• The Korean Journal of Mycology
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• v.47 no.3
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• pp.187-197
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• 2019

Ascomycota is the largest phylum of the Fungi, including approximately 6,600 genera. They are often isolated from soils, indoor air, and freshwater environments, but also from plants as pathogens or endophytes. In this study, four species of Ascomycota (two of Cladosporium and one of each Daldinia and Nigrospora) were collected from the leaves of four woody plants (Camellia japonica, Ginkgo biloba, Quercus sp., Vitis vinifera). Their cultural characteristics were investigated on five different media (PDA, V8A, CMA, MEA, CZA) at 3 days after incubation at $25^{\circ}C$ in darkness. BLASTn search and phylogenetic analysis were performed using the internal transcribed spacer (ITS) rDNA sequences, in addition to tef1 gene sequences for Cladosporium species. Based on the cultural, morphological, and phylogenetic data, the isolates were identified as Cladosporium anthropophilum, Cladosporium pseudocladosporioides, Daldinia eschscholtzii, and Nigrospora chinensis. Previously, some members of Cladosporium and Nigrospora have been recorded as endophytes inhabiting the leaves and stems of various plants, whereas Daldinia eschscholtzii is a wood-inhabiting endophyte or wood-decaying fungus. To our knowledge, this is the first report of these four ascomycetes in Korea.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.1
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• pp.125-131
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• 2015

Currently, taxol is mainly extracted from the bark of yews; however, this method can not meet its increasing demand on the market because yews grow very slowly and are a rare and endangered species belonging to first-level conservation plants. Recently, increasing efforts have been made to develop alternative means of taxol production; microbe fermentation would be a very promising method to increase the production scale of taxol. To determine the activities of the taxol extracted from endophytic fungus N. sylviforme HDFS4-26 in inhibiting the growth and causing the apoptosis of cancer cells, on comparison with the taxol extracted from the bark of yew, we used cellular morphology, cell counting kit (CCK-8) assay, staining (HO33258/PI and Giemsa), DNA agarose gel electrophoresis and flow cytometry (FCM) analyses to determine the apoptosis status of breast cancer MCF-7 cells, cervical cancer HeLa cells and ovarian cancer HO8910 cells. Our results showed that the fungal taxol inhibited the growth of MCF-7, HeLa and HO8910 cells in a dose-and time-dependent manner. IC50 values of fungal taxol for HeLa, MCF-7 and HO8910 cells were $0.1-1.0{\mu}g/ml$, $0.001-0.01{\mu}g/ml$ and $0.01-0.1{\mu}g/ml$, respectively. The fungal taxol induced these tumor cells to undergo apoptosis with typical apoptotic characteristics, including morphological changes for chromatin condensation, chromatin crescent formation, nucleus fragmentation, apoptotic body formation and G2/M cell cycle arrest. The fungal taxol at the $0.01-1.0{\mu}g/ml$ had significant effects of inducing apoptosis between 24-48 h, which was the same as that of taxol extracted from yews. This study offers important information and a new resource for the production of an important anticancer drug by endofungus fermentation.

• Biomolecules & Therapeutics
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• v.24 no.2
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• pp.147-155
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• 2016

Chaetominine is a quinazoline alkaloid originating from the endophytic fungus Aspergillus fumigatus CY018. In this study, we showed evidence that chaetominine has cytotoxic and apoptotic effects on human leukemia K562 cells and investigated the pathway involved in chaetominine-induced apoptosis in detail. Chaetominine inhibited K562 cell growth, with an $IC_{50}$ value of 35 nM, but showed little inhibitory effect on the growth of human peripheral blood mononuclear cells. The high apoptosis rates, morphological apoptotic features, and DNA fragmentation caused by chaetominine indicated that the cytotoxicity was partially caused by its pro-apoptotic effect. Under chaetominine treatment, the Bax/Bcl-2 ratio was upregulated (from 0.3 to 8), which was followed by a decrease in mitochondrial membrane potential, release of cytochrome c from mitochondria into the cytosol, and stimulation of Apaf-1. Furthermore, activation of caspase-9 and caspase-3, which are the main executers of the apoptotic process, was observed. These results demonstrated that chaetominine induced cell apoptosis via the mitochondrial pathway. Chaetominine inhibited K562 cell growth and induced apoptotic cell death through the intrinsic pathway, which suggests that chaetominine might be a promising therapeutic for leukemia.

• Mycobiology
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• v.45 no.3
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• pp.178-183
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• 2017

Diabetes mellitus is a chronic disorder which affects millions of population worldwide. Global estimates published in 2010 reported the world diabetic prevalence as 6.4%, affecting 285 million adults. Foot ulceration and wound infection are major forms of disabilities arising from diabetic diseases. This study was aimed to develop a natural antimicrobial finishing on medical grade textile that meets American Association of Textiles Chemists and Colorists (AATCC) standard. The textile samples were finished with the ethanolic extract of Penicillium amestolkiae elv609, an endophytic fungus isolated from Orthosiphon stamineus Benth (common name: cat's whiskers). Endophyte is defined as microorganism that reside in the living plant tissue, without causing apparent disease symptom to the host. The antimicrobial efficacy of the ethanolic extract of P. minioluteum was tested on clinical pathogens isolated from diabetic wound. The extract exhibited significant inhibitory activity against 4 bacteria and 1 yeast with the minimal inhibitory concentration ranged from 6.25 to 12.5 mg/mL. The results indicate different susceptibility levels of the test microorganism to the ethanolic extract. However, the killing activity of the extract was concentration-dependent. The finished medical textile showed excellent antimicrobial efficacy on AATCC test assays. All the microbial cultures treated with the textile sample displayed a growth reduction of 99.9% on Hoheinstein Challenge Test. The wash durability of the finished textile was found good even after 50 washes with commercial detergent. Besides, the gas chromatography mass spectrometry analysis showed that 6-octadecenoic acid and diethyl phthalate were the main bioactive constituents of the extract. In conclusion, the developed medical textile showed good antimicrobial efficacy on laboratory tests. This work can be extended to in vivo trials for developing healthcare textile products for antimicrobial applications.

• Mycobiology
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• v.51 no.4
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• pp.230-238
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• 2023

Glabridin is a well-known active isoflavone found in the root of licorice (Glycyrrhiza glabra L.) that possess a wide range of biological activity. Plant cells, hairy roots, and fungal endophytes cultures are the most important alternative methods for plant resources conservation and sustainable production of natural compounds, which has received much attention in recent decades. In the present study, an efficient culture condition was optimized for the biomass accumulation and glabridin production from fungal endophyte Aspergillus eucalypticola SBU-11AE isolated from licorice root. Type of culture medium, range of pH, and licorice root extract (as an elicitor) were tested. The results showed that the highest and lowest biomass production was observed on PCB medium (6.43 ± 0.32 g/l) and peptone malt (5.85 + 0.11 g/l), respectively. The medium culture PCB was produced the highest level of glabridin (7.26 ± 0.44 mg/l), while the lowest level (4.47 ± 0.02 mg/l) was obtained from the medium peptone malt. The highest biomass (8.51 ± 0.43 g/l) and glabridin (8.30 ± 0.51 mg/l) production were observed from the PCB medium adjusted with pH = 6, while the lowest value of both traits was obtained from the same medium with pH = 7. The highest production of total glabridin (10.85 ± 0.84 mg/l) was also obtained from the culture medium treated with 100 mg/l of the plant root extract. This information can be interestingly used for the commercialization of glabridin production for further industrial applications.

• Journal of Life Science
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• v.20 no.9
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• pp.1373-1377
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• 2010

Coastal sand-dune plants can survive very effectively with the help of various microbes, especially ecto- and endomycorrihizae. Penicillium citrinum KACC43900 was screened according to growth promoting activity on sand-dune plants Calystegia soldanella and Ischaemum anthephoroides. In this study, coastal sand-dune plants were treated with a fungal culture filtrate of endophytic fungus P. citrinum KACC43900 to confirm the promotion of plant growth. C. soldanella and I. anthephoroides were used as representative coastal sand-dune plants, and their shoot length, plant length, and dry weight were analyzed. The shoot lengths of control C. soldanella and I. anthephoroides were 8.60 cm and 8.12 cm, and those of samples treated with fungal culture filtrates of C. soldanella and I. anthephoroides for 20 days were 16.30 cm and 10.56 cm, respectively. The plant lengths of control C. soldanella and I. anthephoroides were 14.90 cm and 14.80 cm, and those of samples treated with fungal culture filtrates of C. soldanella and I. anthephoroides for 20 days were 24.06 cm and 17.06 cm, respectively. The dry weight of C. soldanella and I. anthephoroides treated with fungal culture filtrates of C. soldanella and I. anthephoroides for 20 days were 0.163 g and 0.032 g, respectively. It was shown that the growth of shoots in C. soldanella and I. anthephoroides was promoted, 89.54% and 29.60%, by culture filtrate of P. citrinum KACC43900.

• The Korean Journal of Mycology
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• v.32 no.1
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• pp.8-15
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• 2004

This study was conducted to discover new phytotoxins which may be used as lead molecules for the development of new herbicides. A total of 187 endophytic fungi were isolated from 11 plant species, which were collected from 8 locations in Korea. Their herbicidal activities were screened in vivo by herbicidal and duckweed bioassays after they were cultured in potato dextrose broth and rice solid media. Both fermentation broth and solid culture extract of Gliocladium catenulatum F0006 isolated from red pine (Pinus densiflora) showed 70% herbicidal activity only against cocklebur (Xanthium strumarium) out of the 10 weeds tested. Solid culture extract of F0034 isolated from arrowroot (Pueraria thunbergiana) exhibited 20 to 100% herbicidal activities against all of the weeds. Especially, shattercane (Sorghum bicolor), barnyardgrass (Echinochloa crus-galli), large crabgrass (Digitaria sanguinalis), and fall pauicum (Panicum dichtomiflorum) were sensitive to the solid culture extract of F0034. In addition, solid culture extract of F0043 isolated from red pine displayed 20% to 70% herbicidal activities only against 5 grass species, but not against 5 broad-leaf plant species. On the other hand, as the results of duckweed assay, 8 fermentation broths showed 100% growth inhibitory activity at concentrations less than 5.0% of culture supernatants and 12 solid cultures had a potent inhibitory activity against duckweed growth. A toxic metabolite was purified from the solid cultures of G. catenulatum F0006 by repeated column chromatography and bioassay. It caused a phytotoxic syndrome only on cocklebur out of the 10 weeds tested; it completely killed cocklebur seedlings at $500\;{\mu}g/ml$ and showed 85% herbicidal activity against cocklebur at $100\;{\mu}g/ml$. The molecular weight of the toxic metabolite is 238 daltons and its structure determination is underway.

• Journal of Microbiology and Biotechnology
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• v.27 no.8
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• pp.1428-1440
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• 2017

Phomopsis sp. XP-8 (an endophytic fungus) was previously found to produce pinoresinol diglucoside (PDG), a major antihypertensive compound of Tu-Chung (the bark of Eucommia ulmoides Oliv.), which is widely used in Chinese traditional medicines. In the present study, two bioconversion systems were developed for the production of PDG in Tris-HCl buffer containing glucose and Phomopsis sp. XP-8 cells (both resting and freeze-dried). When other factors remained unchanged, the bioconversion time, glucose concentration, cell ages, cell dosage, pH, temperature, and stirring speed influenced PDG production in a similar and decreasing manner after an initial increase with increasing levels for each factor. Considering the simultaneous change of various factors, the optimal conditions for PDG production were established as 70 g/l cells (8-day-old), 14 g/l glucose, $28^{\circ}C$, pH 7.5, and 180 rpm for systems employing resting cells, and 3.87 g/l cells, 14.67 g/l glucose, $28^{\circ}C$, pH 7.5, and 180 rpm for systems employing freeze-dried cells. The systems employing freeze-dried cells showed lower peak PDG production ($110.28{\mu}g/l$), but at a much shorter time (12.65 h) compared with resting cells (23.62 mg/l, 91.5 h). The specific PDG production levels were 1.92 and $24{\mu}g$ per gram cells per gram glucose for freeze-dried cells and resting cells, respectively. Both systems indicated a new and potentially efficient way to produce PDG independent of microbial cell growth.

• Research in Plant Disease
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• v.14 no.2
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• pp.112-116
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• 2008

Powdery mildew of Paprika caused by Leveillula taurica has been a serious problem in greenhouse. It is an unusual endophytic powdery mildew because the mycelia grow inside the leaf, such that the pale yellow lesions on adaxial surfaces appear first and the white powdery lesion/signs develop later on the corresponding to the spots of the abaxial leaf surface, where the conidiophores are typically emerge through the stomatal opening. Although one foliar application of cooking oil and yolk mixture(COY) to the foliage was not practically effective enough, two or three, weekly application of COY to the foliage at either 0.3 or 0.5% concentration resulted in excellent control against powdery mildew with disease index less/lower than 1, respectively. This treatment could provide protection for three weeks, which, we believe, is not only cost-effective, but also environment-friendly. Powdery mildew fungus was affected by COY treatment quickly which is recognizable in three days. Net photosynthesis and evapotranspiration was remarkably reduced by powdery mildew infection compared to healthy leaves, suggesting that prevention and early protection is the most critical strategy for peak paprika fruit yield. Moreover, COY treatment did not adversely affect the photosynthesis and evapotranspiration of foliages.