• Title/Summary/Keyword: Penicillium isolates

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Isolation and Cytotoxic Potency of Endophytic Fungi Associated with Dysosma difformis, a Study for the Novel Resources of Podophyllotoxin

  • Hoa Thi Tran;Giang Thu Nguyen;Hong Ha Thi Nguyen;Huyen Thi Tran;Quang Hong Tran;Quang Ho Tran;Ngoc Thi Ninh;Phat Tien Do;Ha Hoang Chu;Ngoc Bich Pham
    • Mycobiology
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    • v.50 no.5
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    • pp.389-398
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    • 2022
  • Endophytic fungi are promising sources for the production of podophyllotoxin-an important anticancer compound, replacing depleted medical plants. In this study, the endophytes associated with Dysosma difformis-an ethnomedicinal plant species were isolated to explore novel sources of podophyllotoxin. Fifty-three endophytic fungi were isolated and identified by morphological observation and ITS-based rDNA sequencing, assigning them to 27 genera in 3 divisions. Fusarium was found the most prevalent genus with a colonization frequency of 11.11%, followed by Trametes (9.26%) and Penicillium (7.41%). Phylogenetic trees were constructed for the endophytic fungi community in two collection sites, Ha Giang and Lai Chau, revealing the adaptation of the species to the specific tissues and habitats. Cytotoxic activity of endophytic fungal extracts was investigated on cancer cell lines such as SK-LU-1, HL-60, and HepG2, demonstrating strong anti-cancer activity of six isolates belonging to Penicillium, Trametes, Purpureocillium, Aspergillus, and Ganoderma with IC50 value of lower than 10 ㎍/mL. The presence of podophyllotoxin was indicated in Penicillium, Trametes, Aspergillus and for the first time in Purpureocillium and Ganoderma via high-performance liquid chromatography, which implied them as a potential source of this anticancer compound.

Characterisation of fungal contamination sources for use in quality management of cheese production farms in Korea

  • Kandasamy, Sujatha;Park, Won Seo;Yoo, Jayeon;Yun, Jeonghee;Kang, Han Byul;Seol, Kuk-Hwan;Oh, Mi-Hwa;Ham, Jun Sang
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.6
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    • pp.1002-1011
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    • 2020
  • Objective: This study was conducted to determine the composition and diversity of the fungal flora at various control points in cheese ripening rooms of 10 dairy farms from six different provinces in the Republic of Korea. Methods: Floor, wall, cheese board, room air, cheese rind and core were sampled from cheese ripening rooms of ten different dairy farms. The molds were enumerated using YM petrifilm, while isolation was done on yeast extract glucose chloramphenicol agar plates. Morphologically distinct isolates were identified using sequencing of internal transcribed spacer region. Results: The fungal counts in 8 out of 10 dairy farms were out of acceptable range, as per hazard analysis critical control point regulation. A total of 986 fungal isolates identified and assigned to the phyla Ascomycota (14 genera) and Basidiomycota (3 genera). Of these Penicillium, Aspergillus, and Cladosporium were the most diverse and predominant. The cheese ripening rooms was overrepresented in 9 farms by Penicillium (76%), while Aspergillus in a single farm. Among 39 species, the prominent members were Penicillium commune, P. oxalicum, P. echinulatum, and Aspergillus versicolor. Most of the mold species detected on surfaces were the same found in the indoor air of cheese ripening rooms. Conclusion: The environment of cheese ripening rooms persuades a favourable niche for mold growth. The fungal diversity in the dairy farms were greatly influenced by several factors (exterior atmosphere, working personnel etc.,) and their proportion varied from one to another. Proper management of hygienic and production practices and air filtration system would be effective to eradicate contamination in cheese processing industries.

Tolerance of Apple Blue Mold (Penicillium expansum) to Various Fungicides (사과 푸른곰팡이병균의 각종 살균제에 대한 내성)

  • Lee Chang Un
    • Korean Journal Plant Pathology
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    • v.1 no.2
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    • pp.128-135
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    • 1985
  • Responses to the 14 fungicides added in PSA of penicillium expansum isolated froms the four major apple growing areas in Korea were examined. Degrees of the fungal tolerance were various with little difference by area and with great difference by kind and concentration of fungicide. Fall isolates of the fungus grew at higher fungicide concentration than did the spring isolates. $ED_{50}$ and MIC to mycelial growth of genomyl, garbenda, polydong, polyoxin and thiophanate methyl were $0.1{\mu}g-41.7{\mu}g/ml\;and\;100{\mu}g-2,250{\mu}g/ml,$, respectively, indicating high fungicide effects on the fungus isolated from decayed apples in storages and infected apples in orchards with low fungal tolerance; of captafol, captan, iprodione and mancozeb were $2.6{\mu}g - 750{\mu}g/ml\;and\;638{\mu}g-40,500{\mu}g/ml$, respectively, indicating medium degree of both fungicidal effects and fungal tolerance; and of chlorothalonil, folpet, oxidong, propineb and triademefon were $27.8{\mu}g-8,354{\mu}g/ml\;and\;2,625{\mu}g-150,357{\mu}g/ml$, respectively, indicating low fungicide effects and high fungal tolerance. Fungicidal inhibitory effects on conidia formation exhibited a similar trend as on mycelial growth whereas those on conidia germination showed an opposite trend to the mycelial growth with the exception of polyoxin and triademefon.

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Diversity of Endophytic Fungi from Different Verticillium-Wilt-Resistant Gossypium hirsutum and Evaluation of Antifungal Activity Against Verticillium dahliae In Vitro

  • Li, Zhi-Fang;Wang, Ling-Fei;Feng, Zi-Li;Zhao, Li-Hong;Shi, Yong-Qiang;Zhu, He-Qin
    • Journal of Microbiology and Biotechnology
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    • v.24 no.9
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    • pp.1149-1161
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    • 2014
  • Cotton plants were sampled and ranked according to their resistance to Verticillium wilt. In total, 642 endophytic fungi isolates representing 27 genera were recovered from Gossypium hirsutum root, stem, and leaf tissues, but were not uniformly distributed. More endophytic fungi appeared in the leaf (391) compared with the root (140) and stem (111) sections. However, no significant difference in the abundance of isolated endophytes was found among resistant cotton varieties. Alternaria exhibited the highest colonization frequency (7.9%), followed by Acremonium (6.6%) and Penicillium (4.8%). Unlike tolerant varieties, resistant and susceptible ones had similar endophytic fungal population compositions. In three Verticillium-wilt-resistant cotton varieties, fungal endophytes from the genus Alternaria were most frequently isolated, followed by Gibberella and Penicillium. The maximum concentration of dominant endophytic fungi was observed in leaf tissues (0.1797). The evenness of stem tissue endophytic communities (0.702) was comparatively more uniform than the other two tissues. Eighty endophytic fungi selected from 27 genera were evaluated for their inhibition activity against highly virulent Verticillium dahliae isolate Vd080 in vitro. Thirty-nine isolates exhibited fungistasis against the pathogen at varying degrees. Seven species, having high growth inhibition rates (${\geq}75%$), exhibited strong antifungal activity against V. dahliae. The antifungal activity of both volatile and nonvolatile metabolites was also investigated. The nonvolatile substances produced by CEF-818 (Penicillium simplicissimum), CEF-325 (Fusarium solani), CEF-714 (Leptosphaeria sp.), and CEF-642 (Talaromyces flavus) completely inhibited V. dahliae growth. These findings deepen our understanding of cotton-endophyte interactions and provide a platform for screening G. hirsutum endophytes with biocontrol potential.

Isolation and Characterization of Airborne Bacteria and Fungi in Indoor Environment of Elementary Schools (초등학교 실내환경에서 공기 중 세균과 진균의 분리 및 특성)

  • Kim, Na-Yeong;Kim, Young-Ran;Kim, Min-Kyu;Cho, Du-Wan;Kim, Jong-Seol
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.193-200
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    • 2007
  • Indoor airborne bacterial and fungal concentrations were examined at classrooms and corridors of 3 elementary schools in Ulsan. Airborne microorganisms were collected with an impaction-type air sampler using plate count agar and dichloran rose bengal chloramphenicol agar. During the semester, concentrations of bacteria ranged $168{\sim}3,887 MPN/m^3$ at classrooms and $168{\sim}6,339 MPN/m^3$ at corridors, while those of fungi ranged $34{\sim}389 MPN/m^3$ at classrooms and $91{\sim}507 MPN/m^3$ at corridors. The bacterial concentrations showed larger variations between situations and schools compared to those of fungi. When airborne bacteria were isolated and identified, 84% were observed as Gram-positive, and Micrococcus spp. was the most abundant group with 61% of tested isolates, followed by genus Staphylococcus with 10%. The Micrococcus spp. isolates, of which 75% were identified as M. luteus, appeared to be from human origins. The protective pigments and substantial cell wall of Micrococcus may provide selective advantage for their survival in the air. We also isolated and identified 15 genera of filamentous fungi. The most common culturable fungi were Cladosporium, Aspergillus and Penicillium, and these 3 genera were 69% of fungal isolates. Genus Stachybotrys, of which S. chartarum is a well known producer of many potent mycotoxins, was also detected from one of the schools. further systematic studies are necessary with an emphasis on species identification and mycotoxin production of isolated fungal genera, including Aspergillus, Penicillium, and Stachybotrys.

Distribution and Antifungal Activity of Endophytic Fungi in Different Growth Stages of Chili Pepper (Capsicum annuum L.) in Korea

  • Paul, Narayan Chandra;Deng, Jian Xin;Sang, Hyun-Kyu;Choi, Young-Phil;Yu, Seung-Hun
    • The Plant Pathology Journal
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    • v.28 no.1
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    • pp.10-19
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    • 2012
  • This study investigated the distribution of endophytic fungi obtained from the surface sterilized tissues of leaves, stems and roots of chili pepper ($Capsicum$ $annuum$ L.) plants in seedling, flowering and fruiting stages in Korea and their antifungal activity. A total of 481 isolates were recovered and were identified using molecular techniques. Based on rDNA ITS gene sequence and phylogenetic analysis, 21 fungal genera were characterized, belonging to 16 Ascomycota and 5 Basidiomycota. $Penicillium$ in seedling stage, $Fusarium$ in flowering stage, $Colletotrichum$ followed by $Fusarium$, $Alternaria$ and $Xylaria$ in fruiting stage was predominant and $Alternaria$, $Cladosporium$ and $Fusarium$ were common in all growth stages. Among 481 endophytes 90 phenotypes were evaluated for the antimicrobial activity against three major pathogens ($Phytophthora$ $capsici$, $Colletotrichum$ $acutatum$ and $Fusarium$ $oxysporum$) of chili pepper. Among them 16 isolates inhibited the growth of at least one test microorganisms. Three strains showed a broad spectrum antifungal activity and displayed strong inhibition against chili pepper pathogenic fungi.

Diversity of Fungi Isolated from Soil of Jeollabuk-do and Chungcheongbuk-do, Korea

  • Yadav, Dil Raj;Adhikari, Mahesh;Kim, Sang Woo;Kim, Hyun Seung;Um, Yong Hyun;Lee, Youn Su
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.30-30
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    • 2015
  • This study was conducted aiming with the assessment of fungal diversity in soil samples collected from different locations of Jeollabuk-do and Chungcheongbuk-do, Korea. Forty soil samples were collected in 2015 and fungi were isolated through serial dilution technique. Isolated fungi were purified and differentiated according to their morphological and microscopic characteristics. In total, 150 different representative isolates were recovered and the genomic DNA of each isolate was extracted by using QIAGEN$^{(R)}$ Plasmid Mini Kit (QIAGEN Sciences, USA) and the identification of fungi was carried out by sequence analysis of internal transcribed spacer (ITS) region of the 18S ribosomal DNA (18S rDNA). Recovered isolates belonged to 37 family, 67 genera and 108 species. Aspergillus spp., Penicillium spp., Trichoderma spp., Chaetomium spp. And Fusarium spp. were the most dominant taxa in this study. Out of total species, 20 species were identified as new records for Korea.

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Genetic Diversity of Culturable Endophytic Fungi Isolated from Halophytes Naturally Growing in Muan Salt Marsh (무안 염습지에 자생하는 염생식물(halophyte)의 뿌리로부터 분리된 내생진균(endophytic fungi)의 유전학적 다양성)

  • You, Young-Hyun;Yoon, Hyeok-Jun;Seo, Yeong-Gyo;Kim, Mi-Ae;Kang, Myung-Suk;Kim, Chang-Mu;Ha, Sang-Chul;Cho, Ga-Youn;Kim, Jong-Guk
    • Journal of Life Science
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    • v.22 no.7
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    • pp.970-980
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    • 2012
  • Native halophytes, such as Suaeda maritima, Limonium tetragonum, S. japonica, Zoysia sinica, and Phragmites australis were collected from the Muan salt marsh. Ninety endophytic fungi were isolated from the roots of the collected halophytes. Molecular insights inferred by internal transcribed spacer containing ITS1, 5.8s, and the ITS2 region showed that all the fungal strains belong to ten orders, i.e., Capnodiales (4.44%), Cystofilobasidiales (1.11%), Dothideales (3.33%), Eurotiales (53.33%), Glomerellales (3.33%), Hypocreales (8.89%), Mucorales (1.11%), Pleosporales (15.56%), Sordariales (1.11%), and Trichosphaeriales (1.11%). The rest (6.67%) of all fungal isolates were not identified. Ninety fungal strains were confirmed at the genus level, containing Acremonium, Alternaria, Aspergillus, Aureobasidium, Cephalosporium, Chaetomium, Cladosporium, Colletotrichum, Cryptococcus, Didymella, Dothideomycete, Emericellopsis, Epicoccum, Eupenicillium, Fusarium, Gibberella, Gongronella, Macrophoma, Microsphaeropsis, Nigrospora, Paecilomyces, Paraconiothyrium, Penicillium, Phaeomyces, Phoma, Pleosporales, Purpureocillium, and Talaromyces. Of all the endophytic fungi identified from the various halophytes, Aspergillus and Penicillium of Eurotiales had the highest abundance.

Diversity of Culturable Soil Micro-fungi along Altitudinal Gradients of Eastern Himalayas

  • Devi, Lamabam Sophiya;Khaund, Polashree;Nongkhlaw, Fenella M.W.;Joshi, S.R.
    • Mycobiology
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    • v.40 no.3
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    • pp.151-158
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    • 2012
  • Very few studies have addressed the phylogenetic diversity of fungi from Northeast India under the Eastern Himalayan range. In the present study, an attempt has been made to study the phylogenetic diversity of culturable soil fungi along the altitudinal gradients of eastern Himalayas. Soil samples from 24 m above sea level to 2,000 m above sea level altitudes of North-East India were collected to investigate soil micro-fungal community structure and diversity. Molecular characterization of the isolates was done by PCR amplification of 18S rDNA using universal primers. Phylogenetic analysis using BLAST revealed variation in the distribution and richness of different fungal biodiversity over a wide range of altitudes. A total of 107 isolates were characterized belonging to the phyla Ascomycota and Zygomycota, corresponding to seven orders (Eurotiales, Hypocreales, Calosphaeriales, Capnodiales, Pleosporales, Mucorales, and Mortierellales) and Incertae sedis. The characterized isolates were analysed for richness, evenness and diversity indices. Fungal diversity had significant correlation with soil physico-chemical parameters and the altitude. Eurotiales and Hypocreales were most diverse and abundant group of fungi along the entire altitudinal stretch. Species of Penicillium (D=1.44) and Aspergillus (D=1.288) were found to have highest diversity index followed by Talaromyces (D=1.26) and Fusarium (D=1.26). Fungal distribution showed negative correlation with altitude and soil moisture content. Soil temperature, pH, humidity and ambient temperature showed positive correlation with fungal distribution.

Microbial Diversity in Korean Traditional Fermenting Starter, Nuruk, Collected in 2013 and 2014

  • Seo, Jeong Ah
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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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.

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