• 제목/요약/키워드: In vitro inoculation

검색결과 204건 처리시간 0.027초

Dual Biocontrol Potential of the Entomopathogenic Fungus, Isaria javanica, for Both Aphids and Plant Fungal Pathogens

  • Kang, Beom Ryong;Han, Ji Hee;Kim, Jeong Jun;Kim, Young Cheol
    • Mycobiology
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    • 제46권4호
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    • pp.440-447
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    • 2018
  • Dual biocontrol of both insects and plant pathogens has been reported for certain fungal entomopathogens, including Beauveria bassiana and Lecanicillum spp. In this study, we demonstrate, for the first time, the dual biocontrol potential of two fungal isolates identified by morphological and phylogenetic analyses as Isaria javanica. Both these isolates caused mortality in the greater wax moth, and hence can be considered entomopathogens. Spores of the isolates were also pathogenic to nymphs of the green peach aphid (Myzus persicae), with an $LC_{50}$ value of $10^7spores/mL$ 4 days after inoculation and an $LT_{50}$ of 4.2 days with a dose of $10^8spores/mL$. In vitro antifungal assays also demonstrated a strong inhibitory effect on the growth of two fungi that are pathogenic to peppers, Colletotrichum gloeosporioides and Phytophthora capsici. These results indicate that I. javanica isolates could be used as novel biocontrol agents for the simultaneous control of aphids and fungal diseases, such as anthracnose and Phytophthora blight, in an integrated pest management framework for red pepper.

Escherichia coli에서 발현된 재조합 인간 상피세포 증식인자의 정제 및 특성

  • 박세철;유광현
    • 한국미생물·생명공학회지
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    • 제24권4호
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    • pp.478-484
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    • 1996
  • Recombinant human epidermal growth factor (rhEGF) was produced by E. coli BL21 harboring a plasmid pYHB101. The maximum production was 68.7 mg/l when the E. coli strain was cultured at 25$\circ$C for 48 hours in the modified MBL medium containing 10 g/l glucose with 1 mM IPTG induction at 2 hours after inoculation. The rhEGF was purified upto 267 folds by Amberlite XAD- 7 chromatography, ultrafiltration, and DEAE Sepharose fast flow ion exchange chromatography with an overall yield of 66.6%. The purified rhEGF was further separated into two fractions by HPLC. The N-terminal amino acid sequence of the second fraction was Asn-Ser-Asp-Ser-Glu-Cys-Pro-Leu-Ser-His. The effect of rhEGF on the DNA synthesis was examined using in vitro biological assay based on the incorporation of 5'-bromo-2'- deoxy-uridine (BrdU). The purified rhEGF shows no difference with natural human epidermal growth factor (nhEGF) in N-terminal amino acids residues and biological activity. From the results, we concluded that rhEGF produced from E. coli harboring the plasmid pYHB101 was apparently the same as nhEGF.

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Proliferation Effect of Conditioned Medium Produced by Lymph Node Stromal Cells

  • Lee, Sang-Han;Moon, Sung-Hoon;Ha, Sang-Chul;Ma, Jin-Yeul;Kim, Min-Sook;Lee, Jin-Man;Kwon, O-Yu
    • Journal of Life Science
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    • 제12권1호
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    • pp.22-26
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    • 2002
  • CS21 lymphoma cells that preferentially metastasize to lymph nodes after s.c. inoculation into BALB/c mice were grown in vitro in the presence of CA12 stromal cells isolated from lymph nodes. To obtain fundamental data on identification and characterization of the soluble factor(s) produced by CA12 stromal cells, we investigated the biological profile of the conditioned medium produced by CAl2 stromal cells. CAl2 conditioned medium has no affinity with Con A. CAl2 conditioned medium is associated with the proliferation of splenic T- and thymic T-cells with-out adding mitogen, although the conditioned medium cannot induce the differentiation of thymocytes. Additionally, we showed that H-7, not HA-1004 inhibits CS21 cell proliferation. These results suggest that CAl2 conditioned medium has a specific soluble factor(s) produced by lymph node stromal cells.

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Lactobacillus plantarum Improves the Nutritional Quality of Italian Ryegrass with Alfalfa Mediated Silage

  • Ilavenil, Soundarrajan;Arasu, Mariadhas Valan;Vijayakumar, Mayakrishnan;Jung, Min-Woong;Park, Hyung Soo;Lim, Young Cheol;Choi, Ki Choon
    • 한국초지조사료학회지
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    • 제34권3호
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    • pp.174-178
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    • 2014
  • The present study was planned to analyze the nutritional quality, microbial counts and fermentative acids in Italian ryegrass (IRG) 80% and alfalfa 20% (IRG-HV) mediated silage inoculated with lactic acid bacteria (LAB) as a probiotic strain for 3 months. Crude protein (CP), acid detergent fiber (ADF), and neutral detergent fiber (NDF), total digestible nutrient (TDN) and In-vitro dry matter digestibility (IVDMD), lactic acid bacteria (LAB), yeast and fungi counts and fermentation metabolites such as lactic acid, acetic acid and butyric acids were analyzed. The result shows that the nutritional quality and metabolite profiles of silage were significantly improved with LAB. For microbial counts, LAB showed dominant followed by yeast as compared with control silage. The pH of the silage also reduced significantly when silage inoculated with LAB. The result confirmed that silage preparation using different crops with L. plantarum inoculation is most beneficial for the farmers.

약용식물 추출물에 의한 사과 저장병 방제 효과 (Effect of Medicinal Plant Extracts on Apple Storage Diseases)

  • 백수봉;정일민
    • 한국식물병리학회지
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    • 제13권1호
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    • pp.57-62
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    • 1997
  • This experiment was conducted to test the control effect of methanol extracts of 10 medicinal plants on apple storage diseases caused by Botryosphaeria berengeriana, Glomerella cingulata and Penicillium expansum. Out of the 10 medicinal plants, methanol extracts of Coptis japonica and Anemarrhena asphodeloides inhibited effectively the mycelial growth of B. berengeriana, G. cingulata and P. expansum in vitro, for which the inhibition ratios of the two plant extracts were 100.0% and 89.3%, 73.7% and 94.1%, and 100.0% and 51.6%, respectively. Spore germination of the three fungi was inhibited 100% only by C. japonica extract, but only P. expansum was inhibited 100% by A. asphodeloides extract. No lesion was formed y the fungi at 5$^{\circ}C$ up to 2 weeks after inoculation. Lesion sizes produced by the three fungi at the temperature ranges of 1$0^{\circ}C$ to $25^{\circ}C$ and infection of B. berengeriana and G. cingulata were inhibited by C. japonica extract, but not by A. asphodeloides extract, while no lesion was formed by the fungi at 5$^{\circ}C$. Infections of the fungi on apples were somewhat stimulated by A. asphodeloides extract.

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구강 편평세포암종에서 Taxol과 Cyclosporin A의 세포사멸 상승 작용 효과 (SYNERGISTIC APOPTOTIC EFFECT OF TAXOL ON ORAL SQUAMOUS CELL CARCINOMA BY CYCLOSPORIN A)

  • 서민정;한세진;이재훈
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제29권5호
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    • pp.394-404
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    • 2007
  • Oral squamous cell carcinoma is the most prevalent oral cancer, which is characterized by its high metastasis and recurrent rates and poor prognosis. Taxol is an anticancer agent which is microbial products extracted from jew tree. It combines with the tubulin and induces apoptosis by inhibiting mitosis of cell with microtubule stabilization. Recently, it was reported to be effective in various solid tumors, but only very slight effect has been seen in oral squamous cell carcinomas due to its cell-specific potencies. Cyclosporin A is used as immune suppressant and is being applied in anticancer therapy as its mechanism of induction of change of apoptotic process in various cells have been known. In this study, oral squamous cell carcinoma HN22 cell line was used for in vitro experiment and as for the experimental group taxol and cyclosporin A were applied alone and to observe the synergistic effect of apoptosis, Taxol and cyclosporin A were coadministered with different concentration of taxol for comparison. The results were obtained as follow: 1. There was no difference in Bcl-2, Bax, caspase 3, 8, 9 mRNA expression when cyclosprin A or taxol was applied alone to HN 22 cell line. 2. Caspase 3, 9 mRNA expression was prominently increased when cyclosprin A and taxol were applied together to cancer cell. 3. No significant difference was observed when cyclosporin A and taxol($1{\mu}g/ml$ and $3{\mu}g/ml$) were applied together to cancer cell line. 4. No significant difference was seen in Bcl-2, Bax, and caspase 8 mRNA expression in all the groups of in vitro experiments. 5. When cyclosporin A was applied alone in vivo study on the nude mice, histopathologi cal findings was similar to those of the control group. Oral squamous cell carcinoma induced by inoculation of HN 22 cell line was not reduced after treatment of cyclosporin A. 6. When taxol was applied alone, the islands of squamous cell carcinoma still remained, which meant insignificant healing effect. There was a lesser volume increase compared with the cyclosporin A alone. 7. When taxol and cyclosporin A were applied together, the connective tissue and calcification were seen in the histopathologic findings. Oral squamous cell carcinoma was decreased and cancer cell was disappeared. In observing the tumor mass change with time, there was a gradual decreased size and healing features. As the results of the in vitro experiment, it could conclud that only when the two agents are applied together, mitochondria-mediated apoptosis occurred by considerable increase of caspase 3, 9 mRNA expression, irrespectable of the concentration of taxol. In vivo experiment, there was a discrete synergistic effect when the two agents were applied together. But single use of cyclosporin A was not effective in this study. Based on the results of this experiment, if further clinical studies are done, taxol and cyclosporin A could be effectively used in treatment of oral squamous cell carcinomas.

고려인삼의 항암효과에 관한 연구 (A Study on the Antitumor Activity of Panax ginseng)

  • Hwang, Woo-lk
    • Journal of Ginseng Research
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    • 제17권1호
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    • pp.52-60
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    • 1993
  • Panax ginseng has been extensively used in the traditional oriental medicine as a restorative, tonic and Prophylactic agent. Recently, several reports regarding to anticancer effects of Panax ginseng has accumulated. These studies emphasized the fact that the anticancer activities might be due to a glycoside group called ginsenoside or pan.u saponin which has a water soluble characteristic. However, the authors and collaborates demonstrated that a highly lipid soluble component in extract of Panax ginseng roots contains a considerable cytotoxic activities against marine leukemic cells (L1210, P388) and human censer cells (HRT-18, HT-29, HCT48). This study was devised to observe the cytotoxic activities of Petroleum-ether extract of Panax giuseng roots (crude GBD and its Partially Purified fraction from silicic acid column chromatography (7 : 3 GX) against sarcoma-180 (5-180) and Walker carcinosar- coma 256 (Walker 256) in vivo, and murine leukemic Lymphocytes (L1210) and human rectal cancer cells (HRT-18) and human colon cancer cells (HT-29 and HCT48) in vitro. Each cell-line was cultured in medium containing serial concentration 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 have 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 L1210 cells being equivalent to 2.54 Ug and 058 Ug for the crude GX and 7 : 3 GX, respectively. The Ri 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 so lvent was 053. While, the Panaxydol and Panaxynol as active compounds were purified from Petroleum-ether extract of Panax ginseng root by Drs. Ahn and Kim, and author found out that the one unit of cytotoxic activity of the Panaxydol and Panaxynol against L1210 cells being equivalent to 056 Ug and 0.3918 respectively. 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 Gt 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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Comparison of in vitro digestibility and chemical composition among four crop straws treated by Pleurotus ostreatus

  • Nie, Haitao;Wang, Ziyu;You, Jihao;Zhu, Gang;Wang, Hengchang;Wang, Feng
    • Asian-Australasian Journal of Animal Sciences
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    • 제33권1호
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    • pp.24-34
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    • 2020
  • Objective: The effects of Pleurotus ostreatus on the feed utilization of broad bean stalks (BBS), rape straw (RS), paddy straw (PS), and corn stalk (CS) was examined. Methods: The four roughages were co-cultured with Pleurotus ostreatus. The chemical composition; enzyme activities of laccase, carboxymethylcellulase (CMCase) and xylanase; carbohydrate and protein fractions (based on The Cornell Net Carbohydrate and Protein System [CNCPS]) were assessed at different days after inoculation (7, 14, 21, 28 d) and un-inoculated roughages (control, 0 d). The digestibility of nutrient components and the gas production of roughage with various incubation times were monitored at 0, 2, 4, 6, 9, 12, 24, 36, 48, 60, and 72 h using an in vitro ruminal fermentation method. Results: A higher CMCase activity (0.1039 U/mL) and earlier time to peak (14 d) were detected in Pleurotus ostreatus cultured with CS (p<0.05). Significantly, the incubation length-dependent responses of cumulative gas production were observed from 24 to 72 hours post fermentation (p<0.05), and these incubation length-dependent effects on cumulative gas production of PS and CS appeared earlier (24 h) for PS and CS than those (48 h) for BBS and RS (p<0.05). The fast-degradable carbohydrate (CA) content for all four roughages significantly increased over time (p<0.05). Nonetheless, increased degradation efficiency for CA treated with Pleurotus ostreatus was detected at both 21 and 28 days of incubation (p<0.05). With the exception of PS (p<0.05), there were no significant difference among the roughages (p>0.05) in slowly-degradable carbohydrate (CB2) at different incubation times (p<0.05). Conclusion: Assessment of the alterations in chemical composition, CNCPS system fractions, and the fermentation kinetics after biological pretreatment may yield a valuable database for evaluating the biological pretreatment of Pleurotus ostreatus in ruminant feed.

고랭지 배추 무름병 방제를 위한 우수약제 선발 (Screening of effective control agents against bacterial soft rot on Chinese cabbage in alpine area)

  • 정은경;장현철;용영록;김병섭
    • 농약과학회지
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    • 제7권1호
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    • pp.32-37
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    • 2003
  • Erwinia carotovora subsp. carotovora에 의한 배추 무름병은 배추에서 가장 큰 문제가 되고 있는 병 중 하나이다. 본 연구는 배추 무름병 방제에 우수한 약제를 선발하여 무름병을 효과적으로 방제하고자 실시하였다. 항생제, 식물활성제 및 비병원성 Erwinia의 효과를 실험실내 약효 검정과 유묘 검정, 포장시험으로 수행하였다. 실험실내 약효 검정 방법으로는 paper disc 방법과 potato slice 방법을 수행하였다. 유묘 검정은 mineral oil 접종법을 사용하여 병발생을 균일하게 하여 수행하였다. 실험실내 약효 검정 결과 streptomycin과 oxolinic acid, bronopol 과 copper hydroxide 가 병원균의 생장 및 감자 절편의 부패를 크게 억제하였다. 그러나 식물 활성제들은 실험실내 약효 검정에서는 병원균의 생장 및 감자 절편의 부패를 크게 억제하지 못하였다. 배추 (강력여름배추) 유묘를 이용한 생물 검정에서 비병원성 Erwinia 는 83.5%의 방제가를 나타냈고, oxolinic acid, 항생제 streptomycin, validamycin 은 각각 95.2%, 91.2% 그리고 57.5% 의 방제가를 나타냈으며 구리제는 79.9% 의 방제가를 나타냈다. 그러나 acibenzolar-S-methyl 은 약해를 유발하여 방제가가 낮게 나타났다. 포장시험 결과 acibenzolar-S-methyl은 2000년 포장 시험 (산촌배추) 에서는 통계적으로 유의성 (p=0.05) 있는 방제 효과를 나타내었으나, 2001년 포장 시험 (강력여름배추) 에서는 약해를 일으켜 방제 효과가 낮게 나타났다. 이러한 차이는 배추 품종에 따른 약제에 대한 반응 차이에 기인한 것으로 사료된다. 2002 년 포장 시험에서는 streptomycin+copper가 79.7% 의 방제가를 나타냈고 copper hydroxide 와 비병원성 Erwinia 는 각각 71.9% 와 60.9% 의 방제가를 나타냈다.

살균제 효과 검정을 위한 고추 탄저병의 실내 열매 검정법 (In vitro Fruit Assay for the Evaluation of Fungicide Activity Against Pepper Anthracnose)

  • 이수민;장호선;김홍태
    • 농약과학회지
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    • 제18권2호
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    • pp.115-121
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    • 2014
  • 고추 탄저병균을 고추 열매에 무상처 분무 접종할 경우, 접종원의 밀도, 습실처리 기간, 발병 온도 등이 열매 상에서 병원균의 포자 발아, 부착기 형성과 발병 정도에 미치는 영향을 조사하였다. 병원균의 접종 밀도와 발병온도는 포자 발아와 부착기 형성에는 유의성 있는 영향을 미치지 못하였지만, 발병도에는 영향을 미쳐서 $1{\times}10^6$$mL^{-1}$의 밀도와 $30^{\circ}C$의 발병 온도에서 병 발생이 가장 높았다. 습실처리 기간은 포자 발아, 부착기 형성과 발병도에 모두 영향을 미쳤는데, 처리 기간이 길어질수록 포자 발아율, 부착기 형성율과 발병도가 증가하였다. 하지만 7일 이상 기간이 길어지면, 열매에서 균사의 생장이 활발하여 포장에서 보는 것과 같은 전형적인 병징을 관찰하기가 어려웠다. 병원균 밀도를 $1{\times}10^6$$mL^{-1}$로, 습실처리 기간은 5일로, 발병온도는 $30^{\circ}C$로 결정하고 병원균을 무상처 접종과 상처 접종하여 살균제의 효과를 비교하였다. 예방 살균제인 propineb는 병원균 접종 1일 전에 처리한 예방효과가 가장 우수하였으며, 병원균을 접종하고 1일 후에 처리한 치료 처리에서는 무상처 접종의 경우에는 효과가 유지되었지만, 상처 접종 시에는 효과가 급감하였다. 치료 살균제인 tebuconazole은 두 가지의 접종 방법 모두에서 치료 효과가 예방효과보다 우수하였다. 예방과 치료 효과를 모두 지니고 있는 trifloxystrobin은 무상처와 상처 접종 모두에서 예방 및 치료 효과가 우수하였다. 하지만 병원균을 접종하고 5일 후에 살균제를 처리하였을 경우에는 3종의 살균제 모두 효과가 아주 미미 하였다. 본 실험을 통하여 표준화한 검정 방법은 병원균을 무상처와 상처 접종하면서 살균제의 효과를 정확하게 검정할 수 있을 뿐만 아니라, 살균제의 작용 특성을 조사하는데도 유용하리라고 생각한다.