• 식물병연구
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• 제12권3호
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• pp.168-177
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• 2006

한국농업미생물자원센터(Korean Agricultural Culture Collection, KACC)에 보존되어 있는 Colletotrichum gloeosporioides 39 균주와 C. acutatum 5 균주를 ribosomal DNA-ITS와 $\beta$-tubulin 부분염기서열과 PDA와 Benomyl-PDA배지에서의 생장 특성에 의하여 재동정 하였다. 그 결과 13 균주가 Talhinhas 등의 C. acutatum A2 그룹으로, 5 균주가 C. acutatum A3 그룹으로, 1 균주가 C. acutatum A4 그룹으로, 18 균주가 Moriwaki 등의 C. gloeosporioides ribosomal DNA group(RG) 4로 2 균주가 C. gioeosporioides RG6로 2 균주가 c. boninense RG5로 2 균주는 C. coccodes RG2 그리고 1 균주는 c. dematium RG12로 재동정되었다. 이들 중에서 한국에서 분리된 31 균주는 12 균주가 C. acutatum A2 그룹으로, 1 균주가 C. acutatum A3 그룹으로, 14 균주가 C. gloeosporioides RG4 로 2 균주가 C. gloeosporioides RG6로 1 균주가 C. boninense RG5로 그리고 1 균주는 C. dematium RG12로 동정되었다. 더불어 C. acutatum/C. gloeosporioides의 분류적인 특징과 국내 주요 기주별 분포 등에 대하여 고찰하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제33권12호
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• pp.1975-1984
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• 2020

Objective: The study was conducted to determine the effects of body weight (BW) and fiber sources on nutrient digestibility, fiber fermentation and short chain fatty acids (SCFA) concentration in different intestinal segments of growing pigs fed high-fiber diets. Methods: Nine barrows with initial BW of 25.17±0.73 kg and 9 barrows with initial BW of 63.47±2.18 kg were allotted to a duplicate 9×2 Youden Square design with 3 dietary treatments and 2 periods. The dietary treatments were formulated with 3 different high-fiber ingredients: corn bran, sugar beet pulp, and soybean hulls, respectively. Each diet was fed to 3 barrows with different stage of BW in each period. Results: There were no differences in the apparent ileal digestibility (AID) of most nutrients between pigs at different BW stages. Pigs at 60 kg had greater (p<0.05) apparent total tract digestibility (ATTD) of total dietary fiber (TDF), soluble dietary fiber (SDF) and insoluble dietary fiber (IDF), and had greater (p<0.05) hindgut disappearance of IDF and cellulose than pigs at 25 kg. The acetate, propionate and total SCFA concentrations in ileal digesta and feces of pigs at 60 kg were greater (p<0.05) than those of pigs at 25 kg. In addition, fiber sources affected (p<0.05) the AID of gross energy (GE), organic matter (OM), ether extract (EE), crude protein, SDF and hemicellulose, the hindgut disappearance and ATTD of dietary fiber components, the lactate and propionate concentrations in ileal digesta and the butyrate, valerate and total SCFA concentrations in feces. There were interactions (p<0.05) between BW and fiber sources on the AID of GE, OM, EE, SDF, hemicellulose, the ATTD of EE, TDF, and IDF, and the hindgut disappearance of SDF and hemicellulose. Conclusion: Increasing BW mainly improved the digestibility of dietary fiber fractions, and the dietary fiber sources influenced the digestibility of almost all the dietary nutrients in growing pigs.

• Asian Pacific Journal of Cancer Prevention
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• 제17권7호
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• pp.3309-3315
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• 2016

Helicobacter pylori as the second most common cause of gastric cancer in the world infects approximately half of the developed countries population and 80% of the population living in developing countries. Integrons as genetic reservoirs play major roles in dissemination of antimicrobial resistance genes. To the best of our knowledge, this is the first study to report carriage of class 1 and 2 integrons and associated gene cassettes in H. pylori isolates from Iran. This cross-sectional study was conducted in Tehran among 110 patients with H. pylori infection. Antimicrobial susceptibility testing (AST) for H. pylori strains were assessed by the micro broth dilution method. Class 1 and 2 integrons were detected using PCR. In order to determine gene cassettes, amplified fragments were subjected to DNA sequencing of both amplicon strands. The prevalence of resistance to clarithromycin, metronidazole, clarithromycin, tetracycline, amoxicillin, rifampin, and levofloxacin were 68.2% (n=75), 25.5% (n=28), 24.5% (n=27), 19.1% (n=21), 18.2% (n=20) and 16.4% (n=18), respectively. Frequency of multidrug resistance among H. pylori isolates was 12.7%. Class 2 integron was detected in 50 (45.5%) and class 1 integron in 10 (9.1%) H. pylori isolates. The most predominant gene cassette arrays in class 2 integron-bearing H. pylori were included sat-era-aadA1, dfrA1-sat2-aadA1, blaoxa2 and, aadB whereas common gene cassette arrays in class 1 integron were aadB-aadA1-cmlA6, aacA4, blaoxa2, and catB3. The high frequency of class 2 integron and multidrug resistance in the present study should be considered as a warning for clinicians that continuous surveillance is necessary to prevent the further spread of resistant isolates.

• Mycobiology
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• 제40권3호
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• pp.181-188
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• 2012

This study was initiated in order to investigate the anticancer and immunomodulating activities of crude polysaccharides extracted in methanol, neutral saline, and hot water (hereinafter referred to as Fr. MeOH, Fr. NaCl, and Fr. HW, respectively) from the fruiting bodies of Panellus serotinus. Content of ${\beta}$-glucan and protein in Fr. MeOH, Fr. NaCl, and Fr. HW extracts of P. serotinus ranged from 22.92~28.52 g/100 g and 3.24~3.68 g/100 g, respectively. In vitro cytotoxicity tests, none of the various fractions of crude polysaccharides were cytotoxic against sarcoma 180, HT-29, NIH3T3, and RAW 264.7 cell lines at the tested concentration. Intraperitoneal injection with crude polysaccharides resulted in a life prolongation effect of 23.53~44.71% in mice previously inoculated with sarcoma 180. Treatment with Fr. HW resulted in an increase in the numbers of spleen cells by 1.3 fold at the concentration of $50{\mu}g/mL$ compared with control. Treatment with Fr. NaCl resulted in improvement of the immuno-potentiating activity of B lymphocytes by increasing the alkaline phosphatase activity by 1.4 fold, compared with control, at the concentration of $200{\mu}g/mL$. Among the three fractions, maximum nitric oxide ($13.48{\mu}M$) was recorded at $500{\mu}g/mL$ in Fr. HW. Production of tumor necrosis factor alpha, interleukin-$1{\beta}$, and interleukin-6 was significantly higher, compared to the positive control, concanavalin A, at the tested concentration. Therefore, treatment with crude polysaccharides extracted from the fruiting body of P. serotinus could result in improvement of antitumor activity.

• Mycobiology
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• 제37권1호
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• pp.53-61
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• 2009

The process of biodegradation in lingo-cellulosic materials is critically relevant to biospheric carbon. The study of this natural process has largely involved laboratory investigations, focused primarily on the biodegradation and recycling of agricultural by-products, generally using basidiomycetes species. In order to collect super white rot fungi and evaluate its ability to degrade lingo-cellulosic material, 35 fungal strains, collected from forests, humus soil, livestock manure, and dead trees, were screened for enzyme activities and their potential to decolorize the commercially used Poly-R 478 dye. In the laccase enzymatic analysis chemical test, 33 white rot fungi and 2 brown rot fungi were identified. The degradation ability of polycyclic aromatic hydrocarbons (PAHs) according to the utilized environmental conditions was higher in the mushrooms grown in dead trees and fallen leaves than in the mushrooms grown in humus soil and livestock manure. Using Poly-R 478 dye to assess the PAH-degradation activity of the identified strains, four strains, including Agrocybe pediades, were selected. The activities of laccase, MnP, and Lip of the four strains with PAH-degrading ability were highest in Pleurotus incarnates. 87 fungal strains, collected from forests, humus soil, livestock manure, and dead trees, were screened for enzyme activities and their potential to decolorize the commercially used Poly-R 478 dye on solid media. Using Poly-R 478 dye to assess the PAHdegrading activity of the identified strains, it was determined that MKACC 51632 and 52492 strains evidenced superior activity in static and shaken liquid cultures. Subsequent screening on plates containing the polymeric dye poly R-478, the decolorization of which is correlated with lignin degradation, resulted in the selection of a strain of Coriolus versicolor, MKACC52492, for further study, primarily due to its rapid growth rate and profound ability to decolorize poly R-478 on solid media. Considering our findings using Poly-R 478 dye to evaluate the PAH-degrading activity of the identified strains, Coriolus versicolor, MKACC 52492 was selected as a favorable strain. Coriolus versicolor, which was collected from Mt. Yeogi in Suwon, was studied for the production of the lignin-modifying enzymes laccase, manganese-dependent peroxidase (MnP), and lignin peroxidase (LiP).

• Mycobiology
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• 제47권4호
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• pp.449-456
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• 2019

Invasive fungal infections caused by Cyberlindnera fabianii have recently increased. However, biochemical kits such as API 20 C AUX and Vitek-2C have misidentified this species as other Candida spp. such as C. pelliculosa or C. utilis due to no information of Cy. fabianii in yeast database. During our 2016-2017 surveys, eleven isolates of Cy. fabianii were obtained in International St. Mary's Hospital in Korea. Here, we describe its morphological and molecular characteristics and tested its antifungal susceptibility against nine antifungal agents. The sequences of the ITS region and the D1/D2 region of LSU revealed 100% identity with the sequences of Cy. fabianii. In comparison with the results from MALDI-TOF mass spectrometry, we found that Cy. fabianii can be distinguished from other species. In antifungal susceptibility test, voriconazole and echinocandins exhibited good antifungal activities against the majority of Cy. fabianii isolates despite the absence of standard criteria.

• Journal of Microbiology and Biotechnology
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• 제23권12호
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• pp.1654-1663
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• 2013

Rice straw is closely associated with meju fermentation and it is generally known that the rice straw provides meju with many kinds of microorganisms. In order to elucidate the origin of meju fungi, the fungal diversity of rice straw was examined. Rice straw was collected from 12 Jang factories where meju are produced, and were incubated under nine different conditions by altering the media (MEA, DRBC, and DG18), and temperature ($15^{\circ}C$, $25^{\circ}C$, and $35^{\circ}C$). In total, 937 strains were isolated and identified as belonging to 39 genera and 103 species. Among these, Aspergillus, Cladosporium, Eurotium, Fusarium, and Penicillium were the dominant genera. Fusarium asiaticum (56.3%), Cladosporium cladosporioides (48.6%), Aspergillus tubingensis (37.5%), A. oryzae (31.9%), Eurotium repens (27.1%), and E. chevalieri (25.0%) were frequently isolated from the rice straw obtained from many factories. Twelve genera and 40 species of fungi that were isolated in the rice straw in this study were also isolated from meju. Specifically, A. oryzae, C. cladosporioides, E. chevalieri, E. repens, F. asiaticum, and Penicillium polonicum (11.8%), which are abundant species in meju, were also isolated frequently from rice straw. C. cladosporioides, F. asiaticum, and P. polonicum, which are abundant in the low temperature fermentation process of meju fermentation, were frequently isolated from rice straw incubated at $15^{\circ}C$ and $25^{\circ}C$, whereas A. oryzae, E. repens, and E. chevalieri, which are abundant in the high temperature fermentation process of meju fermentation, were frequently isolated from rice straw incubated at $25^{\circ}C$ and $35^{\circ}C$. This suggests that the mycobiota of rice straw has a large influence in the mycobiota of meju. The influence of fungi on the rice straw as feed and silage for livestock, and as plant pathogens for rice, are discussed as well.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2015년도 추계학술대회 및 정기총회
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• pp.37-37
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• 2015

dsRNA was found in malformed cultures of Lentinula edodes strain FMRI0339, one of the three most popular sawdust cultivated commercial strains of shiitake, and was also found in healthy-looking fruiting bodies and actively growing mycelia. Cloning of the partial genome of the dsRNA revealed the presence of the RdRp sequence of a novel L. edodes mycovirus (LeV), and sequence comparison of the cloned amplicon showed an identical sequence to known RdRp genes of LeV found in strain HKA. The meiotic stability of dsRNA was examined by measuring the ratio of the presence of dsRNA among sexual monokaryotic progeny. More than 40% of the monokaryotic progeny still contained the dsRNA, indicating the persistence of dsRNA during sexual reproduction. Comparing the mycelia growth of monokaryotic progeny suggested that, although variations in the growth rate existed among progeny and virus infection was observed in highly actively growing progeny, there appeared to be a tendency toward a lower frequency of virus incidence in actively growing progeny. This study attempted to cure the edible mushroom L. edodes strain FMRI0339 of the L. edodes mycovirus (LeV) in order to obtain an isogenic virus-free fungal strain as well as a virus-infected strain for comparison. Mycelial fragmentation, followed by being spread on a plate with serial dilutions resulted in a virus-free colony. Viral absence was confirmed with gel electrophoresis after dsRNA-specific virus purification, Northern blot analysis, and PCR using reverse transcriptase (RT-PCR). Once cured, all of fungal cultures remained virus-free over the next two years. Interestingly, the viral titer of LeV varied depending on the culture condition. The titer from the plate culture showed at least a 20-fold higher concentration than that grown in the liquid culture. However, the reduced virus titer in the liquid culture was recovered by transferring the mycelia to a plate containing the same medium. In addition, oxygen-depleted culture conditions resulted in a significant decrease of viral concentration, but not to the extent seen in the submerged liquid culture. Although no $discernable phenotypic changes in colony morphology were observed, virus-cured strains showed significantly higher growth rates and mycelial mass than virus-infected strains. We were also explored effects of LeV on fruiting body formation and mushroom yield. The fruiting body formation yield of virus-free L. edodes was larger than virus-infected L. edodes. These results indicate that LeV infection has a deleterious effect on mycelial growth and fruiting body formation. In addition, we have been investigated host-parasite interaction between L. edodes and its mycovirus interaction to study viral mechanism by establishment of proteomics.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2015년도 추계학술대회 및 정기총회
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• pp.41-41
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• 2015

Oomycetes belong to the kingdom Straminipila, a remarkably diverse group which includes brown algae and planktonic diatoms, although they have previously been classified under the kingdom Fungi. These organisms have evolved both saprophytic and pathogenic lifestyles, and more than 60% of the known species are pathogens on plants, the majority of which are classified into the order Peronosporales (includes downy mildews, Phytophthora, and Pythium). Recent phylogenetic investigations based on DNA sequences have revealed that the diversity of oomycetes has been largely underestimated. Although morphology is the most valuable criterion for their identification and diversity, morphological species identification is time-consuming and in some groups very difficult, especially for non-taxonomists. DNA barcoding is a fast and reliable tool for identification of species, enabling us to unravel the diversity and distribution of oomycetes. Accurate species determination of plant pathogens is a prerequisite for their control and quarantine, and further for assessing their potential threat to crops. The mitochondrial cox2 gene has been widely used for identification, taxonomy and phylogeny of various oomycete groups. However, recently the cox1 gene was proposed as a DNA barcode marker instead, together with ITS rDNA. To determine which out of cox1 or cox2 is best suited as universal oomycete barcode, we compared these two genes in terms of (1) PCR efficiency for 31 representative genera, as well as for historic herbarium specimens, and (2) in terms of sequence polymorphism, intra- and interspecific divergence. The primer sets for cox2 successfully amplified all oomycete genera tested, while cox1 failed to amplify three genera. In addition, cox2 exhibited higher PCR efficiency for historic herbarium specimens, providing easier access to barcoding type material. In addition, cox2 yielded higher species identification success, with higher interspecific and lower intraspecific divergences than cox1. Therefore, cox2 is suggested as a partner DNA barcode along with ITS rDNA instead of cox1. Including the two barcoding markers, ITS rDNA and cox2 mtDNA, the multi-locus phylogenetic analyses were performed to resolve two complex clades, Bremia lactucae (lettuce downy mildew) and Peronospora effuse (spinach downy mildew) at the species level and to infer evolutionary relationships within them. The approaches discriminated all currently accepted species and revealed several previously unrecognized lineages, which are specific to a host genus or species. The sequence polymorphisms were useful to develop a real-time quantitative PCR (qPCR) assay for detection of airborne inoculum of B. lactucae and P. effusa. Specificity tests revealed that the qPCR assay is specific for detection of each species. This assay is sensitive, enabling detection of very low levels of inoculum that may be present in the field. Early detection of the pathogen, coupled with knowledge of other factors that favor downy mildew outbreaks, may enable disease forecasting for judicious timing of fungicide applications.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2015년도 추계학술대회 및 정기총회
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• pp.11-11
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• 2015

A total of sixty-six samples of Nuruk, a fermention starter used to make the Korean traditional rice wine, Makgeolli, were collected from central and southern regions of Korea in 2013 and 2014. We classified two groups of the Nuruk samples, "commercial" and "home-made", according to the manufacturing procedure and purpose of use. Commercial Nuruks were made in a controlled environment where the temperature and humidity are fixed and the final product is supplied to Makgeolli manufacturers. Home-made Nuruks were made under uncontrolled conditions in the naturally opened environment and were intended for use in the production of small amounts of home-brewed Makgeolli. We obtained more than five hundred isolates including filamentous fungi and yeasts from the Nuruk samples followed by identification of fungal species. Also we stored glycerol stocks of each single isolate at $-70^{\circ}C$. We identified the species of each isolate based on the sequences of ITS regions amplified with two different universal primer pairs. We also performed morphological characterization of the filamentous fungi and yeast species through observations under the microscope. We investigated the major fungal species of commercial and home-made Nuruks by counting the colony forming units (CFU) and analyzing the occurrence tendency of fungal species. While commercial Nuruks contained mostly high CFU of yeasts, home-made Nuruks showed relatively high occurrence of filamentous fungi. One of the representative Nuruk manufacturers used both domestic wheat bran and imported ones, mainly from US, as raw material. Depending on the source of ingredient, the fungal diversity was somewhat different. Another commercial Nuruk sample was collected twice, once in 2013 and again in 2014, and showed different diversity of fungal species in each year. Nuruks obtained from the southern regions of Korea and Jeju island showed high frequency of yeast such as Saccharomycopsis fibuligera and Pichia species as well as unique filamentous fungus, Monascus species. S. fibuligera was easily found in many Nuruk samples with high CFU. The major filamentous fungi were Aspergillus, Lichtheimia, Mucor and Penicillium species. In order to further our understanding of the isolates and their potential industrial applications, we assayed three enzymes, alpha amylase, glucoamylase and acid protease from 140 isolates out of about five hundred isolates and selected about 10 excellent strains with high enzyme activities. With these fungal isolates, we will perform omics analyses including genomics, transcriptomics, metabolic pathway analyses, and metabolomics followed by whole genome sequencing of unique isolates associated with the basic research of Nuruk and that also has applications in the Makgeolli making process.