• Applied Biological Chemistry
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• v.11
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• pp.137-141
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• 1969

In order to obtain amino acids and nucleic acid derivatives from adenine auxotrophic mutants of Bacillus subtilis and Escherichia coli, vitamin, nucleic acid analogue, streptomycin as well as ultraviolet light were adopted for the production of adenine auxotrophic mutants and the results showed efficient production of desired mutants. 1. Ultraviolet ray $(2530\;{\AA}\;2080\;erg/mm^2)$ irradiation to Bacillus subtilis and E. coli at a distance of 30 cm for 80-90 sec. and for 15-20 sec. respetively induced four and eight strains of auxotrophic mutants. 2. Treatment of aminopterine$(200\;{\mu}g/ml)$ inhibited the growth of Bacillus sultilis significantly but a subsequent irradiation of ultraviolet light at the above mentioned conditions induced six times as much mutants as compared to the irradiation alone. In case of E. coli a similar tendency was observed with treatment of streptomycin$(200\;{\mu}g/ml)$ with doubled induction rate of adenine auxotrophic mutants as compared to the irradiation alone.

• Korean Journal of Microbiology
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• v.24 no.2
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• pp.91-97
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• 1986

Intra and interspecfic fusants were produced by the protoplast fusion of auxotrophic mutants from Trichoderma koningii ATCC 26113 and Trichoderma reesei QM 9414. It was found that 0.6M $MgSO_4\;and\;0.6M\;NH_4Cl$ was the best osmotic stabilizer for the preparation of protoplasts from the mycelium of T. koningii and T. reesei respectively. However, $MgSO_4$ was the most suitable one for the regeneration of the protoplasts from both species. The intraspecific protoplast fusion frequencies between the auxotrophic mutants from T. reesei were $1.8{\times}10^{-2}\;to\;5.1{\times}10^{-1}$. Interspecific protoplast fusion frequencies between the auxotrophic mutants from T. koningii and T. reesei were $3.6{\times}10^{-3}$\;to\;8.4{\times}10^{-2}. Interspecific complementing fusants, however, were not alwats produced. Fusants obtained from interspecific potoplast fusion were spontaneously segregated into various strains including parental types, non-parental auxotrophic hybrids, and prototrophic hybrids on complete plate. Interspecific hybrids revealed to have partially enhanced celluloytic activities.

• The Korean Journal of Mycology
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• v.25 no.1 s.80
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• pp.26-29
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• 1997

Pyrimidine auxotrophic basidiospores of Pleurotus sajor-caju were selected using positive selection method. Wild type basidiospores could not grow on minimal medium containing the pyrimidine analog 5'-fluoro-orotic acid (5'-FOA) whereas pyrimidine auxotrophs grew normally. After treatment of basidiospores with ultraviolet light, a total of 13 pyrimidine auxotrophic basidiospores were isolated among 24 5'-FOA resistant mutants. These mutants require the pyrimidine such as uracil, cytosine, thymine. Mating type group and growth rate of their mutants were determined.

• Korean Journal of Microbiology
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• v.22 no.2
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• pp.103-110
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• 1984

The conditions for the protoplast fusion of auxotrophic mutants of Trichoderma koningii were determined. A preparation of commercial enzyme Driselase was used successfully to isolate protoplasts from the 18 hr old mycelium of T. koningii. The yields of protoplasts production were ranged from $0.3{\times}10^8$ to $2.5{\times}10^8$ protoplasts per mg of damp mycelium of various auxotrophic mutant strains. The regeneration frequencies from $9.3{\times}10^{-3}\;to\;2.0{\times}10^{-1}$ were obtained when the protoplasts from auxotrophic mutants were plated on the malt extract medium containing 0.6M $MgSO_4$, and 2% agar, and the optimal concentration of PEG for protoplst fusion was 30%. Exposure of protoplasts to PEG for 10 min was found to be sufficient to induce high frequency heterokaryon formation. Optimal pH of fusion mixture was determined as 5.5, and 1 mM of calcium chloride in fusion mixture was found to be sufficient to enhance protoplast fusion frequency. Under optimal condition, the fusion frequency of the cross between protoplasts from various auxotrophic mutants were $1.6{\times}10^{-2}\;and\;4.1{\times}10^{-2}$.

• Korean Journal of Microbiology
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• v.20 no.1
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• pp.1-10
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• 1982

From Aspergillus nidulans, six suppressor mutants, MSI-MS6, were isolated, and their characteristics were analysed. These were the suppressor mutants for acriflavin resistant and nicotinamide auxotrophic mutant phenotypes. MS1, MS2, MS5 and MS6, were linked to the chromosome IV, I, II respectively, and MS2 was linked to one of the rest chromosomes, MS3 and MS6 mutants had both suppressors for acriflavin resistant marker and for nicotinamide auxotrophic marker. In order to know the stability and efficiency of the suppressors, their reversion frequencies, that is, frequencies of losing the suppressibility, were analysed. Only MS3 and MS5 were reversed with high frequency. The four mutants didn't lose their suppressibilities, and this meant that the suppressors of these four were very stable and highly effcient. The suppressor specificities of these mutants were tested for other mutant's phenotype marker. One of the six suppressors, MS1, had the suppressor specificity for acriflavin resistant marker of 163 strain.

• The Korean Journal of Mycology
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• v.21 no.3
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• pp.195-199
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• 1993

This experiments was designed for development of Fusarium oxysporum strains with enhanced activity of polygalacturonase by means of mutants and protoplasts fusion. Six auxotrophic mutants of F. oxysporum were induced by treatment of MNNG. Protoplasts from mutants were formed from early exoponential mycelium after treatment with driselase(10 mg/ml) using 0.6 M KCl as osmotic stabilizer. Fusion experiments between protoplasts of several auxotrophic mutants were done using polyethyleneglycol 8,000 and $CaCl_2$. The frequency of fusion was $5.0{\times}10^{-3}$ as determined from several experiments. Activity of polygalacturonase was determined by the methods of modified DNS. Consequently, the polygalacturonase activities of mutants and fusant derived F. oxysporum were 1.4 to 3.5 times greater than that of the parental strain, and mutant Fx-2 seemed to be the best strain. Thus, the method we used seemed to be favorable for the improvement of strains.

• The Korean Journal of Mycology
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• v.14 no.2
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• pp.185-188
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• 1986

After treatment of basidiospores of P. cornucopiae with ultraviolet light, 84 putative mutants from 4671 isolates were obtained. The highest proportion of auxotrophic mutants was obtained from the isolates irradiated to give $0.4{\sim}1.0%$ survival. Fourteen auxotrophs were selected for protoplast fusion and each genetic marker was identified.

• The Korean Journal of Mycology
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• v.13 no.2
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• pp.75-78
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• 1985

Mutagenesis of protoplast could serve a great potential tool for improvement of strains and genetics in higher fungi. For the isolation of auxotrophic mutants from protoplasts of Pleurotus ostreatus and Pleurotus florida, viability levels of ultraviolet lights were determined. Seven auxotrophs were obtained from protoplasts irradiating UV to give $0.83{\sim}15%$ survival. The mutants showed a single requirement for each of Arg, Ribo-l, Ribo-2 or Phen for growth. Some of them showed two or three kinds of requirements, Gly Ser, Ade Hypo or Ala Om Tryp for growth.

• Proceedings of the Zoological Society Korea Conference
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• 1994.10a
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• pp.251.2-251
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• 1994

No Abstract, See Full Text

• Korean Journal of Microbiology
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• v.29 no.2
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• pp.123-129
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• 1991

In order to develop the protoplast fusion method of the strains of Fusarium, the interspecific protoplast fusion was attempted between Fusarium poae and F. sporotrichioides. Various auxotrophic mutants were isolated by the treatment of N-Methyl-N'-Nitro-N-Nitrosoguanidine. The optimal conditions for the formation and regeneration of protoplasts were examined and the characteristics of a fusant were studied. As a results, protoplasts were readily obtained from 18 hours cultured mycelia by the treatment of driselase for 3 hours and 0.6 M KCl as a best osmotic stabilizer at pH 6.0 for the formation of protoplast. Sucrose was the most suitable for the regeneration. Polyetylene glycol (M.W. 8,000) in $CaCl_{2}$-glycine solution was used to induce the protoplast fusion. The interspecific fusion frequency between protoplasts among the auxotrophic mutants of the two strains ranged from $2.7*10^{-2}$ to $5.7*10^{-3}$ . DNA content and cellulase activity were rather increased in the interspecific fusant. The lag phase of growth curve was slightly elongated in the fusant.