• Title/Summary/Keyword: Single spore isolate

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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.

Strain improvement of Pleurotus ostreatus using self-fertility monospore isolate (느타리 단핵 임성체를 이용한 균주개발)

  • Yoo, Young-Bok;Kim, In-Yeup;Kong, Won-Sik;Jang, Kab-Yeul;Oh, Se-Jong;Jhune, Chang-Sung
    • Journal of Mushroom
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    • v.4 no.2
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    • pp.48-52
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    • 2006
  • Oyster mushroom, Pleurotus ostreatus is bifactorial heterothallism. Single basidiospore isolates from fruiting bodies are homokaryotic and self-sterile. However, we found that homokaryons derived from some strains of P. ostreatus could develop fruiting bodies with two different types. One hundred and two isolates out of 155 monospore isolates formed fruiting bodies (65.8%). First group did not have only mature or sporulating fruiting bodies but also clamp connections, which initial isolate also did not present clamp connections (Abortive homokaryotic fruiting, AHF). Second group had developed fruiting bodies with clamp connections even though initial homokaryotic colony did not form clamp connections (Pseudo- homokaryotic fruiting, PHF). The mycelial colonies derived from PHF by tissue culture formed clamp connections, while mycelial colonies of AHF lacked them. We obtained 535 PHF and 79 AHF inter-strain hybrids among 8 strains of P. ostreatus by hyphal anastomosis. The fruiting body yield of PHF group is higher than that of AHF group in bottle cultivation. A preselection of single spore isolates for fertility would save labour in strain improvement of P. ostreatus.

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A study of morphological characteristics and hybridization on Lepista nuda (민자주방망이버섯(Lepista nuda )의 형태적 특성 및 교배에 관한 연구)

  • Jung, Won Soon;Kim, Jong Bong
    • Journal of Mushroom
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    • v.11 no.1
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    • pp.1-8
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    • 2013
  • In this study, twelve of Lepista nuda were collected from various localities in Korea. Also thirteen exotic L. nuda species were collected from Japan, France, Switzerland and Portugal. Spores were isolated under optical microscope. These spores were placed on the surface of YM medium for inducing to germination. Eleven mating-groups were selected by morphological characters of fruit body such as size, color and stipe patterns. Intra-isolate crosses were made between two single-spore isolates derived from mating-groups. Also, dikaryotic crossing using the isolates from L. nuda were carried out to evaluated tetrakaryon formation. Cross-mating compatibility tests also verified its dikaryotic state by microscopic or molecular genetic observation of clamp connection and Random Amplified Polymorphic DNA (RAPD) band pattern. To analyze the growth rate of hybrids and parents mycelium in dikaryons obtained from compatible mating groups were placed on PDA medium. Intra-isolate crosses determined eleven mating-groups within L. nuda. The typical clamp connection were mostly observed in mating-groups of Korean L. nuda in $K1{\times}K2$, $K1{\times}K3$, $K1{\times}K4$, $K1{\times}K6$, $K1{\times}K5$, $K2{\times}K4$, $K2{\times}K3$, $K2{\times}K6$, $K3{\times}K4$, $K4{\times}K5$, and $K4{\times}K6$. Korean L. nuda type of dikaryon, shown to cross-incompatibility with L. sordida, it seemed that mating induce more rapidly than wild types in a view of growth rate. In conclusion, it would be useful to improve mass production with better morphological characteristics through a special mating of L. nuda.

Monitoring for the Resistance of Strobilurin Fungicide Against Botrytis cinerea Causing Gray Mold Disease (Strobilurin계 살균제에 대한 잿빛곰팡이병균 Botrytis cinerea의 저항성 검정)

  • Kim, Ah Hyeong;Kim, Seon Bo;Han, Ki Don;Kim, Heung Tae
    • The Korean Journal of Pesticide Science
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    • v.18 no.3
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    • pp.161-167
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    • 2014
  • This study was conducted to investigate the resistance of Botrytis cinerea to azoxystrobin, which belongs to strobilurin fungicides. The sensitivity of B. cinerea isolates, which were collected from infected pepper, strawberry, cucumber and tomato by a single spore isolation, to the fungicide was tested through a agar dilution method on PDA amended with fungicides and $100{\mu}g\;mL^{-1}$ of salicylhydroxamic acid (SHAM). All isolates of B. cinerea tested in this study were classified as a sensitive and a resistant group by $1.0{\mu}g\;mL^{-1}$ of $EC_{50}$ value to azoxystrobin. While the sensitive isolates accounted for 46.5% of B. cinerea population, the resistant ones did for 53.5%. According to the regions isolating B. cinerea, the highest isolation frequency was showed as 81.1% in Chungnam among the all. Among 4 host plants as pepper, strawberry, cucumber and tomato, the highest isolation frequency was obtained in strawberry, while the lowest was done in pepper. The isolate resistant to azoxystrobin showed the cross resistance to other fungicides included into strobilurins as kresoxim-methyl and trifloxystrobin. In spite of an excellent efficacy of strobilurins, it should be taken care to use them in the field, because of the high risk in the fields.

Studies on the nuclear cytology of Pyricularia oryzae CAV (도열병균의 핵학적 연구)

  • Lee S. C.;Shim J. S.;Lee E. J.
    • Korean journal of applied entomology
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    • v.5_6
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    • pp.19-26
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    • 1968
  • Results obtained from the observation of 1,000 matured resting conidia of Pyricularia oryzae CAV. were as fellows. The percentage of a condium conitaining was 95.5, while that of a multinucleate conidium was 4.5. Formation of the three-celled mature conidium containing a nucleus in each cell, which was derived from the immatured conidium, could be recognized. Chromosomes could be observed in the conidium when the nuclear division took place. The number of chromosomes was n=3, 4, 5 or 6 but rile majority was 5. The nucleus was moved into the germ-tube from the conidium by following ways: (a) a resting nucleus divided into two nuclei in the conidium and one of them was moved into the germ-tube. The other nucleus remained in the conidium (43 per cent): (b) a nucleus was moved into the germ-tube from the conidium without nuclear division (57 Per cent). The appressorium without a nucleus was abundantly observed when stain was made. However, tile number of a mononucleate appressorium was 476 out of 500 appressoria which had nuclear sap. On the ground of this experiment, we could support the conidium of blast fungus contained mononucleus and also homokaryon seemed to b: appealed on the conidia even though those were multilocular. As the results, it could be concluded that use of inoculum derived from single spore isolate was reasonable for any experiments in the rice blast fungus.

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