• Title/Summary/Keyword: antagonistic gene

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Transfer of Bacillus thuringiensis toxin gene into Bacillus subtilis and its inoculation effects (식물 생장촉진 미생물의 외부 유전자 도입과 그 접종효과)

  • Rhee, Young-Hwan;Kim, Kwang-Sik;Kim, Yong-Woong;Kim, Yeong-Yil
    • Applied Biological Chemistry
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    • v.35 no.5
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    • pp.361-366
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    • 1992
  • The antagonistic bacteria, showing distinguished effect against Fusarium oxysporum and Rhizoctonia solani were isolated from the rhizosphares of horticultural plants and identified as Bacillus subtilis. The strains were studied for their chracteristics of biochemistry, physiology, antagonistic effect against plant pathogenic fungi, and growth promoting effect on horticultural plants. The Bacillus thuringiensis(BT) HD-1 toxin gene was introduced into these B. subtilis. The BT toxin genes on chromosome of the bacteria were identified by southern blotting, but its proteins were not detected by SDS-PAGE. These transformed bacteria showed growth promoting effect and showed also insecticidal and antagonistic effects against Bombix mori and fungi F. oxysporum and R. solani but not against nematode Bursaphelenchus xylophilus.

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Diversity and Characterization of Endophytic Bacteria Associated with Tidal Flat Plants and their Antagonistic Effects on Oomycetous Plant Pathogens

  • Bibi, Fehmida;Yasir, Muhammad;Song, Geun-Cheol;Lee, Sang-Yeol;Chung, Young-Ryun
    • The Plant Pathology Journal
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    • v.28 no.1
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    • pp.20-31
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    • 2012
  • Endophytic bacterial communities of tidal flat plants antagonistic to oomycete plant pathogens were studied by the isolation of 256 root colonizing endophytic bacteria from surface-disinfected root tissues of six plants ($Rosa$ $rugosa$, $Suaeda$ $maritima$, $Vitex$ $rotundifolia$, $Carex$ $scabrifolia$, $Glehnia$ $littoralis$ and $Elymus$ $mollis$) growing in a tidal flat area of Namhae Island, Korea. To understand the antagonistic potential, an $in$ $vitro$ antagonistic assay was performed to characterize and identify strains that were antagonistic to the oomycete plant pathogens $Phytophthora$ $capsici$ and $Pythium$ $ultimum$ from the total population. Nine percent of the total number of isolated bacteria exhibited in vitro inhibitory activity against target plant pathogenic oomycetes. Taxonomic and phylogenetic placement of the antagonistic bacteria was investigated by analysis of the 16S rRNA gene sequences. The sequence analysis classified the antagonistic strains into four major classes of the domain bacteria ($Firmicutes$, ${\alpha}-Proteobacteria$, ${\gamma}-Proteobacteria$ and $Actinomycetes$) and 10 different genera. Further production of secondary metabolites, hydrolytic enzymes and plant growth promoting traits were determined for the putative new species of antagonistic endophytic bacteria. These new strains could not be identified as known species of ${\alpha}-Proteobacteria$, and so may represent novel bacterial taxa. The unexpected high antagonistic bacterial diversity associated with the tidal flat plants may be indicative of their importance in tidal flat plants as a promising source of novel antimicrobial compounds and biocontrol agents.

Isolation and Characterization of Burkholderia cepacia strain YJK2, Antagonistic Microorganism of Paprika Pathogens (파프리카 병원균들에 대한 길항미생물, Burkholderia cepacia strain YJK2의 분리 및 특성)

  • Yang, Soo-Jeong;Kim, Hyung-Moo;Ju, Ho-Jong
    • Korean Journal of Organic Agriculture
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    • v.23 no.1
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    • pp.133-148
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    • 2015
  • Although several adverse effects have been increased in recent years, synthetic agro-chemicals have been widely used to control diseases on paprika. This research was conducted to isolate and to characterize the antagonistic microorganism to control major paprika diseases, gray mold rot, fruit and stem rot, phytophthora blight, sclerotium rot, and wilt disease. Analysis of the fatty acid and analysis of the 16S rDNA gene sequence revealed that YKJ2 isolated in this research belongs to a group of Burkholderia cepacia. Specially, 16S rDNA gene sequence of YKJ2 showed 99% of sequence similarity with B. cepacia. Observation through the optical microscope revealed that YKJ2 was effective on suppression of the spore germination and the hyphal growth of pathogens. YKJ2 treatment on pathogens induced marked morphological changes like hyphal swelling and degradation of cell wall. In the case of phytophthora blight, the zoosporangium formation was restrained. On the basis of the results of this study, we propose that an antagonistic microorganism, B. cepacia, found in this study naming as "B. cepacia strain YKJ2" and has great potential as one of biological control agents against major diseases of paprika.

Purification of Chitinase from an Antagonistic Bacterium Bacillus sp.7079 and Pro-Inflammatory Cytokine Gene Expression by PCTC

  • Han, Ok-Kyung;Lee, Eun-Tag;Lee, Young-Sun;Kim, Sang-Dal
    • Journal of Microbiology and Biotechnology
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    • v.13 no.1
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    • pp.77-84
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    • 2003
  • Chitinase was purified from an antagonistic bacterium Bacillus sp. 7079 by ammonium sulfate precipitation, QAE-Sephadex anion exchange chromatography, Sephadex G-100 gel filtration, and SP-Sephadex cation exchange chromatography. The molecula. weight of purified chitinase (PC-1) was approximately 66.5 kDa on SDS-PACE. PC-1 exhibited optimum pH and temperature of pH 7.5 and $45^{\circ}C$, respectively. More than $80\%$ of PC-1 was stable at pH 5.0 to 9.0, and more than $90\%$ at $40^{\circ}C$. $Fe^2+\;and\;Ca^2+$ inhibited the chitinase activity about $20\%$, and EDTA and p-CMB by about $30\%$, whereas $Ag^+$ inhibited the activity up to $65\%$. The $K_m$ value of PC-1 was 1.215 mg/ml with colloidal chitin as a substrate. We also investigated the effect of PC-1 treated chitin (PCTC) on the pro-inflammatory cytokine gene expression in macrophage RAW 264.7 cells. The expression of IL-$1{\alpha}$ and IL-$1{\beta}$ mRNA gene was investigated using reverse transcriptase polymerase chain reaction (RT-PCR). IL-$1{\alpha}$ and IL-$1{\beta}$ mRNA were induced by the treatment of PCTC and chitin only in RAW 264.7 cells. These expressions were induced as early as 2 h and sustained up to 24 h in RAW 264.7 cells. IL-$1{\alpha}$ and IL-$1{\beta}$ mRNA were more strongly expressed by the treatment of PCTC than chitin treatment alone in RAW 264.7 cells.

Studies on Development of Antagonistic Microorganism by Cell Fusion - Biological control of disease - ) (세포융합에 의한 신 길항미생물 육종에 관한 연구 - 목초 병해의 생물학적 방제 -)

  • 최기춘;이영환;전우복
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.15 no.1
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    • pp.1-12
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    • 1995
  • This study was to investigate an effective biological control of forage diseases and provide a basic data and a model in improving variety of antagonistic bacteria, with growth promoting effect on forage, through cell fusion. The results obtained were summarized as follows; 1. The antagonistic himbacterium against soil-borne phathogenic fungi Fusarium oxysporum and Rhizoctonia solani was isolated from continuous cropping himsphere soil of forage, and its biological and physiological characteristics were investigated. This bacterium was identified as Bacillus subrilis and named BS 101. Another strain for cell fusion was Bacillus thur ingiensis ssp. kurstaki HD-I(BT 37669) with insecticidal crystal. 2. The auxotropic mutants of BS 101 and BT 37669 were derived after mutagenesis using N-methyl-N'nitro- Nitrosoguanidine(NTG) to give amino acid requirement marker. n e s e auxotropic mutants of BS 101 and BT 37669 were named BS 1013(his-) and BT 69(asp-), respectively. 3. The best protoplast requirement was obtained using DM 3 medium, containing 5% casamino acid, 1 M $MgCI_2$ and 2% bovine semm albumin, to give Fusant 3, 7 and 8. BT toxin gene was not identified with fusants by Southern blotting. However, SDS-PAGE analysis of strains showed various protein patterns among fusants. 4. From the dark culture experiment, growth of forage in inoculated soil with antagonistic bacteria was delayed than that of non-inoculated soil with antagonistic bacteria in each continuous cropping soil and in each sterilized soil. On the other hand, growth duration of forage was different between continuous cropping soil and sterilized soil. 5. Seed germination of Alfalfa, Italian ryegrass and Orchardgrass were significantly improved by inoculation of antagonistic bacteria(p< 0.05).

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Suppression of Fusarium Wilt Caused by Fusarium oxysporum f. sp. lactucae and Growth Promotion on Lettuce Using Bacterial Isolates

  • Yadav, Dil Raj;Adhikari, Mahesh;Kim, Sang Woo;Kim, Hyun Seung;Lee, Youn Su
    • Journal of Microbiology and Biotechnology
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    • v.31 no.9
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    • pp.1241-1255
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    • 2021
  • This study was carried out to explore a non-chemical strategy for enhancing productivity by employing some antagonistic rhizobacteria. One hundred eighteen bacterial isolates were obtained from the rhizospheric zone of various crop fields of Gangwon-do, Korea, and screened for antifungal activity against Fusarium wilt (Fusarium oxysporum f. sp. lactucae) in lettuce crop under in vitro and in vivo conditions. In broth-based dual culture assay, fourteen bacterial isolates showed significant inhibition of mycelial growth of F. oxysporium f. sp. lactucae. All of the antagonistic isolates were further characterized for the antagonistic traits under in vitro conditions. The isolates were identified on the basis of biochemical characteristics and confirmed at their species level by 16S rRNA gene sequencing analysis. Arthrobacter sulfonivorans, Bacillus siamensis, Bacillus amyloliquefaciens, Pseudomonas proteolytica, four Paenibacillus peoriae strains, and Bacillus subtilis were identified from the biochemical characterization and 16S rRNA gene sequencing analysis. The isolates EN21 and EN23 showed significant decrease in disease severity on lettuce compared to infected control and other bacterial treatments under greenhouse conditions. Two bacterial isolates, EN4 and EN21, were evaluated to assess their disease reduction and growth promotion in lettuce in field conditions. The consortium of EN4 and EN21 showed significant enhancement of growth on lettuce by suppressing disease caused by F. oxysporum f. sp. lactucae respectively. This study clearly indicates that the promising isolates, EN4 (P. proteolytica) and EN21 (Bacillus siamensis), can be commercialized and used as biofertilizer and/or biopesticide for sustainable crop production.

Megakaryocyte-Derived IL-8 Acts as a Paracrine Factor for Prostate Cancer Aggressiveness through CXCR2 Activation and Antagonistic AR Downregulation

  • Sadan, Dahal;Prakash, Chaudhary;Yi-Sook, Jung;Jung-Ae, Kim
    • Biomolecules & Therapeutics
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    • v.31 no.2
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    • pp.210-218
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    • 2023
  • Prostate cancer is the fifth leading cause of cancer-related mortality in men, primarily because of treatment resistance, recurrence, and metastasis. In the present study, we investigated the role of paracrine interleukin-8 (IL-8) in the antagonistic expression of IL-8 and androgen receptor (AR), and the contribution of IL-8 to prostate cancer aggressiveness. In hormone-responsive LNCaP cells that do not express IL-8, recombinant IL-8 treatment significantly increased expressions of IL-8, CXC chemokine receptor 2 (CXCR2), matrix metalloproteinase (MMP)-2/9, Snail, and vimentin. IL-8 treatment significantly decreased AR and E-cadherin expression. IL-8-induced gene expression changes were suppressed by navarixin, a CXCR1/2 inhibitor, and gallein, a Gβγ inhibitor. In PC-3 androgen-refractory prostate cancer cells, IL-8 knockdown reduced expressions of CXCR2, MMP-2/9, Snail, and vimentin, and increased AR and E-cadherin expressions at the mRNA and protein levels. Co-culture with MEG-01 human megakaryocytic cells secreting high levels of IL-8 induced gene expression changes in both LNCaP and PC-3 cells, similar to those induced by IL-8 treatment. The altered gene expressions were accompanied by significant activation of transcription factor Snail in LNCaP and PC-3 cells. Treatment with the CXCR blocker navarixin inhibited the invasion of PC-3 cells but not LNCaP cells. However, invasion induced by MEG-01 was inhibited by navarixin in both LNCaP and PC-3 cells. The collective findings demonstrate that IL-8 enhances CXCR2 expression, which antagonistically regulates AR expression. More importantly, through changes in IL-8/CXCR2-regulated gene expression, IL-8 induces antiandrogen therapy resistance and epithelial-mesenchymal transition in prostate cancer.

Isolation and Identification of Newly Isolated Antagonistic Streptomyces sp. Strain AP19-2 Producing Chromomycins

  • Wu, Xue-Chang;Chen, Wei-Feng;Qian, Chao-Dong;Li, Ou;Li, Ping;Wen, Yan-Ping
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.499-504
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    • 2007
  • A new antagonistic strain of actinomycete, designated AP19-2, was isolated from the feces of giant pandas inhabiting the Foping National Nature Reserve in China. Cultural characteristic studies strongly suggested that this strain is a member of the genus Streptomyces. The nucleotide sequence of the 16S rRNA gene of strain AP19-2 evidenced profound similarity (97-99 %) with other Streptomyces strains. Two pure active molecules were isolated from a fermentation broth of Streptomyces sp. strain AP19-2 via extraction, concentration, silica gel G column chromatography, and HPLC. The chemical structures of the two related compounds (referred to as chromomycin $A_2$ and chromomycin $A_3$) were established on the basis of their Infrared spectra (IR), High Resolution Electrospray Ionization Mass Spectrometry (HR-ESI-MS), and Nuclear Magnetic Resonance (NMR) data, and by comparison with published data.

Assessment of the Contribution of Antagonistic Secondary Metabolites to the Antifungal and Biocontrol Activities of Pseudomonas fluorescens NBC275

  • Dutta, Swarnalee;Yu, Sang-Mi;Lee, Yong Hoon
    • The Plant Pathology Journal
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    • v.36 no.5
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    • pp.491-496
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    • 2020
  • An understanding of the contribution of secondary metabolites (SMs) to the antagonistic and biocontrol activities of bacterial biocontrol agents serves to improve biocontrol potential of the strain. In this study, to evaluate the contribution of each SM produced by Pseudomonas fluorescens NBC275 (Pf275) to its antifungal and biocontrol activity, we combined in silico analysis of the genome with our previous study of transposon (Tn) mutants. Thirteen Tn mutants, which belonged to 6 biosynthetic gene clusters (BGCs) of a total 14 BGCs predicted by the antiSMASH tool were identified by the reduction of antifungal activity. The biocontrol performance of Pf275 was significantly dependent on 2,4-diacetylphloroglucinol and pyoverdine. The clusters that encode for arylpolyene and an unidentified small linear lipopeptide influenced antifungal and biocontrol activities. To our knowledge, our study identified the contribution of SMs, such as a small linear lipopeptide and arylpolyene, to biocontrol efficacy for the first time.

Molecular Cloning of Antagonistic Genes in Pseudomonas maItophiliQ B-14 (토양병해 길항성 Pseudomonas maltophilia B-14의 길항유전자탐색)

  • 구본성;서영우;윤상홍;박경수;은무영;김용환;오상우;류진창;은무영
    • Microbiology and Biotechnology Letters
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    • v.20 no.6
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    • pp.619-624
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    • 1992
  • Tn5 lac 삽입으로 채소입고병원균에 길항력이 약화된 T-67 및 고추역병균과 참깨역병균에 길항력이 약화된 T-81의 Tn5 lac 유전자 일부와 오른쪽 말단에 있는 길항관련 유전자의 flanking sequence가 cloning된 pAG67 및 pAG81 clone을 선발하였고, pAG67 및 pAG81 clone된 길항관련 유전자의 flanking sequence를 야생 길항균 Pseudomonas maltophilia B-14의 DNA를 probe로 사용하여 Southern hybridization으로 확인하였으며, 제한효소 지도를 작성하여 8Kb 및 4Kb 크기의 flanking sequence가 cloning되었음을 확인하였다. pAG6 및 pAG81의 flanking sequence를 EcoRi-BglII와 EcoRI-MpaI으로 분리하여 유전자 은행으로부터 길항관련 유전자가 cloning된 cosmid clone 7개주를 선발하였다.

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