• Title/Summary/Keyword: Pathogenicity

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Stability of pUC-Derived Plasmids with a Fluorescence Marker in Pectobacterium carotovorum subsp. carotovorum and subsp. betavasculorum

  • Hur, Woon-Yung;Roh, Eun-Jung;Oh, Chang-Sik;Han, Man-Wi;Lee, Seung-Don;Kim, Doo-Ho;Heu, Sung-Gi
    • The Plant Pathology Journal
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    • v.25 no.3
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    • pp.286-290
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    • 2009
  • The stability of three different kinds of pUC-derived plasmids, pDsRed, pZsYellow, and pGFPuv, was investigated in Pectobacterium strains to utilize those plasmids as tracers. All three plasmids pDsRed, pZsYellow and pGFPuv showed their specific colors in Pectobacterium strains. Especially, the plasmid pDsRed conferred bright pink colonies on the Pectobacterium strains. When the bacteria lost the plasmid pDsRed, the colonies turned white, suggesting that the plasmid could be a good marker system for Pectobacterium strains on different environmental conditions. The effect of the antibiotic pressure on the stability of the plasmid was different depending on the host bacteria. P. carotovorum subsp. betavasculorum was more sensitive to the antibiotic pressure than P. carotovorum subsp. carotovorum Pcc21. However, temperature change significantly affected plasmid stability on both Pectobacterium strains. Almost all strains lost the plasmids with the shift in temperature from $28^{\circ}C$ to $37^{\circ}C$. Presence of the plasmids did not affect bacterial pathogenicity on their own host plants. Among three plasmids, pZsYellow was not useful as a marker because the yellow fluorescent proteins from pZs Yellow were interfered with the yellow natural fluorescence of the plant tissues induced by the defense system. Since the red color of DsRed can be seen with naked eyes, plasmid pDsRed was applicable as a marker. However, the color change was slow so that additional manipulation to increase the expression speed was necessary. Plasmid pGFPuv could serve as a perfect marker without any problem, tracing the reproduction and spread of the plant pathogens perfectly.

Environmental Factors Affecting on Shrimp Cultivation and Bacterial Examination in Shrimp Aquaculture

  • Chun, Jae-Woo;Ma, Chae-Woo;Oh, Kye-Heon
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.775-779
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    • 2003
  • Cultivation environment for effective production of shrimp in shrimp aquaculture, Institute of Marine & Fishery, Soonchunhyang University located at Tae-An, Chung-Nam Province, with the ultimate aim of probiotics development, was monitored. Several environmental factors including dissolved oxygen, water body temperature, pH, salinity, $NH_4-N$, $NO_2-N$, $NO_3-N$, chlorophyll a affecting on the growth of shrimp were examined. Concentrations of $NH_4-N$, $NO_2-N$, and $NO_3-N$ dissolved in water samples were determined to 0.024-0.034 mg/L, 0.03-0.02, and 0.004-0.009 mg/L, respectively. Chlorophyll a content was examined in the range of 0.002-0.118 $ug/m^3$. In order to understand the distribution of different bacteria in water samples collected in shrimp aquaculture, bacteria were isolated and enumerated on the marine agar plates. Total number of bacteria were increased to approximately $6.5\;{\times}\;10^4$. Thirteen predomonant bacteria were isolated and identified. As the results of BIOLOG test of the isolates, these bacteria were identified as Corynebacterium nirilophilus, Clavibacter agropyri, Sphingomonas adhaesiva, Brevundimonas vesicularis, Vibrio parahaemolyticus, Pseudomonas bathycetes, Vibrio tubiashii, Sphingomonas macrogoltabidus, Rhodococcus, rhodochrous, Burkholderia glumae, Corynebacterium urealyticus, Rhodococcus fascians, Psychrobacter immobilis, respectively. Further work will stimulate the elucidation of pathogenicity, corresponding bacteria related to environment and probiotics, providing good information for effective production of shrimp.

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Diversity of VC and incidence of hypovirulence-associated ds-RNAs in the chestnut blight fungus Cryphonectria parasitica in Korea

  • Byeongjin Cha;Jinyoung Lim;Ju, Young-Jik;Kim, Dae-Hyuk
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.23-23
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    • 2003
  • Chestnut blight, caused by Cryphonectria parasitica, is the most destructive disease of American and European chestnut trees. A total of 672 C prasitica was isolated from blight lesion on chestnut twigs, which were collected from major chestnut plantations all over Korea in 1999. Isolation rates were over 30% in Kyunggj-, Kyongnam-, and Chonnam-do. The highest isolation rate was 37.4% and recorded in Kyongnam-do. On the other hand, Chonbuk-do had the lowest isolation rate as 13.5%. In grouping of C parasitica by colony shape and color, yellow colony with irregular margin were the most dominant colony type with a frequency of 65.2%. When the 672 isolates were inoculated on the chestnut twigs, 380 isolates (56.5%) caused lesions larger than the standard virulent isolate EP155-2, while 158 isolates (23.4%) caused smaller lesions than the standard hypovirulent isolate UEP-1. In Bavendamm test that determines phenol oxidase activity, 97.1% of all the isolates resulted the same or darker discoloration than EP155-2, and only 12.2% resulted the same or lighter discoloration than UEP-1. In the vegetative compatibility (VC) tests, total 670 isolates were divided into 121 VC groups (VCGs). Kyongnam-, Chonnam-, and Chungnam-do, the three principal chestnut plantation area, had 49, 33, and 27 VCGs, respectively. Among the VCGs, the biggest VCG, KR-VC104, was composed of 164 isolates and the second biggest VCG had 62 isolates. But, 64 of 121 VCGs consisted of sole member. More than 65.8% of KR-VC104, was isolated from the three provinces, Kyongnam-, Kangwon-, and Chungbuk-do. In KR-VC104, 62.8%, 59.1%, and 85.9% of the isolates looked like virulent in colony type, pathogenicity test, and Bavendamm test. In ds-RNA detection tests using cellulose chromatography, 77 of total 650 isolates were ds-RNA positive and detected ds-RNA segments were approximately 12kb, 3kb, 2.7kb, 2kb, and 1.8kb in size. Among the 77 isolates, 46 isolates had 12kb and 25 isolates had 12kb and 2.7kb. Other 6 Isolates had small ds-RNA segments. Kyongnam-, Chonnam-, and Chungnam-do had 43, 16, and 5 ds-RNA positive isolates, respectively. Among the 121 VCGs, only 29 VCGs had ds-RNA positive isolates.(중략)

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Biological control of Lettuce Sclerotinia rot using Bacillus mojanvinensis Pro-EB 15 strain.

  • Bak, Joung-Woo;Kim, Hyun-Ju;Park, Jong-Young;Lee, Kwang-Youll;Gang, Jun-Ho;Lee, Jin-Woo;Jung, Soon-Je;Moon, Byung-Ju
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.102.2-103
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    • 2003
  • This studies were investigated the occurrence of Sclerotinia rot by Sclerotinia Sclerotiorum at the lettuce field in Uiryeong-Gun, Gyeongsangnam-Do and were isolated the most effective microorganism for the biological control to the pathogen, S. sclerotiorum YR-1 strain from diseased soil and lettuce leaves. For the pathogenicity test, the most suitable inoculmn density of YR-1 strain was selected as the mycelial suspension of 40m1 showing disease incidence of 80%, and the symptom showed as same as at the fields, the leaves and stem had rotten and developed white downy mycelial at the diseased lesion on the leaves and stems, and produced black and irregular sclerotinia. On the PDA dual test, about 300 isolates were examined the antifungal activity to the pathogen, YR-1 strain, and among them, A-2, A-7, and RH-4 strain were selected most effective antagonistic bacteria. At pots test, the control value of A-7 strain showed the highest value as 85% which was more effective than that of others in a growth chamber. For the promotion of control effect, the selected 3 isolates were spayed on the lettuce leaves as a sole and/or mixed treatments in a growth chamber, the mixed treatment of A-7 and RH-4 strain showing the control value of 90% was most effective than that of sole treatment with A-7 or RH-4 strain showing the control value of 80%, respectively and mixed treatment with A-2 and A-7 strain and A-2, A-7 and RH-4 strain. In addition, 3 bacteria re-isolated from diseased soils, and all of the selected 6 isolates investigated the control effect at pots in a growth chamber, According to the results, A-7 and Pro-EB 15 strain showed the control value of 91.0% and 90.1% respectively, and they were selected most effectual antagonistic bacteria to control lettuce sclerotinia rot and identified as the Bacillus mojanuinensis by 16s RNA analysis. This is the first report on the biological control using by B. mojanvinensis to the lettuce Sclerotinia rot.

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Influence of Temperature, Wetness Duration and Fungicides on Fungal Growth and Disease Progress of Soybean Anthracnose Caused by Colletotrichum spp. (콩 탄저병균의 생장 및 병 진전에 미치는 온도, 수분 존재시간, 살균제의 영향)

  • Oh, Jeung-Haing;Kim, Gyu-Hong
    • Research in Plant Disease
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    • v.9 no.3
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    • pp.131-136
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    • 2003
  • The effects of temperature, duration of wetness period, and fungicides on the spore germination, appressorium formation, acervulus formation and lesion development by Colletotrichum spp., cause of soybean anthracnose, and their pathogenicity were assessed in controlled environment. C. gloeosporioides was highly pathogenic on inoculated soybean seeds as high as C. truncatum, whereas remarkably low on the soybean leaves. Spore germination, appressorium formation and mycelial growth of C. gloeosporioides were best at $25^{\circ}C$, but C. truncatum was best at $30^{\circ}C$. It has also done at $15^{\circ}C$, even though it was much retarded. C. truncatum showed high sensitivity to the fungicides, fluazinam and benomyl, meanshile C. gloeosporioides showed to fluazinam and triflumizole. At least 8 hrs. of wetness period was requird for the pathogen to develop lesions at $30^{\circ}C$. When the wetness period was 32 hrs. lesion size of was larger at $25^{\circ}C$ than $30^{\circ}C$, however it was traceable at $20^{\circ}C$. Different sensitivity of Colletotrichum spp. to fungicides suggests that proper fungicide is required to effective control of soybean anthracnose ingected multiply with Colletotrichum spp.

Characterization of the Causal Fungus of Citrus Melanose, Diaphorthe citri Isolated from Blighted Twigs of Citrus in Jeju (감귤나무 가지에서 분리한 검은점무늬병균 Diaporthe citri의 균학적 특성)

  • Kwon, Hyeog-Mo;Nam, Ki-Woong;Kim, Kwang-Sik;Kim, Dong-Hwan;Lee, Seong-Chan;Hyeon, Jae-Wook
    • Research in Plant Disease
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    • v.9 no.3
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    • pp.153-158
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    • 2003
  • Mycological characteristics of Phomopsis citri isolates obtained from blighted twigs of citrus cultivation areas in Jeju island were all identified as Phomopsis citri known as the anamorph stage of Diaporthe citri. Pycnidia the isolates sized from 112.2 to 614.4${\mu}m$ (av. 328.8${\mu}m$) and shaped conical to lenticulata in black. Two types of phialospores, ${\alpha}$-and ${\beta}$-spores were observed from pycnidia. The ${\alpha}$-spores were hyaline, unicellula, fusiform to ellipsoidal and sized 4.7~8.7 1.9 ${\times}$ 3.5${\mu}m$ (av. 6.7 ${\times}$ 2.3 ${\mu}m$). The ${\beta}$-spores were hyaline, unicellula, filiform, curved and often strongly hooked and sized 13.2~27.1 ${\times}$ 0.8~1.6 ${\mu}m$ (av. 22.1 ${\times}$ 1.0 ${\mu}m$). All isolates grew well and produced abundant pycnidia and spore horns on PDA. In addition, all isolates showed strong pathogenicity to citrus fruit inducing melanose symptoms when artificially inoculated with the pycnidial spores.

Phytophthora Diseases of Apple in Korea: II. Occurrence of an Unusual Fruit Rot Caused by P. cactorum and P. cambivora (사과의 역병: II. Phytophthora cactorum과 P. cambivora에 의한 사과 과실역병의 발생)

  • Jee, Hyeong-Jin;Cho, Weon-Dae;Kim, Wan-Gyu
    • Korean Journal Plant Pathology
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    • v.13 no.3
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    • pp.145-151
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    • 1997
  • An unusual young fruit rot of apple caused by two species of Phytophthora was epidemic from late May to early July of 1996 in Andong, Uisung and Chungwon areas of Korea. The disease spread to over 30 apple orchards in the areas and percent of the infected tree and fruit was ca. 10~90% and 1`15%, respectively. Water soaking lesions or spots on leaves and shoot blight were also developed by the pathogen. Among 39 isolates collected, 25 were identified as P. cactorum and the others were as zp. cambivora on the basis of their distinctive morphological characters. While the former fungus was homothallic, all isolates of the latter were A1 mating types. Koch's postulate was fulfilled. Both fungi showed strong pathogenicity not only to young fruits, leaves and shoots of apple but also to those of pear and peach. Several vegetables tested did not show symptoms even by wound inoculation. An Occurrence of young fruit rot of apple caused by Phytophthora has not been reported in Korea, especially, P. cambivora has not been recorded previously as the causal agent of the disease in the world.

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Effect of Nitrogen Sources on Mycelial Growth and Sclerotial Formation of Rhizoctonia solani Causing Rice Sheath Blight (벼 잎집무늬마름병균의 균사생장 및 균핵형성에 미치는 질소원의 효과)

  • Park Kyung Seok;Chung Bong Koo
    • Korean Journal Plant Pathology
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    • v.1 no.1
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    • pp.44-50
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    • 1985
  • Effect of nitrogen sources on mycelial growth and sclerotial formation of Rhizoctonia solani causing rice sheath blight was studied by using sixty isolates from diseased samples of different localities in Chungbuk area. Based on the cultural characters, pathogenicity and sclerotial formation, three isolates of the fungus were selected. The virulence of the three isolates varied based on the percent of seeds germination, seedling blights and length and lesion number of lesions. As nitrogen sources, arginine, alanine, urea, ammonium sulfate and sodium nitrate were more effective for mycelial growth and sclerotial formation, than were cystine, methionine, lysine, histidine, tryptophan and thyrosine. The mycelium of R. so/ani grew well in a lower concentration of urea, sodium nitrate, arginine, ammonium sulfate and lysine, but in a higher concentration of proline, histidine, and alanine. In all nitrogen sources tested, high concentration of nitrogen nutrients greatly stimulated sclerotial formation of the fungus. Arginine, urea, and ammonium sulfate increased virulence of the fungus. All the isolates grown on the basic medium plus proline produced large sclerotia. Mycelial growth of the fungus was not well in the tryptophan-added medium, showing brown discoloration of medium.

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Studies on the effects of culture filtrates of sesame-wilt organism(Fusarium oxysprum f. vasinfectum) on the germination of sesame seeds and the growth of sesame seedlings (참깨$\cdot$시들음병균(Fusarium oxysporum)의 배양여액이 참깨의 종자발아 및 유묘기의 생육에 미치는 영향)

  • PARK Jong Seong
    • Korean journal of applied entomology
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    • v.1
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    • pp.3-10
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    • 1962
  • 1) The purpose of the present study is to investigate the effects of culture filtrates of Fusarium oxysporum f. vasinfectum which is known to produce fusaric acid (wilt toxin) on the germination of sesame seeds and the growth of sesame seedlings. 2) Culture filtrates of Fusarium oxysporum 1. vasinfectum used in this study strongly or weakly inhibited the germination and bring about necrosis accompanying black discoloration of sesame seeds. 3) Varietal difference of sesame in the germination response on the culture filtrates of Fnsarium oxysporum f. vasinfectum is not shown in this study. 4) This study reveals that differential five strains of Fnsarium oxysoprum f. vasinfectum used in this study differ greatly in the toxicity of culture filtrates inhibiting the germination of sesame seeds. 5) In the seedling bed added with culture filtrates of Fnsarium oxysporum f. vasinfectum, the growth of shoot as well as root system of sesame seedlings are notably inhibited and necrotic black discoloration appear on both shoot and root system. But in the seedling beds added with weaker concentration of culture filtrates $/(10\%)$ the growth of shoot is slightly promoted. 6) In culture of sesame seedlings with Knop's solution containing 1 to 3 per cent culture filtrates, the growth of shoot as well as root system are slightly retarded" and till the time of development of the third leaves the whole stem and leaf petiole tissue are weakened so that they become thread like accompanying brown discoloration, interveinal light brown area appear in the second leaves, and the third leaves curl from both sides towards the middle with necrotic brown discoloration, especially symptoms of injury on the third leaves are nearly similar that of the leaves of wilted sesame in the field. 7) A pararell relationship is not found between toxicity of culture filtrates and pathogenicity of five differential strains of Fnsarium oxysporum f. vasinfectum used in this study.

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Biological Control of Clover by Penicillium sp. (Penicillium sp.를 이용한 토끼풀의 생물학적 방제)

  • Kim, Pan-Kyung;Park, Dong-Jin;Choi, Jung-Sub;Hwang, In-Taek;Hong, Kyung-Sik;Kim, Chang-Jin
    • Applied Biological Chemistry
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    • v.40 no.1
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    • pp.65-70
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    • 1997
  • Among 980 fungal strains isolated from the lesions of Trifolium repens L. and various weeds, three Penicillium sp. F40362, F40496, F40497 were selected by the first selection test and a pathogenicity test. The spores of these strains germinated readily 90 to 100% and readily infected the respective plant. The wheat bran-corn starch formulation of F40362 strain showed 100% mycoherbicidal activity against clover plant at $4{\times}10^8$ spores/pot. The same formulations of F40496 and F40497 strains showed 100% mycoherbicidal activity against clover plant at $12{\times}10^8{\sim}4{\times}10^8$ spores/pot. The same formulations of tee strains showed over 30% mycoherbicidal activity against Leguminosae plants. This method of pelletiation was potentially useful for the production of inoculum formulation as mycoherbicides and it was effective enough to treat $2{\sim}2.5\;g$ formulation($4.5{\times}10^7\;spores/mg$) to a $350\;cm^2$ pot. The three strains, F40362, F40496 and F40497 have selective mycoherbicidal activity between Trifolium repens L. and Zoysia japonica and nonselective mycoherbicidal activities against some other crop plants and weeds.

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