• Title/Summary/Keyword: In vitro inoculation

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A Cytotoxic Activity of Panax Ginseng Extract Against Bome Cancer Cells In Vivo and In Vitro.

  • Hwang, Woo-Ik;Park, Gil-Hong;Paik, Jeong-Mi
    • Proceedings of the Ginseng society Conference
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    • 1987.06a
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    • pp.29-37
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    • 1987
  • This study was devised to observe the cytotoxlc activities of petroleum-ether extract of Panax ginseng root(crude Gx) and its partially purified fraction from silicon acid column chromatography(7:3 CX) against sarcoma-180(5-180) and Walker carcinosarcoma 256(Walker 256) in vivo, and murine leukemic lymphocytes(L1210) and human rectal cancer cell(HRT-18) and human colon cancer cells(HT-29 and HCT-48) in vitro . Each cell-line was cultured in medium containing serial concentrations of the crude Gx or 7:3 Gx in vitro. A highly lipid soluble compound in the extract of Panax ginseng root was cytocidal to murine leukemic cells and human colon and rectal cancer cells in vitro In the meantime, ginseng saponin derivatives did not cytotoxic effects at its corresponding concentration. The growth rates of the cancer cells in medium containing ginseng extracts were inhibited gradually to a significant degree roughly in proportion to the increase of the extract concentration. The cytotoxic activity of 7:3 Gx was about 3 times more potent than that of crude Gx, one unit of cytotoxic activity against L121f cells being equivalent to 2.54$\mu\textrm{g}$ and 0.88 $\mu\textrm{g}$ for the crude Gx and 7:3 Gx, respectively. The Rf value of the active compound on silica -gel thin layer chromatography with petroleum-ether/ethyl ether/acetic acid mixture (90:10:1, v/v/v) as a developing solvent was 0.23. The survival times of mice inoculated with S-180 cells were extended about 1.5 to 2 times by the 7:3 Gx treatment compared with their control group. The significantly decreased hemoglobin values of rats after inoculation with Walker 256 were recovered to normal range by oral administration of the crude Gx. The synthetic levels of protein, DNA and RNA in human colon and rectal cancer cells were significantly diminished by treatment with the crude Gx, which can explain a part of the origin of its anticancer activity.

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The Effect of Seed-borne Mycoflora from Sorghum and Foxtail Millet Seeds on Germination and Disease Transmission

  • Yago, Jonar I.;Roh, Jae-Hwan;Bae, Soon-Do;Yoon, Young-Nam;Kim, Hyun-Ju;Nam, Min-Hee
    • Mycobiology
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    • v.39 no.3
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    • pp.206-218
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    • 2011
  • The seed-borne mycoflora of sorghum and foxtail millet collected from different growing areas in South Korea were isolated and taxonomically identified using dry inspection, standard blotter and the agar plate method. We investigated the in vitro and in vitro germination rates of disinfected and non-disinfected seeds of sorghum and foxtail millet using sterilized and unsterilized soil. The percent recovery of seed-borne mycoflora from the seed components of sorghum and foxtail millet seeds was determined and an infection experiment using the dominant species was evaluated for seedling emergence and mortality. A higher number of seed-borne fungi was observed in sorghum compared to that of foxtail millet. Eighteen fungal genera with 34 fungal species were identified from the seeds of sorghum and 13 genera with 22 species were identified from the seeds of foxtail millet. Five dominant species such as Alternaria alternata, Aspergillus flavus, Curvularia lunata, Fusarium moniliforme and Phoma sp. were recorded as seed-borne mycoflora in sorghum and 4 dominant species (Alternaria alternata, Aspergillus flavus, Curvularia lunata, Fusarium moniliforme) were observed in foxtail millet. The in vitro and in vitro germination rates were higher using disinfected seeds and sterilized soil. More seed-borne fungi were recovered from the pericarp compared to the endosperm and seed embryo. The percent recovery of seed-borne fungi ranged from 2.22% to 60.0%, and Alternaria alternata, Curvularia lunata and 4 species of Fusarium were isolated from the endosperm and embryo of sorghum and foxtail millet. Inoculation of the dominant seed-borne fungi showed considerable mortality of seedlings. All the transmitted seed-borne fungi might well be a primary source of infection of sorghum and foxtail millet crops.

Effect of Lactic Acid Bacteria Treatment on Nutritive Value and In Vitro Ruminal Fermentation of Italian Ryegrass (Lolium multiflorum L.) Silage

  • Lee, Kihwan;Marbun, Tabita Dameria;Kim, Suyeon;Song, Jaeyong;Kwon, Chan Ho;Yoon, Duhak;Kang, Jungsun;Lee, Chanho;Cho, Sangbuem;Kim, Eun Joong
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.40 no.3
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    • pp.182-189
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    • 2020
  • This study was conducted to evaluate the effect of lactic acid bacteria (LAB) inoculation to domestically-cultivated Italian ryegrass (IRG) on silage fermentation and in vitro ruminal fermentation. There were six treatments based on the LAB inoculants: 1) no addition of LAB (negative control: NC), additions of 2) commercially-available LAB (positive control: PC), 3) Lactobacillus plantarum (LPL), 4) L. paracasei (LPA), 5) L. acidophilus (LA), and 6) L. pentosus (LPT). All treatments were inoculated at a concentration of 106 CFU/g and ensiled for 3, 7, 21, and 42 days in triplicate and analyzed for nutritive values when ensiling was terminated. Day 42 silage from all treatments were also examined for in vitro ruminal fermentation. After 42 days, LAB-inoculated silages had higher (P<0.05) lactic acid concentration compared to the NC. In terms of nutritive values, the silages treated with LPA, LA, and LPT showed higher (P<0.05) crude protein and lower (P<0.05) neutral detergent fiber and acid detergent fiber content compared to the rest of the treatment. In vitro ruminal dry matter degradability was not affected by LAB addition. However, LAB-treated IRG had shown higher (P<0.05) ammonia-N compared with that of the NC. LPA had shown the highest (P<0.05) volatile fatty acid concentration among the LAB examined. In conclusion, the addition of a single strain of LAB appeared to produce a quality IRG silage compared with the NC and the PC. Among the strains examined, LPA seemed to be superior to the others.

In vivo Anti-fungal Activity of the Essential Oil Fraction from Thymus Species and in vitro Synergism with Clotrimazole

  • Kim, Ji-Hyun;Shin, Seung-Won
    • Natural Product Sciences
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    • v.13 no.3
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    • pp.258-262
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    • 2007
  • The antifungal activity of the essential oil fraction from Thymus magus, and its major component thymol, against Candida albicans was investigated in vitro and in vivo. The combined effects of the oils and clotrimazole, a commonly used antifungal drug for treatment of external candidiasis, were evaluated in this study. In experimental vaginal candidiasis the essential oil fraction of T. magnus resulted in relatively milder inhibition of fungal growth following the inoculation of test mice compared to clotrimazole. However, new fungal growth was not detected up to 12 days after cessation of treatment. In contrast, in a similar experiment using clotrimazole, C. albicans was detected in the $12^{th}$ day post-treatment with the sample. This result indicates that T. magnus oil could be a promising drug to control vaginal candidiasis. In checkerboard titer tests, the combination of clotrimazole with the essential oil fraction of T. magus or T. quinquecostatus resulted in significant synergism, with FIC indices between 0.14 and 0.27 against C. albicans, while clotrimazole combined with thymol, the major component of these oils, produced only an additive effect, with FIC indices ranging between 0.50 and 1.00. Thus, the prominent synergistic effects of clotrimazole combined with T. magus essential oil indicate that these compounds may be an effective treatment for C. albicans infections.

Induced Resistance in Tomato Plants Against Fusarium Wilt Invoked by Nonpathogenic Fusarium, Chitosan and Bion

  • Amini, J.
    • The Plant Pathology Journal
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    • v.25 no.3
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    • pp.256-262
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    • 2009
  • The potential of. nonpathogenic Fusarium oxysporum strain Avr5, either alone or in combination with chitosan and Bion, for inducing defense reaction in tomato plants inoculated with F. oxysporum f. sp lycopersici, was studied in vitro and glasshouse conditions. Application Bion at concentration of 5, 50, 100 and $500{\mu}g$/ml, and the highest concentration of chitosan reduced in vitro growth of the pathogen. Nonpathogenic F. oxysporum Avr5 reduced the disease severity of Fusarium wilt of tomato in split plants, significantly. Bion and chitosan applied on tomato seedlings at concentration $100{\mu}g$ a.i./plant; 15, 10 and 5 days before inoculation of pathogen. All treatments significantly reduced disease severity of Fusarium wilt of tomato relative to the infected control. The biggest disease reduction and increasing tomato growth belong to combination of nonpathogenic Fusarium and Bion. Growth rate of shoot and root markedly inhibited in tomato plants in response to tomato Fusarium wilt as compared with healthy control. These results suggest that reduction in disease incidence and promotion in growth parameters in tomato plants inoculated with nonpathogenic Fusarium and sprayed with elicitors could be related to the synergistic and cooperative effect between them, which lead to the induction and regulation of disease resistance. Combination of elicitors and non-pathogenic Fusarium synergistically inhibit the growth of pathogen and provide the first experimental support to the hypothesis that such synergy can contribute to enhanced fungal resistance in tomato. This chemical could provide a new approach for suppression of tomato Fusarium wilt, but its practical use needs further investigation.

First Report of Pyrenophora tritici-repentis in Wheat (Triticum aestivum L.) in Korea and in vitro Selection of an Effective Fungicide

  • Min-Hye Jeong;Eu Ddeum Choi;Seol-Hwa Jang;Sang-Min Kim;Sook-Young Park
    • The Korean Journal of Mycology
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    • v.51 no.4
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    • pp.277-286
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    • 2023
  • Tan spot, caused by Pyrenophora tritici-repentis, is a major foliar disease in wheat worldwide. In April 2021, tan spot symptoms were observed in a commercial wheat field in Suncheon, Jeonnam Province, Korea, with over 5% of the wheat leaves exhibiting symptoms. These symptoms included oval-shaped tan necrosis surrounded by a bright halo. The three representative isolates exhibited irregular mycelial growth on V8-potato dextrose agar and produced pseudothecia. Based on the concatenated sequence datasets of four multi-genes, including the internal transcribed spacer, large subunit ribosomal RNA, glyceraldehyde-3-phosphate dehydrogenase, and RNA polymerase II second-largest subunit genes, phylogenetic analysis revealed that these three isolates clustered in the same clade as P. tritici-repentis. Results of pathogenicity test indicated that the initial symptoms appeared 5 days post-inoculation (dpi), with typical tan spot symptoms developing at 7 dpi. The pathogen was successfully re-isolated from the symptomatic tissues, thus fulfilling Koch's postulates. Furthermore, we selected three fungicides that effectively inhibited the mycelial growth of P. tritici-repentis by more than 90% in vitro. To the best of our knowledge, this is the first report of tan spot disease in wheat in Korea.

In Vitro Mycorrhization and Morphological Characterization of Xanthoconium affine with Pinus densiflora

  • Wang, Eun-Jin;Jang, Yeongseon;Ka, Kang-Hyeon
    • The Korean Journal of Mycology
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    • v.45 no.4
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    • pp.319-327
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    • 2017
  • In this study, we investigated the culture conditions of four ectomycorrhizal fungi, namely, Amanita spissacea NIFoS 2719, Pisolithus arhizus NIFoS 2784, Suillus spraguei NIFoS 2848, and Xanthoconium affine NIFoS 2716, in solid and liquid culture media. In addition, we attempted to induce in vitro mycorrhization of the fungi with Pinus densiflora. Prior to liquid culture, we determined the optimal culture conditions for each species in solid media. The results revealed that all species examined are capable of growth in potato dextrose agar (PDA), malt extract agar (MEA), and modified Melin-Norkran's medium (MMN), although their preferred growth media were different. Liquid culture experiments showed that inorganic nitrogen did not enhance the mycelial growth of all four species. Therefore, we used MMN-based liquid inocula to promote the growth of ectomycorrhizal fungi in our symbiosis culture system. Mycorrhization was observed in Xanthoconium affine NIFoS 2716. Morphological analysis revealed that fungi-inoculated roots of P. densiflora form simple and dichotomous lateral roots with dense mycelia. In addition, inoculation with X. affine NIFoS 2716 promoted root and shoot developments.

Effects of additives on the fermentation quality, in vitro digestibility and aerobic stability of mulberry (Morus alba L.) leaves silage

  • Dong, Zhihao;Wang, Siran;Zhao, Jie;Li, Junfeng;Shao, Tao
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.8
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    • pp.1292-1300
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    • 2020
  • Objective: To explore feed resources capable of replacing regular poor-quality fodder, this study was conducted to evaluate the effects of additives on the fermentation quality, in vitro digestibility and aerobic stability of mulberry leaves silage. Methods: The mulberry leaves were ensiled either untreated (control) or treated with 1×106 cfu/g fresh matter Lactobacillus plantarum (L), 1% glucose (G), 3% molasses (M), a combination of 1% glucose and Lactobacillus plantarum (L+G), and a combination of 3% molasses and Lactobacillus plantarum (L+M). The fermentation quality and chemical composition were analyzed after 7, 14, 30, and 60 d, respectively. The 60-d silages were subjected to an aerobic stability test and fermented with buffered rumen fluid to measure the digestibility. Results: Inoculating lactic acid bacteria (LAB) resulted in more rapid increase in lactic acid concentrations and decline in pH of mulberry leaves silage as compared control. Higher acetic acid and lower ethanol and ammonia nitrogen concentrations (p<0.05) were observed in the LAB-inoculated silages as opposed to control during ensiling. The LAB-inoculated silages contained lower water-soluble carbohydrates compared with control during the first 14 d of ensiling, and lower neutral detergent fibre (p<0.05) concentrations as compared with non-LAB inoculated silages. Adding molasses alone increased (p<0.05) the digestibility of dry matter (DM). The aerobic stability of mulberry leaves silage was increased by LAB inoculation, whereas decreased by adding glucose or molasses. Conclusion: The LAB inoculation improved fermentation quality and aerobic stability of mulberry leaves silage, while adding glucose or molasses failed to affect the fermentation and impaired the aerobic stability. Inoculating LAB alone is recommended for mulberry leaves especially when ensiled at a relatively high DM.

Light and Electron Microscopy Studies Elucidating Mechanisms of Tomato Leaf Infection by Pseudocercospora fuligena

  • Zelalem Mersha;Girma Birru;Bernhard Hau
    • The Plant Pathology Journal
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    • v.39 no.2
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    • pp.181-190
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    • 2023
  • The fungal pathogen Pseudocercospora fuligena, known to affect tomatoes in the tropics and subtropics, has been reported from temperate climates including the United States and Turkey in recent years. In this study, an isolate from fresh tomatoes and the disease it causes were characterized and infection mechanisms investigated. Macroscopically, both sides of tomato leaves show indistinct effuse patches but prolific production of fuliginous lesions is conspicuous on the abaxial side first but also on the adaxial side later on as infection progressed. Microscopically, fascicles of conidiophores (11-128 ㎛ × 3.5-9 ㎛) arising from stromata and conidia with up to 12 septations were observed. Molecular characterization of the isolate revealed high homology (99.8%) to other P. fuligena isolated from tomatoes in Turkey. Out of the 10 media tested, P. fuligena grew significantly well and sporulated better on unsealed tomato oatmeal agar and carrot leaf decoction agar, both supplemented with CaCO3. Direct transfer of conidia from profusely sporulating lesions was the easiest and quickest method of isolation for in-vitro studies. Light and scanning electron microscopy on cleared and intact tomato leaves further confirmed stomatal penetration and egress as well as prevalence of primary and secondary infection hyphae. In situ, blocked stomatal aperture areas of 154, 401, and 2,043 ㎛2 were recorded at 7, 12, and 17 days after inoculation, respectively. With the recent expanded horizon of the pathosystem and its consequential impact, such studies will be useful for a proper diagnosis, identification and management of the disease on tomato worldwide.

In Vitro Anti-Oomycete Activity and In Vivo Control Efficacy of Phenylacetic Acid Against Phytophthora capsici

  • Lee, Jung-Yeop;Kim, Hye-Sook;Kim, Ki-Deok;Hwang, Byung-Kook
    • The Plant Pathology Journal
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    • v.20 no.3
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    • pp.177-183
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    • 2004
  • Phenylacetic acid (PAA) was evaluated for in vitro anti-oomycete activity and in vivo control efficacy against Phytophthora capsici. Microscopic observation revealed that the high level of anti-oomycete activity of PAA (10 $\mu\textrm{g}$/ml) against P. capsici is mainly due to the lytic effect on zoospores. Zoospore lysis began in the presence of 5 u$\mu\textrm{g}$/ml of PAA and most of the zoospores were collapsed at 10 $\mu\textrm{g}$/ml. PAA showed inhibitory activity against the zoospore germination and hyphal growth of P. capsici at the concentration of 50 $\mu\textrm{g}$/ml. In the glasshouse, the protective effect of PAA against Phytophthora blight was high on pepper plants when treated just before inoculation with P. capsici. In the artificially infested field, protection of pepper plants against the Phyto-phthora epidemic was achieved at a considerable level by PAA treatment.