• Korean Journal of Microbiology
• /
• v.27 no.2
• /
• pp.98-107
• /
• 1989

Intraspecific fusants, produced by protoplast fusion of auxotrophic mutants from Trichoderma koningii ATCC 26113, were segregated into various strains including parental types, non-parental auxotrophic hybrids, and prototrophic hybrids on complete plate. Interestingly, some of non-parental prototrophic hybrids revealed to have enhanced cellulolytic activity incomparison with other strains of parents or hybrids derived thereafter. It was also evident that prototrophic hybrids of aneuploid could be constructed after the spontaneous segregation of complementing fusants produced through the protoplast fusion.

• Journal of Microbiology
• /
• v.34 no.4
• /
• pp.327-333
• /
• 1996

Yeast, like many other microbes, encounters large variations in ambient pH in their natural environments. Microorganisms capable of growing over a wide pH range require a versatile, efficient pH homeostatic mechanism protecting intracellular processes against extremes of pH. In several organisms, fusions to the bacterial lacZ gene have been extremely useful for the identification of genes expressed at different time during the life cycle or under different growth conditions. In this study, using the lacZ gene screening system, we surveyed a large number of yeast strains with lacZ insertion to identify genes regulated by pH. A yeast genomic library was constructed and inserted with lacZ by a shuttle mutagenesis procedure. The yeast transformants were individually picked up with a toothpick, replica-plated, and grown in alkaline pH medium. Among the 35,000 colonies screened, 10 candidate strains were identified initially by the $\beta$-gal assay. We finally confirmed two yeast strains carrying the genes whose expression are strictly dependent on pH of growth medium. One of the fusions showing a 10-fold induction in expression level in response to alkali pH was selected and further characterized. The pH-regulated gene was cloned by inverse PCR and a partial sequence of the gene was determined. Identification and characterization of the gene is currently under investigation.

• Korean Journal of Soil Science and Fertilizer
• /
• v.45 no.1
• /
• pp.51-58
• /
• 2012

Salinity is one of the most relevant abiotic factor limiting crop yield and its net primary productivity. In addition, salinity induces an increased stress ethylene synthesis in plants which, in turn, exacerbate the responses to the stressor. Bacterial single or co-inoculation effect was tested using previously characterized plant growth promoting (PGP) bacteria Brevibacterium iodinum RS16 and Methylobacterium oryzae CBMB20 on maize and sorghum-sudan grass hybrid under different concentrations of NaCl. Non-inoculated maize and sorghum-sudangrass hybrid showed 33.4% and 20.0% reduction in seed germination under highest NaCl (150 mM) level tested. However, under the same NaCl concentration, co-inoculation with B. iodinum RS16 and M. oryzae CBMB20 PGP strains increased the seed germination in maize (16.7%) and sorghum-sudangrass hybrid (4.4%). In Gnotobiotic growth pouch experiments conducted for maize and sorghum-sudangrass hybrid, co-inoculation of PGP B. iodinum RS16 and M. oryzae CBMB20 mitigated the salinity stress and promoted root length by 22.9% and 29.7%, respectively. Thus the results of this study could help in development of potential bioinoculants that may be suitable for crop production under saline conditions.

• Journal of Mushroom
• /
• v.2 no.2
• /
• pp.109-113
• /
• 2004

Pleurotus eryngii is not edible and medicinal mushrooms indigenous to Korea. To improvement of strain suitable to the geographic setting of Korea, we are mated with 22 dikaryons and 47 monokaryons isolated from Pleurotus eryngii ASI 2547 by Di-Mon mating. 19 strains forming fruit body obtained from clamped 253 bred strains. 7 excellent strains are selected from 19 bred strains by various morphological features of fruit body. Among the selected 7 strains, H6 strain were identified into ASI 2547-like recombinant hybrids with URP uniprimer by RAPD analysis. This suggested that Di-Mon crossing is one of rapid and easy breeding method for strain improvement with molecular techniques.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.29 no.2
• /
• pp.198-206
• /
• 2014

The olive fly Bactrocera oleae (Rossi) is the key pest for olive cultivation worldwide. Substantial effort has been invested in the development of the sterile insect technique (SIT) to control this pest. One of the limitations to develop SIT technology for olive fruit fly is the low ability of wild females to lay eggs in other medium than olive fruits, and their slow adaptation to oviposition in artificial substrates. In the present study, fruit grapes were used as an alternative egg collection medium to harvest eggs and young larvae from freshly colonized wild strains originating from France, Italy, Spain and Croatia. The larvae were allowed to develop into the fruits until the second instar, before they were extracted out and further reared on a standard artificial diet. Furthermore, F1 to F4 female flies were alternatively offered wax bottles to oviposit. Finally, the performance of hybrid strains created from crosses between wild and long colonised flies was assessed. The results showed that females of all 4 wild strains readily oviposited eggs in grapes and from the F2 generation onward, females from all strains were adapted to laying eggs in wax bottles. No difference was observed in eggs and pupae production among all strains tested. The findings are discussed for their implications on SIT application against olive fruit fly.

• The Korean Journal of Mycology
• /
• v.49 no.3
• /
• pp.405-412
• /
• 2021

Wolfiporia cocos is an important medicinal fungus that has been used in regions of Northeast Asia including Korea, Japan, and China. W. cocos is classified in Korea into two types (red bokryeong and white bokryeong) based on the internal colors (yellow orange-pale pink and white) of the sclerotium. Generally, the W. cocos type cultivated on farms produces white sclerotium. In this study, we endeavored to select strains that form sclerotium in sawdust medium using 2-way cross-breeding among two cultivated strains and three wild strains. Monospores were isolated from the fruiting bodies of cultivated and wild strains on potato dextrose agar. Thirty-nine strains of 338 hybrid strains isolated formed sclerotia with white or yellow colors upon culture for 3 months in Pinus densiflora sawdust medium. Selection for sclerotium forming strains using sawdust culture follows a very simple and easy procedure that is presented for the first time in this paper. We plan to test selected strains in the field to aid in developing new varieties for the future.

• Journal of Microbiology and Biotechnology
• /
• v.26 no.1
• /
• pp.207-212
• /
• 2016

PacBio's long-read sequencing technologies can be successfully used for a complete bacterial genome assembly using recently developed non-hybrid assemblers in the absence of second-generation, high-quality short reads. However, standardized procedures that take into account multiple pre-existing second-generation sequencing platforms are scarce. In addition to Illumina HiSeq and Ion Torrent PGM-based genome sequencing results derived from previous studies, we generated further sequencing data, including from the PacBio RS II platform, and applied various bioinformatics tools to obtain complete genome assemblies for five bacterial strains. Our approach revealed that the hierarchical genome assembly process (HGAP) non-hybrid assembler resulted in nearly complete assemblies at a moderate coverage of ~75x, but that different versions produced non-compatible results requiring post processing. The other two platforms further improved the PacBio assembly through scaffolding and a final error correction.

• 한국균학회소식:학술대회논문집
• /
• 2015.05a
• /
• pp.60-60
• /
• 2015

Hybrid histidine kinase is a part of two-component system that is required for various stress responses and pathogenesis of pathogenic fungi. In the present study, Tco1, a homologue of human pathogen Cryptococcus neoformans Tco1 encoding a hybrid histidine kinase, was identified in corn smut pathogen Ustilago maydis by bioinformatic analysis. To explore the role of Tco1 in the virulence of U. maydis, mutants in which the tco1 gene was partially deleted were constructed by allelic exchange. The U. maydis tco1 mutants did show unaltered growth rate on axenic medium but were unable to produce conjugation tubes and develop fuzzy filaments, resulting in impaired mating of compatible strains. The expression levels of prf1, pra1, and mfa1 which are involved in the pheromone pathway significantly decreased in the tco1 mutants. In inoculation tests to host, the tco1 mutants showed significantly reduced ability in the production of anthocyanin pigments and tumor development on maize leaves. Overall, the combined results indicated that Tco1 plays important roles in sexual development and virulence of U. maydis by regulating the expression of the genes involved in the pheromone pathway.

• Proceedings of the Korean Society for Applied Microbiology Conference
• /
• 2002.10a
• /
• pp.18-25
• /
• 2002

The pikromycin biosynthetic system in Streptomyces venezuleae is unique for its ability to produce two groups of antibiotics that include the 12-membered ring macrolides methymycin and neomethymycin, and the 14-membered ring macrolides narbomycin and pikromycin. The metabolic pathway also contains two post polyketide-modification enzymes, a glycosyltransferase and P450 hydroxylase that have unusually broad substrate specificities. In order to explore further the substrate flexibility of these enzymes a series of hybrid polyketide synthases were constructed and their metabolic products characterized. The plasmid-based replacement of the multifunctional protein subunits of the pikromycin PKS in S. venezuelae by the corresponding subunits from heterologous modular PKSs resulted in recombinant strains that produce both 12- and 14-membered ring macrolactones with predicted structural alterations. In all cases, novel macrolactones were produced and further modified by the DesVII glycosyltransferase and PikC hydroxylase leading to biologically active macrolide structures. These results demonstrate that hybrid PKSs in S. venezuelae can produce a multiplicity of new macrolactones that are modified further by the highly flexible DesVII glycosyltransferase and PikC hydroxylase tailoring enzymes. This work demonstrates the unique capacity of the S. venezuelae pikromycin pathway to expand the toolbox of combinatorial biosynthesis and to accelerate the creation of novel biologically active natural products. The polyketide backbone of rifamycin B is assembled through successive condensation and ${\beta}$-carbonyl processing of the extender units by the modular rifamycin PKS. The eighth module, in the RifD protein, contains nonfunctional DH domain and functional KR domain, which specify the reduction of the ${\beta}$-carbonyl group resulting in the C-21 bydroxyl of rifamycin B. A four amino acid substitution and one amino acid deletion were introduced in the putative NADPH binding motif in the proposed KR domain encoded by rifD. This strategy of mutation was based on the amino acid sequences of the corresponding motif of the KR domain of module 3 in the RifA protein, which is believed dysfunctional, so as to introduce a minimum alteration and retain the reading frame intact, yet ensure loss of function. The resulting strain produces linear polyketides, from tetraketide to octaketide, which are also produced by a rifD disrupted mutant as a consequence of premature termination of polyketide assembly. Much of the structural diversity within the polyketide superfamily of natural products is due to the ability of PKSs to vary the reduction level of every other alternate carbon atom in the backbone. Thus, the ability to introduce heterologous reductive segments such as ketoreductase (KR), dehydratase (DH), and enoylreductase (ER) into modules that naturally lack these activities would increase the power of the combinatorial biosynthetic toolbox. The dehydratase domain of module 7 of the rifamycin PKS, which is predicted to be nonfunctional in view of the sequence of the apparent active site, was replaced with its functional homolog from module 7 of rapamycin-producing polyketide synthase. The resulting mutant strain behaved like a rifC disrupted mutant, i.e., it accumulated the heptaketide intermediate and its precursors. This result points out a major difficulty we have encountered with all the Amycolatopsis mediterranei strain containing hybrid polyketide synthases: all the engineered strains prepared so far accumulate a plethora of products derived from the polyketide chain assembly intermediates as major products instead of just analogs of rifamycin B or its ansamycin precursors.

• Korean Journal of Microbiology
• /
• v.23 no.1
• /
• pp.56-63
• /
• 1985

We have characterized the plasmids of zymomonas, and constructed E. coli-Zymomonas shuttle vector. Plasmids have been detected in four strains of Zymomonas mobilis. All strains tested had at least one plasmid ranging in size from about 1.7 to 46kb. Antibiotics resistances of Z. mobilis were tested to select the host strain. All strains were very sensitive to tetracycline and chloramphenicol. Homology tests between the plasmids in four strains showed that the plasmids of ATCC10988 is highly homologous to those of ZM1, and that there is no homology between plasmids of ZM4 and Agll. The 1.7kb plasmid of ATCC10988, named as pZM886, also has no homology with plasmids of ZM4. A hybrid plasmid, designated to pBZ41, was constructed from pZM886 and pBR322. A restriction map of pBZ41 was established. Replicon of pZM886 didn't operate in E.coli and pBR322 seemed not to replicate in Zymomonas. pBZ41 was transfered from E. coli to Zymomonas by conjugal mobilization. The transconjugants were resistant to tetracycline and maintained pBZ41 stably.