A kind of endophytic fungus producing ergosterol and application thereof
A technology of endophytic fungi and ergosterol, applied in the field of microorganisms, to achieve the effect of great application value
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Embodiment 1
[0048] The endophytic fungus of the present invention is a strain obtained from the leaves of Danshen in Shangluo, Shaanxi.
[0049] The endophytic fungus of the present invention is separated and obtained according to the following steps: washing with tap water for 30 minutes to remove the sediment and then washing with deionized water 3 times. The leaves were surface disinfected according to the following procedures: 75% ethanol, 30s→1.3M sodium hypochlorite (3-5% available chlorine), 1min→75% ethanol, 30s→sterile deionized water wash 3 times. After the filter paper absorbs the remaining water, use a sterilized scalpel to cut the leaves into 1cm×1cm size tissue pieces, and place them on PDA containing 100mg / L penicillin (potatoes 200g / L, glucose 20g / L, agar, 15g / L) Cultured on the medium at 28°C. Observe the growth of the fungus in the sample every day. After 5-7 days, the hyphae grow out, and the tips of the hyphae are picked and transferred to a new PDA medium for purifica...
Embodiment 2
[0051] (1) Take the endophytic fungus of the present invention, pick a small amount of hyphae with an inoculation needle under aseptic conditions, connect them to a sterilized solid PDA medium plate, and activate and cultivate them at 28°C for 72 hours;
[0052] (2) Take the activated and cultivated strains, transfer them to sterilized liquid PDB seed medium under aseptic conditions, and culture them on a shaker at 180 rpm at 28°C for 72 hours to obtain seeds;
[0053] (3) The prepared PDB liquid medium is divided into 250ml Erlenmeyer flasks (about 100ml per bottle), sterilized and cooled for later use. Under aseptic conditions, the seeds were inoculated at 10% of the inoculum, and cultured on a shaker at 180 rpm at 28°C for 7 days.
[0054] (4) After fermentation is completed, the culture is collected and filtered, the mycelium is suspended in 5 times the volume of methanol, homogenized for 5 minutes, and ultrasonicated for 60 minutes; then filtered, vacuum-evaporated, and reconsti...
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