• Mycobiology
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• v.51 no.5
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• pp.281-287
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• 2023

The symbiotic association between fungus-gardening termites Macrotermes and its fungal symbiont has a moderate degree of specificity-although the symbiotic fungi (Termitomyces) form a monophyletic clade, there is not a one-to-one association between termite species and their fungus-garden associates. Here, we aim to determine the origin and phylogenetic relationships of Termitomyces in Oman. We used sequences of the internal transcribed spacer region (ITS) and the nuclear large subunit ribosomal RNA (LSU rRNA, 25S) gene and analyzed these with sequences of Termitomyces from other geographic areas. We find no evidence for more than a single colonization of Oman by Termitomyces. Unexpectedly, we find Termitomyces in Oman is most closely related to the symbiont of M. subhyalinus in West Africa rather than to those of geographically closer populations in East Africa.

• Environmental Engineering Research
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• v.11 no.4
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• pp.173-180
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• 2006

Autotrophic nitrogen removal and its microbial community from a laboratory scale upflow anaerobic sludge bed reactor were characterized with dynamic behavior of nitrogen removal and sequencing result of molecular technique (DNA extraction, PCR and amplification of 16S rDNA), respectively. In the experiment treating inorganic wastewater, the anaerobic granular sludge from a full-scale UASB reactor treating industrial wastewater was inoculated as seed biomass. The operating results revealed that an addition of hydroxylamine would result in lithoautotrophic ammonium oxidation to nitrite/nitrate, and also hydrazine would play an important role for the success of sustainable nitrogen removal process. Total N and ammonium removal of 48% and 92% was observed, corresponding to nitrogen conversion of 0.023 g N/L-d. The reddish brown-colored granular sludge with a diameter of $1{\sim}2\;mm$ was observed at the lower part of sludge bed. The microbial characterization suggests that an anoxic ammonium oxidizer and an anoxic denitrifying autotrophic nitrifier contribute mainly to the nitrogen removal in the reactor. The results revealed the feasibility on development of high performance lithoautotrophic nitrogen removal process with its microbial granulation.

• Journal of Life Science
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• v.28 no.1
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• pp.99-104
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• 2018

Streptococcus pneumoniae is one of the major pathogens in community-acquired diseases, and it contains several factors that promote its pathogenesis, including pneumolysin (PLY). PLY is a member of the cholesterol-dependent cytolysin family, which attacks cholesterol-containing membranes, thereby forming ring-shaped pores. Thus, it is a major key target for vaccines against pneumococcal disease. We cloned the PLY gene from S. pneumoniae D39 and inserted it into the pQE-30 vector. Recombinant PLY (rPLY) was overexpressed in Escherichia coli M15 and purified by $Ni^{2+}$ affinity chromatography. Similarly, a PLY-EGFP fusion gene was produced by inserting the EGFP gene at the 3' end of the PLY gene in the same vector, and the recombinant protein was purified. Sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) showed that both recombinant proteins were purified. rPLY exhibited significant hemolytic activity against 1% human red blood cells (RBCs). Complete hemolysis was obtained at 500 ng/ml, and 50% hemolysis was found with a 240 ng/ml concentration. In contrast, rPLY-EGFP did not show hemolytic activity. However, rPLY-EGFP did bind the RBC membrane, indicating that rPLY-EGFP lost hemolytic activity via EGFP fusion, while retaining its membrane-binding ability. These data suggest that PLY's C terminus is important for its hemolytic activity. Therefore, these two recombinant proteins can be extremely useful for investigating the toxin mechanism of PLY and cell damage during pneumonia.

• The Korean Journal of Mycology
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• v.45 no.3
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• pp.246-250
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• 2017

Sclerotium rot was observed on mungbean (Phaseolus radiatus L.) plants cultivated in the exhibition field of Gyeongsangnam-do Agricultural Research and Extension Services in September 2015. The progression of rot was initially observed as water-soaked lesions on several parts of the affected plant. Severely infected plants were blighted and eventually died. White mycelial mats spread over the lesions and numerous sclerotia formed on stems near the soil line. The sclerotia were globoid in shape, 1~3 mm in size, and white to brown in color. The optimum temperature for mycelial growth and sclerotia formation on potato dextrose agar (PDA) was $30^{\circ}C$ and the hyphal width was $4{\sim}8{\mu}m$. Typical clamp connections were observed on the hyphae of fungus grown on PDA. For molecular identification, the complete internal transcribed spacer (ITS) ribosomal DNA (rDNA) of the causal fungus was sequenced and analyzed. Based on the mycological characteristics, ITS rDNA sequence analysis, and pathogenicity to host plants, the fungus was identified as Sclerotium rolfsii. This is the first report of Sclerotium rot on mungbean caused by S. rolfsii in Korea.

• Journal of Microbiology and Biotechnology
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• v.12 no.4
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• pp.544-552
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• 2002

Several microorganisms from rat and human feces and lumen fluid of cows were screened for their ability to decolorize the synthetic dyes. Consequently, a novel dye-degrading strain AB&J was isolated. Taxonomic identification including 165 rDNA sequencing and phylogenetic analysis indicated that the isolate had 99.9% homology in its 165 rDNA base sequence with Clostridium perfringens. After 27 h Incubation with the strain, brilliant blue R, bromophenol blue, crystal violet, malachite green, methyl green, and methyl orange were decolorized by about 69.3%, 97.7%, 96.3%, 97.9%, 75.1%, and 97.2%, respectively. The triphenlmethane dye, bromophenol blue, was decolorized extensively by growing Clostridium perfringens AB&J cells in liquid cultures under anaerobic condition, although their growth was strongly inhibited in the initial stage of incubation. This group of dyes is toxic, depending on the concentration used. The dye was significantly decolorized at a relatively lower concentration of below 50 $\mu g \;ml^{-1}$, however, the growth of the cells was mostly suppressed at a dye concentration of 100 $\mu g \;ml^{-1}$. The decolorization activity in cell-free extracts was much higher in cytoplasm than in periplasm and cytoplasmic membrane. Therefore, the enzyme related uptake of bromophenol blue seemed to be localized in cytoplasm. The optimal pH and temperature of bromophenol blue uptake fur decolorization activities were 7.0 and 4$0^{\circ}C$, respectively.

• Journal of Microbiology and Biotechnology
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• v.25 no.5
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• pp.620-628
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• 2015

A fungal strain, R9, was isolated from the South Atlantic sediment sample and identified as Aspergillus fumigatus. An antifungal protein, AfAFP_R9, was purified from the culture supernatant of Aspergillus fumigatus R9. AfAFP_R9 was identified to be restrictocin, which is a member of the ribosome-inactivating proteins (RIPs), by MALDI-TOF-TOF-MS. AfAFP_R9 displayed antifungal activity against plant pathogenic Fusarium oxysporum, Alternaria longipes, Colletotrichum gloeosporioides, Paecilomyces variotii, and Trichoderma viride at minimum inhibitory concentrations of 0.6, 0.6, 1.2, 1.2, and 2.4 μg/disc, respectively. Moreover, AfAFP_R9 exhibited a certain extent of thermostability, and metal ion and denaturant tolerance. The iodoacetamide assay showed that the disulfide bridge in AfAFP_R9 was indispensable for its antifungal action. The cDNA encoding for AfAFP_R9 was cloned from A. fumigatus R9 by RT-PCR and heterologously expressed in E. coli. The recombinant AfAFP_R9 protein exhibited obvious antifungal activity against C. gloeosporioides, T. viride, and A. longipes. These results reveal the antifungal properties of a RIP member (AfAFP_R9) from marine-derived Aspergillus fumigatus and indicated its potential application in controlling plant pathogenic fungi.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.sup3
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• pp.219-224
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• 2016

Hepatocellular carcinoma (HCC) is the most common primary malignancy of the liver making up more than 80 percent of cases. It is known to be the sixth most prevalent cancer and the third most frequent cause of cancer related death worldwide. Epigenetic regulation constitutes an important mechanism by which dietary components can selectively activate or inactivate target gene expression. The miR-34 family members including mir-34a, mir-34b and mir-34c are tumor suppressor micro RNAs, which are expressed in the majority of normal tissues. Several studies have indicated silencing of miR-34 expression via DNA methylation in multiple types of cancers. Bioactive nutrients like curcumin (Cur) have excellent anticarcinogenic activity and minimal toxic manifestations in biological systems. This compound has recently been determined to induce epigenetic changes. However, Cur is lipophilic and has a poor systemic bioavailability and poor absorption. Its bioavailability is increased through employing dendrosome nanoparticles. The aim of the current study was to investigate the effect of dendrosomal nanocurcumin (DNC) on expression of mir-34 family members in two HCC cell lines, HepG2 and Huh7. We performed the MTT assay to evaluate DNC and dendrosome effects on cell viability. The ability of DNC to alter expression of the mir-34 family and DNA methyltransferases (DNMT1, DNMT3A and 3B) was evaluated using semi-quantitative and quantitative PCR. We observed the entrance of DNC into HepG2 and Huh7 cells. Gene expression assays indicated that DNC treatment upregulated mir34a, mir34b and mir34c expression (P<0.05) as well as downregulated DNMT1, DNMT3A and DNMT3B expression (P<0.05) in both HepG2 and Huh7 cell lines. DNC also reduced viability of Huh7 and HepG2 cells through restoration of miR-34s expression. We showed that DNC could awaken the epigenetically silenced miR-34 family by downregulation of DNMTs. Our findings suggest that DNC has potential in epigenetic therapy of HCC.

• Korean Journal of Breeding Science
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• v.42 no.1
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• pp.50-55
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• 2010

The R-r:standard (R-r:std) allele of maize R gene complex consists of S subcomplex and P component; the S subcomplex regulates anthocyanin pigmentation of seed aleurone layer, and the P component confers pigmentation of the other plant parts. The S subcomplex contains two functional genes, S1 and S2 components. In the presence of Pl gene some alleles of R gene induce anthocyanin pigmentation of pericarp. In the present study, the effects of different R alleles on the anthocyanin pigmentation of pericarp in the presence of Pl gene were analyzed in order to identify the R gene component responsible for pericarp pigmentation. The results show that R-ch and r-ch alleles condition similar degrees of pericarp pigmentation, and that R-r:Ecuador (R-r:Ec) conditions stronger pigmentation. The r-ch allele, which is inferred that its S subcomplex has lost function but the P component is normal, induces pericarp pigmentation in the presence of Pl gene. On the contrary, the R-g:g1111 allele, derived from R-r:Ec and inferred that its S subcomplex functions normal but the P component has lost its function, did not induce pericarp pigmentation in the presence of Pl gene. Moreover, PCR analysis of genomic DNA's of R-ch and r-ch indicate that R-ch maintains both P and S1 components, whereas r-ch lacks for the S1 component. Taken together, The results suggest that the P components of R alleles inducing pericarp pigmentation in the presence of Pl gene are responsible for pericarp pigmentation.

• Research in Plant Disease
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• v.17 no.3
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• pp.386-390
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• 2011

In June 2011, walnuts (Juglans regia) from orchards in Kimcheon and Muan, in southern Korea, were found to exhibit downy mildew-like symptoms of a foliar disease. Whitish polygonal efflorescence was produced on the abaxial surface of affected leaves and discolored light green blotches on the corresponding adaxial surfaces. In the later stage of disease development, diseased tissues collapsed and became necrotic. Based on morphological and cultural characteristics, the causal fungus was identified as Microstroma juglandis. The sequence of ITS rDNA of the present isolate showed 100% similarity with those of M. juglandis obtained from GenBank databases, thus confirming its identity. Pathogenicity tests were conducted on leaves of walnut seedlings, fulfilling Koch's postulates. The disease has been previously reported in North America, Europe, Oceania and some western Asia. This is the first report of downy leaf spot on walnuts in East Asia.

• Journal of Microbiology and Biotechnology
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• v.19 no.3
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• pp.314-322
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• 2009

Four different bacterial colonies were isolated from an old stock of an entomopathogenic nematode, Steinernema monticolum. They all showed entomopathogenicity to final instar larvae of beet armyworm, Spodoptera exigua, by hemocoelic injection. However, they varied in colony form, susceptibility to antibiotics, and postmortem change of the infected host insects. Biolog microbial identification and 16S rDNA sequence analyses indicate that these are four different species classified into different bacterial genera. Owing to high entomopathogenicity and a cadaver color of infected insect host, Serratia sp. was selected as a main symbiotic bacterial species and analyzed for its pathogenicity. Although no virulence of Serratia sp. was detected at oral administration, the bacteria gave significant synergistic pathogenicity to fifth instar S. exigua when it was treated along with a spore-forming entomopathogenic bacterium, Bacillus thuringiensis. The synergistic effect was explained by an immunosuppressive effect of Serratia sp. by its high cytotoxic effect on hemocytes of S. exigua, because Serratia sp. caused septicemia of S. exigua when the bacterial cells were injected into S. exigua hemocoel. The cytotoxic factor(s) was present in the culture medium because the sterilized culture broth possessed high potency in the cytotoxicity, which was specific to granular cells and plasmatocytes, two main immune-associated hemocytes in insects.