Preparation of peltate yam endophytic fungi spiro-dinaphthyl compound and application as germicide thereof
A technology of endophytic fungi, Dioscorea scutellaria, applied in the field of microorganisms, can solve the problems such as the separation of spirobinaphthyl compounds that have not yet been seen
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Embodiment 1
[0037] Example 1: Isolation of endophytic fungus Dzf12
[0038] Collect fresh rhizomes of Dioscorea scutellaria, treat the surface with 70% ethanol for 30 seconds, then treat with 0.2% mercuric chloride for 20 minutes to carry out thorough surface sterilization, remove the epidermis under aseptic conditions, and divide the rhizomes into about 0.5 Place the pieces of cm×0.5cm×0.5cm on the PDA medium, 1 piece per dish, and cultivate them until the 7th to 20th day at 25°C under light conditions, and pick one piece from the edge of each colony. A small piece of hyphae was inoculated on PDA medium for purification and culture, and the purification and culture were continued for 4 to 5 times until the colony morphology was consistent. The purified strains were preserved on the PDA slant, and one of the endophytic fungi was numbered Dzf12.
Embodiment 2
[0039] Embodiment 2: the antibacterial activity of endophyte Dzf12 extract
[0040] The antibacterial active components and their activities of the n-butanol extract and the acetone extract of the endophytic fungus Dzf12 bacterial liquid and the acetone extract of the endophytic fungus Dzf12 were preliminarily detected by thin-layer chromatography bioautographic method and thiazolium blue (MTT) as the chromogen .
[0041] (1) A small amount of preparation of the extract: what adopted was shaker fermentation, two 500mL volume Erlenmeyer flasks were filled with 250mL of PDB liquid medium respectively, and the pre-cultured Dzf12 mycelium was inoculated into the sterilized medium, At 25° C. and 150 rpm, culture in dark for 15 days to obtain a fermentation broth, and filter under reduced pressure to obtain mycelium and bacterial liquid. Bacterial solution was extracted with n-butanol of equal volume (repeated 3 times), combined with n-butanol extract, and concentrated to obtain n-...
Embodiment 3
[0049] Example 3: Observation of Morphological Characteristics of Endophytic Fungus Dzf12
[0050] The endophytic fungus Dzf12 was cultured on PDA medium at 25°C, and the colonies were dense, fluffy, gray, with obvious water droplets on the surface, and produced a large amount of dark brown pigment on the back. The hyphae were separated, some of them were dense, and had a structure similar to chlamydospores, but no conidia were found. Morphological characteristics see Figure 1 to Figure 4 . Under culture conditions, no sexual generation of Dzf12 was found.
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