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Studies on Antifugal Activity from a Strain of Gliocladium Virens

Author: ZhuZuoZuo
Tutor: FanMeiZhen；HuFengLin
School: Anhui Agricultural University
Course: Microbiology
Keywords: Gliocladium virens Antifungal activity Liquid fermentation Antibacterial Mechanism Pot experiment Structural identification
CLC: S476
Type: Master's thesis
Year: 2009
Downloads: 91
Quote: 1
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Abstract


In this study of a Provincial Key Laboratory of Microbial Control, Anhui Agricultural University, has a higher antibacterial activity of green is Gliocladium RCEF4099 strains secondary metabolites for research object, the system the study of RCEF4099 the fermentation broth antibacterial activity and antibacterial ingredients. The main contents include: the strain Some biological parameters were measured; antibacterial test; conducive to optimization of fermentation conditions generated by the active ingredient; Strains Mechanism of; indoor pot experiment; Isolation and structure identification of the active substance. On the growth rate of RCEF4099 strains and chitinase enzyme activity were measured. As can be seen by measuring the diameter of the strains RCEF4099 colonies grow faster, including the fourth day of the fastest growing, the fifth day that reached 9cm diameter. A total of eight days the strains RCEF4099 chitin enzyme activity was measured chitinase expression in two days, seven days reached a peak, and then begin to decline. The Gliocladium the new strains RCEF4099 respectively and pepper gray mold wheat gibberelic, small patches of corn, wheat, Rhizoctonia, cotton withered apple anthrax, rape Sclerotial, rice grain dry the 8 plant pathogenic fungi confrontation between the test and the cylinder plate method suppression bacteria test. Comprehensive the two antibacterial experimental methods, derived Gliocladium virens RCEF4099 pair of pepper gray mold of antibacterial effect. In this study, the antibacterial activity indicators (inhibition zone size), comparing different nutritional factors, including carbon and nitrogen sources, inorganic salts and different non-nutritional factors including seed age, inoculum size, medium volume, shake flask speed on impact of active substances, the final screening RCEF4099 strains produce antimicrobial substances were optimum fermentation conditions were as follows: RCEF4099 medium volume 40/100 ml (V / V), inoculum size 10% (V / V), incubation time 5d shaker at 160 r / min, the most active; fermentation medium ingredients: white sugar of 4% glucose, 0.5%, 1.5% dry yeast, 0.02% K2HPO4. The stability test showed that the fermentation broth RCEF4099 strains Strains adapter 6 generation before the activity is stable, the 10th generation of the inhibition zone diameter of only 1.7mm less than the starting strain. RCEF4099 fermentation broth has better thermal stability and pH stability. Both the impact of the fermentation broth of Gliocladium pepper gray mold spores and hyphae growth morphology to to explore green of Gliocladium RCEF4099 the antibacterial mechanism. RCEF4099 strain metabolites pepper germination of Botrytis cinerea spores strong inhibitory effect on spore germination after treatment, a higher rate of malformations, the pathogen mycelium also have teratogenic effects. Indoor potted Experimental results show that high protective and therapeutic effect: RCEF4099 fermentation broth pepper gray mold, preventive and therapeutic role in more than 76%. Processing and related physical and chemical testing by comparing different extraction methods to establish the optimum extraction process of the active substance: 50 ℃ fermentation broth was concentrated to 1/5 of its original volume, three times the volume of 95% ethanol (ethanol final concentration of 75% ) alcohols Shen discarded twice precipitated and dealcoholated after acetone extraction reagent, 2:1 volume Scores twice extracted taken acetone phase, and concentrated to dryness to give the crude products of the active substance. This test uses a high-performance liquid chromatography analysis of crude extract, stars while collecting flow activity was measured to determine the substantially part of the active ingredient, and then the active fraction to HPLC-MS analysis. The active ingredient is 1,3,5,7 - cyclooctatetraene preliminary fermentation broth can be inferred, and a molecular weight of 104, of the formula C8H8.

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CLC: > Agricultural Sciences > Plant Protection > A variety of control methods > Biological control
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