Active And Structure Identification Of Alternaria Alternate Mount Tai Bacterial Strain Metabolites

Because the soil fungi can produce rich variety of has a variety of bioactive secondary metabolites,has important ecology function in nature,and got extensive attention worldwide and has made great progress.At present,the chain's spore fungus(Alternaria alternata)structure and its active secondary metabolites of prophase research at home and abroad related reports rarely own laboratory collection separation evaluation chain spore fungus taishan variant,and prove their fermented liquid has a promoting effect on plant growth On the basis of previous research,this study on the chain,spore fungus taishan variant separation structure identification of metabolites and a series of activity test,for the fermented liquid of metabolites in promoting the growth of plants Improve the plant resistance,etc,to provide the theory and application.This study by cultivating chain spore fungus taishan variant independent strains,the liquid medium,using the optimal fermentation condition and its metabolites;And by adopting the combination of purification and biological activity tracking way,through to a series of separation and purification of metabolites,column chromatography separation get raw metabolites;Preparation of high performance liquid chromatography(HPLC)with preparation,further get metabolites-TS001;With ultra-high resolution finally liquid MS(HPLC-MS)and nuclear magnetic resonance(NMR)to identify the structure.The main result:1,The optimal fermentation conditions: medium component is peptone 5g/L,glucose 20g/L,yeast powder 2g/L,dipotassium phosphate 1g/L,magnesium sulfate 0.5g/L,rose-bengal 0.1g/L,optimal cultivation temperature 27?.2,Metabolites of coarse product preparation: column chromatography conditions for filler in a 200-300 mesh chromatography silica gel,the solvent of ethyl acetate,eluent for ethyl acetate:methanol=20:1.Systematically,adding fermented liquid mixtures extracted chromatography column,collect the eluent,decompression evaporation in the rotary evaporation apparatus to dry,antibiotic crude product was obtained.The coarse product with drawal rate was 1.5 ‰.3,TS-001 Isolation and identification: After the high performance liquid chromatographic separation,natural products to get preliminary separation,in high performance liquid chromatography(HPLC)analysis can accurately see crude extract contains three main elements,retention time is respectively 1.004min?4.727min?7.939 min.Previous studies and related literature data show that in the laboratory,the retention time for 1.004 min is solvent peak.After HPLC-MS and NMR analysis,get the retention time for 4.727 min is known substance.Retention time of 7.939 min for the molecular weight of material.For the new compound,the concrete structure still need further analysis.4,Metabolites,crude extract and TS001 biological activity and biological safety to the environment:(1)For the growth of the cucumber and promoting role: In different stages of separation material processing,under the same concentration,the cucumber seedlings of plant height,root length,stem weight,root weight,stem diameter no major changes occurred,the concentration under the condition of cucumber,growth are presented,within a certain range,with the increase of concentration,promote role.The fermented liquid 1000 times the best promoting effect,tasted 8000 times promoting effect is poorer.(2)Effect on cucumber in vivo biochemical indexes: metabolites and TS001 processing cucumber,PPO,POD,SOD in the body,are to promote and PAL activity induced effect,under the condition of various concentrations increased its activity,in the certain concentration range,the enzyme activity with the increase of the concentration,the higher enzyme activity,including flower seedlings treated by fermented liquid 1000 times the highest enzyme activity,cucumber seedlings were treated by single product 8000 times the lowest enzyme activity.The above results show that exists in the fermented liquid to promote secondary metabolism substance of cucumber seedling growth.(3)The safety of the fermented liquid of environmental biological effect: compare the fermented liquid and TS-001 and other 6 kinds of pesticide preparations for the acute toxicity of adult follows,results show that the fermented liquid and TS-001 on the environment of adult follows non-toxic,high safety.