• Title/Summary/Keyword: artificial inoculum

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Further Evidence that Ramularia-type Conidia in vivo Plays a Role as a Secondary Inoculum of Mycosphaerella nawae (감나무 둥근무늬낙엽병균 Mycosphaerella nawae 분생포지의 2창 전염원으로서 역할)

  • 권진혁;강수웅;박창석;김희규
    • Korean Journal Plant Pathology
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    • v.14 no.5
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    • pp.393-396
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    • 1998
  • The characteristic Ramularia type conidia of Mycosphaerella nawae were formed on naturally infected leaves. Artificial inoculation with the conidia induced typical symptom on leaves, which was not distinguishable from those of ascospore infection, which has been considered as a primary inoculum source in nature. Also the morphology of the conidia produced on PDA was not different from those formed on artificially inoculated leaves or on naturally infected leaves at later stage of symptom development. Accordingly, we report the role of the conidia as a secondary inoculum of the circular leaf spot pathogen of persimmon for the first time.

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Occurrence of Bottom Rot of Crisphead Lettuce Caused by Rhizoctonia solani and Its Pathogenicity (Rhizoctonia soiani에 의한 결구상추 밑둥썩음병(Bottom rot)의 발생과 병원성)

  • 김현주;박종영;백정우;이진우;정순재;문병주
    • Journal of Life Science
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    • v.14 no.4
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    • pp.689-695
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    • 2004
  • This study was investigate the occurrence of bottom rot caused by Rhizoctonia solani at the crisphead lettuce fields in Uiryeong-gun, Gyeongsangnam-do from November to December in 2003. Incidence of bottom rot on crisphead lettuce was up to 5.3% at the six plastic houses. A total of 30 isolates of R. solani were obtained from diseased leaves of plants and were tested by artificial inoculation to the host. Among them, PY-1 isolate was selected showing highly virulent on the whole plant and was identified as R. solani AG1 (IB) based on the anastomosis test, morphological and cultural characteristics. Symptoms of bottom rot by PY-1 isolate produced small dark brown, depressed and elliptical spots on the lower part of leaves in the early stage as same as at the fields, were enlarged onto the upper part of leaves later, and the infected plant wilted and ultimately died in the end. For the pathogenicity test, triturated mycelia-inoculum (A$_{550}$=1.0) of PY-1 isolate was selected the most effective inoculum showing disease incidence of 51.1% for the mycelial inoculation at pot assay. Otherwise, WSRP media-inoculum (wheat brane : sawdust : rice brane : PDB media=30 g : 10 g : 10 g : 100 ml, w/w/w/v) of PY-1 isolate was effectual inoculum showing disease incidence of 61.6% for soil inoculation at the plastic house. Also, in selection of density and amount of inoculum, most suitable density of triturated mycelia-inoculum and amount of WSRP media- inoculum were determined as $A_{550}$=1.0 and 40 ml, respectively. This is the first report on the pathogenicity test using by WSRP media-inoculum of R. solani PY-1 isolate for the bottom rot of crisphead lettuce.

Optimization of Culture Conditions and Bench-Scale Production of $_L$-Asparaginase by Submerged Fermentation of Aspergillus terreus MTCC 1782

  • Gurunathan, Baskar;Sahadevan, Renganathan
    • Journal of Microbiology and Biotechnology
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    • v.22 no.7
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    • pp.923-929
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    • 2012
  • Optimization of culture conditions for L-asparaginase production by submerged fermentation of Aspergillus terreus MTCC 1782 was studied using a 3-level central composite design of response surface methodology and artificial neural network linked genetic algorithm. The artificial neural network linked genetic algorithm was found to be more efficient than response surface methodology. The experimental $_L$-asparaginase activity of 43.29 IU/ml was obtained at the optimum culture conditions of temperature $35^{\circ}C$, initial pH 6.3, inoculum size 1% (v/v), agitation rate 140 rpm, and incubation time 58.5 h of the artificial neural network linked genetic algorithm, which was close to the predicted activity of 44.38 IU/ml. Characteristics of $_L$-asparaginase production by A. terreus MTCC 1782 were studied in a 3 L bench-scale bioreactor.

Proliferation of Tricholoma matsutake Mycelial Mats in Pine Forest Using Mass Liquid Inoculum

  • Lee, Won-Ho;Han, Sang-Kuk;Kim, Beom-Seok;Shrestha, Bhushan;Lee, Soo-Yong;Ko, Cheol-Soon;Sung, Gi-Ho;Sung, Jae-Mo
    • Mycobiology
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    • v.35 no.2
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    • pp.54-61
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    • 2007
  • Two isolates of Tricholoma matsutake T-008 and T-034, preserved in Entomopathogenic Fungal Culture Collection (EFCC) of Korea, were used in the present study. The isolates had 100% Bootstrap homology with Tricholoma matsutake U62964 and T. matsutake AB188557 and AF309538 preserved in Gene Bank of NCBI. Mycelial growth of T. matsutake was highest in TMM and MYA at $25^{\circ}C$. The highest dry wt. of mycelium was obtained after 65 days of culture, when 6 mycelial discs were inoculated in 100 ml of broth in 250 ml shaking flask. Mycelial mats were observed in clumped condition at the inoculation sites of pine forest after two weeks of inoculation. After 5 months of inoculation, mycelia mats were observed growing inside soil and walls of a few inoculation sites, while mycelial mats growth up to $5{\sim}8$ cm were observed in the roots of pine tree after 6 months. The survival rate of the inoculum was about 40% of the total inoculation sites. The survival rate was found below 20% when the mycelium was inoculated in the summer. The reasons for low survival rates of the mycelium were mainly due to dry season and the soil-borne small animals such as earthworm and mole. After one year of inoculation, no external difference was observed between the artificially inoculated mycelia and the naturally existing mycelia of T. matsutake. The present study showed that fruiting bodies of T. matsutake could be produced by artificial inoculation under the appropriate environmental conditions.

Occurrence of Clubroot Caused by Plasmodiophora brassicae in Baecheongchae

  • Kim, Wan-Gyu;Oh, Sang-Keun;Semunyana, Marc;Han, Man-Jong;Lee, Gyo-Bin;Cho, Weon-Dae
    • The Korean Journal of Mycology
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    • v.48 no.4
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    • pp.499-503
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    • 2020
  • Clubroot symptoms were frequently observed on the roots of Baecheongchae plants grown in vinyl greenhouses of a farmer located in Yangpyeong area of Korea during a disease survey in June 2019. The incidence of diseased Baecheongchae plants ranged from 30 to 90% in the vinyl greenhouses investigated. Many resting spores were found in the tissue of root galls collected. The resting spores were hyaline and spherical and measured 2.5-4.2 ㎛ in diameter. Three inoculum suspensions of resting spores prepared from the root galls were inoculated to the roots of healthy Baecheongchae plants. All the inoculum suspensions caused clubroot symptoms to appear on the roots of the inoculated Baecheongchae plants. The symptoms on the roots induced by artificial inoculation were similar to those observed in the plants of the vinyl greenhouses during the disease survey. Resting spores of the pathogen were recovered from the root galls of the inoculated plants. Three root gall isolates obtained from the inoculated plants were used for molecular identification. Comparing the isolates to the Plasmodiophora brassicae strains in GenBank, the amplification products demonstrated 100% similarity with the internal transcribed spacer (ITS2) sequences. The clubroot pathogen was identified as P. brassicae according to its morphological, pathological, and molecular characteristics. This is the first report of P. brassicae causing clubroot in Baecheongchae.

Estimation of In vitro Digestibility of Barley Straw by Using a Homogenized Rumen Fluid and Artificial Saliva Mixed with Nitrogen and Energy Sources

  • Chaudhry, Abdul S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.11 no.1
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    • pp.13-16
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    • 1998
  • A $2{\times}2{\times}4$ factorial study was conducted to examine the possibility of improving estimates of in vitro digestibility, using untreated (UBS) and ammonia-treated (ABS) barley straw, through homogenization of rumen fluid (RF) and by additions of urea (U) and casein (C) as N sources and Xylose + Glucose (XG) as energy sources into artificial saliva. Digestibiltiy of ABS was significantly greater than that of UBS (p < 0.001). There was a significant decrease in digestibility when additions (U, UC, UCXG) were compared with the control (p < 0.001). A 2-way interaction between RF and straw type was significant (p < 0.05) for dry matter digestibility (DMD). Homogenization of RF increased DMD of ABS (p < 0.05) whereas it decreased DMD of UBS (p > 0.05). The study showed that addition of N alone or in combination with energy sources was not better than control, rather the reverse, digestion was inhibited by a combination of U and casein (UC). It was concluded that sufficient N and branched chain fatty acids were supplied in the inoculum from sheep fed high protein grass cubes to support the growth of cellulolytic microbes during in vitro incubation. Further studies are, however, required to explore the potential of homogenization in improving the in vitro method to estimate digestibility of cereal straws.

Artificial Microparticle Diets for Culturing Rotifer, Brachionus plicatilis

  • Hwang Joo-Ae;Kim Sung-Koo
    • Fisheries and Aquatic Sciences
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    • v.1 no.1
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    • pp.30-34
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    • 1998
  • Rotifer culture fed on five types of artificial microparticle diets were evaluated to substitute the natural diets such as Chlorella or w-yeast. These microparticle diets including solidified blood using squid oil (SBSO), solidified blood using soybean oil (SBSB), nylon protein walled particle (NPW) simple coacervation oil capsule (SCO), complex coacervation oil capsule(CCO), were tested for the evaluation of feeding efficiency. The prepared micro particle diets had diameters ranging from 3 to 30 Jim. Rotifer culturing experiments were carried out in 3-liter beakers for 13-16 days. The initial inoculum density of rotifers was 10 ind./ml. The rotifers fed on Chlorella or $\omega-yeast$ showed maximal densities of 2,000 ind./ml in 9 days or 500 ind./ml in 7 days, respectively. Those fed on SBSO, SBSB or NPW showed maximal densities of 1568 ind./ml, 586 ind./ml or 503 ind./ml, respectively and the reproductive rates for those diets were equivalent to or better than w-yeast. However, the coacervated oil capsule showed lower maximal densities of 400 ind./ml for SCO and less than 100 ind./ml for CCO due to the unbalanced diet formulation and indigestibility.

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Improved Method to Increase Conidia Production from Isolates of Different Pathotypes of Citrus Scab Pathogen Elsinoe spp.

  • Hyun, Jae-Wook;Paudyal, Dilli Prasad;Hwang, Rok-Yeon
    • Research in Plant Disease
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    • v.21 no.3
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    • pp.231-234
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    • 2015
  • Elsinoe fawcettii and E. australis are two currently recognized scab pathogens of citrus. E. fawcettii has at least six pathotypes while E. australis has at least two pathotypes. Colonies of E. fawcettii and E. australis do not sporulate in artificial media including potato dextrose agar (PDA). Whiteside's method has been widely used for preparing conidial inoculum in vitro. This study was carried out to develop efficient method for conidia production from artificial media. We developed a shaking method which included the following steps: 1) Colony grown on PDA was mashed with a steel spatula; 2) Mycelia fragments were cultured in 50 ml sterilized rain water in a rotary shaker-incubator (180 rpm) at $25^{\circ}C$ for 24 h: 3) The conidia suspension was filtered through two layers of cheesecloth. Average conidia production of all isolates tested using this shaking method was approximately 13.1 times higher than that from Whiteside's method in this study.

Influence of Cultivated Conditions and Fungicides on Development of Rhizoctonia Diseases of Zoysiagrass and Bentgrass (한국들잔디와 벤트그라스의 라이족토니아 마름병 발생에 미치는 재배조건 및 살균제의 영향)

  • 이재홍;이두형
    • Asian Journal of Turfgrass Science
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    • v.9 no.3
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    • pp.199-206
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    • 1995
  • This study is aimed to clarify on the influence of cultivated conditions and fungicides on development of Rhizoetonia diseases of zoysiagrass and creeping bentgrass and results are as follows. Infection rates of disease were observed more serious when inoculum amounts were higher than low. Effects of the watering intervals on the severity of disease with artificial inoculating conditions, infection rates were higher when two days watering interval for creeping bentgrass and one day interval for zoysiagrass. Disease severity of creeping bentgrass incited by Rhizoetonia solani AG 1 (1A) was grater under conditions of double amounts of nitrogen fertilization. A posi- tive correlation was founded between the pH and disease development. The pots with pH 5 and 8 were significantly more diseased grass than those in which the pH was in 6.5. Mepronil and myclobutanil were extremely effective against brown patch of creeping bentgrass and large patch of zoysiagrass and followed by capro, and tebeonazole. Fenari was inferior effect to both diseases.

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Screening of Resistance Melon Germplasm to Phytotpthora Rot caused by Phytophthora Capsici

  • Kim, Min-Jeong;Shim, Chang-Ki;Kim, Yong-Ki;Jee, Hyeong-Jin;Hong, Sung-Jun;Park, Jong-Ho;Lee, Min-Ho;Han, Eun-Jung
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.57 no.4
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    • pp.389-396
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    • 2012
  • Melon (Cucumis melo) is an annual herbaceous plant of the family Cucurbitaceae. Phytophthora rot, caused by Phytophthora capsici is a serious threat to cucurbits crops production as it directly infects the host plant, and it is difficult to control because of variable pathogenicity. This study investigated the resistance of 450 accessions of melon germplasm against Phytophthora rot by inoculating the seedlings with sporangial suspension ($10^{5\;or\;6}$ zoosporangia/ml) of P. capsici. Disease incidence of Phytophthora rot was observed on the melon germplasm at 7-day intervals for 35 days after inoculation. Susceptible melon germplasm showed either severe symptoms of stem and root rot or death of the whole plant. Twenty out of 450 tested accessions showed less than 20% disease incidence, of which five accessions showed a high level of resistance against Phytopthtora rot. Five resistant accessions, namely IT119813, IT138016, IT174911, IT174927, and IT906998, scored 0% disease incidence under high inoculum density of P. capsici ($10^6$ zoosporangia/mL). We recommend that these candidate melon germplasm may be used as genetic resources in the breeding of melon varieties resistant to Phytophthora rot.