The Studies On Isolation And Identification Endophytic Fungi And Antibacterial Activity Of Honeysuckle

Bioactive compounds produced by medicinal plant endophyte have antibacterial properties, which inhibit on pathogens such as pathogenic bacterium, viruses and a number of mycoplasma. Endophytes of medicinal plants is a new resource screening resistance bioactive substances in microbial. This study focused on that the strains had been isolated with resistant poultry cause-disease bacteria from medicine plant Honeysuckle. To lay down substantial foundation for efficient and fast screening drug resistance from medicinal plant endophyte, complete, systemic, various studies had been carried out.38 strains of endophytic fungi had been isolated from honeysuckle. These endophytic fungi attributed to 5 orders, 5 families, 7 genus by morphological identification. ITS sequence analysis of the antibacterial strains JY2 and JY9 by molecular identification show: JY2 strains with powerful antibacterial activity belongs to Fusarium, JY9 strains is deduced to be a unknown species because of not in the same branch with similar sequences, very lower homology.Optimized the composition of fermentation medium of high-efficient antibacterial JY2 and JY9 to produce antibacterial active substances, it was found that 7% and 5% honeysuckle juice was added in fermentation medium of JY2 and JY9 could be better promote the production of antimicrobial bioactive substances. Contemporary, optimized the fermentation conditions by the orthogonal test, the optimum fermentation conditions had determined in JY9 which inoculation of bacterial suspension was 5%, pH of initial medium was 7.0, medium capacity was 100 mL/250 mL flask, fermentation temperature was 28℃, shaking speed was 160r/min, and antibacterial activity substances had the most capability on 8th of fermentation.The properties of two bioactive material produced from JY2 and JY9 fermentations suggested that favorable heat-endurance range was among 30℃～121℃, pH range of powerful inhibitory was pH3～9. The JY2 mainly composed of polysaccharides and organic acids, and the polysaccharide content was 48.97μg/mL; bioactive compounds of JY9 mainly came from polysaccharides, and the polysaccharide content was 55.28μg/mL.Characterization of bioactive antibacterial substances of JY2 and JY9 had analyzed by continuous extracting. The antibacterial substances to coli. and staphylococcus. main remain in raffinate phase of butanol in JY2, and in ethyl acetate, and in raffinate phase of butanol in JY9.Two effective bioactive components have been continuity and synergy by silica gel chromatography. There are hydroxyl and carboxyl functional groups in JY2-50 and JY9-80, which selected among these effective substances for structural analysis, and, should be consist of pentose forming polysaccharide by IR and LC-MS analysis.In conclusion, Two high-bioactive antimicrobial components had been isolated and purified from endophytic fungi in Honeysuckle. Structure Analysis by IR and LC-MS suggested that polysaccharide composing by pentose may be the main the bioactive substances.