Bolete extract, preparation and application thereof
A bolete, extract technology, applied in the directions of botanical equipment and methods, applications, chemicals used for biological control, etc., can solve problems such as no reports on the effect, achieve low cost, simple production method, promote effect of growth
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Embodiment 1
[0026] The preparation of embodiment 1 black boletus extract
[0027] (1) Dry the commercially available black boletus at 40°C, weigh 10g, smash it with a tissue grinder, and pass through a 60-mesh sieve to obtain black boletus dry powder;
[0028] (2) Add 100ml of phosphate buffer solution (pH8.0, 25mmol / L) to the black boletus dry powder obtained in step (1) at 4°C, oscillate and mix well, and all the operations are carried out on ice to obtain a mixed solution;
[0029] (3) Centrifuge the mixture obtained in step (2) at 4° C. and 12,000 rpm for 20 minutes; discard the precipitate, and collect the supernatant to obtain 90 ml of Boletus nigella extract, which should be frozen at -20° C. for future use.
[0030] The protein concentration of the black boletus extract was detected with the BCA protein concentration kit (Pierce company, U.S.), and the assay method was carried out according to the instructions of the kit. As a result, the protein concentration of the above-mention...
Embodiment 2
[0031] The preparation of embodiment 2 yellow boletus extract
[0032] (1) Dry the commercially available Boletus cerevisiae at 40°C, weigh 10g, crush it with a tissue grinder, and pass through a 60-mesh sieve to obtain a dry powder;
[0033] (2) Add 100ml of phosphate buffer (PH8.0, 25mmol / L) to the boletus dry powder obtained in step (1) at 4°C, shake and mix, and operate on ice to obtain a mixed solution;
[0034] (3) Centrifuge the mixture obtained in step (2) at 4°C and 12000rpm for 20min; discard the precipitate, and collect the supernatant to obtain 90ml of Boletus flavus extract, which should be frozen at -20°C for future use.
[0035] The BCA protein concentration kit (Pierce, USA) was used to detect the protein concentration of the Boletus flavus extract, and the assay method was carried out according to the instructions of the kit. As a result, the protein concentration of the Boletus flavus extract was 7.3425mg / ml.
Embodiment 3
[0036] Example 3 Antiviral Activity Test of Boletus nigra Extract to Tobacco Mosaic Virus
[0037] (1) Processing: the black boletus extract prepared in Example 1 is diluted with water successively to three concentrations of protein concentrations of 10.00 μg / ml, 20.00 μg / ml, and 40.00 μg / ml, and compared with water;
[0038] (2) When the leaves of the same seedling age are grown to 8-10 true leaves, the topping treatment is carried out, and the 5th to 8th true leaves are reserved, and the whole plant of the dead-spotted Sansheng tobacco is sprayed. Do three replicates, three plants per replicate, three leaves per plant;
[0039] (3) After 72 hours of induction, remove the main veins from the tobacco leaves of common tobacco infected with tobacco mosaic virus, mix with 0.01M phosphate buffer solution of pH 7.0 according to the weight-to-volume ratio of 1:20, add 0.015g corundum per 20ml, and Grind the leaves, inoculate the induced tobacco with dry spot by rubbing, rinse with ...
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