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An Endophytic Actinomycetes from Camellia Sinensis and Screening of Its Antifungal Metabolites

Author: ChenDan
Tutor: YangMinHe
School: Fujian Normal University
Course: Biochemistry and Molecular Biology
Keywords: Streptomyces Classification and identification The inhibitory spectrum Antibacterial substances Separation and purification
CLC: S476.1
Type: Master's thesis
Year: 2010
Downloads: 98
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Abstract


Students from the tea tree the fungal mango tee bacteria (Guignardia mangiferae) colonies separated from the surface to the actinomycetes named as BF-01 strain BF-01 and the following aspects: 1, the use of morphological and molecular methods to identify the strain BF-01; 2, strain BF-01 Determination of the inhibitory spectrum; 3, the separation and purification of antibacterial substances. By studying the following results: 1, the Institute of isolated strains BF-01 colonies cultured at 28 ° C under 30 to 40d of tea endophytic fungi mango tee bacteria (Guignardia mangiferae) surface. Study of morphological characteristics of colonies observed and electron microscopy of the spores and spores wire strain BF-01, to determine the actinomycetes of the genus Streptomyces. Extract strain BF-01 genomic DNA amplification its 16SrDNA conserved sequences, sequence alignment and phylogenetic tree was constructed after sequencing. The results showed the test strains BF-01 and Ka Wuer Streptomyces (Streptomyces cavourensis subsp) in the same branch of the evolutionary tree, that is, the affinity of both relatively close. Morphological observation, determine the initial strain BF-01 belongs to the Kawu Er Streptomyces (the Streptomyces cavourensis subsp). 2, strain BF-01 series of antibacterial activity and spore suppression activity assay: (1) plate confrontation trials have shown that the strain BF-01 endophytic fungi in the test of seven tea and seven plant pathogenic fungi good inhibitory effect of strain TN15, TN5, TN13, TN2-2 inhibition rate of more than 90%; (2) fermentation broth bacteriostatic test strains of BF-01 fermentation undiluted solution of the 7 strains tested tea tree Endophytic fungi and 13 of the 7 strains of phytopathogenic fungi have different degrees of inhibition, only Trichoderma showed no suppression capability; (3) the fermentation broth was extracted with chloroform solution of 5 kinds of pathogenic bacteria and Candida albicans significantly inhibited, SPSS software significant difference analysis showed significant and significant. The foregoing three different antibacterial test showed that the strain BF-01 has a broad spectrum of antimicrobial activity. (4) fermentation broth of the chloroform extract of corn stalk rot bacteria (Fusarium gramimearum) and banana spot Fusarium (Fusarium stoveri) spore germination have good inhibitory effect, and with the increasing concentration of the extract, the rate of spore germination in gradually decreases, i.e. spore germination inhibition rate gradually increases. 3, the fermentation broth of strains of BF-01 alcohol precipitation, chloroform extraction, thin layer chromatography, silica gel column chromatography and the like methods active substance initially isolated and purified to obtain a higher activity component, the component Ⅱ component Ⅲ component VIII component Ⅳ and component Ⅷ, further dextran gel component Ⅷ LH-20 column chromatography to obtain 2 has a higher antibacterial activity. Component VIII by Gas Chromatography / Mass Spectrometry (GC / MS) analysis showed that the separation of the active compound is 2-Methyl-4-ethoxycarbonyl-3H-imidazo [] pyridazine-5, 7 - (6H)-dione (2 - methyl-4 - ethoxycarbonyl-3H-imidazo [1,5-b] pyridazine-5 ,7 - (6H) - dione), named compound A, and 1-Phenyl-3-(2-pyridyl) -5-pyrazolon (1 - phenyl-3 - (2 - pyridyl) -5 - pyrazolone) named compound B. They are two alkaloids heterocyclic compound, a basic skeleton of these two compounds very similar in structure, but the these two basic skeleton structure has a large number of reports in the previous literature, having a high value of pesticides and pharmaceutical and other applications.

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CLC: > Agricultural Sciences > Plant Protection > A variety of control methods > Biological control > The use of microbial pathogens
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