• YAKHAK HOEJI
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• v.58 no.1
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• pp.7-11
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• 2014

To study the resistance mechanism of E. faecalis to LCB01-0371, several resistant mutants to LCB01-0371 or linezolid were isolated by step-wise selection. The frequency of spontaneous mutations resistant to LCB01-0371 was lower than that of linezolid in E. faecalis. The genetic variations in resistant mutants were analyzed by DNA sequencing of domain V of 23S rRNA in each mutant. The first-step mutant to LCB01-0371 had a G2576T point mutation in V domain of 23S rRNA. However, no resistant mutant to LCB01-0371 was isolated in second-step mutant selection.

• Journal of Microbiology and Biotechnology
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• v.34 no.5
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• pp.1178-1187
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• 2024

Cordyceps militaris is a significant edible fungus that produces a variety of bioactive compounds. We have previously established a uridine/uracil auxotrophic mutant and a corresponding Agrobacterium tumefaciens-mediated transformation (ATMT) system for genetic characterization in C. militaris using pyrG as a screening marker. In this study, we constructed an ATMT system based on a dual pyrG and hisB auxotrophic mutant of C. militaris. Using the uridine/uracil auxotrophic mutant as the background and pyrG as a selection marker, the hisB gene encoding imidazole glycerophosphate dehydratase, required for histidine biosynthesis, was knocked out by homologous recombination to construct a histidine auxotrophic C. militaris mutant. Then, pyrG in the histidine auxotrophic mutant was deleted to construct a ΔpyrG ΔhisB dual auxotrophic mutant. Further, we established an ATMT transformation system based on the dual auxotrophic C. militaris by using GFP and DsRed as reporter genes. Finally, to demonstrate the application of this dual transformation system for studies of gene function, knock out and complementation of the photoreceptor gene CmWC-1 in the dual auxotrophic C. militaris were performed. The newly constructed ATMT system with histidine and uridine/uracil auxotrophic markers provides a promising tool for genetic modifications in the medicinal fungus C. militaris.

• Journal of Microbiology and Biotechnology
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• v.13 no.1
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• pp.90-98
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• 2003

In vivo $^31p$ Nuclear Magnetic Resonance ($^31p$NMR) and metabolic studies were carried out on an acetic acid tolerant mutant, Zymomonas mobilis $ZM4/Ac^R$, and compared to those of the parent strain, Z. mobilis ZM4, to evaluate possible mechanisms of acetic acid resistance. This investigation was initiated to determine whether or not the mutant strain might be used as a suitable recombinant host far ethanol production from lignocellulose hydrolysates containing various inhibitory compounds. $ZM4/Ac^R$ showed multiple resistance to other lignocellulosic toxic compounds such as syringaldehyde, furfural, hydroxymethyl furfural, vanillin, and vanillic acid. The mutant strain was resistant to higher concentrations of ethanol or lower pH in the presence of sodium acetate, compared to ZM4 which showed more additive inhibition. in vivo $^31p$ NMR studies revealed that intracellular acidification and de-energization were two mechanisms by which acetic acid exerted its inhibitory effect. For $ZM4/Ac^R$, the internal pH and the energy status were less affected by sodium acetate compared to the parent strain. This resistance to pH change and de-energization caused by acetic acid is a possible explanation for the development of resistance by this strain.

• Journal of Microbiology and Biotechnology
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• v.26 no.9
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• pp.1570-1578
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• 2016

Commercial application of laccase is often hampered by insufficient enzyme stocks, with very low yields obtained from natural sources. This study aimed to improve laccase production by mutation of a Coriolopsis gallica strain and to determine the biological properties of the mutant. The high-yield laccase strain C. gallica TCK was treated with N-methyl-N-nitro-N-nitrosoguanidine and ultraviolet light. Among the mutants isolated, T906 was found to be a high-production strain of laccases. The mutant strain T906 was stabilized via dozens of passages, and the selected ones were further processed for optimization of metallic ion, inducers, and nutritional requirements, which resulted in the optimized liquid fermentation medium MF9. The incubation temperature and pH were optimized to be 30℃ and 4.5, respectively. The mutant strain T906 showed 3-times higher laccase activity than the original strain TCK under optimized conditions, and the maximum laccase production (303 U/ml) was accomplished after 13 days. The extracellular laccase isoenzyme 1 was purified and characterized from the two strains, respectively, and their cDNA sequence was determined. Of note, the laccase isoenzyme 1 transcription levels were overtly increased in T906 mycelia compared with values obtained for strain TCK. These findings provide a basis for C. gallica modification for the production of high laccase amounts.

• Microbiology and Biotechnology Letters
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• v.23 no.1
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• pp.47-52
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• 1995

Since the original productivity of new aminopeptidase M inhibitors MR-387A and B by Streptomyces sp. SL-387 (KCTC 0102BP) was not enough for further chemical and biological evaluation, mutation of parent strain by the treatment of N-methyl-N'-nitro-N-nitrosoguanidine was performed in order to obtain a clone with greater inhibitory activity. Mutant N-3 was selected due to a 6-fold greater productivity (40 $\mu$g/ml) than that of the wild type(6.7 $\mu$g/ml). This mutant was resistant to 3,4-dehydro-DL-proline, an antimetabolite of proline, with 25 $\mu$g/ml of minimum inhibitory concentration. Furthermore, the characteristic morphological change from spiral spore chain in wild type to straight in mutant was observed. An aminopeptidase M nhibitor different from MR-387A and B was isolated from the culture broth of the mutant. This inhibitor was composed of 2 proline, 1 valine, and an unknown amino acid which is presumably 3-amino-4-phenylbutanoic acid. IC$_{50}$ value (89.1 $\MU$g/ml) of the purified inhibitor was lower than that of other inhibitors, which may be due to the absence of 2(S)-hydroxyl group within the structure of 3-amino-4-phenyl- butanoic acid.

• Journal of Crop Science and Biotechnology
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• v.11 no.3
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• pp.181-186
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• 2008

To assess the potential as biofortified rice varieties, new endosperm and grain mutant lines were selected from $M_4$ generation seeds of the rice cultivar Shindongjin, which were either $\gamma$-irradiated or treated with N-methyl-N-nitrosourea(MNU) and lipid, sugar, and tocopherol content were analyzed. Amylose content in non-waxy mutants ranged from 8.8% in SM-4, a dull-type mutant, to 29.5% in SM-51, compared to 18.9% in the parental variety, Shindongjin. SM-23, a floury-type mutant, contained 0.09 ${\mu}g/g$ $\alpha$-tocopherol(40.9% of total tocopherol), was three times higher than in the parental variety. SM-32, a giant embryo-type mutant, had a 2.2-fold higher total tocopherol content, 2.1-fold higher $\alpha$-tocopherol, and 5.5-fold higher $\delta$-tocopherol content(47.3% of total tocopherol) than the parental variety(0.13 ${\mu}g/g$). Total free sugar content was elevated in all selected mutants and 1.2-8.6 times higher than in the parental variety(11.38 ${\mu}g/g$). These increased sugar levels were due to increase in sucrose concentration. SM-23(floury-type mutant) and SM-51(high amylose-type mutant) had 4.6 and 7.0 times more sugar, respectively, than the parental variety(11.38 ${\mu}g/g$). With relatively high concentrations, most mutants showed elevated fatty acid content in the SM 32(giant embryo-type) and SM-51(high amylose-type) mutants, at 124.56 and 89.59 mg/g, respectively. All selected mutants displayed valuable characteristics for the development of new varieties in rice-breeding programs.

• Journal of the Korean Society of Tobacco Science
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• v.17 no.1
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• pp.27-32
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• 1995

A new flue-cured tobacco mutant line BU 8832-85 was developed at Taegu Experiment Station, Korea Ginseng and Tobacco Research Institute in 1994. KF 8832-85 was resulted from a cross of flue-cured cultivars NC 95$\times$NC 2326, and developed by a pedigree system of breeding ; initial selection was made by plant type and resistance to bacterial wilt(BW) disease(heudomonas solanaceamm) in the F2 generation under the natural field conditions infested with the pathogen. One white flowered plant was occurred by spontaneous mutation in a certain line among the F3 generatioin while the others were pink. Six plants from the seeds by selfing were selected at the field infested with the pathogen among 240 populations with white flowering in the F4, KF 8832-85 was selected based on yield and leaf quality trials among 6 lines in Fs generation. BCF 8832-85 was compared with its Parent for certain agronomic and chemical characteristics at Taegu Experiment Station in 1993 and 1994. The results showed that KF 8832-85 have white flower, the stalk height was approximately that of NC 2326 but averaged about loom taller than NC 95. It produced ground suckers as much as NC 95, and did not breakdown leaf at the same as WC 2326. KF 8832-85 have high resistance to bacterial wilt disease. Yield of KF 8832-85 was 10 and 18% higher then that of NC 2326 and WC 95, respectively. Price per Kg was equal to that of NC 2326. The contents of nicotine and reducing sugar did not differ significantly from NC 95, while total nitrogen was significantly lower than NC 95. Therefore, the new mutant line is genetically stable for agronomic and chemical characteristics and provides a source of bacterial wilt disease resistance for use in breeding resistant flue-cured cultivars. Key words : Mutant line, White flower, Spontaneous mutation.

• Korean Journal of Breeding Science
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• v.41 no.4
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• pp.510-514
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• 2009

Cymbidium is one of the largest genus in the orchid family and a number of hybrids have been bred in the world. During mass-propagating the Cymbidium "Dongyang" using the meristem culture technology, a useful leaf mutant was selected from the protocom like bodies. The new Cymbidium variety by in vitro mutangesis from "Dongyang" was named as 'Daegook' in 1998. Compared to Dongyang, "Daegook" mutant has white or yellow stripes along the margin of leaves and flowers. The plant length of "Daegook" was shorter than "Dongyang" and the mean length and width of leaf in "Daegook" was 40 cm and 1.6 cm, respectively. The new variety, "Daegook", is expected to be a popular Cymbidium variety among consumer as a ornamental orchid mutant for pot culture by its fine and unique stripes and growth characters.

• Korean Journal of Breeding Science
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• v.40 no.3
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• pp.223-227
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• 2008

In rice, tillering is an important trait determining yield. To study tillering at the agricultural and molecular aspects, we have examined a spontaneous rice mutant that showed reduction in the number of culms. The mutant was derived from a $F^6$ line of the cross of Junambyeo*4 / IR72. It could produce, on average, 4 tillers per hill in the paddy field while wild-type plants usually have 15. Except the reduced culm numbers, they also show pale green phenotypes. The phenotypes of this mutant were co-segregated as the monogenic Mendelian ratio (${\chi}^b=0.002$, p=0.969). In order to locate a gene responsible for the rcn phenotype, the mutant with the japonica genetic background was crossed with Milyang21 of the indica background. Bulked segregant analysis was used for rapid determination of chromosomal location. Three SSR markers (RM551, RM8213, and RM16467) on chromosome 4 were genetically associated with the mutant phenotype. Each of the 217 $F_2$ plants was genotyped with simple sequence length polymorphisms. The data showed that RM16572 on chromosome 4 was the closest marker that showed perfect co-segregation among the $F_2$ population. We suggest the new rcn gene studied here name as $rcn10^t$ because there was no report which exhibit a rcn phenotype with a pleiotropic effect of pale green (chlorophyll deficiency), and mapped at same position on chromosome 4.

• Korean Journal of Microbiology
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• v.15 no.4
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• pp.145-153
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• 1977

Penicillin amidase (EC 3.5.1.11) was produced by a mutant strain of Bacillius megaterium ATCC 14945. Hydroxylamine assay method for the determination of 6-APT was modified by using "HCl addition techniques" in order to simplify the time consuming orginal assay method without sacrifice of accuracy. Using the new mutant strain, the effects of fermentation conditions on enzyme production were studied.e studied.