• Current Research on Agriculture and Life Sciences
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• 제31권4호
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• pp.245-249
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• 2013

In 2008, leaf blight symptoms were observed on several Chinese chive farms in Sangju. The Pathogenicity of the isolate was confirmed by artificial inoculation, where the pathogen exhibited a strong pathogenicity toward healthy plants. Morphological classification identified the isolate as from the Fusarium genus. For further analysis, PCR and phylogenetic classification were performed with ITS region and 28S rRNA gene which are commonly used for fungal identification. However, the results provided a poor resolution. To solve this problem, we analyzed translation elongation factor 1-alpha (TEF-$1{\alpha}$) gene. The analyzed results using TEF-$1{\alpha}$ gene indicated that the isolate was F. proliferatum. Therefore, it is assumed that TEF-$1{\alpha}$ gene is important when Fusarium sp. was identified using molecular classification method.

• Journal of Microbiology and Biotechnology
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• 제20권11호
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• pp.1563-1570
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• 2010

To construct a highly sensitive detection system for endocrine disruptors (EDs), we have compared the activity of promoters with the n-alkane-inducible cytochrome P450 gene (ALK1), isocitrate lyase gene (ICL1), ribosomal protein S7 gene (RPS7), and the translation elongation factor-1${\alpha}$ gene (TEF1) for the heterologous gene in Yarrowia lipolytica. The promoters were introduced into the upstream of the lacZ or hERa reporter genes, respectively, and the activity was evaluated by ${\beta}$-galactosidase assay for lacZ and Western blot analysis for hER${\alpha}$. The expression analysis revealed that the ALK1 and ICL1 promoters were induced by n-decane and by EtOH, respectively. The constitutive promoter of RPS7 and TEF1 showed mostly a high level of expression in the presence of glucose and glycerol, respectively. In particular, the TEF1 promoter showed the highest ${\beta}$-galactosidase activity and a significant signal by Western blotting with the anti-estrogen receptor, compared with the other promoters. Moreover, the detection system was constructed with promoters linked to the upstream of the expression vector for the hER${\alpha}$ gene transformed into the Y. lipolytica with a chromosome-integrated lacZ reporter gene under the control of estrogen response elements (EREs). It was indicated that a combination of pTEF1p-hER${\alpha}$ and CXAU1-2XERE was the most effective system for the $E_2$-dependent induction of the ${\beta}$-galactosidase activity. This system showed the highest ${\beta}$-galactosidase activity at $10^{-6}\;M\;E_2$, and the activity could be detected at even the concentration of $10^{-10}\;M\;E_2$. As a result, we have constructed a strongly sensitive detection system with Y. lipolitica to evaluate recognized/suspected ED chemicals, such as natural/synthetic hormones, pesticides, and commercial chemicals. The results demonstrate the utility, sensitivity, and reproducibility of the system for identifying and characterizing environmental estrogens.

• The Plant Pathology Journal
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• 제26권4호
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• pp.409-416
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• 2010

Fusarium species, a large group of plant pathogens, potentially pose quarantine concerns worldwide. Here, we focus on the development of a method for detecting four Fusarium species in quarantined plants in Korea: F. solani f. sp. cucurbitae, F. stilboides, F. redolens, and F. semitectum var. majus. Species-specific primers were designed from the nucleotide sequences of either the translation elongation factor-1 alpha (TEF1) gene or RNA polymerase II subunit (RPB2) gene. Two different primer sets derived from TEF1, all specific to F. solani f. sp. cucurbitae, were able to differentiate the two races (1 and 2) of this species. A set of nested primers for each race was designed to confirm the PCR results. Similarly, two primer sets derived from RPB2 successfully amplified specific fragments from five F. stilboides isolates grouped within a single phylogenetic clade. A specific TEF1 primer set amplified a DNA fragment from only four of the 12 F. redolens strains examined, which were grouped within a single phylogenetic clade. All of the F. semitectum var. majus isolates could be specifically detected with a single RPB2 primer set. The specificity of the primer sets developed here was confirmed using a total of 130 Fusarium isolates.

• Mycobiology
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• 제47권4호
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• pp.408-414
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• 2019

Crinum asiaticum and Hymenocallis littoralis, commonly known as spider lilies are bulbous perennial and herbaceous plants that widely planted in Malaysia as ornamental. During 2015-2016, symptom of leaf blight was noticed on the hosts from several locations in Penang. The symptom appeared as irregular brown to reddish lesions surrounded by yellow halos. As the disease progressed, the infected leaves became blighted, dried, and fell off with the presence of black microsclerotia and pycnidia on the lesions parts. The present study was conducted to investigate the causal pathogen of leaf blight on C. asiaticum and H. littoralis. Based on morphological characteristics and DNA sequences of internal transcribed spacer (ITS) region and translation elongation factor 1-alpha (TEF1-α) gene, the causal pathogen was identified as Macrophomina phaseolina. Phylogenetic analysis of combined dataset of ITS and TEF1-α grouped the isolates studied with other isolates of M. phaseolina from GenBank. The grouping of the isolates was supported by 96% bootstrap value. Pathogenicity test proved the role of the fungus in causing leaf blight on both hosts.

• Mycobiology
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• 제51권3호
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• pp.122-138
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• 2023

Elsinoë are plant pathogenic fungi that cause scabs, spotted anthracnose, and some morphological distortions on various plants, including woody plants, economically important crops, and ornamental plants. Taxonomical reexamination of Elsinoë species in Japan has not yet been conducted based on the modern species criteria. In this study, several Japanese isolates were reexamine based on the morphological and molecular-phylogenetic analysis of the internal transcribed spacer region (ITS), large subunit gene (LSU)m and protein-coding gene such as RNA polymerase II subunit (rpb2) and Translation elongation factor 1-alpha (tef). Japanese isolates were divided into four clades and three new species, Elsinoë hydrangeae, E. sumire, and E. tanashiensis were proposed. One species, Sphaceloma akebiae, was transferred to the genus Elsinoë.

• Mycobiology
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• 제49권3호
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• pp.267-274
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• 2021

Two Diaporthe species isolated from fruit of Citrus sinensis in China were characterized based on morphology and multilocus phylogeny of ITS, tef1, and tub2 gene sequences. The phylogeny indicated that the two species match Diaporthe taoicola and D. siamensis. A critical examination of phenotypic characteristics confirmed the phylogenetic results. Diaporthe taoicola was morphologically characterized by producing Alpha conidia with tapering toward both ends. Meanwhile, D. siamensis produced cylindrical or ellipsoidal Alpha conidia with two oil drops. Pathogenicity tests revealed that both species were pathogenic to fruit of C. sinensis. To our knowledge, the two species were firstly reported on Citrus sinensis in China.

• Mycobiology
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• 제50권2호
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• pp.99-103
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• 2022

A new fungus isolated from the leaves of Eleutharrhena macrocarpa in southern Yunnan, China is described using morphological and molecular evidence. Phylogenetic trees based on the combined nuclear ribosomal DNA internal transcribed spacer (ITS), translation elongation factor-1α (TEF1), and β-tubulin gene (TUB2) sequences showed that Diaporthe eleutharrhenae sp. nov. is sister to Diaporthe chinensis N.I. de Silva, Lumyong & K.D. Hyde and morphologically differs in shorter alpha conidia (5-8.5× 1.5-2 ㎛) and the presence of beta conidia. This study also resolves a nomenclatural problem, as two taxa were published using the same name. To avoid confusion, the unrelated D. chinensis H. Dong, J. W. Xia & X. G. Zhang is here renamed as D. dongii (H. Dong, J. W. Xia & X. G. Zhang) S. J. Song & Landrein, sp. nov. in honor of the author that described this species. Study and description of fungi associated with threatened tropical species could help to understand their ecology as well as the potential spread of fungi onto cultivated crop species.

• 한국균학회지
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• 제50권3호
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• pp.183-194
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• 2022

Two fungal strains, designated KNUF-21-006 and KNUF-21-028, were isolated from soil samples collected from Gyeongbuk Province, Korea. The strain KNUF-21-006 was similar to other Pestalotiopsis species in terms of morphological characteristics, including whitish to pale brown mycelium, conidial shape, and size. The isolate had aerial hyphae that produced black fruiting bodies on the mycelium. The conidia were fusoid to ellipsoid, four-septate, and appendage-bearing. Phylogenetic analysis using the internal transcribed spacer (ITS) region, translation elongation factor 1-alpha (TEF), and β-tubulin (TUB) gene sequences confirmed that the closest relationship of the isolate at the species level was with Pestalotiopsis clavata. The strain KNUF-21-028 exhibits similar morphological characteristics to other Botryotrichum species, including white aerial mycelium with sulcate and irregular margins, conidial shape, and size. The conidia were globose, single, and hyaline. Upon molecular analysis-using the ITS region, large subunit (LSU) rRNA gene, and TUB gene sequences-the fungus was identified as Botryotrichum iranicum. This is the first record of these fungal species in Korea.

• 한국균학회지
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• 제49권3호
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• pp.417-423
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• 2021

Apple decline symptoms were frequently observed on cv. Fuji apple orchards located in Gyeonggi, Gyeongbuk, and Gangwon provinces during surveys conducted from May until the end of September 2020. Three fungal strains were isolated from the margins of internal lesions of diseased apple trees, and their morphological characteristics were considered similar to Botryosphaeria sinensis. Phylogenetic analysis using internal transcribed spacer (ITS) regions, translation elongation factor 1-alpha (tef1), beta-tubulin (tub2), and the second largest subunit of RNA polymerase II (rpb2) gene sequences confirmed the closest relationship of isolates with B. sinensis at the species level. According to a pathogenicity test, the appearance of dark-brown discolorations and vascular necrosis on apple branches inoculated with the isolated strain KNUF-20-014 was observed. To the best of our knowledge, this is the first report of B. sinensis as the causal agent of apple disease in Korea.

• Mycobiology
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• 제38권1호
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• pp.7-12
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• 2010

A green mold species that has not previously been reported in Korea was isolated from oak log beds used for shiitake (Lentinula edodes) cultivation that were infested by mushroom flies. In this study, we identify the mold species as Gliocladium viride (an anamorph of Hypocrea lutea) and describe its mycological properties. The fungus was cottony on both potato dextrose agar (PDA) and Czapek yeast extract agar (CYA), but was colored white on PDA and became yellowish green and brown on CYA. Mycelial growth on PDA attained a diameter of 73 mm at $30^{\circ}C$ after 5 days. The fungus grew faster on malt extract agar (> 80 mm, 5 days at $25^{\circ}C$) compared to CYA and PDA (< 68 mm, 5 days at $25^{\circ}C$). Penicillate conidiophores of the fungus are hyaline, smooth walled, branching above typically in four stages, and $120\sim240\;{\mu}m$ in length. Club-shaped or slender phialides are formed on the metulae. Conidia of the fungus were ovate and elliptic, yellowish brown and green, and $2.5\sim3.0\;{\mu}m\times1.8\sim2.3\;{\mu}m$ in size. Typically, slimy conidia are formed in a mass and colored brown to dark green to almost black. The internal transcribed spacer rDNA and translation elongation factor 1 alpha gene sequences of the fungus isolated here show 99% identity with previously identified G. viride strains.