• Title/Summary/Keyword: in vitro cultivation

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Characteristics of the Oxygen Uptake Rate of Entomopathogenic Nematodes Steinernema spp. (곤충병원성 선충 Steinernema spp.의 산소 요구도 특성)

  • 김도완;박선호
    • Korean journal of applied entomology
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    • v.38 no.2
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    • pp.123-128
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    • 1999
  • Recently, entomopathogenic nematodes have received a considerable attention as biologicalcontrol agents. For in vitro cultivation, storage and transportation of nematodes, oxygen supply isextremely impotant due to its limited solubility and mass transfer problem. The oxygen uptake rates(OURs) of four different Steinernema species were measured in a 5L bioreactor at varying temperatures.The OURs of the Steinernema spp. were below 0.5 x mmolO'||'&'||' . min in the range of 13-17$^{\circ}$C. TheOURs (mmo102/L - min) of S. glaseri Dongrae and S. carpocapsae Pocheon strains were 0.4 x lo-', 0.75x lo-\ulcorner at 21$^{\circ}$C, 1.5 x lo-\ulcorner, 3.2 x 10-2 at 25"C, and 2.8 x lo-', 5.8 x lo-\ulcorner at 29"C, respectively. However,the OURs were not significantly altered by the agitation speed of 50-150 rpm. The specific oxygenuptake rates (qol) of S. glaseri NC, S. glaseri Dongrae, S. glaseri Mungyeong and S. carpocapsaePocheon strains were 0.3 x 0.5 x 0.3 x and 0.2 x mmolO~/cell min at 25"C,respectively. As the nematode size and temperature were increased, the qo, was also increased.the qo, was also increased.

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Bacillus spp. as Biocontrol Agents of Root Rot and Phytophthora Blight on Ginseng

  • Bae, Yeoung-Seuk;Park, Kyungseok;Kim, Choong-Hoe
    • The Plant Pathology Journal
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    • v.20 no.1
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    • pp.63-66
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    • 2004
  • Ginseng (Panax ginseng) is one of the most widely cultivated medicinal herbs in Korea. However, yield losses reached up to 30-60% due to various diseases during 3 or 5 years of ginseng cultivation in the country. Therefore, successful production of ginseng roots depends primarily on the control of diseases. The objective of this study was to select potential biocontrol agents from rhizobacteria isolated from various plant internal root tissues for the control of multiple ginseng diseases as an alternative to fungicides. Among 106 Bacillus strains, two promising biocontrol agents, Bacillus pumilus strain B1141 and Paenibacillus lentimobus strain B1146, were selected by screening against root rot of ginseng caused by Cylindrocarpon destructans in a greenhouse. Pre-inoculation of selected isolates to seed or l-year-old root of ginseng resulted in stimulation of shoot and/or root growth of seedlings, and successfully controlled root rot caused by C. destructans (P<0.05). Furthermore, drenching of cell suspension of the selected isolates on seedling-growing pots reduced the incidence of Phytophthora blight after the seedlings were challenged with zoospores of Phytophthora cactorum (P<0.05). P. lentimorbus strain B1146 showed antifungal activity against various soil-borne pathogens in vitro, while B. pumilus strain B1141 did not show any. Results of this study suggest that some rhizobacteria can induce resistance against various plant diseases on ginseng.

Fungicide selections for control of chili pepper stem rot caused by Sclerotium rolfsii using an agar dilution method

  • Lee, Soo Min;Min, Jiyoung;Kim, Heung Tae
    • Korean Journal of Agricultural Science
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    • v.44 no.3
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    • pp.339-347
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    • 2017
  • Sclerotium rolfsii causing southern blight on numerous vegetable and fruit crops was isolated from stems of chili peppers showing wilting symptoms. The pathogen was identified by morphological observation and DNA sequencing analysis of ITS region. To select an effective fungicide for control of southern blight, we investigated the inhibition efficacy of thirty fungicides included in nine groups of fungicides with different mechanisms of action. A fungal growth inhibition assay was conducted through an agar dilution method by using mycelial discs and sclerotia of the pathogen as inoculum, respectively. When mycelial discs were used as an inoculum, several fungicides showed good inhibitory activity against the mycelial growth of S. rolfsii 12-6. All DMI fungicides tested had a good inhibition except for prochloraz which had low inhibitory effect. All strobilurin fungicides tested except for kresoxim-methyl and all SDHI fungicides tested except for boscalid and fluopyram, had a good inhibition. Also, fludioxonil, a protective fungicide and fluazinam had a good inhibitory effect. Interestingly, when sclerotia were used as an inoculum, inhibition efficacy was increased for fluopyram, a SDHI fungicide, and for some protective fungicides such as propineb, chlorothalonil, dithianon, and folpet. All the fungicides selected in this study should be tested in the field for their control activities against stem rot for practical use in chili pepper cultivation.

Biological characteristics of Scuticociliate with a polymorphic life cycle of ciliate and cyst isolated from cultured olive flounder, Paralichthys olivaceus (양식넙치, Paralichthys olivaceus에서 분리된 Ciliate와 Cyst의 생활사를 갖는 스쿠티카섬모충의 생물학적 특성)

  • Jin, Chang-Nam;Kang, Hyun-Sil;Park, Geun-Tae;Lee, Je-Hee;Lee, Young-Don;Choi, Woo-Bong;Heo, Moon-Soo
    • Journal of fish pathology
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    • v.18 no.1
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    • pp.19-28
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    • 2005
  • Scuticociliatosis has badly settled one of most damaging diseases during the seedling production process of olive flounder. Paralichthys olivaceus in Korea. We isolated a new type of Scuticociliate from flounder. The parasite metamorphoses to ciliate and cyst phases with each other by environmental changes and survive for a relatively long span. The ciliate was measured average 41.8 ${\mu}m$ in length and 21.0 ${\mu}m$ in width, and cyst was 17.0 ${\mu}m$ and 13.5 ${\mu}m$, respectively. Nutritional condition was determined as a major parameter of metamorphosing between ciliate and cyst stages. The ciliate transforms to a cyst stage because of food shortage, and the cyst returns to a ciliate stage with a favorite environmental condition and shows active growth and reproduction. The ciliate multiplied at the maximal density of $2.9 {\times} 10^5 {m\ell}^{-1}$cells in vitro cultivation at $15 ^{\circ}C$temperature using MS BHI medium and bacterial food sources. The ciliate could be proliferated at a 2.5 to $30 ^ \circ}C$ temperature range, pH 6 to 9, and 1 to 55 ppt salinity. Particularly, it survived over one week at $0 ^{\circ}C$temperature showing a high resistance against unfavorable environmental conditions. And the cyst survived for 320 days in the condition of $5 ^{\circ}C$with no feeding, but its survival period was markedly shortened in higher temperature conditions. The chemotherapeutants (formalin and hydrogen peroxide) were clarified as effective chemicals against the ciliate during in vitro trials, but the effect of therapeutants differed in proportion, depending upon the density and the bathing time of chemical compounds.

Growth Inhibition of In Vitro Plantlets and Improvement of Survival Rate of Acclimated Plant of Strawberry according to Polyethylene Glycol during Bioreactor Culture (딸기 조직배양묘의 생물반응기 배양 시 Polyethylene Glycol 처리에 따른 기내 생육억제 효과 및 순화율 향상)

  • Kim, Hye Jin;Lee, Jong Nam;Kim, Ki Deog;Lim, Hak Tae;Yeoung, Young Rok
    • Horticultural Science & Technology
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    • v.33 no.6
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    • pp.877-882
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    • 2015
  • This study was carried out treatment of polyethylene glycol (PEG) in order to increase of survival rate of acclimated plants of strawberry's in vitro plantlets through bioreactor culture. We used PEG with molecular weight 6000 (PEG 6000) in this study. Concentration of PEG is non-treatment, 5, 10, 15, and $20g{\cdot}L^{-1}$ each bioreactor. $5g{\cdot}L^{-1}$ of PEG was treated at $1^{st}$, $2^{nd}$, $3^{rd}$, $4^{th}$, and $5^{th}$ week during culture. We investigated growth characteristics of in vitro plantlets after 6 weeks cultivation. Growth amount of all PEG treatment decreased as compared to non-treatment. The more concentration increased, the more plant growth decreased. In $5g{\cdot}L^{-1}$ of PEG, shoot length was shorter than non-treatment that shoot length was 7.9 cm and especially fresh weight that is 1.6 g was more decrease than non-treatment. Shoot length was ranged 3.0-3.9 cm at $1^{st}$ week treatment to $4^{th}$ week treatment of $5g{\cdot}L^{-1}$ PEG. The shoot length was not significant by 5.3 cm at $5^{th}$ week treatment. The survival rate was improved 5.4% at the treatment of $4^{th}$ week and was improved 8.7% at the $5^{th}$ week as compared to non-treatment. In order to improve of survival rate of strawberry' in vitro plantlets through bioreactor culture, the method is suitable that adding $5g{\cdot}L^{-1}$ of PEG in the medium and $5^{th}$ week's treatment is suitable.

Characteristics of sawdust cultivation of Lentinula edodes with different methods of spawn inoculation

  • Chang, Hyun You;Seo, Geum Hui;Lee, Yong Kuk;Jeon, Sung Woo
    • Journal of Mushroom
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    • v.16 no.2
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    • pp.61-64
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    • 2018
  • This study was carried out to investigate the management characteristics and growth performance of L. edodes from the cooling stage to incubation. Bags of different heights and weights are available for bagging. When the medium size of $17{\times}13cm$ was used and the size of the inoculation hole was changed from 1/3 to 2/3, the browning period was shortened to 30 days. Mycelial growth was evaluated according to the cooling temperature after sterilization. It was observed to be the highest at 122 mm/15 days at $10^{\circ}C$ and 114 mm/15 days and 117 mm/15 days at $15^{\circ}C$ and $20^{\circ}C$, respectively. The contamination rate of the sawdust media before inoculation was measured as 0, $4.5{\times}10$, $1.3{\times}10^2$, $4.0{\times}10^3cfu$ at $5^{\circ}C$, $10^{\circ}C$, $15^{\circ}C$, and $24^{\circ}C$ respectively. The average of $1.6{\times}10^8$ colony forming units (cfu) of microorganisms was observed in the sawdust that had been piled for six months outdoors. In summer, the sawdust has to be used immediately after mixing. The sterilized medium had an average of $4{\times}10^3cfu$ of microorganisms at $24^{\circ}C$ and $1.3{\times}10^2cfu$ at $15^{\circ}C$. After 15 days of inoculation in vitro, the growth conditions of the sawdust was the best at 132 mm, followed by grain and liquid. When inoculated with liquid spawn, the moisture content of the substrate should be adjusted between 50% and 55% in advance.

Two Flavonoid-Based Compounds from Murraya paniculata as Novel Human Carbonic Anhydrase Isozyme II Inhibitors Detected by a Resazurin Yeast-Based Assay

  • Sangkaew, Anyaporn;Samritsakulchai, Nawara;Sanachai, Kamonpan;Rungrotmongkol, Thanyada;Chavasiri, Warinthorn;Yompakdee, Chulee
    • Journal of Microbiology and Biotechnology
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    • v.30 no.4
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    • pp.552-560
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    • 2020
  • Human carbonic anhydrase (CA) isozyme II has been used as protein target for disorder treatment including glaucoma. Current clinically used sulfonamide-based CA inhibitors can induce side effects, and so alternatives are required. This study aimed to investigate a natural CA inhibitor from Murraya paniculata. The previously developed yeast-based assay was used to screen 14 compounds isolated from M. paniculata and identified by NMR analysis for anti-human CA isozyme II (hCAII) activity. Cytotoxicity of the compounds was also tested using the same yeast-based assay but in a different cultivation condition. Two flavonoid candidate compounds, 5, 6, 7, 8, 3', 4', 5'-heptamethoxyflavone (4) and 3, 5, 7, 8, 3', 4', 5'-heptamethoxyflavone (9), showed potent inhibitory activity against hCAII with a minimal effective concentration of 10.8 and 21.5 μM, respectively, while they both exhibited no cytotoxic effect, even at the highest concentration tested (170 μM). The results from an in vitro esterase assay of the two candidates confirmed their hCAII inhibitory activity with IC50 values of 24.0 and 34.3 μM, respectively. To investigate the potential inhibition mechanism of compound 4, in silico molecular docking was performed using the FlexX and SwissDock software. This revealed that compound 4 coordinated with the Zn2+ ion in the hCAII active site through its methoxy oxygen at a distance of 1.60 Å (FlexX) or 2.29 Å (SwissDock). The interaction energy of compound 4 with hCAII was -13.36 kcal/mol. Thus, compound 4 is a potent novel flavonoid-based hCAII inhibitor and may be useful for further anti-CAII design and development.

Effect of Chlorella sp. on Improving Antioxidant Activities and Growth Promotion in Organic Soybean Sprout Cultivation (클로렐라 처리에 의한 유기농 콩나물 생육촉진 및 항산화 능력 증진효과)

  • Kim, Min-Jeong;Shim, Chang-Ki;Kim, Yong-Ki;Hong, Sung-Jun;Park, Jong-Ho;Han, Eun-Jung;Jee, Hyeong-Jin;Lee, Sung-Buk;Kim, Seok-Cheol
    • Korean Journal of Organic Agriculture
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    • v.23 no.4
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    • pp.939-950
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    • 2015
  • The purpose of this study was to estimate the growth promoting effects and improvement of antioxidant activity of the soybean sprouts treated with Chlorella sp. culture solution. The soybean sprout treated with 0.1% and 0.2% Chlorella sp. culture solution was significantly increased the length (more than 43.0%), the thickness (more than 0.5~0.7 mm), fresh weight (more than 2.9~3.7 g) compared to non-treated control in vitro. In organic soybean sprouts farm, the 0.2% chlorella culture solution applied to mass culture of soybean sprout and the fresh weight of soybean sprouts increased by more than 25% and the yield was very high as 598.33% compared to untreated control. In addition of sensory test, there is no fishy odor and better crunchy texture and nutty flavor for the treatment soybean sprouts compared to untreated soybean sprouts. Particularly, free-radical scavenging activity (DPPH) and superoxide dismutase activity (SOD) of the soybean sprouts were significantly increased more than 26.1% and 40.4%, respectively by treated with 0.1% and 0.2% Chlorella culture solution. Consequently, the treatment of chlorella culture solution to grow soybean sprouts is also promoting quality and antioxidant activity as well as promoting the growth of sprouts. Therefore, chlorella is considered to be worth as functional materials for high-quality sprouts grown.

Biological Control of Strawberry Bud Rot Caused by Rhizoctonia solani AG2-1 with Antagonistic Microorganism (길함미생물에 의한 시설재배 딸기 눈마름병의 생물학적 방제)

  • 신동범;소림기언;이준탁
    • Korean Journal Plant Pathology
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    • v.10 no.2
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    • pp.112-118
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    • 1994
  • Forth microbial isolates out of 167 isolates from the soil of controlled cultivation areas inhibited mycelial growth of Rhizoctonia solani AG2-1 causing the strawberry bud rot in vitro. Among the isolates, Kr013 and Kr020 showed suppressive effect to R. solani AG2-1 on seedlings of chinese cabbage treated by root immersion, charcoal carrier granule and drenching on 1.0% infested soil in pot. Furthemore, the corresponding effect was also revealed when the charcoal carrier granule of the isolates were treated on the seedling of strawberry that were planted on the planting hole in pot. To examine the effects of biological control in green house, it had been tested the infection rates by using two different treatments. First, the strawberry runner were planted on the nursery soil mixed with 20% charcoal carrier granule of Kr013 and Kr020 isolate respectively, and grown for 20 days before transplanting. Then the young plants form the mother plant were separated and transplanted on the 1.0% infested soil. Another method was that the charcoal carrier of Kr013 and Kr020 isolates applied to planting hole of 1.0% infested soil just before transplanting. Then the young plants were grown for 20 days on the sterilized nursery soil before transplanting. From the results, the effects of biological control was significantly higher on former treatment (e.g. the infection rates were 7.3 and 5.7%, respectively) than on the latter treatment (e.g. the corresponding value were 16.7 and 15.7%, respectively). The antagonistic isolates of Kr013 and Kr020 were respectively identified as Pseudomonas cepacia with the similarity of 55.0% and 60.0% by using the Biolog GN Microplate system.

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Effect of Seeding Dates and Rates on the Productivity and Nutritive value of Hairy Vetch(Vicia villosa Roth) (파종시기 및 파종량이 헤어리 벳치의 수량 및 사료가치에 미치는 영향)

  • Kim Jong Geon;Chung Eui Soo;Kim Meng Jung;Seo Sung;Lee Jong Kyung;Kim Jong Duk;Seo Jong Ho
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.25 no.1
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    • pp.17-22
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    • 2005
  • This experiment was conducted to investigate the effect of seeding dates and rates on the productivity and nutritive value of hairy vetch(Vicia villosa Roth) at experimental field of Grassland and Forage Crops Division, National Livestock Research Institute, RDA, Suwon from 1999 to 2000. The experimental design was consisted of split-plot design with three replications. The main plot was seeding dates(25 Aug., 15 Sep. and 5 Oct.) and the subplot was seeding rates(20, 30 and 40 kg/ha). Late seeding dates increased the plant height and delayed the flowering stage as 1 or 2 days. The dry matter(DM) content was decreased with low seeding rates, and the trend was same in the all plots. Average crude protein(CP) content was $24.1\%$ and increased with delayed seeding date and lower seeding rate. The content of acid detergent fiber(ADF) and neutral detergent fiber(NDF) was increased with higher seeding rate, but seeding date did not affect. In vitro dry matter digestibility(IVDMD) of hairy vetch tended to decrease with high seeding rate, and total digestible nutrient(TND) did not show significant difference among seeding rates. Average relative feed value(RFV) of hairy vetch showed 140 which means very high quality. Dry matter and CP yield was decreased with delayed seeding date. The results of this experiments indicated that seeding in middle-August with 40 kg/ha would be recommended to produce the highest yield and quality f3r the cultivation of hairy vetch in middle part of Korea