Application of Schizophyllan in Controlling Tobacco Mosaic Virus Disease
A technology of Schizophyllum polysaccharide and Schizophyllum, which is applied in the directions of application, disinfectant, chemical used for biological control, etc., can solve the problem that the application of anti-plant virus has not yet been reported, and achieves low cost, simple preparation method, good control effect
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Embodiment 1
[0021] The cultivation of embodiment 1 schizophyllum and the preparation of mycelium powder
[0022] Proceed as follows:
[0023] (1) activation of the mother species: the mother species of the Schizophyllum is transferred to the PDA solid medium (the composition and weight percentages of the PDA solid medium are: potato starch 20%, glucose 20%, agar 20%, and all the other are water ) and cultured at 28°C for 10 days;
[0024] (2) Preparation of the primary strain: the plate mycelium obtained in step (1) is cut into small pieces less than 1mm, and the Schizophyllum mycelium is transferred to a 100ml liquid fermentation tank according to the inoculation amount of 0.5g mycelium / 100ml Culture medium (the composition and weight percent of liquid fermentation medium are: soluble starch 4%, glucose 4%, wheat flour 2%, corn flour 2%, yeast extract 0.3%, soybean flour 0.5%, peptone 0.2%, KH 2 PO 4 1%, MgSO 4 0.5%, VB 1 0.01%, and the rest is water) in a 500ml shake flask, at 28°C...
Embodiment 2
[0026] The preparation of embodiment 2 schizophyllan extract
[0027] Proceed as follows:
[0028] (1) Add 10 g of Schizophyllum mycelium powder to 100 mL of hot water at 98° C., extract in a water bath at 100° C. for 8 hours, and filter to obtain filtrate and filter residue;
[0029] (2) Add the filter residue obtained in step (1) into 100 mL of hot water at 98°C, extract in a water bath at 100°C for 8 hours, filter, and collect the filtrate;
[0030] (3) The 2 filtrates of steps (1) and (2) were combined, concentrated in vacuo to 5 mL, fully mixed with 25 mL volume of 95% ethanol, left to stand overnight at 4 ° C, centrifuged at 5000 rpm for 10 min, poured off the supernatant, and the precipitate was White fibrous, the precipitate was washed twice with 75% ethanol, dried at 60°C until constant weight, weighing 1.12g.
[0031] Determination of polysaccharide content: enone-sulfuric acid assay
[0032] Preparation of reagents:
[0033] Glucose standard solution: 100ug / mL (...
Embodiment 3
[0040] The preparation of embodiment 3 schizophyllan extract
[0041] Proceed as follows:
[0042] (1) Add 12 g of Schizophyllum mycelium powder into 180 mL of hot water at 95° C., extract in a water bath at 100° C. for 9 hours, and filter to obtain filtrate and filter residue;
[0043] (2) Add the filter residue obtained in step (1) into 180 mL of hot water at 95°C, extract in a water bath at 100°C for 9 hours, filter, and collect the filtrate;
[0044] (3) Combine the two filtrates of steps (1) and (2), concentrate in vacuo to 6 mL, mix the filtrate with 30 mL of 95% ethanol, let it stand overnight at 4 ° C, centrifuge at 5000 rpm for 10 min, pour off the supernatant, and the precipitate is white The fibrous crude polysaccharide was washed twice with 75% ethanol, dried at 60°C until constant weight, and weighed 1.41g. The polysaccharide content in the sample was calculated to be 0.452 g by the standard curve prepared by the enone-sulfuric acid assay.
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