• Title/Summary/Keyword: In vitro inoculation

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Effect of Harvesting Time and Storage Duration of Viscum album Seeds on in vitro and ex vitro Germination on the Branch of Prunus mume

  • Lee, Hyun Woo;Ghimeray, Amal Kumar;Lee, Bo-Duk;Sharma, Pankaja;Shim, Ie Sung;Park, Cheol Ho
    • Korean Journal of Plant Resources
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    • v.27 no.3
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    • pp.256-262
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    • 2014
  • Viscum album var. coloratum (mistletoe) is considered as one of the endangered plant species in Korea. Our objective is to restore its population and multiplication of plant by ex situ method. In this research we explored the maximum germination (in vitro) of freshly harvested and stored seeds of mistletoe collected in different time intervals. Ex vitro germination after artificial inoculation on the branches of Prunus mume in different physiological conditions was also monitored. The research revealed that the lately harvested seeds (Feb. and March 2014) were superior over early harvested seeds (Nov. 2013 and Jan. 2014) of mistletoe due to the higher percentage of germination (above 93%). According to the data, it is also revealed that the survival and germination rate of mistletoe seeds decreased with the increase in storage duration. In ex vitro germination, the fluctuated temperature of a glass house in natural condition enhanced (four fold) the rate of germination on the branches of Prunus mume than the constant temperature condition in the glass house.

Control Efficacy of Bordeaux Mixture against Pepper Anthracnose (고추 탄저병에 대한 보르도액의 방제 효과)

  • Park, Se-Jung;Lee, Soo-Min;Gwon, Hyeon-Wook;Lee, Hotaek;Kim, Heung Tae
    • The Korean Journal of Pesticide Science
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    • v.18 no.3
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    • pp.168-174
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    • 2014
  • It was conducted to evaluate the control efficacy of Bordeaux mixture on pepper anthracnose in vitro and in vivo. The suppressive efficacy of Bordeaux mixture on mycelial growth and spore germination of Colletotrichum acutatum was investigated on PDA, cellophane membranes and pepper fruits, respectively. Furthermore, its control value was evaluated on detached pepper fruits inoculated with C. acutatum by wound and non-wound inoculation method, and in fields. The mycelial growth of C. acutatum JC24 was inhibited by 90.0% on PDA amended with $937{\mu}g\;mL^{-1}$ of Bordeaux mixture. While the spore germination of C. acutatum JC24 was inhibited perfectly on cellophane membranes treated at $187{\mu}g\;mL^{-1}$ of Bordeaux mixture, that on fruits inoculated with the pathogen by wound inoculation and non-wound inoculation method was inhibited by 88.1% and 91.3%, respectively. Although the control value on fruits treated with $937{\mu}g\;mL^{-1}$ of Bordeaux mixture was 17.6% in wound inoculation method, it was 58.8% in non-wound inoculation method. In fields, when Bordeaux mixture was sprayed five times at 14 day-intervals, it showed 55.7% and 61.7% of control value in 2012 and 2013, respectively. We think Bordeaux mixture was able to use as an eco-friendly organic farming material to control pepper anthracnose based on the above-mentioned results.

In vitro and In vivo Activities of a Biocontrol Agent, Serratia plymuthica A2l-4, Against Phytophthora capsici

  • Shen, Shun-Shan;Park, Ok-Hee;Lee, Sun-Mi;Park, Chang-Seuk
    • The Plant Pathology Journal
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    • v.18 no.4
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    • pp.221-224
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    • 2002
  • In vitro and in vivo activities of a biocontrol agent, Serratia plymuthica strain A2l-4, was evaluated for the control of Phytophthora blight of pepper, Strain A2l-4 inhibited mycelial growth, germination of zoosporangia and cystospores, and formation of zoospore and zoosporangia of Phytophthora capsici in vitro. In the pot experiment, incidence of Phytophthora blight of pepper in non-treated control was 100% at 14 days after inoculation, while no disease was observed in the plot treated with S. plymuthica A2l-4. In the greenhouse test, infection rate of pepper in the non-treated plots was 74.5%, while it was only 12.6% in the plots treated with A2l-4. Results indicate that S. plymuthica A2l-4 is a potential biocontrol agent for Phytophthora blight of pepper.

Mycorrhizal Formations and Seedling Growth of Pinus desiflora by in vitro Synthesis with the Inoculation of Ectomycorrhizal Fungi

  • Chung, Hung-Chae;Kim, Dong-Hun;Lee, Sang-Sun
    • Mycobiology
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    • v.30 no.2
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    • pp.70-75
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    • 2002
  • The mycelia were directly isolated from eight species of fungal basidiocarps, confirmed to the ectomycorrhiza in the roots from the fields(forestry); Suillus bovinus, Paxillus involutus, Lactarius hysginus, Russula fragilis, Lepista nuda, Lyophyllum shimeji, Tricholoma matsutake, and Russula integra. The mycelia were pure-cultured with several transferring in various agars, and inoculated to the roots of pine(Pinus densiflora) seedling by in vitro method. After ten months growth under artificially aseptic conditions, all pine seedlings inoculated were stimulated at the growth-height, whereas those not inoculated were nearly dead. Also, the ramifications of ectomycorrhizal pine roots formed in the synthetic in vitro systems and were various according to the different mycelia. Synthesis of ectomycorrhiza were clearly confirmed in ten months growth, but not distinguished at this moment. It was clearly proved that the mycelia isolated caused the ectomycorrhizae in the roots of pine seedlings.

Detection of Plant RNA Viruses by Hybridization Using In Vitro Transcribed RNA Probes (In Viro 전사 RNA Probe를 이용한 식물 바이러스병의 진단)

  • 최장경;이종희;함영일
    • Korean Journal Plant Pathology
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    • v.11 no.4
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    • pp.367-373
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    • 1995
  • The cDNAs derived from the coat protein (CP) genes of six plant RNA viruses, tobacco mosaic virus-pepper strains (TMV-P) and -ordinary strain (TMV-OM), potato virus Y (PVY), turnip mosaic virus (TuMV), cucumber mosaic virus (CMV) and potato leafroll virus (PLRV), were subcloned into the transcription vector, pSPT18, containing SP6 and T7 promoters. The digoxigenin (DIG)-labeled RNA polymerase after linearlization of the cloned pSPTs with XbaI or SacI, and were tested for their sensitivities for the detection of the six viruses. In slot-blot hybridization, dilution end points for the detection of TMV-P and TMV-OM were 10-4, while those of PVY, TuMV and CMV were 10-3. PLRV was detected at the dilution of 10-2. When each RNA probe was applied for the detection of the viruses in the preparations from the leaf disks (8 mm in diameter, and 12 to 15 mg in weight) of infected natural host plants, TMV-P, TMV-OM and TuMV could be detected from one disk, while PVY from 1 or 2 disks. CMV was detected in the preparation from two disks, and PLRV from three disks. With DIG-labeled RNA probe, PVY was detected at 5 days after inoculation, but with ELISA the virus was detected at 8 days after inoculation to tobacco (Nicotiana tabacum cv. Xanthi nc) plants on which symptoms appeared at 9 days after inoculation. No difference was observed in cross reaction between the RNA probes for the detection of TMV-P and TMV-OM.

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Seasonal Occurrence and Infection Site of Strawberry Anthracnose (딸기탄저병의 시기별 발생과 감염경로)

  • 김승한;최성용;임양숙;윤재탁;최부술
    • Research in Plant Disease
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    • v.8 no.1
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    • pp.45-49
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    • 2002
  • Seasonal occurrence of strawberry anthracnose in greenhouses caused by Colletotrichum sp. was examined from 1997 through 1999 at three locations, Kyeongju, Goryeong, and Cheongdo in Kyungbuk province, Korea. Also some factors related to the anthracnose infection such as initial infection sites, inoculation methods, and soil nature were studied through in vitro and field experiments. The anthracnose disease begun to occur from 15 days after transplanting in early October, and continued but gradually decreased thereafter for 2 months until December, After transplanting, initial infection mainly occurred through the runner of which the tissue was more susceptible to the anthracnose than those of the leaf and petiole when the fungal mycelial disk was inoculated. Postplanting inoculation by irrigation with spore suspension was much more effective in inducing the anthracnose disease than preplanting soil mix. However without inoculation, no or little anthracnose occurred regardless of commercial, non-cultivated or diseased field soils when healthy seedlings were planted. This suggests that occurrence of strawberry anthracnose in fields may be related to contamination of plant propagules with the anthracnose pathogen.

Identification of Novel Source of Resistance and Differential Response of Allium Genotypes to Purple Blotch Pathogen, Alternaria porri (Ellis) Ciferri

  • Nanda, Satyabrata;Chand, Subodh Kumar;Mandal, Purander;Tripathy, Pradyumna;Joshi, Raj Kumar
    • The Plant Pathology Journal
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    • v.32 no.6
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    • pp.519-527
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    • 2016
  • Purple blotch, caused by Alternaria porri (Ellis) Cifferi, is a serious disease incurring heavy yield losses in the bulb and seed crop of onion and garlic worldwide. There is an immediate need for identification of effective resistance sources for use in host resistance breeding. A total of 43 Allium genotypes were screened for purple blotch resistance under field conditions. Allium cepa accession 'CBT-Ac77' and cultivar 'Arka Kalyan' were observed to be highly resistant. In vitro inoculation of a selected set of genotypes with A. porri, revealed that 7 days after inoculation was suitable to observe the disease severity. In vitro screening of 43 genotypes for resistance to A. porri revealed two resistant lines. An additional 14 genotypes showed consistent moderate resistance in the field as well as in vitro evaluations. Among the related Allium species, A. schoenoprasum and A. roylei showed the least disease index and can be used for interspecific hybridization with cultivated onion. Differential reaction analysis of three A. porri isolates (Apo-Chiplima, Apn-Nasik, Apg-Guntur) in 43 genotypes revealed significant variation among the evaluated Allium species (P = 0.001). All together, the present study suggest that, the newly identified resistance sources can be used as potential donors for ongoing purple blotch resistance breeding program in India.

Astudy on the Anticancer Activies of Lipid Soluble Ginseng Extract and Ginseng Sapongin DErivatives Against Some Cancer Cells (인삼의 지용성 성분과 사포닌 유도체의 항암작용 연구)

  • 항우익;오수경
    • Journal of Ginseng Research
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    • v.8 no.2
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    • pp.153-166
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    • 1984
  • The anticancer activities of petroleum ether extract of Panax ginseng root(crude GX) and its partially purified fraction from silicic acid column chromatography (7:3 GX) were studied with Sarcoma 180(S-180) or Walker carcinosarcoma 256 (Walker 256) in vivo and with L1210 leukemic lympocyte in vitro. Potential cytotoxic activities of the crude GX and against L1210 cells were compared with those of 5-Fluorouracil (5-FU) and saponin derivatives (Panax-diol, Panax-triol, Diol saponin, Triol saponin) in vitro. In order to observe the physiological effects of the crude GX and 7:3 GX on the animals with cancer, hemoglobin(Hb), red blood cell(R.B.C) and white blood cell after treatment with each GX in comparison with corresponding control groups, respectively. The anticancer effects of the crude GX and 7:3 GX were estimated by measuring the survival time of S-180 bearing mice after treatment with them. The experimental results obtained are summarized as follows; 1. The one unit of cytotoxic activity against L1210 cells was equivalent to 2.54$\mu\textrm{g}$ and 0.88$\mu\textrm{g}$of the crude GX and 7:3 GX per ml of culture medium, respectively. 2. The cytotoxic activities of Panax-diol, Panax=triol, Diol saponin and triol saponin against L1210 cells were not detected. 3. The anticancer activities of 5-FU against L1210, S-180 and Walker 256 were very effective in vivo and vitro tests. 4. The significantly increased W.B.C values of mice after inoculation with S-180 cells were reduced to normal range by the crude GX treatment. 5. The significantly decreased Hb values of rats after inoculation with Walker 256 were recovered to normal range by oral administration of the crude GX. 6. 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.

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Antitumor activity of Acanthopanax senticosus extract and its possible immunological mechanism

  • Shin, Kwang-Soon;Yoo, Yung-Choon;Yoon, Taek-Joon
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.129.2-129.2
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    • 2003
  • Antitumor and immunomodulatory activities of an aqueous extract (GF100) of Acanthopanax senticosus was examined. In experimental lung metastasis of colon26-M3.l carcinoma cells, intravenous (i.v.) administration of GF100 2 days before tumor inoculation significantly inhibited lung metastasis in a dose-dependant manner. The i.v. administration of GF100 also exhibited the therapeutic effect on tumor metastasis of colon26-M3.1 cells, when it was injected 1 day after tumor inoculation. In an in vitro cytotoxicity analysis, GF100 enhanced the responsiveness to a mitogen, concanavalin A (ConA), of splenocytes in a dose-dependent manner. (omitted)

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Development of an Efficient In Vitro Screening Method for Selection of Resistant Lily Cultivars Against Fusarium oxysporum f. sp. lilii (백합 구근부패병 저항성 검정을 위한 기내 간편 검정법 개발)

  • Jang, Ji-Young;Moon, Ki-Beom;Ha, Jang-Ho;Park, Ji-Sun;Kim, Mi-Jin;Jeon, Jae-Heung;Lee, Geung-Joo;Kim, Hyun-Soon
    • Horticultural Science & Technology
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    • v.33 no.6
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    • pp.883-890
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    • 2015
  • The soilborne fungus Fusarium oxysporum f. sp. lilii (Fol) is a serious threat to all lily cultivars, especially infecting bulbs and flowers. It has become increasingly important to develop varieties resistant against the bulb rot disease. Genetic diversity of cultivars and reliable screening methods are required for this purpose. Here, an efficient in vitro screening system for evaluating resistance to Fol in 38 in vitro-grown lily plants was investigated. Various factors including culture conditions of Fol, inoculum density, appropriate plant materials, inoculation method and duration, and incubation period of plant materials after inoculation were combined to optimize the screening method. As a result, we suggest optimal conditions for an in vitro screening system for the selection of Fol-resistant lily cultivars as follows. Fol was grown on potato dextrose agar (PDA) medium for 6 days at $25^{\circ}C$ in darkness and used as working inoculation. Spore suspensions were prepared (inoculum density: $1.0{\times}10^4$ $spores{\cdot}mL^{-1}$), and then leaf segments $1.5{\times}2.0cm^2$ were inoculated by dipping for 22 hours at $25^{\circ}C$ in dark. Later, leaves were cultured on 0.6% agar plates at $25^{\circ}C$ and 50% humidity with a photoperiod of 16 hours light/8 hours dark (fluence rate of $40{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$) to examine the progress of bulb rot. After 7 days, disease levels were classified into indices 1 (no symptom) to 6 (serious bulb rot). Soil inoculation of Fol carried out with resistant or susceptible lily cultivars that had been selected through in vitro screening confirmed the reproducibility of results. Therefore, the in vitro screening method established in this study is efficient and reliable for selection of lily cultivars resistant against bulb rot disease.