1.Genetic Analysis of Pigmentation in Cordyceps militaris.
Bhushan SHRESTHA ; Sung Keun CHOI ; Ho Kyoung KIM ; Tae Woong KIM ; Jae Mo SUNG
Mycobiology 2005;33(3):125-130
Pigmentation of ascospore-derived isolates from seven different natural specimens of Cordyceps militaris EFCC C-5888, EFCC C-7159, EFCC C-7833, EFCC C-7991, EFCC C-8021, EFCC C-8023 and EFCC C-8179 was observed on the plates of Sabouraud Dextrose agar plus Yeast Extract at 25degrees C under continuous illumination (500 lux). Pigmentation of the wild-type isolates of C. militaris was diverse ranging from yellowish white to orange, while white color was believed as a mutant. Inheritance of pigmentation was found to be controlled by both parental isolates when F1 progeny were analyzed. Pigmentation and mating type were shown to be either independent or distantly linked each other due to the high percentage of non-parental phenotypes among F1 progeny. Crosses between white mutant isolates of C. militaris yielded progeny with wild type pigmentations, indicating that the albino mutations in the parents were unlinked to each other.
Agar
;
Citrus sinensis
;
Cordyceps*
;
Glucose
;
Humans
;
Lighting
;
Parents
;
Phenotype
;
Pigmentation*
;
Wills
;
Yeasts
2.Immobilization of Keratinase from Aspergillus flavus K-03 for Degradation of Feather Keratin.
Mycobiology 2005;33(2):121-123
Extracellular keratinase isolated from Aspergillus flavus K-03 was immobilized on calcium alginate. The properties and reaction activities of free and immobilized keratinase with calcium alginate were characterized. The immobilized keratinase showed proteolytic activity against soluble azo-casein and azo-keratin, and insoluble feather keratin. Heat stability and pH tolerance of keratinase were greatly enhanced by immobilization. It also displayed a higher level of heat stability and an increased tolerance toward alkaline pHs compared with free keratinase. During the durability test at 40degrees C, 48% of the original enzyme activity of the immobilized keratinase was remained after 7 days of incubation. The immobilized keratinase exhibited better stability, thus increasing its potential for use in industrial application.
Animals
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Aspergillus flavus*
;
Aspergillus*
;
Calcium
;
Feathers*
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Hot Temperature
;
Hydrogen-Ion Concentration
;
Immobilization*
3.First Report of Penicillium brasilianum and P. daleae Isolated from Soil in Korea.
Hye Sun CHO ; Seung Beom HONG ; Seung Joo GO
Mycobiology 2005;33(2):113-117
In this study, a total of 300 isolates of Penicillium and related teleomorphic genera were collected from soils of 17 locations in Korea from April to May, 2004. Ninety four isolates were identified as the species of Penicillium subgenus Furcatum based on cultural and morphological characteristics and beta-tubulin gene sequences. Among the species, Korean isolates of P. brasilianum Bat. and P. daleae K. M. Zalessky were phylogenetically identical to the reference species based on DNA sequence of the beta-tubulin gene. Here we described and illustrated P. brasilianum and P. daleae that are new in Korea.
Base Sequence
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Korea*
;
Penicillium*
;
Soil*
;
Tubulin
4.Sequence Analysis of the Internal Transcribed Spacer of Ribosomal DNA in the Genus Rhizopus.
Mycobiology 2005;33(2):109-112
The internal transcribed spacer (ITS) regions including the 3'-end of 18S rRNA gene, 5.8S rRNA gene and the 5'-end of the 28S rRNA gene of Rhizopus spp. were amplified by PCR and analyzed by DNASIS program. Length polymorphism of these region ranged from 564 bp in R. oryzae to 789bp in R. stolonifer. The length and sequence of 5.8S was very conserved with 154~155 bp. The sequence of ITS2 was more variable than that of ITS1. The base substitution rates were ranged from 0 to 0.6069 per site, and higher rate was found in R. stolonifer. In general, transition was usually more frequent than transversion. On the basis of sequencing results, four groups were clustered with value of 61.9% similarity; R. oryzae, R. microspores, R. homothallicus, and R. stolonifer groups.
DNA, Ribosomal*
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Genes, rRNA
;
Oryza
;
Polymerase Chain Reaction
;
Rhizopus*
;
Sequence Analysis*
5.Development of Species-specific Primers for Rapid Detection of Phellinus linteus and P. baumii.
Mun Ok KIM ; Gi Young KIM ; Byung Hyouk NAM ; Cheng Yun JIN ; Ki Won LEE ; Jae Min PARK ; Sang Joon LEE ; Jae Dong LEE
Mycobiology 2005;33(2):104-108
Genus Phellinus taxonomically belongs to Aphyllophorales and some species of this genus have been used as a medicinal ingredients and Indian folk medicines. Especially, P. linteus and morphological-related species are well-known medicinal fungi that have various biological activities such as humoral and cell-mediated, anti-mutagenic, and anti-cancer activities. However, little is known about the rapid detection for complex Phellinus species. Therefore, this study was carried out to develop specific primers for the rapid detection of P. linteus and other related species. Designing the species-specific primers was done based on internal transcribed spacer sequence data. Each primer set detected specifically P. linteus (PL2/PL5R) and P. baumii (PB1/PB4R). These primer sets could be useful for the rapid detection of specific-species among unidentified Phellinus species. Moreover, restriction fragment length polymorphism analysis of the ITS region with HaeIII was also useful for clarifying the relationship between each 5 Phellinus species.
Fungi
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Phylogeny
;
Polymorphism, Restriction Fragment Length
;
Polyporales
6.Antifungal Activity of Nor-securinine Against Some Phytopathogenic Fungi.
Sangita SAHNI ; S MAURYA ; U P SINGH ; A K SINGH ; V P SINGH ; V B PANDEY
Mycobiology 2005;33(2):97-103
Crude extracts and active principles from medicinal plants have shown potential role in controlling plant diseases in glasshouses as well as in fields as one of the safest and ecofriendly methods. The effect of nor-securinine (an alkaloid) isolated from Phyllanthus amarus has been seen against spore germination of some fungi (Alternaria brassicae, A. solani, Curvularia pennisetti, Curvularia sp., Erysiphe pisi, Helminthosporium frumentacei) as well as pea powdery mildew (Erysiphe pisi) under glasshouse conditions. The sensitivity of fungi to nor-securinine varied considerably. Nor-securinine was effective against most of the fungi. H. frumentacei was more sensitive even at the lowest concentration (1,000 microg/ml). Likewise conidia of E. pisi were also inhibited in partially or completely appressorium formation. Pre-inoculation treatment showed greater efficacy than post-inoculation in inhibiting powdery mildew development on pea plants in a glasshouse. Maximum inhibition occurred at 2000 microg/ml
Brassica
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Complex Mixtures
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Fungi*
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Germination
;
Helminthosporium
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Peas
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Phyllanthus
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Plant Diseases
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Plants, Medicinal
;
Spores
;
Spores, Fungal
7.Production of Xylanolytic Enzyme Complex from Aspergillus flavus using Agricultural Wastes.
Mycobiology 2005;33(2):84-89
Five types of agricultural wastes were used for the production of xylanolytic enzyme by Aspergillus flavus K-03. All wastes materials supported high levels of xylanase and beta-xylosidase production. A high level of proteolytic activity was observed in barley and rice bran cultures, while only a weak proteolytic activity was detected in corn cob, barley and rice straw cultures. Maximum production of xylanase was achieved in basal liquid medium containing rice barn as carbon source for 5 days of culture at pH 6.5 and 25degrees C. The xylanolytic enzyme of A. flavus K-03 showed low thermostability. The times required for 50% reduction of the initial enzyme activity were 90 min at 40degrees C, 13 min at 50degrees C, and 3 min at 60degrees C. Xylanolytic activity showed the highest level at pH 5.5~10.5 and more than 70% of the original activity was retained at pH 6.5 and 7.0. The higher stability of xylanolytic enzymes in the broad range of alkaline pH is useful for utilization of the enzymes in industrial process requiring in alkaline conditions. Moreover, the highest production of xylanolytic enzyme was obtained when 0.5% of rice bran was supplied in basal liquid medium. SDS-PAGE analysis revealed a single xylanase band of approximately 28.5 kDa from the culture filtrates.
Aspergillus flavus*
;
Aspergillus*
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Carbon
;
Electrophoresis, Polyacrylamide Gel
;
Hordeum
;
Hydrogen-Ion Concentration
;
Zea mays
8.Occurrence of Fungal Species and Mycotoxins from Decayed Sugarcane (Saccharrum officinarum) in Egypt.
Gamalat A ABD-ELAAH ; Samya SOLIMAN A
Mycobiology 2005;33(2):77-83
Seventy-three fungal species belonging to forty-three genera were isolated from 40 samples of Saccharrum officinarum (collected from Naage-Hamadi canal in Qena Governorate, Egypt). Aspergillus, Trichoderma, Mucor and Pythium were the most common genera on the two isolation media. The dominant species of Aspergillus were A. niger, A. flavus, A. ustus, A. terreus and A. wentii. Some species were dominant on 40 g/l sucrose such as Aspergillus niger, A. flavus, Emericella nidulans, Trichoderma viride, Torula herbarum and Mamaria echinoeotryoides, while the dominant species on 10 g/l glucose were Mucor circinelloides, Aspergillus niger, Torula herbarum and Trichoderma viride. Mycotoxins including aflatoxins B1, B2, G1 and G2, zearalenone and diacetoxyscirpenol were detected in the examined samples of Saccharrum officinarum. The mycelial growth of A. flavus, A. niger, Fusarium moniliforme and Torula herbarum decreased with the increase in Dimethoate concentrations, although 25 ppm was less effective than the higher levels of the insecticide (75~200 ppm). Dimethoate stimulated the activity of Go-T in A. niger, F. moniliforme and T. harbarum, while the Go-T activity was inhibited in A. flavus with the Dimethoate treatments.
Aflatoxins
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Aspergillus
;
Aspergillus niger
;
Cryptococcus
;
Dimethoate
;
Egypt*
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Emericella
;
Fusarium
;
Glucose
;
Mucor
;
Mycotoxins*
;
Niger
;
Pythium
;
Saccharum*
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Sucrose
;
Trichoderma
;
Zearalenone
9.Morphological Characteristics of Conidiogenesis in Cordyceps militaris.
Bhushan SHRESTHA ; Sang Kuk HAN ; Kwon Sang YOON ; Jae Mo SUNG
Mycobiology 2005;33(2):69-76
Conidial development of Cordyceps militaris was observed from germinating ascospores and vegetative hyphae through light and scanning electron microscopy (SEM). Ascospores were discharged from fresh specimens of C. militaris in sterile water as well as Sabouraud Dextrose agar plus Yeast Extract (SDAY) plates. We observed ascospore germination and conidial formation periodically. Under submerged condition in sterile water, most part-spores germinated unidirectionally and conidia were developed directly from the tips of germinating hyphae of part-spores within 36 h after ascospore discharge, showing microcyclic conidiation. First-formed conidia were cylindrical or clavate followed by globose and ellipsoidal ones. Germination of ascospores and conidial development were observed on SDAY agar by SEM. Slimy heads of conidia on variously arranged phialides, from solitary to whorl, developed 5 days after ascospore discharge. Besides, two distinct types of conidia, elongated pyriform or cylindrical and globose, were observed in the same slimy heads by SEM. Conidia were shown to be uninucleate with 4,6-diamidino-2-phenylindole staining. Conidiogenous cells were more slender than vegetative hyphae, having attenuated tips. Microcyclic conidiation, undifferentiated conidiogenous hyphae (phialides), polymorphic conidia and solitary, opposite to whorled type of phialidic arrangement are reported here as the characteristic features of asexual stage of C. militaris, which can be distinguished from other Cordyceps species.
Agar
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Cordyceps*
;
Germination
;
Glucose
;
Head
;
Hyphae
;
Microscopy, Electron, Scanning
;
Spores, Fungal
;
Water
;
Yeasts
10.Four Newly Recorded Amanita Species in Korea: Amanita sect. Amanita and sect. Vaginatae.
Chang Sun KIM ; Jong Won JO ; Young Nam KWAG ; Junsang OH ; Bhushan SHRESTHA ; Gi Ho SUNG ; Sang Kuk HAN
Mycobiology 2013;41(3):131-138
We collected nearly 70 specimens of Amanita species during a diversity study of Korean mushrooms conducted in 2012. In this study, we primarily investigated 23 Amanita specimens belonging to sections Amanita and Vaginatae. Based on sequence data of the internal transcribed spacers and partial large subunit of ribosomal RNA and morphological characteristics, we identified the following 15 phylogenetic species: A. alboflavescens, A. ceciliae, A. farinosa, A. fulva, A. griseofolia, A. ibotengutake, A. melleiceps, A. orientifulva, A. pantherina, A. rubrovolvata, A. sinensis, A. subglobosa, A. vaginata, A. cf. vaginata f. alba, and an undescribed Amanita species. In this study, four of the identified Amanita species (A. griseofolia, A. ibotengutake, A. orientifulva, and A. sinensis) were reported for the first time in Korea.
Agaricales
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Amanita*
;
Korea
;
Phylogeny
;
RNA, Ribosomal