• Title/Summary/Keyword: Single conidial isolates

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Stable Formation of Fruiting Body in Cordyceps bassiana

  • Lee, Je-O;Shrestha, Bhushan;Kim, Tae-Woong;Sung, Gi-Ho;Sung, Jae-Mo
    • Mycobiology
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    • v.35 no.4
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    • pp.230-234
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    • 2007
  • In order to breed a Cordyceps bassiana isolate that stably forms fruiting body in artificial cultivation, isolates derived from subculturing and single spores were tested through mating. From C. bassiana EFCC 783, three subcultured isolates EFCC 2830, EFCC 2831 and EFCC 2832 were obtained and fourteen single conidial isolates were obtained from these three subcultured isolates. Two different morphological types were found in the fourteen single conidial isolates. One type was able to form synnemata and another type was not able to form synnemata. Since switch of morphological type was not observed despite their continuous subculturing, cross was performed between the two types and the formation of fruiting body was examined. Ascospores were obtained from a selected fruiting body formed by hybrid of the cross. Self-cross and combinational cross of the ascospore-derived isolates generated hybrids that stably produce high quality fruiting body in artificial media.

Successful Development of Cordyceps bassiana Stromata from Beauveria bassiana

  • Lee, Je-O;Shrestha, Bhushan;Sung, Gi-Ho;Han, Sang-Kuk;Sung, Jae-Mo
    • Mycobiology
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    • v.38 no.1
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    • pp.13-16
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    • 2010
  • A specimen of Beauveria bassiana was collected from Yang-yang of Gangwon province, Korea in October 2006. Conidial isolates were prepared from the specimen by the dilution method and inoculated in brown rice medium for fruiting body production. After nearly two months incubation for perithecial stromata developed from single isolates as well as from their combinations. They were determined as Cordyceps bassiana by observing the stromatal characters and their conidial structures. This is the first report of the development of C. bassiana from B. bassiana cultures.

Isolation and Identification of Fusicoccum Species from Quercus dentata

  • Kim, Ki Woo;Kim, Pan-Gi;Lee, Myung-Bo
    • Journal of Korean Society of Forest Science
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    • v.96 no.5
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    • pp.515-519
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    • 2007
  • An imperfect fungus Fusiococcum species was isolated from Quercus dentata. A naturally infected Daimyo oak tree was collected and showed elongate wounds on the stem. The fungal cultures were initially white and cottony, and later turned dark gray. Numerous solitary pycnidia were developed on the medium surface, and typically spherical. Yellowish conidial masses were exuded from pycnidia on the culture plates. Conidial masses were swollen and measured as approximately 100 to $300{\mu}m$ in length. It appeared that conidia were usually held together in globose to oval drops. Conidia were hyaline, single-celled (nonseptate), ellipsoid to fusoid, and measured as approximately $8.0{\times}2.7{\mu}m$. Based on these cultural and morphological characteristics, the fungal isolate was identified as a species of Fusicoccum Corda. To preserve and examine fungal spores exuded from pycnidia on the medium surface, a vapor fixation procedure for scanning electron microscopy was employed in this study. The specimens were exposed to the vapor of 2% (v/v) glutaraldehyde and 2% (w/v) osmium tetroxide each for 2 h. With the vapor fixation we obtained excellent retention of conidial masses in this study. The simple and versatile procedure for demonstrating fungal spores and their exudation from fruiting bodies would facilitate characterization of diverse pathological and environmental isolates as they are in native environments.

Genetic Stability of Magnaporthe oryzae during Successive Passages through Rice Plants and on Artificial Medium

  • Park, Sook-Young;Chi, Myoung-Hwan;Milgroom, Michael G.;Kim, Hyo-Jung;Han, Seong-Sook;Kang, Seog-Chan;Lee, Yong-Hwan
    • The Plant Pathology Journal
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    • v.26 no.4
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    • pp.313-320
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    • 2010
  • Genetic instability of the rice blast fungus Magnaporthe oryzae has been suggested as a major factor underlying the rapid breakdown of host resistance in the field. However, little information is available on the mechanism of genetic instability. In this study, we assessed the stability of repetitive DNA elements and several key phenotypic traits important for pathogenesis after serially transferring two isolates though rice plants and an artificial medium. Using isolate 70-15, we obtained a total of 176 single-spore isolates from 10 successive rounds of culturing on artificial medium. Another 20 isolates were obtained from germ tubes formed at the basal and apical cells of 10 three-celled conidia. Additionally, 60 isolates were obtained from isolate KJ201 after serial transfers through rice plants and an artificial medium. No apparent differences in phenotypes, including mycelial growth, conidial morphologies, conidiation, conidial germination, appressorium formation, and virulence, or in DNA fingerprints using MGR586, MAGGY, Pot2, LINE, MG-SINE and PWL2 as probes were observed among isolates from the same parent isolate. Southern hybridization and sequence analysis of two avirulence genes, AVR-Pita1 and AVR-Pikm, showed that both genes were also maintained stably during 10 successive generations on medium and plants. However, one reversible loss of restriction fragments was found in the telomere-linked helicase gene (TLH1) family, suggesting some telomere regions may be more unstable than the rest of the genome. Taken together, our results suggest that phenotype and genotype of M. oryzae isolates do not noticeably change, at least up to 10 successive generations on a cultural medium and in host plants.

Occurrence of Gray Mold on Blueberry Trees Caused by Botrytis cinerea in Korea (Botrytis cinerea에 의한 블루베리 잿빛곰팡이병의 한국 내 발생)

  • Hong, Sung-Kee;Choi, Hyo-Won;Lee, Young-Kee;Lee, Sang-Yeob;Kim, Wan-Gyu
    • The Korean Journal of Mycology
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    • v.39 no.3
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    • pp.213-216
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    • 2011
  • Gray mold symptoms were frequently observed on green twigs, blossoms, leaves, and fruits of blueberry trees grown in greenhouses in Cheongyang, Dangjin, Daejeon, and Jeju during disease survey in eight locations of Korea from 2007 to 2010. The disease symptoms were not observed in the fields of the other locations investigated. The disease incidence ranged 1~30% in the greenhouses investigated. A total of 27 single spore isolates of Botrytis species were obtained from the gray mold symptoms, and all the isolates were identified as Botrytis cinerea based on their morphological and cultural characteristics. Four isolates of the fungus were tested for pathogenicity to leaves of four varieties of blueberry trees by artificial inoculation with conidial suspensions. All the tested isolates caused gray mold symptoms on the leaves, which were similar to those observed in the greenhouses. This is the first report that B. cinerea causes gray mold of blueberry trees grown in greenhouses in Korea.

Isolation and Characterization of Colletotrichum Isolates Causing Anthracnose of Japanese Plum Fruit (자두 탄저병균의 분리 및 동정)

  • Lee, Yong-Se;Ha, Da-Hee;Lee, Tae-Yi;Park, Min-Jung;Chung, Jong-Bae;Jeong, Byeong-Ryong
    • Korean Journal of Environmental Agriculture
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    • v.36 no.4
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    • pp.299-305
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    • 2017
  • BACKGROUND: Although the filamentous fungal pathogen Colletotrichum species causing anthracnose disease on various fruits including peach, apple, persimmon and grape, there is no report on Japanese plum in Korea. METHODS AND RESULTS: In 2016, diseased fruits showing typical anthracnose symptoms of Japanese plum were collected in market and ochards. Diseased tissue was cut off and disinfected subsequently with 70% ethanol for 1 min, and in 1% sodium hypochloride solution for 1 min, followed by three washes with sterile distilled water. The disinfected tissues were placed onto potato dextrose agar (PDA), and incubated at $25^{\circ}C$ in the dark for 5 to 7 days. For single-spore isolation, conidia were scraped off the plate using a loop, and suspended with 10 mL sterile distilled water. One hundred microliter of the conidial suspension was spread on PDA plates and incubated at $25^{\circ}C$. Finally, one germinated conidium was transferred onto PDA plates. Morphological and cultural characteries of colonies and spores of isolated Colletotrichum were observed after 7 to 10 days incubation on PDA. Molecular identification of isolates were analyzed by comparing rDNA-ITS gene sequences with NCBI GeneBank. CONCLUSION: Of eleven isolates of Colletotrichum isolated from anthracnose diseased Japanese plum fruits, six were identified as C. acutatum, and five as C. gloeosporioides based on diagnostic characteristics such as colony growth rate, shape and size of conidia, and rDNA-ITS sequences. This is the first report of Colletotrichum causing the anthracnose on Japanese plum in Korea.

Integration of Bological and Chemical Methods for the Control of Pepper Gray Mold Rot Under Commercial Greenhouse Conditions

  • Park, Seon-Hee;Bae, Dong-Won;Lee, Joon-Taek;Chung, Sung-Ok;Kim, Hee-Kyu
    • The Plant Pathology Journal
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    • v.15 no.3
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    • pp.162-167
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    • 1999
  • Integration of microbial antagonists with fungicides was tried to control the gray mold caused by Botrytis cinerea on pepper in greenhouse conditions and to reduce fungicide uses. All of the selected bacterial antagonists, Bacillus amyloliquefaciens BL3, Paenibacillus polymyxa BL4, and Pseudomonas putida Cha94, completely inhibited the conidial germination of B. cinerea until 30 days after treatment. However, bacterial colonization of pepper phylloplane was poor in BL4, while the other bacterial isolates and the fungal antagonist Trichoderma harzianum TM colonized well on the phylloplane, maintaining the population density of 104-105 cfu/g until 15 days after microbial treatments. Out of 13 kinds of selected fungicides used for gray mold diseases, polyoxin B and BKF 1995 showed the most discriminatory activity on the fungal growth between B. cinerea and TM. TM grew readily on the media containing those fungicides, while B. cinerea showed poor or no mycelial growth on them. The selected fungicides and antagonists alone reduced incidence of gray mold on pepper, showing disease indices of about 2.4 to 3.0, while its was increased up to 4.2 in the untreated control. Alternate treatments with the antagonists and 2-fold diluted fungicides inhibited the disease incidence as much as the antagonists or fungicides alone, and reduced the secondary inoculum more than the single treatments. This suggests that integration of antagonists and fungicides may be an efficient way to reduce fungicide sprays with reliable control efficacy of the disease. However, there was not much difference in the early and mid-term disease progress among the treatments and the untreated control, probably due to extremely favorable environmental conditions for the disease development in this experiment.

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