• Title/Summary/Keyword: primary inoculum

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Ecological Characteristics and Unique Diagnostic Techniques of Apple Blotch Disease Caused by Marssonina coronaria in Korea

  • Back, Chang-Gi;Lee, Seung-Yeol;Jung, Hee-Young
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.36-36
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    • 2014
  • Apple blotch, caused by Marssonina coronaria, induce early defoliation in apple and leading to critical economic losses in apple orchards in Korea. Since M. coronaria is difficult to culture, we developed isolation and cultural method. We collected M. coronaria isolates from Gyeongbuk Province and then constructed phylogentic tree based on ITS regions. As the results, phylogenetic relationship indicated that all Korean isolates formed a same cluster and closely related to Chinese isolates [1]. Ecological characteristic of M. coronaria have been observed in apple orchards which located in Gyeongbuk Province from 2011 to present. As the results, the typical apple blotch symptoms were observed from July, and then the infected leaves were discolored and formed acervuli on the leaves. After rainfall, severe infection of symptoms such as discoloration and early defoliation were continuously observed until October. Also overwintered conidia were observed in next March on the fallen diseased leaves [2]. In the last 5 years, ascopores of M. coronaria were not observed in apple orchards which were severely infected by M. coronaria in Korea. Thus, it is assumed that overwintered conidia could be a primary inoculum of M. coronaria. Meanwhile, apple blotch has long latent periods compare to other apple disease. During the latent period, early diagnosis of apple blotch is the most important to control the disease by spray fungicide. In this reason, we developed novel diagnostic method to detect M. coronaria during latent period using optical coherence tomography (OCT) and Loop-mediated isothermal amplification (LAMP) method [2, 3]. In this presentation, it will introduce ecological characterization of M. coronaria in Korea and unique detection technique of M. coronaria in apple. It will be helpful to develop new strategies to control apple blotch in Korea.

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Bacterial Community Profiling during the Manufacturing Process of Traditional Soybean Paste by Pyrosequencing Method (Pyrosequencing을 이용한 전통된장 제조과정 중 세균군집구조의 분석)

  • Kim, Yong-Sang;Jeong, Do-Yeon;Hwang, Young-Tae;Uhm, Tai-Boong
    • Korean Journal of Microbiology
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    • v.47 no.3
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    • pp.275-280
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    • 2011
  • In order to evaluate the diversity and change of bacterial population during the manufacturing process of traditional soybean paste (doenjang), bacterial communities were analyzed using 16S rRNA gene-based pyrosequencing. In rice straw, the most important inoculum source for fermentation, the bacterial sequences with a relative abundance greater than 1% were assigned to four phyla, Proteobacteria (71%), Actinobacteria (20.6%), Bacteroidetes (4.2%), and Firmicutes (1.3%). Unlike bacterial community composition of rice straw, a different pattern of bacterial population in meju was observed with predominantly high abundance (99.1%) of Firmicutes. Phylum composition in young doenjang was almost same as that of meju. Major genera in young doenjang were Bacillus (81.3%), Clostridium (6.9%) and Enterococcus (6.3%) and the predominant species among bacterial population was B. amyloliquefaciens (63.6%). Abundance of the phylum Firmicutes in mature doenjang was 99.98%, which was even higher value than those in meju and young doenjang. Predominant species in mature doenjang were B. amyloliquefaciens (67.3%), B. atrophaeus (12.7%), B. methylotrophicus (6.5%), B. mojavensis (3.2%), and B. subtilis. (2.5%), which were also identified as major species of the microbial flora in meju. These results suggested that rice straw was a primary source for supplement of Bacillus species in manufacturing the traditional doenjang and that some species of Bacillus strains were mainly involved in the fermentation process of traditional doenjang.

Ginseng anthracnose in Korea Factors affecting primary inoculum, growth of the pathogen, disease development and control (인삼${\cdot}$탄저병에 관한 연구 전염원, 병원균의 생태, 발병요인 및 방제)

  • Chung Hoo-Sup;Bae Hyo-Won
    • Korean journal of applied entomology
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    • v.18 no.1 s.38
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    • pp.35-41
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    • 1979
  • Four to $17\%$ of the seeds of ginseng (Panax ginseng Meyer) collected from seemingly healthy plants carried Colletotrichum panacicola Nakata et Takimoto whereas the seeds from the plants with anthracnose sympotoms carried $42\%$ of the same fungus. Prevalent organisms isolated other than C. panacicola from seeds of both kinds of plants were Fusarium, Alternaria, Phoma, Trichoderma and others, ana in that order on acidified potato sucrose agar. C. panacicola also was isolated from 18 months old herbarium specimens. The fungus in the infected tissues also survived during the Korean winter months either on the soil surface or in the soil at 10 and 30 em in depth. When conidial suspensions of C. panacicola were inoculated on detached ginseng leaves, anthracnose symptoms occurred from 25 to $35^{\circ}C$. No symptoms occurred at temperatures below $17^{\circ}C$. Direct sunlight increased significantly the number of anthracnose lesions over those obtained in leaves inoculated in darkness or in 400 lux of fluorescent light. The lesions decreased as age of the leaves increased or as the number of conidia applied decreased. Optimum temperature for mycelial growth and conidial formation of C. panacicola was $25^{\circ}C$. Optimum pH for the mycelial growth was at $pH\;2.8\~4.6$ while the most conidial formation occurred at $pH\;5.2\~5.8.$. When fungicides were applied in the field to ginseng plants with a conidial suspension of C. panacicola, the most effective control of the anthracnose disease was by spraying with difolatan, and followed by maneb, zineb, captan and phaltan; Bordeaux mixture and ferbam were significantly less effective but significantly better than the inoculated control plants.

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Effects of Multiple Stress Factors Including Iron Supply on Cell Growth and Lipid Accumulation in Marine Microalga Dunaliella tertiolecta (해양 미세조류 Dunaliella tertiolecta에서 철 공급을 포함한 다중스트레스 인자가 세포성장 및 지질생산에 미치는 영향)

  • Rizwan, Muhammad;Mujtaba, Ghulam;Lee, Kisay
    • Applied Chemistry for Engineering
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    • v.28 no.3
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    • pp.306-312
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    • 2017
  • Changes in the cell growth and lipid accumulation of marine microalga Dunaliella tertiolecta were investigated in response to the combination of different stress factors including the variation of iron supply as a primary stress factor and different options in light irradiation and $CO_2$ supply as a secondary stress factor. High or limited Fe conditions could act as a stress for lipid synthesis. As a secondary stress factor, non-$CO_2$ condition was good for lipid accumulation, but the overall cell growth was sacrificed significantly after a long-time cultivation. Dark condition as a secondary stress factor also favored lipid accumulation and the extent of cell density reduction at the early period in the dark was small compared to other stress conditions. The two-stage cultivation strategy was necessary to maximize lipid production because tendencies of the cell growth and lipid content were not identical under the chosen stress condition. The first stage was for preparing a high cell density under the normal growth-favoring condition and the second stage was the stress condition to induce lipid accumulation in a short time. The short-term (12 h) incubation under the 5X Fe (3.25 mg/L) and dark conditions resulted in the best lipid productivity of 1.44 g/L/d providing 2 g/L inoculum at the second stage.

Disinfection of Phytophthora spp. in Recycling Nursery Irrigation Water by Ozone Treatment (오존처리에 의한 폐양액내 Phytophthora spp. 멸균)

  • Lee, Jung-Sup;Han, Kyoung-Suk;Park, Jong-Han;Cheong, Seung-Ryong;Jang, Han-Ik
    • Research in Plant Disease
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    • v.12 no.3
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    • pp.272-277
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    • 2006
  • Recycled irrigation water is a primary inoculum source of Phytophthora spp. and is capable of spreading propagules throughout nursery cultivation. Ozonation is commonly used to disinfest the recycled irrigation water; however, ozone has not been fully researched as a disinfectant for this purpose. In this study, zoospores of four species of Phytophthora were exposed for $1{\sim}9$ min to free available ozone at $0.1{\sim}0.3,\;0.5{\sim}0.7,\;0.9{\sim}1.2,\;1.4{\sim}1.7\;and\;1.9{\sim}2.2mg/l$. Zoospores, mycelial fragments, and culture plugs of P. nicotianae also were exposed to ozone concentrations ranging from 0.1 to 2.2 mg/l for periods ranging from 1 to 9 min. In addition, ozonated water was assayed monthly in 2004 and 2005 at two commercial nurseries, and quarterly in the first year at two other nurseries in Suwon, for ozone and survival of pythiaceous species using a selective medium. No zoospores of any species tested survived endpoint free ozone at 1.4 mg/l while limited mycelial fragments of P. nicotianae survived at 1.9 mg/l, and mycelial plugs treated at the same level of ozone were able to produce few sporangia. Phytophthora spp. were recovered only from nursery irrigation water with levels of free ozone at 0.3 mg/l or lower. The results of this study are essential for improving current ozonation sterilization.

Effect of Lime Sulfur on Changes of Fungal Diversity in Pear Fallen Leaves (석회유황합제가 배나무 낙엽의 진균 다양성 변화에 미치는 영향)

  • Min, Kwang-Hyun;Song, Jang Hoon;Cho, Baik Ho;Yang, Kwang-Yeol
    • The Korean Journal of Mycology
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    • v.43 no.4
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    • pp.281-285
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    • 2015
  • This study was conducted to examine changes in the fungal community on fallen leaves of pear by treatment with lime sulfur. Although the lime sulfur could reduce the primary inoculum of several pathogens on spring season, the effect of lime sulfur has not been well determined scientifically. Fallen leaves infected by pear diseases in pear orchards in Naju were collected and treated with lime sulfur or water as a control. To determine the fungal diversity from each treatment, rDNA internal transcribed spacer (ITS) regions were analyzed after extraction of fungal genomic DNA from lime sulfur-treated or water-treated fallen leaves, respectively. The most common fungal species were Ascomycota and Basidiomycota in both treated leaves. However, the population dynamics of several fungal species including Alternari sp., Cladosporium sp., and Phomopsis sp., which are known as pear pathogens for skin sooty dapple disease, were quite different from each treated leaves. These results indicated that lime sulfur treatment led to changes of fungal communities on pear fallen leaves and could be applicable as a dormant spray.

Ultimate Anaerobic Biodegradability and Multiple Decay Rate Coefficients of Organic Wastes (유기성 폐자원의 최종생분해도 및 다중 분해속도 평가)

  • Kim, Sun-Woo;Kang, Ho;Jeong, Ji-Hyun
    • Journal of Korean Society of Environmental Engineers
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    • v.37 no.7
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    • pp.387-395
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    • 2015
  • Anaerobic mesophilic batch test of several organic wastes were carried out by a graphical statistic analysis (GSA) to evaluate their ultimate biodegradability and two distinctive decay rates ($k_1$ and $k_2$) with their corresponding degradable substrate fractions ($S_1$ and $S_2$). Each 3 L batch reactor was operated for more than 100 days at the substrate to inoculum ratio (S/I) of 0.5 as an initial total volatile solids (TVS) mass basis. Their Ultimate biodegradabilities were obtained respectively as follow; 69% swine waste, 45% dairy cow manure, 66% slaughterhouse waste, 79% food waste, 87% food waste leachate, 68% primary sludge and 39% waste activated sludge. The readily biodegradable fraction of 89% ($S_1$) of Swine Waste BVS ($S_o$) degraded with in the initial 31 days with $k_1$ of $0.116day^{-1}$, where as the rest 11% slowly biodegradable fraction ($S_2$) of BVS degraded for more than 100 days with the long term batch reaction rates ($k_2$) of $0.004day^{-1}$. For the Food Waste and Waste Activated Sludge, their readily biodegradable portions ($S_1$) appeared 89% and 80%, which degrades with $k_1$ of $0.195day^{-1}$ and $0.054day^{-1}$ for an initial 15 days and 28 days, respectively. Their corresponding long term batch reaction rates ($k_2$) were $0.003day^{-1}$ and $0.002day^{-1}$. Results from other organic wastes are addressed in this paper. The theoretical hydraulic retention times (HRTs) of anaerobic digesters treating organic wastes are easily determined by the analysis of multiple decay rate coefficients ($k_1$ and $k_2$) and their corresponding biodegradable substrate fractions ($S_1$ and $S_2$).

Disinfection of Fusarium-infected Rice Seeds by Prochloraz and Gaseous Chlorine Dioxide

  • Jeon, Young-ah;Lee, Young-yi;Lee, Ho-sun;Sung, Jung-sook;Lee, Seokyoung
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.25-25
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    • 2014
  • Three species of Fusarium, F. fujikuroi, F. verticillioides and F. proliferatum, are known to be associated with bakanae disease of rice [1, 2]. F. fujikuroi infects rice flowers and survive in endosperm and embryo of the seeds. Infected seed is an important source of primary inoculum of pathogens [3]. Seeds of rice (Oryza sativa cv. Boramchan) collected from bakanae-infected field were found to be 96% infected with Fusarium sp., 52% with F. fujikuroi, 42% with F. verticillioides, and 12% with F. proliferatum as determined by incubation method and species-specific PCR assays. F. fujikuroi was detected at lemma/palea, endosperm and embryo whereas F. verticillioides and F. proliferatum were recovered only from lemma/palea by means of component plating test. Seed disinfection methods have been developed to control bakanae disease and prochloraz has been most widely used for rice seeds. Two chemicals formulated with prochloraz (PC 1) and prochloraz + hexaconazole (PC 2) that inhibit biosynthesis of ergosterol strongly reduced the incidence of Fusarium spp. on selective media to 4.7% and 2.0%, respectively. Disease symptoms of rice seedlings in nursery soil were alleviated by chemical treatment; seedlings with elongated leaves or wide angle between leaf and stem were strikingly reduced from 15.6 to 3.2% (PC 1) and 0 (PC 2), stem rots were reduced from 56.9 to 26.2% (PC 1) and 32.1% (PC 2), and normal seedling increased from 0.4 to 13.3% (PC 2). Prochloraz has some disadvantages and risks such as the occurrence of tolerant pathogens [4] and effects on the sterol synthesis in animals and humans [5]. For these reasons, it is necessary to develop new disinfection method that do not induce fungal tolerance and are safe to humans and animals. Chlorine dioxide ($ClO_2$), that is less toxic, produces no harmful byproducts, and has high oxidizing power, has been reported to be effective at disinfection of several phytopathogenic fungi including Colletotrichum spp. and Alternaria spp. [6]. Gaseous $ClO_2$ applied to rice seeds at a concentration of 20 ppm strongly suppressed mycelial growth of Fusarium fujikuroi, F. verticillioides and F. proliferatum. The incidence of Fusarium spp. in dry seed with 8.7% seed moisture content (SMC) tended to decrease as the concentration of $ClO_2$ increased from 20 to 40 ppm. Applying 40 ppm $ClO_2$ at 90% relative humidity, incidence was reduced to 5.3% and resulted in significant reduction of disease symptoms on MS media. In nursery soil, stem rot was reduced from 56.9 to 15.4% and the number of normal seedlings increased from 0.4 to 25.5%. With water-soaked seeds (33.1% SMC) holding moisture in the endosperm and embryo, the effectiveness of disinfection using $ClO_2$ increased, even when treated with only 20 ppm for four hours. This suggests that moisture was a key element for action of $ClO_2$. Removal of the palea and lemma from seeds significantly decreased the incidence of Fusarium spp. to 3.0%. Seed germination appeared to decrease slightly by water-soaking at $30^{\circ}C$ because of increased SMC and by physical damage of embryos from hulling. These results indicate that the use of gaseous $ClO_2$ was effective as a means to disinfect rice seeds infected with Fusarium spp. and that moisture around the pathogens in the seed was an important factor for the action of $ClO_2$. Further investigations should be conducted to ascertain the best conditions for complete disinfection of Fusarium spp. that infect deep site of rice seeds.

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CELL CULTURE STUDIES OF MAREK'S DISEASE ETIOLOGICAL AGENT (조직배양(組織培養)에 의한 Marek 병(病) 병원체(病原體)의 연구(硏究))

  • Kim, Uh-Ho
    • Korean Journal of Veterinary Research
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    • v.9 no.1
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    • pp.23-62
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    • 1969
  • Throughout the studies the following experimental results were obtained and are summarized: 1. Multiplication of agents in primary cell cultures of both GF classical and CR-64 acute strain of Marek's disease infected chicken kidneys was accompanied by the formation of distinct transformed cell foci. This characteristic nature of cell transformation was passaged regularly by addition of dispersed cell from infected cultures to normal chicken kidney cell cultures, and also transferred was the nature of cell transformation to normal chick-embryo liver and neuroglial cell cultures. No cytopathic changes were noticed in inoculated chick-embryo fibroblast cultures. 2. The same cytopathic effects were noticed in normal kidney cell monolayers after the inoculation of whole blood and huffy coat cells derived from both forms of Marek's disease infected chickens. In these cases, however, the number of transformed cell foci appearing was far less than that of uninoculated monolayers prepared directly from the kidneys of Marek's disease infected chickens. 3. The change in cell culture IS regarded as a specific cell transformation focus induced by an oncogenic virus rather than it plaque in slowly progressing cytopathic effect by non-oncogenic viruses, and it is quite similar to RSV focus in chick-embryo fibroblasts in many respects. 4. The infective agent (cell transformable) were extremely cell-associated and could not be separated in an infective state from cells under the experimental conditions. 5. The focus assay of these agents was valid as shown by the high degree of linear correlation (r=0.97 and 0.99) between the relative infected cell concentration (in inoculum) and the transformed cell foci counted. 6. No differences were observed between the GF classical strain and the CR-64 acute strain of Marek's disease as far as cell culture behavior. 7. Characterization of the isolates by physical and chemical treatments, development of internuclear inclusions in Infected cells, and nucleic acid typing by differential stainings and cytochemical treatments indicated that the natures of these cell transformation agents closely resemble to those described fer the group B herpes viruses. 8. Susceptible chicks inoculated with infected kidney tissue culture cells developed specific lesions of Marek's disease, and in a case of prolonged observation after inoculation (5 weeks) the birds developed clinical symptoms and gross lesions of Marek's disease. Kidney cell cultures prepared from those inoculated birds and sacrificed showed a superior recovery of cell transformation property by formation of distinct foci. 9. Electron microscopic study of infected kidney culture cells (GF agent) by negative staining technique revealed virus particles furnishing the properties of herpes viruses. The particle was measured about $100m{\mu}$ and, so far, no herpes virus envelop has been seen from these preparations. 10. No relationship of both isolates to avian leukosis/sarcoma group viruses and PPLO was observed.

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