• Mycobiology
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• v.40 no.2
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• pp.100-106
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• 2012

Interest in commercial cultivation and product development of Cordyceps species has shown a recent increase. Due to its biochemical and pharmacological effects, Cordyceps militaris, commonly known as orange caterpillar fungus, is being investigated with great interest. Cultivation of C. militaris has been practiced on a large scale in order to fulfill a demand for scientific investigation and product development. Isolates of C. militaris can be easily established from both spores and tissue. For isolation of spores, ascospores released from mature stromata are trapped in sterile medium. Multi-ascospore isolates, as well as combinations of single ascospore strains, are used for production of fruiting bodies. Progeny ascospore strains can be isolated from artificial fruiting bodies, thus, the cycle of fruiting body production can be continued for a long period of time. In this study, we examined fruiting body production from multi-ascospore isolates and their progeny strains for three generations. $F_1$ progeny strains generally produced a larger number of fruiting bodies, compared with their mother multi-ascospore isolates; however, $F_2$ and $F_3$ progeny strains produced fewer fruiting bodies. Optimum preservation conditions could help to increase the vitality of the progeny strains. In order to retain the fruiting ability of the strains, further testing of various methods of preservation and different methods for isolation should be performed.

• Korean Journal Plant Pathology
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• v.11 no.4
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• pp.344-347
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• 1995

Factors affecting on the ascospore release of Mycosphaerella nawae, the causal organism of persimmon leaf casting disease, were investigated. The ascospore release of the pathogen occurred following the precipitation of rain. The ascospore release started from the beginning of raining, reached maximum at 2 hours after the precipitation of rain, and then decreased abruptly. When the inoculum source (a file of infected leaves) was submerged in water to imitate raining conditions, 92.5% of the total ascospores were released within 1 hour after submerging, 5.8% were after 2 hours, 1.4% were after 4 hours, 0.1% were after 8 hours, and none detected after 10 hours. The inoculum source overwintered in the field released ascospores much more and earlier than the inoculum source kept in the greenhouse. The first ascospore release was about 10 days earlier, and the amount of the total liberated spores was 3∼4 times higher in the field inoculum source than the greenhouse nioculum source. The early defoliated leaves (in early October) in the previous year produced ascospores twice more than the late defoliated leaves (in early November) produced.

• The Plant Pathology Journal
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• v.27 no.1
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• pp.20-25
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• 2011

Gibberella zeae (anamorph: Fusarium graminearum), a homothallic (self-ferile) ascomycete with ubiquitous geographic distribution, causes serious diseases in several cereal crops. Ascospores (sexual spores) produced by this fungal pathogen have been suggested as the main source of primary inoculum in disease development. Here, we report the function of a gene designated GzRUM1, which is essential for ascospore formation in G. zeae. The deduced product of GzRUM1 showed significant similarities to the human retinoblastoma (tumor suppressor) binding protein 2 and a transcriptional repressor, Rum1 in the corn smut fungus (Ustilago maydis). The transcript of GzRUM1 was detected during the both vegetative and sexual stages, but was more highly accumulated during the latter stage. In addition, no GzRUM1 transcript was detected in a G. zeae strain lacking a mating-type gene (MAT1-2), a master regulator for sexual development in G. zeae. Targeted deletion of GzRUM1 caused no dramatic changes in several traits except ascospore formation. The ${\Delta}$GzRUM1 strain produced perithecia (sexual fruit bodies) but not asci nor ascospores within them. This specific defect leading to an arrest in ascospore development suggests that GzRUM1, as Rum1 in U. maydis, functions as a transcriptional regulator during sexual reproduction in G. zeae.

• The Plant Pathology Journal
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• v.19 no.3
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• pp.152-158
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• 2003

Stromata of the white root rot fungus, Rosellinia necatrix, were produced on diseased roots although they were reported to develop rarely in nature. Forty-two (42) out of 47 samples produced synnemata while 23 developed stromata. Forty-seven (47) isolates obtained from diseased root samples were divided into 24 mycelium compatibility groups (MCGs). Sixteen (16) out of 24 MCGs produced stromata. Single ascospore isolates from 10 stroma samples produced dsRNA-containing isolates from diseased tissue beneath stromata. The frequency of synnema production on axenic culture varied among isolates with different origin. The dsRNA was not transmitted vertically to the ascospore offspring despite the infection of various dsRNA in the parental isolates. The dsRNA was absent in 35 ascospore isolates in two stroma samples that originated from the isolates, in which dsRNA was not eliminated by hyphal tip isolation. Consequently, sexual reproduction in the white root rot fungus was suggested to produce propagules as a new infection source and to have the function to eliminate infectious factors such as mycoviruses.

• Mycobiology
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• v.38 no.4
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• pp.282-285
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• 2010

Cordyceps cardinalis successfully produced its fruiting bodies from multi-ascospore isolates. However, subcultures of multiascospore isolates could not produce fruiting bodies after few generations. Fruiting body production also differed from sector to sector of the same isolate. Single ascospore isolates were then co-inoculated in combinations of two to observe the fruiting characteristics. Combinations of certain isolates produced perithecial stromata formation, whereas other combinations did not produce any fruiting bodies. These results show that C. cardinalis is a heterothallic fungus, requiring two isolates of opposite mating types for fruiting body production. It was also shown that single ascospore isolates are hermaphrodites.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.6
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• pp.440-446
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• 2004

Interest in in vitro study of entomopathogenic fungi, including Cordyceps species, has been increasing due to their valuable bioactive compounds and biocontrol effects. Among Cordyceps species, in vitro stromata of C militaris has been successfully produced and cultivated for industrial purposes. However, genetic study on in vitro stromata formation of C militaris has not been carried out yet. Here, relationship between mating system and perithecial stromata formation of C militaris is reported. Mating system was determined by observing perithecial stromata formation from mono-ascospore cultures and their pair-wise combinations. Certain combinations of mono-ascospore strains produced perithecial club-shaped stromata, whereas other combinations produced either no stromata or only abnormal non-perithecial stromata. Similarly, mono­ascospore cultures without combination produced either no stromata or only abnormal non­perithecial stromata. Despite obvious heterothallism, self-fertility was occasionally observed in few strains of C militaris. These observations indicated that C militaris behaves as a bipolar het­erothallic fungus and requires two mating compatible strains in order to produce regular club­shaped perithecial stromata, a fundamental requirement for its industrial cultivation.

• Mycobiology
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• v.33 no.2
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• pp.69-76
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• 2005

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.

• Mycobiology
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• v.34 no.4
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• pp.196-199
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• 2006

Effects of various preservation periods and subcultures on fruiting body formation of Cordyceps militaris were investigated using EFCC C-10995 single ascospore strains. Fruiting body formation by original strains was profuse when preserved at $4^{\circ}C$ for $5{\sim}6$ months. Fruiting from subcultures was stable till second to sixth subcultures, after which it decreased sharply. The more the colony color of subcultures changed, the less the fruiting bodies formed. Liquid inoculum preparation of single ascospore strains in the same or separate broths did not affect fruiting body formation. Similarly, two strains C-10995-3 and C-10995-6 in different numbers during liquid inoculum preparation produced similar fruiting bodies.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.151.2-151
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• 2003

One fungus was isolated from lesions on imported chinese cabbage leaves in process of quarantine inspection from China. The fungus was identified as Mycosphaerlla brassicicola, based on morphology of perithecia, asci, ascospore, and curtural characteristics. In Korea, this fungus has been quarantine fungus, and not yet report to occur. Perithecia of the fungus were globose, dark brown with apical papilate ostioles. The size was 90-100 ${\times}$ 130-135$\mu\textrm{m}$. Asci were bitunicate, 8-spored and 38-43 ${\times}$ 15-l9$\mu\textrm{m}$. Ascospore were irregularly biseriate, hyaline, cylindrical, 2-celled, and rounded at the ends. Optium growth temperature of the fungus was at 20$^{\circ}C$ on PDA but did rarely grow over 24$^{\circ}C$. Colony on PDA was of black aerial mycelia.

• Research in Plant Disease
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• v.7 no.1
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• pp.16-19
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• 2001

The cultural caracteristics of Monosporascus cannonballus causing root rot of Cucurbitaceae plants were examined in vitro, and population density of the fungal ascospores were measured and compared aoming diferent host species and soil depths. Potato-dextrose agar(PDA) was the most appropriate medium for the mycelial growth and perithecial formation among the 5 media tested. Corn-meal agar(CMA), oat-meal agar (OMA) and V-8 juice agar were moderate media for the mycelial growth and perithecial formation, whereas water agar(WA) was poor medium. Perithecia were not formed on WA. Optimum temperature for the formation of perithecia was about 25 to 30$^{\circ}C$. distribution of ascospores in the infested fields was variable amount the curcubitaceae plants and within the same plant species, ranging from 1.7 to 14.6 ascospores in 20 g of soil, but no ascospore was detected in the uninfested field soils. Ascospores were distributed more at 20cm of soil depth than at 10cm or 30cm of soil depth.