• 제목/요약/키워드: In vitro inoculation

검색결과 204건 처리시간 0.026초

눈꽃동충하초(Paecilomyces tenuipes)의 분생자경속 형성 (Optimal medium and cultural conditions for synnemata formation of Paecilomyces tenuipes)

  • 박기범;박기병;부샨 쓰레스타;성재모
    • 한국버섯학회지
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    • 제2권2호
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    • pp.60-68
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    • 2004
  • 눈꽃동충하초 분생자경속을 형성하는데 액체배양을 이용한 접종원의 배지는 PDB가 가장 우수하였으며, 분생자경속 형성에 유리한 배양온도는 $20^{\circ}C$로 나타났다. 조명은 500lx.에서 가장 양호한 성장을 하였으며 광조사 시간은 18hr이 가장 유리한 분생자경형성 조건을 나타내었다. 알맞은 배지 조건은 번데기와 현미를 50 : 50의 비율에 증류수를 현미에 1 : 1비율로 첨가한 혼합배지에서 높게 나타났다. 번데기배지는 원형상태와 분쇄상태 중에서는 분쇄된 상태에서 번데기배지의 공극률이 높아 균사의 침입이 용이하여 분생자경속 회수율과 균사의 배지내에 형성이 원형상태의 번데기보다 더욱 높게 나타났으며, 균사의 생장도 역시 분쇄한 번데기에서 더욱 빠르고 고르게 나타났다. 종균 접종량은 $15m{\ell}$의 종균을 접종하면 분생자경속이 잘 형성되었다. 눈꽃동충하초 재배에 가장 안정하게 재배하려면 분쇄한 번데기에 현미와 혼합한 배지에 종균 $15m{\ell}$를 접종하여 24 에서 발이를 시킨 후 $20^{\circ}C$의 배양실에서 재배하는 것이 가장 좋은 것으로 나타났다.

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고사리 전엽체의 증식에 미치는 배지구성물질과 배양방법의 영향 (Effect of Medium Components and Culture Methods on Prothallus Propagation of Pteridium aquilinum var. latiusculum (Desv.) Underw. ex Hell.)

  • 신소림;이무열;최재선;이철희
    • 한국자원식물학회지
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    • 제22권4호
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    • pp.337-342
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    • 2009
  • 본 연구는 식용 양치식물인 고사리의 대량번식체계를 확립하기 위하여 기내에서 전엽체 증식에 미치는 배지종류, 당의 종류와 농도, agar 농도, 전엽체 접종량 및 배양기간의 영향을 구명하기 위하여 수행되었다. MS배지를 기본으로 하여 배지구성물질을 1/4${\sim}$2배 첨가한 배지와 Hyponex를 0.1${\sim}$0.4%에서 전엽체를 배양한 결과 2MS배지에서 전엽체의 생육이 가장 왕성하였으며, MS배지 구성물질들 중 인산염이 고사리 전엽체의 증식에 가장 영향이 큰 것으로 나타났다. 탄소원의 종류 및 농도별로는 sucrose와 glucose 공히 1% 이상을 첨가한 구에서 전엽체의 생육이 비슷한 경향을 보였다. Agar의 첨가량은 전엽체의 증식에 큰 영향을 미치지 않았으며, agar를 첨가하지 않은 액체배지에서도 전엽체 증식이 왕성하였다. 액체배지 100 mL에 전엽체를 각각 100과 200 mg 접종하여 22일 동안 진탕배양한 결과, 전엽체는 20일 이후에는 증가속도가 완만해지는 경향을 보였으며, 초기 접종량이 20일 후 전엽체 증식에 큰 영향을 미치지 않으므로 100 mg을 곱게 다져 진탕배양 한 후, 20일 후 계대배양 하여 증식시키거나, 기외로 이식하여 포자체를 형성하는 것이 유리한 것으로 사료되었다.

The In Vitro and In Vivo Efficacy of Hen IgY Against Vibrio parahaemolyticus and Vibrio vulnificus

  • Kassim, Neema;Mtenga, Adelard B.;Shim, Won-Bo;Chung, Duck-Hwa
    • Journal of Microbiology and Biotechnology
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    • 제22권10호
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    • pp.1423-1431
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    • 2012
  • The inhibitory effect of IgY against Vibrio parahaemolyticus and Vibrio vulnificus responsible for seafood-borne diseases was investigated in this study. Water-soluble fractions (WSF) of protein containing IgYs were isolated from the egg yolk of hens initially immunized with formalin-inactivated V. parahaemolyticus or V. vulnificus. Protein, total and specific IgY contents of the WSF were determined. The inhibitory and protective effects of IgYs on the growth of V. parahaemolyticus and V. vulnificus were assayed in liquid medium and in mice. IgYs showed high affinity to their corresponding antigens with high titer from day 28 onwards. Protein contents and total IgY concentrations remained stable throughout the immunization period, whereas specific IgY concentrations increased steadily and reached a plateau at day 49. Specific IgY powder (150 mg/ml) significantly inhibited further multiplication of both V. parahaemolyticus and V. vulnificus in liquid medium as compared with the control IgY. The bacteria count in mice feces was lower in mice pretreated with specific IgYs than in those pretreated with PBS or control IgY. Higher survival of mice was observed in the experimental groups pretreated with either anti-V. parahaemolyticus (75% survival) or anti-V. vulnificus (87% survival) IgYs, compared with those in the control groups pretreated with PBS or nonspecific IgY. All mice in the control groups died within three days after bacteria inoculation; hence, the protective effect of specific IgYs against infection caused by V. parahaemolyticus and V. vulnificus was demonstrated.

Colony Age of Trichoderma azevedoi Alters the Profile of Volatile Organic Compounds and Ability to Suppress Sclerotinia sclerotiorum in Bean Plants

  • Lincon Rafael, da Silva;Leonardo Luis de Barros, Rodrigues;Amanda Silva, Botelho;Bruna Sartorio, de Castro;Paulo Henrique Pereira Costa, Muniz;Maria Carolina Blassioli, Moraes;Sueli Correa Marques, de Mello
    • The Plant Pathology Journal
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    • 제39권1호
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    • pp.39-51
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    • 2023
  • Common bean (Phaseolus vulgaris L.) is one of the most important crops in human food production. The occurrence of diseases, such as white mold, caused by Sclerotinia sclerotiorum can limit the production of this legume. The use of Trichoderma has become an important strategy in the suppression of this disease. The aim of the present study was to evaluate the effect of volatile organic compounds (VOCs) emitted by Trichoderma azevedoi CEN1241 in five different growth periods on the severity of white mold in common bean. The in vitro assays were carried out in double-plate and split-plate, and the in vivo assays, through the exposure of the mycelia of S. sclerotiorum to the VOCs of T. azevedoi CEN1241 and subsequent inoculation in bean plants. Chemical analysis by gas chromatography coupled to mass spectrometry detected 37 VOCs produced by T. azevedoi CEN1241, covering six major chemical classes. The profile of VOCs produced by T. azevedoi CEN1241 varied according to colony age and was shown to be related to the ability of the biocontrol agent to suppress S. sclerotiorum. T. azevedoi CEN1241 VOCs reduced the size of S. sclerotiorum lesions on bean fragments in vitro and reduced disease severity in a greenhouse. This study demonstrated in a more applied way that the mechanism of antibiosis through the production of volatile compounds exerted by Trichoderma can complement other mechanisms, such as parasitism and competition, thus contributing to a better efficiency in the control of white mold in bean plants.

박과류 검은점뿌리썩음병의 발생분포 및 분리병원균의 병원성 (Occurrence and Distribution of Monosporascus Root Rot and Pathogenicity of Monosporascus cannonballus on Cucurbitaceae Plants)

  • 허노열;류경열;현익화;권진혁
    • 식물병연구
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    • 제7권1호
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    • pp.11-15
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    • 2001
  • 최근 박과류 재배 주산단지에서 발생이 증가하고 있는 검은점뿌리썩음병의 방제에 필요한 기초자료를 얻기 위해서 지역적인 발생분포와 발생소장을 조사하고 병원성을 검정하였다. Monosporascus cannonballus 에 의한 검은점뿌리썩음병은 멜론, 참외, 수박 및 오이에서 발생하였으며, 발생포장 수는 \\`97년 10개 포장에서 \\`98년 32개 포장으로 확대되었고, 발생지역은 김해, 진주, 남해, 광양, 광주, 여주, 이천 등이었다. 경남 남해 멜론 재배포장에서 발생소장을 조사한 결과, 생육중기까지는 발병을 관찰할 수 없었으나, 과실 성숙기에 급격히 발병이 증가하여 50%이상의 식물체 덩굴이 시들어 고사하였다. 시들은 증상을 나타내는 기주에서 M. cannonballus의 분리비율은 36.7%이었고, Fusarium oxysporum과 Rhizoctonia solani도 높은 비율로 분리되었다. 물한천 시험관법에 의한 박과류 유묘에 대한 병원성은 접종후 7∼14일 사이에 대부분의 식물체가 시들고 21일후에는 뿌리에 자낭각이 형성되었다. 특히 오이품종 중에서 백봉과 은화는 저항성 반응을 나타내었다.

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Comparison of respiratory pathogenesis of porcine reproductive and respiratory syndrome virus isolates in vitro and in vivo

  • Park, Bong-kyun;Collins, James E.;Goyal, Sagar M.;Joo, Han-soo
    • 대한수의학회지
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    • 제39권2호
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    • pp.318-326
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    • 1999
  • Respiratory pathogenic effects of several porcine reproductive and respiratory syndrome virus(PRRSV) isolates were examined in swine tracheal ring(STR) cultures by examining their effect on ciliary activity. One high and one low pathogenic PRRSV isolates were then selected and their pathogenicity investigated in 3-week-old conventional PRRSV-seronegative pigs. Ten pigs each were inoculated intranasally with the high or low pathogenic PRRSV isolate and 6 pigs were sham inoculated as negative controls. Two pigs each from the inoculated group and one pig each from negative control group were killed on 4, 7, 14, 21 and 28 days postinoculation(pI). At necropsy, degrees of gross lung lesion was determined. Turbinate, tonsil, trachea and lung samples were collected for virus isolation or histopathology. Gross lung lesions were observed mainly on 14 days PI with high and low pathogenic isolates inducing moderate diffuse and mild gross lung lesions, respectively. Inoculation of either the high or low pathogenic virus resulted in loss of cilia in ciliated epithelium of turbinates and trachea between 7 and 28 days PI. High pathogenic virus caused increased number of Goblet cells in the tracheal epithelial layer between 4 and 21 days PI whereas the low pathogenic virus did it between 14 and 28 days PI and with a lesser degree. Although both viruses produced interstitial pneumonia, the lesion was less severe with the low pathogenic virus. The isolation of high pathogenic virus from tissues and sera was earlier and more consistent than that of the low pathogenic virus. The agreement between in vitro and in vivo tests indicates that STR cultures may be used as a routine method to determine the respiratory pathogenicity of PRRSV isolates.

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Impact of Rhizosphere Competence of Biocontrol Agents upon Diseases Suppression and Plant Growth Promotion

  • Park, Chang-Seuk-
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 1994년도 Proceedings of International Symposium on BIOLOGICAL CONTROL OF PLANT DISEASES Korean Society of Plant Pathology
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    • pp.27-49
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    • 1994
  • Root colonization of biocontrol agents via seed treatment was investigated and a compatible combination, Gliocladium virens G872B and Pseudomonas putida Pf3, in colonizing cucumber rhizosphere was confirmed through the study. Much higher number of fungal and bacterial propagules were detected when two isolates were inoculated together. The presence of Pf3 in root system was greatly helpful to G872B to colonize at root tip. The mechanism of this phenomenon is partially elucidated through the results of in vitro experiments and the observations of scanning electron and fluorescence microscope. Addition of Pf3 cells resulted earlier germination of G872B conidia and increased mycelial growth. And the more number of germinated conidia on seed coat, the more vigorous hypal streching and sporulation on the root surface were observed in coinoculated treatment. The propagules of G872B on the cucumber root when they were challenged against the pathogenic Fusarium oxysporum, were even higher than that of G872B treated alone, and the magnitude of such a difference was getting grater toward the root ip and the population of F. oxysporum on the root was reduced by seed inoculation of G872B. The rhizosphere competence was obviously reflected to disease suppression and plant growth promotion that induced by the given isolate. Green house experiments revealed that the combined treatment provided long-term disease suppression with greater rate and the larger amount of fruit yield than single treatments. Through this study the low temperature growing Pseudomonas fluorescens M45 and MC07 were evaluated to apply them to the winter crops in field or plastic film house. In vitro tests reveal that M45 and MC07 inhibited the mycelial growth of Pythium ultimum, Rhizoctona solani and Phytophthora capsici and enhanced growth of cucumber cotyledon in MS agar. This effect was more pronounced when the bacteria were incubated at 14$^{\circ}C$ than at 27$^{\circ}C$. And disease suppression and plant growth promotion in green house were also superior at low temperature condition. Seed treatment of M45 or soil treatment of MC07 brought successful control of damping-off and enhanced seedling growth of cucumber. The combined treatment of two isolates was more effective than any single treatment.

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Pteris cretica 'Wilsonii'의 전엽체 기내 대량번식 및 형태형성에 미치는 배지 구성물질 및 배양 방법 (Medium Composition Affecting In Vitro Masspropagation and Morphogenesis in Prothalli of Pteris cretica 'Wilsonii')

  • 신소림;황주광;이철희
    • 화훼연구
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    • 제17권2호
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    • pp.114-120
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    • 2009
  • Pteris cretica 'Wilsonii'의 대량생산에 적합한 배양환경을 구명하기 위하여 연구를 수행하였다. 포자는 7주 안에 모두 발아하였다. Knop과 Hyponex배지에서 전엽체 증식이 왕성하였지만, 전엽체의 생육에는 Hyponex 배지가 더 유용하였다. MS배지에서는 전엽체가 괴사하였으며, 질소급원과 sucrose의 농도를 조절한 경우에도 sucrose 무첨가구를 제외한 모든 첨가구에서 전엽체가 괴사하였다. Sucrose 1%와 agar 0.6%를 첨가한 Hyponex배지가 전엽체의 증식과 생육에 가장 적합한 것으로 나타났다. 접종방법을 달리한 결과, Hyponex배지에서는 다져서 접종하는 전엽체의 증식 및 생육에 효과적이었지만, MS배지에서는 전엽체의 군집을 4등 분하여 접종한 처리구에서 전엽체의 증식 및 생육이 우수하였다. 고체배지에서 배양하는 것이 액체배지에서 배양하는 것보다 효과적이었다. 액체배양은 전엽체의 괴사를 유도하였다. 액체진탕배양한 전엽체는 생육은 우수하였으나 고체배양한 전엽체에 비하여 증식이 억제되었다.

A Study on the Reduction of Gossypol Levels by Mixed Culture Solid Substrate Fermentation of Cottonseed Meal

  • Zhang, Wenju;Xu, Zirong;Sun, Jianyi;Yang, Xia
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권9호
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    • pp.1314-1321
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    • 2006
  • The objective of this work was to study the effect of mixed culture solid substrate fermentation of C. tropicalis ZD-3 with A. niger ZD-8 on detoxification of cottonseed meal (CSM), and to investigate the effect of fermentation period, proportion of CSM in substrate, sodium carbonate, minerals and heat treatment on the reduction of free gossypol levels during mixed culture solid substrate fermentation of CSM. Experiment 1: Three groups of disinfected CSM substrate were incubated for 48 h after inoculation with either of the fungi C. tropicalis ZD-3, A. niger ZD-8 or mixed culture (C. tropicalis ZD-3 with A. niger ZD-8). One non-inoculated group was used as the control. Levels of initial and final free gossypol (FG), CP and in vitro CP digestibility were assayed. The results indicated that mixed culture fermentation was far more effective than single strain fermentation, which not only had higher detoxification rate, but also had higher CP content and in vitro digestibility. Experiment 2: CSM substrates were treated according to experimental variables including fermentation period, proportion of CSM in substrate, sodium carbonate, minerals and heat treatment, Then, the treated CSM substrates were inoculated with mixed culture (C. tropicalis ZD-3 with A. niger ZD-8) and incubated at $30^{\circ}C$ for 36 h in a 95% relative humidity chamber. After fermentation ended, FG and CP content of fermented CSM substrate was assayed. The results showed that the appropriate fermentation period was 36 h, and the optimal proportion of CSM in substrate was 70%. Addition of sodium carbonate to CSM substrate was beneficial for fermentative detoxification. Heat treatment could facilitate fermentative detoxification, and supplementation with minerals was instrumental in reducing gossypol levels during mixed culture solid substrate fermentation of CSM.

상홍백출산(桑紅白朮散)이 생쥐 대장암세포의 간전이억제와 면역활성화에 미치는 효과 (Effect of Sanghongbaekchul-san on Anti-metastatic and Immunopotentiating Activities)

  • 오세순;강희;심범상;김성훈;최승훈;안규석
    • 동의생리병리학회지
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    • 제22권2호
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    • pp.282-289
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    • 2008
  • We evaluated the effect of SHBCS on adhesion and invasion of colon L5-26 adenocarcinoma cell line in vitro in vitro and experimental liver metastasis in vivo. SHBCS showed little inhibitory effect on colon 26-L5 cell proliferation. At the concentration of up to 500 mg/ml of SHBCS 80% of cells were viable. SHBCS showed no inhibitory effect on adhesion and invasion of colon 26-L5 cells, which were placed on matrigel. In a dose dependent manner, oral administration of SHBCS showed a significantly inhibitory effect on liver metastasis from colon 26-L5 injected mice. When mice were depleted of NK cells or macrophages before tumor inoculation, SHBCS significantly decreased liver metastasis fromf the tumor injected mice. Compared with the control mice, SHBCS increased the populations of macrophages and NK cells by 30%, 18%(10 mg/mouse, 50 mg/mouse) and 5%, 1% (10 mg/mouse, 50 mg/mouse) respectively. Compared with the control mice, SHBCS increased the populations of CD4 cells by 5%, 18% (10 mg/mouse, 50 mg/mouse) respectively. Spelenocytes from mice administerd with SHBCS were stimulated with LPS plus ConA, proliferation of splenocytes from mice administerd with SHBCS was 140%, 146%(10 mg/mouse, 50 mg/mouse) compared with th control mice. In conclusion, the present study suggests that SHBCS may have an inhibitory effect on liver metastasis through immunopotentiating activity which is associated with macrophages and NK cells.