A kind of Hubei Ophiopogon japonicus endophytic Aspergillus oryzae strain and application thereof
A technology for growing Aspergillus oryzae and Aspergillus oryzae, which is applied in the direction of fungi, microorganisms, microorganisms, etc., which can solve the problems of small planting area, high cost of polysaccharide extraction, and limited production of Ophiopogon japonicus polysaccharides in Hubei, achieving low pollution and environmentally friendly production Effect
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Embodiment 1
[0053] The isolation of embodiment 1 bacterial strain
[0054] (1) The samples were taken from Hubei Maidong produced at the Hubei Maidong GAP Base in Oumiao Town, Xiangyang City, Hubei Province. After collecting the tubers of Ophiopogon japonicus Hubei, use a large amount of water to wash the soil attached to the surface of the tubers of Ophiopogon japonicus Hubei, then put the tubers into a clean beaker, add deionized water, and wash them repeatedly in an ultrasonic cleaner until the water after cleaning become extremely clear;
[0055] (2) Dip the water on the surface of Ophiopogon japonicus root tuber with sterile filter paper, and then process it according to the following steps: first soak the tuber tuber with 75% (V / V) ethanol for 2 minutes, rinse it with sterile water for 3 times; Soak the tubers in sodium hypochlorite solution with an available chlorine content of 4-6% for 2 minutes, rinse them with a large amount of sterile water for 4 times, and then dip them dry w...
Embodiment 2
[0058] Identification of embodiment 2 bacterial strains
[0059] (1) The bacterial strain isolated in Example 1 was cultured on potato dextrose agar medium (PDA) at 28° C. to form colonies, and the characteristics such as colony morphology and color were observed. The result is as figure 1 As shown, the colonies are yellow-green, and multiple colonies are fluffy and flat on the petri dish.
[0060] (2) Perform genome sequencing on the strain isolated in Example 1, and its 18S rRNA gene sequence is shown in SEQ ID NO.1.
[0061] The sequencing results were compared on the NCBI website (http: / / blast.ncbi.nlm.nih.gov / Blast.cgi), and it was found that its sequence had the highest sequence homology with Aspergillus oryzae RIB40, XR_002735719.1, and the similarity up to 99%. Illustrate that the bacterial strain isolated in Example 1 is a fungus of the genus Aspergillus oryzae.
Embodiment 3
[0062] The fermentation of embodiment 3 bacterial strains
[0063] (1) Get the Aspergillus oryzae EF024 bacterial classification that embodiment 1 isolates. Under sterile conditions, use an inoculation needle to pick a small amount of mycelium, insert it into a sterilized PDA medium test tube, and activate it at 28°C for 72 hours;
[0064] (2) Under sterile conditions, inoculate the activated cultured bacterial strains into sterilized potato dextrose liquid medium (PDB), shake and culture at 28° C. and 180 rpm for 72 hours to obtain seed liquid;
[0065] (3) 200g potatoes, 20g glucose, and 1000ml distilled water are mixed with liquid fermentation medium, and 100ml liquid fermentation medium is taken in a 250ml Erlenmeyer flask, sterilized, and cooled for subsequent use; under aseptic conditions, inoculate according to 10% amount, seed liquid was inoculated in potato liquid medium, cultured at 28°C and 180rpm for 7 days to obtain a fermented solid-liquid mixture;
[0066] (4)...
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