 103 strains of Edwardsiella tarda was isolated from eel culture-ponds and examined for drug resistance, distribution and transferabilities of R plasmids. The drugs used were lincomycin(LM), penicillin(PM), sulfamethazine(SF), sulfadimethoxine(SD), cephalosprin(CP), chloramphenicol(CH), streptomycin(SM), oxytetracycline(OT), ampicillin(AP), oxolinic acid(OA), kanamycin(KM), amikacin(AK), gentamicin(GM) and enrofloxacin(EF). Two strains were resistant to the all drugs used, and all isolates were multiply resistant to drugs(at least 8 among 14 drugs), mostly restricted to LM(103 strains), PM(103 strains), SD(103 strains), SF(103 strains), CP(102 strains), CM(101 strains), SM(100 strains), OT(94 strains), AM(92 strains), OA(80 strains), KM(60 strains), AK(30 strains), GM(19 strains) and EF(14 strains), in combination at high degree showing 34 different drug resistant patterns. The most frequently encountered patterns were LM PM SD SF CP CH SM OT AP OA KM(22 strains, 22.4%) followed by LM PM SD SF CP CH SM OT AP OT(12 strains, 11.7%). LM PM SD SF CP CH SM OT AP(7 strains, 6.8%), LM PM SD SF CP CH SM OT AP OA KM AK GM(6 strains, 5.8%) and LM PM SD SF CP CH SM OT AM OX KM AK GM(6 strains, 5.8%). Transfer experiment of drug resistance showed that of 103 resistant strains, 100 strains(97%) transferred part or all resistance to all drugs, indicating that most isolates carried conjugally transferrable R plasmids determining multiple drugs. The most frequently observed transferarble R plasmids were LM PM SD SF CP CH SM OT AP OA(10 strains), LM PM SD SF CP CH SM OT AP OX KM(7 strains) and LM PM SD SF CP CH AP OA(7 strains). These results sugested that eel culture-ponds were highly contaminated with different strains of Edwardsiella tarda, and that contaminated bacteria might be highly multiple resistant strains to drugs, carring transferable R plasmids.

• Korean Journal of Microbiology
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• v.55 no.1
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• pp.69-73
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• 2019

During screening of microbes for compounds having cosmetic benefits, we isolated Marinobacter salarius HL2708#2 from lava seawater on Jeju Island, Republic of Korea. The complete genome sequence was determined. Strain HL27080#2 features a circular chromosome of 4,304,603 bp with 57.21% G+C content and a 244,163 bp plasmid with 53.14% G+C. There were 4,180 protein coding sequences identified, along with 49 transfer RNA and 18 ribosomal RNA noncoding genes. The genome harbored genes for the utilization of alcohol, maltose/starch, and monosaccharide as sole carbon sources. Genes responsible for halophilic characteristics and heavy metal resistance could be annotated, as well as aromatic and alkane hydrocarbons. Contrary to the prior report that M. salarius is negative for nitrate and nitrite reduction, nitrate/nitrite reductase along with nitrate/nitrate transporters and nitronate monooxygenase were evident, suggesting that strain HL2708#2 may be able to denitrify extracellular nitroalkenes to ammonia.

• The Journal of the Korean Society for Microbiology
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• v.14 no.1
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• pp.27-37
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• 1979

Ninety-five strains of Shigella, 70 of Salmonella paratyphi A, and 230 of Salmonella typhi were tested for their resistance to drugs. Also studied was the inhibition and elimination of drug resistance. All except one strain of Shigella consisted of 79 Sh. flexneri and 16 Sh. sonnei were multiply resistant to chloramphenicol, tetracycline, streptomycin, and splfisomidine. Among them, 70 strains were resistant to ampicillin and carbenicillin, 80 to trimethoprim-sulfamethoxazole, 22 to nalidixic acid, and one to kanamycin, but strain resistant to gentamicin, cephaloridine, and rifampin was not encountered. All strains of S. paratyphi A and S. typhi were susceptible to drugs tested, except sulfisomidine and rifampin, for which all S. paratyphi A were slightly resistant to sulfisomidine and the majority of S. paratyphi A and S. typhi were slightly resistant to rifampin. Approximately 80% of multiply drug-resistant Shigella transferred their resistance to E. coli by conjugation, and the resistance was considered to be mediated by R plasmids. The frequency of transfer of drug resistance varied by donor strains and recipients, but not by selecting drugs. Resistance to nalidixic acid was not transferred by conjugation to the recipients. Drug-resistant Shigella strains successively subcultured in nutrient agar stabs contained clones resistant to drugs and those susceptible to drugs, but the ratio of resistant and susceptible clones varied by strains. The multiply drug-resistant S. typhi and Shigella strains were found to not lose completely their drug resistance by subculture in media. Acriflavine has some effect on the elimination of drug resistance mediated by R plasmids, but the effect varied markedly by strains. Atabrine has no effect among strains tested. The combination of drugs increased the drug actions in majority of cases with synergistic or additive effects.