A kind of black fungus chaff extract liquid and its preparation method and application

Abstract

The present invention discloses a black fungus chaff extract liquid, a preparation method thereof, and an application thereof, and belongs to the technical field of chaff recycling. The preparation method of the black fungus chaff extract liquid comprises the following steps: selecting fresh and mildew-free black fungus chaff, naturally air-drying, crushing, and sieving to obtain black fungus chaff powder for standby use; adding the black fungus chaff powder to a citric acid solution, placing the solution in a high-pressure container for pressurized soaking and extraction to obtain a solid-liquid mixture for standby use; centrifuging the solid-liquid mixture, and taking the supernatant to obtain the product. The preparation method of the present invention can better and more fully extract the effective ingredients in the black fungus chaff, and increase the soluble solid content in the black fungus chaff extract liquid. The present invention applies it to the cultivation of winter fungus, purple-backed vegetable, and mint for the first time, and can significantly improve the germination rate and seed vitality of seeds by soaking seeds, regulate the growth of different plants, and achieve good growth effects.

Description

Technical Field

[0001] The invention belongs to the technical field of mushroom dregs recycling and relates to a black fungus mushroom dregs extract liquid and a preparation method and application thereof. Background Art

[0002] The resource development and utilization of mushroom waste remains a bottleneck in the edible fungi industry. On December 24, 2022, the Edible Fungi Branch of the Chinese Society of Agronomy and the National Edible Fungi Industry Technology System jointly released the top ten technical and scientific issues in my country's edible fungi industry, as voted on by industry insiders. Among them, efficient recycling technology for edible fungi waste ranked first, receiving 84.0% of the vote.

[0003] my country is the world's largest producer of black fungus (Auricularia auricula), accounting for over 90% of global production. The black fungus industry currently ranks second nationwide, with a significant area cultivated in Hebei Province. Black fungus waste is a significant source of nutrients and bioactive substances, such as protein, cellulose, and polysaccharides, making it highly valuable for recycling. However, large-scale cultivation of black fungus generates significant amounts of waste. Due to a lack of large-scale disposal methods, much of this waste is haphazardly dumped in fields, with a small amount being incinerated. This can lead to ecological problems such as agricultural non-point source pollution and air pollution, as well as resource waste, which is inconsistent with the concept of green development.

[0004] Although mushroom shavings have many uses, their utilization rate is currently low in actual production, and large-scale processing has not yet been achieved and has not been well promoted. Traditional methods for extracting mushroom shavings include hot water extraction and ultrasonic-assisted extraction, but these methods take a long time to extract and have low extraction efficiency.

[0005] Therefore, how to develop a practical resource utilization method for black fungus chaff, enrich the technical approach of ecological agriculture, and provide a black fungus chaff extract liquid and its preparation method and application are problems that need to be solved urgently by those skilled in the art. Summary of the Invention

[0006] In view of this, the present invention provides a black fungus chaff extract liquid and a preparation method and application thereof.

[0007] In order to achieve the above object, the present invention adopts the following technical solutions:

[0008] A method for preparing a black fungus chaff extract comprises the following steps:

[0009] (1) fresh black fungus chaff without mildew is selected, naturally air-dried, crushed, and sieved to obtain black fungus chaff powder;

[0010] (2) adding the black fungus chaff powder to a citric acid solution and placing the solution in a high-pressure container for pressurized soaking extraction to obtain a solid-liquid mixture;

[0011] (3) centrifuging the solid-liquid mixture and taking the supernatant to obtain the black fungus bran extract.

[0012] The beneficial effects of the present invention are as follows: after the mushroom shavings are crushed, reacted with an acid solvent and pressurized, more contents are released and the extraction efficiency is high. Therefore, the preparation method of the present invention can better and more fully extract the effective ingredients in the black fungus mushroom shavings, increase the soluble solid content in the black fungus mushroom shavings extraction liquid, and compared with the mushroom shavings, the contents of components such as crude protein, crude fat, polysaccharides, calcium, phosphorus, potassium, and total flavonoids in the extraction liquid are all increased.

[0013] Furthermore, in step (1), the sieving is performed through a 60-80 mesh sieve.

[0014] Furthermore, in step (2), the solid-liquid ratio of the black fungus bran powder and the citric acid solution is 1:30 g / mL, and the concentration of the citric acid solution is 0.25 g / L.

[0015] The beneficial effect of adopting the above-mentioned further technical solution is that as the material-to-liquid ratio increases, the soluble solids content tends to first increase and then decrease. This is because the larger the volume of the extracting liquid, the more complete the contact between the solvent and the fungus residue, and more of the contents are dissolved in the same amount of time. When the material-to-liquid ratio exceeds the optimal ratio, the soluble solids content decreases. However, if the material-to-liquid ratio is too high, it will consume more time and energy. If the material-to-liquid ratio is too high, the concentration of the solvent in the extracting liquid will decrease, resulting in reduced extraction efficiency.

[0016] The solvent penetrates the cell membrane of the raw material, dissolving the soluble substances, creating a concentration difference between the inside and outside of the cell, and then seeping through the cell membrane to achieve the purpose of extraction. Increasing the concentration of the medium can promote the release of the extract because higher concentrations can better destroy the biological cell structure, making the extract easier to release.

[0017] Furthermore, in step (2), the pressurization pressure is 0.1 MPa and the pressurization time is 20 min.

[0018] The beneficial effects of adopting the above further technical solution are: pressurization improves the solubility of the solvent and promotes the diffusion of the solvent into the sample matrix, enhances the penetration of the solvent into the solute in the solid, accelerates the transfer of solute molecules from the solid phase to the liquid phase, and thus improves the leaching efficiency.

[0019] The main principle of the influence of pressurization time on extraction effect is that increasing pressure can significantly improve the extraction efficiency of the solvent and the quality of the extract.

[0020] Furthermore, in step (3), the centrifugal speed is 4000 r / min and the centrifugal time is 10 min.

[0021] The invention also provides a black fungus chaff extract liquid prepared by the method.

[0022] The present invention also provides an application of the black fungus husk extract in plant cultivation.

[0023] The beneficial effects of the present invention are as follows: the black fungus chaff extract liquid is applied to plant planting, the black fungus chaff extract liquid can significantly improve the germination rate and seed vitality of plant seeds by soaking them, and spraying fungus chaff extract liquids of different concentrations can regulate the growth of different plants, thereby achieving good growth effects.

[0024] Mushroom bran is the solid culture medium left after edible fungi cultivation. During the cultivation process, it retains a large amount of nutrients and active substances, including mycelium, microbially degraded cellulose, crude protein, crude fat, secondary metabolites of mycelium, and minerals, making it a valuable resource for recycling. Mushroom bran contains a variety of amino acids, which can be absorbed and utilized by plants, promoting their growth and development and enhancing their resistance to stress. It also improves soil quality, increases crop yields, mitigates the toxic effects of heavy metal ions on plants, and improves product quality.

[0025] Mushroom bran contains the metabolites of edible fungi, including sugars, organic acids, enzymes and bioactive substances, a large number of microbial communities and residual mycelium.

[0026] Furthermore, the above-mentioned plant is winter melon, purple-backed vegetable or mint.

[0027] The beneficial effects of adopting the above-mentioned further technical solution are as follows: the present invention applies the black fungus bran extract to the cultivation of winter melon, purple-backed cabbage, and mint for the first time; the black fungus bran extract can significantly improve the germination rate and seed vitality of winter melon, purple-backed cabbage, and mint seeds by soaking them in the black fungus bran extract; spraying different concentrations of the bran extract can regulate the growth of winter melon, purple-backed cabbage, and mint, achieving good growth effects.

[0028] Furthermore, the application of the black fungus chaff extract in plant cultivation comprises the following steps:

[0029] (1) Wash the seeds of winter melon, purple melon or mint with distilled water and set aside;

[0030] (2) diluting the above-mentioned black fungus chaff extracting liquid with water to obtain a dilution of the black fungus chaff extracting liquid for standby use;

[0031] (3) soaking wintergreen seeds, purple-backed cabbage seeds, or mint seeds in the diluted solution of the black fungus husk extract, rinsing them several times with clean water after the soaking, placing the seeds in a culture dish and then culturing them in a constant temperature incubator to allow the seeds to germinate, and keeping the bud bed moist during the culturing period;

[0032] (4) Planting the cultivated winter melon seeds, purple-backed vegetable seeds or mint seeds, and spraying the leaves of the winter melon, purple-backed vegetable or mint with a diluted solution of the black fungus chaff extract when the winter melon, purple-backed vegetable or mint grows two leaves.

[0033] Furthermore, in step (3), the soaking time is 24 hours, the concentration of the dilution of the black fungus bran extract is 20%-100%, and the seeds are placed in a culture dish and cultured in a constant temperature incubator at 20-25° C. to allow the seeds to germinate for 7 days;

[0034] In step (4), when the wintercress grows two leaves, the black fungus husk extract with a concentration of 20%-100% is sprayed on the leaves, and the spraying is done once every two days for a total of 30 days.

[0035] Preferably, in step (3), the wintercress seeds are soaked in a 60% dilution of the black fungus bran extract, or the purple-backed cabbage seeds or mint seeds are soaked in an 80% dilution of the black fungus bran extract;

[0036] In step (4), when the wintercress grows two leaves, the black fungus bran extract with a concentration of 60% is sprayed on the leaves, or when the purple-backed cabbage seeds or mint seeds grow two leaves, the black fungus bran extract with a concentration of 80% is sprayed on the leaves, and the spraying is performed once every two days for a total of 30 days.

[0037] Furthermore, in step (4), the spraying amount is 10-15 mL / plant. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 This is the effect of the material-liquid ratio of test examples 1-7 on the soluble solids content of the black fungus bran extract.

[0039] Figure 2 This is the effect of pressurization time on the soluble solids content of black fungus bran extract in test examples 8-12.

[0040] Figure 3 This is the effect of pressurized pressure on the soluble solids content of black fungus bran extract in Test Examples 13-15.

[0041] Figure 4 This is the soluble solid content of the black fungus bran extract in Test Examples 16-20.

[0042] Figure 5Response surface and contour diagram between factors AB.

[0043] Figure 6 Response surface and contour plots between factors AC.

[0044] Figure 7 Response surface and contour plots between factors AD.

[0045] Figure 8 Response surface and contour plots between factors BC.

[0046] Figure 9 Response surface and contour plots between factors BD.

[0047] Figure 10 Response surface and contour plots between factors CD. DETAILED DESCRIPTION

[0048] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0049] Example 1

[0050] The preparation method of black fungus chaff extract comprises the following steps:

[0051] (1) selecting a first batch of fresh black fungus chaff without mildew, naturally air-drying, crushing, and passing through an 80-mesh sieve to obtain black fungus chaff powder;

[0052] (2) 5 g of black fungus chaff powder was added to a citric acid solution and placed in a high-pressure steam sterilizer for pressurized immersion extraction to obtain a solid-liquid mixture, wherein the solid-liquid ratio of the black fungus chaff powder to the citric acid solution was 1:30 g / mL, the concentration of the citric acid solution was 0.25 g / L, the pressurized pressure was 0.1 MPa, and the pressurization time was 20 min;

[0053] (3) The solid-liquid mixture was centrifuged at a centrifugal speed of 4000 r / min for 10 min, and the supernatant was collected to obtain a black fungus bran extract liquid having a soluble solid content of 2.51%.

[0054] Effect experiment:

[0055] Test Example 1

[0056] The preparation method of black fungus chaff extract comprises the following steps:

[0057] (1) Selecting a second batch of fresh black fungus chaff without mildew, naturally air-drying, crushing, and passing through an 80-mesh sieve to obtain black fungus chaff powder;

[0058] (2) 5 g of black fungus chaff powder was added to a citric acid solution and placed in a high-pressure steam sterilizer for pressurized immersion extraction to obtain a solid-liquid mixture, wherein the solid-liquid ratio of the black fungus chaff powder to the citric acid solution was 1:30 g / mL, the concentration of the citric acid solution was 0.25 g / L, the pressurized pressure was 0.1 MPa, and the pressurization time was 20 min;

[0059] (3) centrifuging the solid-liquid mixture at a centrifugal speed of 4000 r / min for 10 min, taking the supernatant to obtain the black fungus bran extract.

[0060] Test Example 2

[0061] Compared with Experimental Example 1, except that the material-liquid ratio of black fungus bran powder and citric acid solution is 1:15 g / mL, other steps and parameters are exactly the same as those in Experimental Example 1.

[0062] Test Example 3

[0063] Compared with Experimental Example 1, except that the material-liquid ratio of black fungus bran powder and citric acid solution is 1:20 g / mL, other steps and parameters are exactly the same as those in Experimental Example 1.

[0064] Test Example 4

[0065] Compared with Experimental Example 1, except that the material-liquid ratio of black fungus bran powder and citric acid solution is 1:25 g / mL, other steps and parameters are exactly the same as those in Experimental Example 1.

[0066] Test Example 5

[0067] Compared with Experimental Example 1, except that the material-liquid ratio of black fungus bran powder and citric acid solution is 1:35 g / mL, other steps and parameters are exactly the same as those in Experimental Example 1.

[0068] Test Example 6

[0069] Compared with Experimental Example 1, except that the material-liquid ratio of black fungus bran powder and citric acid solution is 1:40 g / mL, other steps and parameters are exactly the same as those in Experimental Example 1.

[0070] Test Example 7

[0071] Compared with Experimental Example 1, except that the material-liquid ratio of black fungus bran powder and citric acid solution is 1:45 g / mL, other steps and parameters are exactly the same as those in Experimental Example 1.

[0072] Test Example 8

[0073] The preparation method of black fungus chaff extract comprises the following steps:

[0074] (1) selecting fresh, mold-free black fungus chaff from the third batch, naturally air-drying, crushing, and passing through an 80-mesh sieve to obtain black fungus chaff powder;

[0075] (2) 5 g of black fungus chaff powder was added to a citric acid solution and placed in a high-pressure steam sterilizer for pressurized immersion extraction to obtain a solid-liquid mixture, wherein the solid-liquid ratio of the black fungus chaff powder to the citric acid solution was 1:30 g / mL, the concentration of the citric acid solution was 0.25 g / L, the pressurized pressure was 0.1 MPa, and the pressurization time was 20 min;

[0076] (3) centrifuging the solid-liquid mixture at a centrifugal speed of 4000 r / min for 10 min, taking the supernatant to obtain the black fungus bran extract.

[0077] Test Example 9

[0078] Compared with Experimental Example 8, except that the pressurization time was 10 min, other steps and parameters were exactly the same as those in Experimental Example 8.

[0079] Test Example 10

[0080] Compared with Experimental Example 8, except that the pressurization time was 15 minutes, other steps and parameters were exactly the same as those in Experimental Example 8.

[0081] Test Example 11

[0082] Compared with Experimental Example 8, except that the pressurization time was 25 min, other steps and parameters were exactly the same as those in Experimental Example 8.

[0083] Test Example 12

[0084] Compared with Experimental Example 8, except that the pressurization time was 30 min, other steps and parameters were exactly the same as those in Experimental Example 8.

[0085] Test Example 13

[0086] The preparation method of black fungus chaff extract comprises the following steps:

[0087] (1) fresh, mold-free black fungus chaff of the fourth batch was selected, naturally air-dried, crushed, and passed through an 80-mesh sieve to obtain black fungus chaff powder;

[0088] (2) 5 g of black fungus chaff powder was added to a citric acid solution and placed in a high-pressure steam sterilizer for pressurized immersion extraction to obtain a solid-liquid mixture, wherein the solid-liquid ratio of the black fungus chaff powder to the citric acid solution was 1:30 g / mL, the concentration of the citric acid solution was 0.25 g / L, the pressurized pressure was 0.1 MPa, and the pressurization time was 20 min;

[0089] (3) centrifuging the solid-liquid mixture at a centrifugal speed of 4000 r / min for 10 min, taking the supernatant to obtain the black fungus bran extract.

[0090] Test Example 14

[0091] Compared with Experimental Example 13, except that the pressurization pressure is 0.05 MPa, other steps and parameters are exactly the same as Experimental Example 13.

[0092] Test Example 15

[0093] Compared with Experimental Example 13, except that the pressurization pressure is 0.15 MPa, other steps and parameters are exactly the same as those of Experimental Example 13.

[0094] Test Example 16

[0095] The preparation method of black fungus chaff extract comprises the following steps:

[0096] (1) Select fresh, mold-free black fungus chaff from the fifth batch, air-dry it, crush it, and pass it through an 80-mesh sieve to obtain black fungus chaff powder;

[0097] (2) 5 g of black fungus chaff powder was added to a citric acid solution and placed in a high-pressure steam sterilizer for pressurized immersion extraction to obtain a solid-liquid mixture, wherein the solid-liquid ratio of the black fungus chaff powder to the citric acid solution was 1:30 g / mL, the concentration of the citric acid solution was 0.25 g / L, the pressurized pressure was 0.1 MPa, and the pressurization time was 20 min;

[0098] (3) centrifuging the solid-liquid mixture at a centrifugal speed of 4000 r / min for 10 min, taking the supernatant to obtain the black fungus bran extract.

[0099] Test Example 17

[0100] Compared with Experimental Example 16, except that the concentration of the citric acid solution was 0.19 g / L, other steps and parameters were exactly the same as those of Experimental Example 16.

[0101] Test Example 18

[0102] Compared with Experimental Example 16, except that the concentration of the citric acid solution was 0.21 g / L, other steps and parameters were exactly the same as those of Experimental Example 16.

[0103] Test Example 19

[0104] Compared with Experimental Example 16, except that the concentration of the citric acid solution was 0.23 g / L, other steps and parameters were exactly the same as those of Experimental Example 16.

[0105] Test Example 20

[0106] Compared with Experimental Example 16, except that the concentration of the citric acid solution was 0.27 g / L, other steps and parameters were exactly the same as those of Experimental Example 16.

[0107] Comparative Example 1

[0108] The preparation method of black fungus chaff extract comprises the following steps:

[0109] (1) Selecting a second batch of fresh black fungus chaff without mildew, naturally air-drying, crushing, and passing through an 80-mesh sieve to obtain black fungus chaff powder;

[0110] (2) 5 g of black fungus chaff powder was added to distilled water and placed in a high-pressure steam sterilizer for pressurized immersion extraction to obtain a solid-liquid mixture, the solid-liquid ratio of black fungus chaff powder to distilled water was 1:30 g / mL, the pressurized pressure was 0.1 MPa, and the pressurization time was 15 min;

[0111] (3) The solid-liquid mixture was centrifuged at a centrifugal speed of 4000 r / min for 10 min, and the supernatant was collected to obtain a black fungus bran extract liquid having a soluble solid content of 2.44%.

[0112] Comparative Example 1 uses distilled water as the medium, which is different from the medium of the present invention, so the optimal process parameters are different. After testing, the soluble solids content of this comparative example is the highest when distilled water is used as the medium.

[0113] Comparative Example 2

[0114] The preparation method of black fungus chaff extract comprises the following steps:

[0115] (1) Selecting a second batch of fresh black fungus chaff without mildew, naturally air-drying, crushing, and passing through an 80-mesh sieve to obtain black fungus chaff powder;

[0116] (2) 5 g of black fungus chaff powder was added to a NaHCO3 solution and placed in a high-pressure steam sterilizer for pressurized immersion extraction to obtain a solid-liquid mixture, wherein the solid-liquid ratio of the black fungus chaff powder to the NaHCO3 solution was 1:35 g / mL, the concentration of the NaHCO3 solution was 8 g / L, the pressurized pressure was 0.1 MPa, and the pressurization time was 20 min;

[0117] (3) The solid-liquid mixture was centrifuged at a centrifugal speed of 4000 r / min for 10 min, and the supernatant was collected to obtain a black fungus bran extract liquid having a soluble solid content of 2.50%.

[0118] Comparative Example 2 uses NaHCO3 solution as the medium, which is different from the medium of the present invention, so the optimal process parameters are different. After testing, the soluble solids content of this comparative example is the highest when NaHCO3 solution is used as the medium.

[0119] Comparative Example 3

[0120] The hot water extraction method for preparing black fungus husk extract comprises the following steps:

[0121] (1) Select fresh, mold-free black fungus chaff from the fifth batch, air-dry it, crush it, and pass it through an 80-mesh sieve to obtain black fungus chaff powder;

[0122] (2) Add 5 g of black fungus chaff powder to citric acid solution and place it in a constant temperature water bath for extraction to obtain a solid-liquid mixture. The solid-liquid ratio of black fungus chaff powder to citric acid solution is 1:30 g / mL, the concentration of citric acid solution is 0.25 g / L, the extraction temperature is 75 ° C, and the extraction time is 3.5 h.

[0123] (3) The solid-liquid mixture was centrifuged at a centrifugal speed of 4000 r / min for 10 min, and the supernatant was collected to obtain a black fungus bran extract liquid having a soluble solid content of 2.34%.

[0124] The hot water extraction method is different from the pressure extraction method of the present invention, so the influencing factors and process parameters of the two are not exactly the same. The influencing factors of the hot water extraction method include the solid-liquid ratio, extraction temperature, extraction time, and extraction medium. After testing, the soluble solids content of this comparative example in the hot water extraction method is the highest.

[0125] Comparative Example 4

[0126] The ultrasonic-assisted extraction method for preparing black fungus husk extract comprises the following steps:

[0127] (1) Select fresh, mold-free black fungus chaff from the fifth batch, air-dry it, crush it, and pass it through an 80-mesh sieve to obtain black fungus chaff powder;

[0128] (2) Add 5 g of black fungus chaff powder to citric acid solution and place it in an ultrasonic cleaner for extraction to obtain a solid-liquid mixture. The solid-liquid ratio of black fungus chaff powder to citric acid solution is 1:30 g / mL, the concentration of citric acid solution is 0.25 g / L, the ultrasonic temperature is 40 ° C, and the ultrasonic time is 20 min.

[0129] (3) The solid-liquid mixture was centrifuged at a centrifugal speed of 4000 r / min for 10 min, and the supernatant was collected to obtain a black fungus bran extract liquid having a soluble solid content of 2.46%.

[0130] The ultrasonic-assisted extraction method is different from the pressurized extraction method of the present invention, so the influencing factors and process parameters of the two are not exactly the same. The influencing factors of the ultrasonic-assisted extraction method include the material-liquid ratio, ultrasonic time, ultrasonic temperature, and extraction medium. After testing, the soluble solids content of this comparative example in the ultrasonic-assisted extraction method is the highest.

[0131] Determination of soluble solids content: Use a handheld refractometer to determine the soluble solids content of the bran extract. The operation method is as follows: First, calibrate the zero. Drop a few drops of distilled water on the light-shielding prism, close the light-inlet plate, and evenly distribute it on the surface of the prism. Aim the light-inlet plate at a bright place, observe the light-dark dividing line through the eyepiece, and rotate the zeroing screw to set it to zero. Then wipe the distilled water clean, take 2 to 3 drops of the test liquid and drop it on the detection prism, close the cover to avoid bubbles. Hold the refractometer in hand and aim the light-inlet plate at a bright light source, observe through the eyepiece, turn the handwheel to adjust the eyepiece, so that the white-blue dividing line in the field of view is clear. The scale value of the dividing line is the soluble solids content of the test liquid.

[0132] (1) Effect of material-liquid ratio on soluble solids content

[0133] from Figure 1 It can be seen that under the conditions of weakly acidic aqueous solution, with the increase of the solid-liquid ratio, the soluble solid content first increases and then decreases, and reaches the maximum of 2.49% when the solid-liquid ratio is 1:35.

[0134] (2) Effect of pressurization time on soluble solids content

[0135] from Figure 2 It can be seen that under the condition of weakly acidic aqueous solution, when the pressurization time is 20 minutes, the soluble solid content is the largest, which is 2.47%.

[0136] (3) Effect of pressure on soluble solids content

[0137] from Figure 3 It can be seen that under weakly acidic aqueous solution conditions, the soluble solid content increases with increasing pressure, and reaches a maximum of 2.42% when the pressure is 0.1 MPa.

[0138] (4) Effect of extraction medium on soluble solids content

[0139] from Figure 4 It can be seen that under the condition of weakly acidic aqueous solution, the soluble solid content increased. When the citric acid concentration was 0.25 g / L, the soluble solid content was the largest, which was 2.51%.

[0140] Box-Behnken design test and verification

[0141] The preparation method of black fungus chaff extract comprises the following steps:

[0142] (1) selecting a first batch of fresh black fungus chaff without mildew, naturally air-drying, crushing, and passing through an 80-mesh sieve to obtain black fungus chaff powder;

[0143] (2) Add 5 g of black fungus husk powder to a citric acid solution and place the mixture in a high-pressure steam sterilizer for pressurized soaking extraction to obtain a solid-liquid mixture;

[0144] (3) centrifuging the solid-liquid mixture at a centrifugal speed of 4000 r / min for 10 min, taking the supernatant to obtain the black fungus bran extract.

[0145] After determining the optimal response region through single-factor experiments, the response surface methodology was used to further optimize the interactions between the various influencing factors and determine the optimal process route. At the same time, after optimization through Box-Behnken design experiments, the optimized process route was verified to test its reliability and accuracy.

[0146] Based on the results of the single-factor experiment, we selected A: pressure, B: material-liquid ratio, C: pressurization time, and D: acid concentration as the influencing factors, and Y: soluble solids content as the response value. A four-factor, three-level Box-Behnken experiment was conducted. The design table is shown in Table 1, and the experimental results are shown in Table 2.

[0147] Table 1 Box-Behnken test factors and levels

[0148]

[0149] Table 2 Box-Behnken design experimental plan and results

[0150]

[0151]

[0152] Box-Behnken test ANOVA

[0153] The experimental data were analyzed and processed using Design-Expert.V8.0.6.1 software, and regression analysis was performed on the interaction between factors. The quadratic multinomial regression model equation was obtained as follows:

[0154] Y=2.53-3.333E-003A+7.500E-003B+4.167E-003C-5.000E-003D-0.048AB-0.035AC-0.048AD-0.047BC-0.028BD-0.040CD-0.057A 2 -0.058B 2 -0.043C 2 -0.055D 2

[0155] According to the regression model variance analysis shown in Table 3, it can be seen that the model P value is less than 0.0001, indicating that the regression equation is significant. The entire model has a good fit in the experimental interval, and the P value of the lack of fit term is 0.1986, which is greater than 0.05 and the difference is not significant, indicating that the selection of variables and response values ​​is reasonable and the experimental operation is credible. Model correlation coefficient R 2 =0.9906, indicating a good correlation, and the corrected determination coefficient AdjR 2 =0.9813, indicating that the model can analyze 98.13% of the experimental data variation. The accuracy of this experiment is 29.71, which is within a reasonable range (greater than 4.0). 2 、B 2 、C 2 、D 2 The P values ​​of the four factors are less than 0.0001, indicating that they have a very significant impact on the response value. The P values ​​of the four-factor interaction terms also indicate that the interaction effects between the different influencing factors are large. Therefore, this model can be used to predict the actual soluble solids content of mushroom bran.

[0156] Table 3 Box-Behnken experimental design regression analysis results

[0157]

[0158]

[0159] Response surface analysis

[0160] The three-dimensional response surface diagram and contour diagram can intuitively show the interaction relationship between different factors, such as Figures 5 to 10 As shown, all 3D response surface plots open downward and have a peak, indicating that the response increases with increasing factor values, then gradually decreases after reaching a maximum. Within the experimental range, the optimal formulation can be determined using response surface plots. The contour plots are all elliptical, indicating a significant interaction between the two factors. The interaction between factors A and B has the greatest impact on the soluble solids content of the bran, resulting in the steepest slope in its 3D response surface plot.

[0161] Verification test results

[0162] Software analysis revealed the optimal process parameters to be: pressure 0.1 MPa, pressurization time 20.28 min, solid-liquid ratio 1:30.39, and citric acid concentration 0.25 g / L. At this point, the theoretical soluble solids content was 2.53%. Based on actual operation, the parameters were adjusted to pressure 0.1 MPa, pressurization time 20 min, solid-liquid ratio 1:30, and citric acid concentration 0.25 g / L. The experiment was repeated under these conditions, yielding a soluble solids content of 2.51%. The relative error from the predicted value was 0.79%, which is within the allowable range, indicating a high degree of model fit and accurate predictions.

[0163] Detection experiment

[0164] 1. The nutritional components of the product of Example 1 were determined, wherein:

[0165] Determination of protein content: The Coomassie brilliant blue method improved by Wang Aiping et al. Wang Aiping, Zhou Liming. Optimization of conditions for determining protein content in tea seed polysaccharides by Coomassie brilliant blue method [J]. Henan Agricultural Science, 2014, 43(03): 150-153;

[0166] Determination of crude fat content: refer to the Soxhlet extraction method in GB / T 5009.6-2016;

[0167] Determination of calcium content: refer to the potassium permanganate method in GB / T 6436-2018;

[0168] Determination of phosphorus content: refer to the spectrophotometric method in GB / T 6437-2018;

[0169] Determination of potassium content: refer to Shen Yaqin's test method and use flame photometer for determination. Shen Yaqin, He Birong. Analysis of preparation technology of sodium and potassium ion standard solutions [J]. Engineering Technology Research, 2020, 5(18): 134-135;

[0170] 2. The active ingredients of the product of Example 1 were measured, wherein:

[0171] Determination of crude polysaccharide content: refer to NY / T 1676-2008;

[0172] The total phenol content was determined by referring to the Folin-Ciocalteus method. After determining the gallic acid standard curve, the OD value of the diluted wine sample was measured and the total phenol content was calculated based on the curve.

[0173] The total flavonoids content was determined by aluminum chloride absorbance method. After the rutin standard curve was determined, the OD value of the diluted wine sample was measured and the total flavonoids content was calculated based on the curve.

[0174] The determination of indoleacetic acid (IAA), gibberellin A3 (GA3), and abscisic acid (ABA) contents was carried out according to the methods in the literature. Zhang Yongmei, Ma Huiling, Zhang Suoke, et al. Determination of four endogenous hormones in Kentucky bluegrass by high performance liquid chromatography [J]. Acta Grasslandiae Sinica, 2016, 24(06): 1384-1387.

[0175] Finally, Microsoft Excel, SPSS20.0, and Origin8.0 software were used for data statistical analysis.

[0176] Table 4 Nutritional component detection and results of Example 1 products

[0177]

[0178]

[0179] Table 5: Functional component detection and results of Example 1 products

[0180]

[0181] The test results of various components of the product of Example 1 are shown in Tables 4 and 5. The results show that the contents of crude protein, crude fat and other components in the extracted liquid are increased, the potassium content can reach 5.77 mg / g, the calcium content can reach 2.99%, the phosphorus content can reach 0.27%, the polysaccharide content can reach 0.55%, the polyphenol content can reach 1.19 mg / g, the flavonoids content can reach 6.57 mg / g, the crude protein content can reach 6.57%, the crude fat content can reach 0.72%, the IAA content can reach 89.75 μg / g, the ABA content can reach 63.23 μg / g, and the GA3 content can reach 50.02 μg / g.

[0182] The crude protein content in the fungus bran is 6.28±0.21%, the crude fat content is 0.67±0.09%, the calcium content is 2.67±0.19%, the phosphorus content is 0.25±0.03%, the potassium content is 6.56±0.23 mg / g, the polysaccharide content is 0.53±0.20%, the total flavonoids content is 1.68±0.19 mg / g, the polyphenol content is 1.24±0.12 mg / g, the gibberellin content is 48.04±3.7μg / g, the abscisic acid content is 65.79±5.2μg / g, and the indoleacetic acid content is 89.52±2.62μg / g.

[0183] After testing, it was found that compared with the mushroom bran, the contents of crude protein, crude fat, polysaccharides, calcium, phosphorus, potassium, total flavonoids and other components in the extracted liquid were increased.

[0184] Example 2

[0185] The application of black fungus husk extract in growing winter vegetables comprises the following steps:

[0186] (1) Wash the wintercress seeds with distilled water for later use;

[0187] (2) diluting the black fungus chaff extracting liquid of described embodiment 1 with water to obtain the dilution of black fungus chaff extracting liquid, for subsequent use;

[0188] (3) Soaking the wintercress seeds in a 60% dilution of the black fungus husk extract for 24 h. After soaking, rinse with clean water several times. Place the seeds in a culture dish and then place them in a constant temperature incubator at 25° C. to allow the seeds to germinate. Keep the bud bed moist during the incubation period. The incubation period is 7 days.

[0189] (4) Plant the cultivated winter melon seeds, and when the winter melon grows two leaves, spray the leaves with a 60% concentration of black fungus bran extract at a rate of 10-15 mL / plant, once every two days for a total of 30 days.

[0190] Example 3

[0191] The application of black fungus husk extract in growing purple-backed vegetable comprises the following steps:

[0192] (1) Wash the purple-backed vegetable with distilled water for later use;

[0193] (2) diluting the black fungus chaff extracting liquid of described embodiment 1 with water to obtain the dilution of black fungus chaff extracting liquid, for subsequent use;

[0194] (3) Soaking the seeds of Auricularia auriculariae in a dilution of 80% black fungus husk extract for 24 h. After soaking, rinse with clean water several times. Place the seeds in a culture dish and then place them in a constant temperature incubator at 25°C for germination. Keep the bud bed moist during the culture period. The culture period is 7 days.

[0195] (4) Planting the cultured purple-backed cabbage seeds, and starting to spray the leaves with 80% black fungus bran extract when the purple-backed cabbage seeds grow two leaves, with a spraying amount of 10-15 mL / plant, once every two days, for a total of 30 days.

[0196] Example 4

[0197] The application of black fungus chaff extract in plant cultivation comprises the following steps:

[0198] (1) Wash mint seeds with distilled water for later use;

[0199] (2) diluting the black fungus chaff extracting liquid of described embodiment 1 with water to obtain the dilution of black fungus chaff extracting liquid, for subsequent use;

[0200] (3) soaking mint seeds in a diluted solution of 80% black fungus husk extract for 24 h, rinsing with clean water several times after soaking, placing the seeds in a culture dish and then culturing in a constant temperature incubator at 25° C. to allow the seeds to germinate, keeping the bud bed moist during the culturing period, and culturing for 7 days;

[0201] (4) Planting the cultured mint seeds. When the mint seeds grow two leaves, spray the leaves with 80% black fungus bran extract at a rate of 10-15 mL / plant, once every two days for a total of 30 days.

[0202] 1. Seed soaking test of wintercress, purple-backed cabbage and mint

[0203] The seeds of wintercress, purple-backed cabbage and mint were repeatedly washed with distilled water for later use. Five treatments were set up for the seed soaking test: the mother liquor was diluted with the bran extract of Example 1 in a gradient manner, with concentrations of 20%, 40%, 60%, 80% and 100% respectively, and soaked in clean water as a control, soaked for 24 hours, and three groups were parallel for each treatment. After the seeds were soaked, they were rinsed with clean water several times, and then the germination test was carried out using the paper bed method. Two layers of qualitative filter paper were laid at the bottom of each culture dish, and the seeds after the seed soaking treatment were placed on the filter paper, and the clean water treatment was used as a control. The culture dish was placed in a constant temperature climate incubator at 25°C for cultivation to keep the bud bed moist. Germination was recorded from the second day, and the germination rate was calculated based on the number of germinations on the 7th day, and the germination index was calculated. The water was absorbed with filter paper, the fresh weight of the seedlings was weighed, and the seed vigor was measured.

[0204] Germination rate (%) = actual number of germinations / total number of tests × 100%

[0205] Seed vigor (%) = germination index × normal seedling fresh weight per plant, germination index = ∑ (number of germinations per day at the end of the germination test / number of germination days).

[0206] Table 6 Effects of seed soaking treatment on germination rate and seed vigor of wintercress, purple-backed cabbage and mint

[0207]

[0208] Note: Data in the same column but different rows without the same lowercase letter indicate significant differences (P<0.05). CK1-CK3 are blank controls.

[0209] The results showed that soaking seeds with mushroom bran extract promoted the germination rate and seed vigor of wintercress, purple-backed radish, and mint. Soaking seeds with a 60% mushroom bran extract had the best germination-promoting effect on wintercress, with a germination rate of 95.3%, a 12.4% increase over the control, and a seed vigor of 95.4%, a 36.1% increase over the control. Soaking seeds with an 80% mushroom bran extract had the best germination effect on purple-backed radish, with a germination rate of 94.2%, an 8.0% increase over the control, and a seed vigor of 88.4%, a 19.0% increase over the control. Soaking seeds with an 80% mushroom bran extract had the best germination effect on mint, with a germination rate of 95.3%, a 10.1% increase over the control, and a seed vigor of 96.1%, a 27.3% increase over the control. Soaking seeds with black fungus bran extract significantly improved the germination rate and seed vigor of wintercress, purple-backed radish, and mint.

[0210] 2. Spraying test on winter cabbage, purple cabbage and mint seedlings

[0211] In the spraying test, the seeds after the cultivation of Examples 2-4 were all planted in the ground. Example 1 The bacterium dregs extract was set to 5 concentration gradients, with the bacterium dregs extract as the mother liquor gradient dilution, respectively 20%, 40%, 60%, 80%, 100% concentration, and the spraying test was started when winter dwarf vegetable, purple back vegetable and mint grew two leaves. The leaves were evenly sprayed, and with clear water as the control, three groups of parallel treatments were performed, sprayed once every two days, and the spraying rate was 10-15mL / plant. After 30 days, its growth index and physiological index were measured.

[0212] (1) Growth indicators

[0213] Root length: After the spraying test, 10 plants were randomly selected from each treatment, and the soil was repeatedly washed off with clean water. The length of the plants from the root base to the root tip was measured with a ruler, and the average value was taken.

[0214] Plant height: After the spraying test, 10 plants were randomly selected from each treatment, and the height of the plants from the ground to the top was measured with a ruler, and the average value was taken.

[0215] Leaf width: After the spraying test, 10 leaves were randomly selected from each treatment, and the widest part of the leaves was measured with a vernier caliper to obtain the average value.

[0216] Aboveground biomass: After the spraying test, 10 plants were randomly selected from each treatment, the soil was repeatedly washed with clean water, and the plants were dried in an oven to a constant weight, and then weighed to obtain the aboveground biomass.

[0217] (2) Physiological indicators

[0218] Before testing, leaves should be carefully rinsed repeatedly with clean water to prevent residual fungus waste extract from affecting the test results. Soluble protein content was determined using Coomassie Brilliant Blue staining. Soluble sugar content was determined using the anthrone colorimetric method. Chlorophyll content was determined using the 95% ethanol grinding and centrifugation method. Malondialdehyde (MDA) content was determined using the thiobarbituric acid method. Superoxide dismutase (SOD) activity was determined using the pyrogallol autoxidation method. Peroxidase (POD) activity was determined using the guaiacol method.

[0219] Results: The effects of spraying different concentrations of mushroom bran extract on the early growth and physiological indicators of wintercress, purple-backed cabbage and mint are shown in Tables 7-9.

[0220] Table 7 Effects of spraying treatment on early growth indicators of wintercress, purple-backed cabbage and mint

[0221]

[0222] Note: Data in the same column but different rows without the same lowercase letters indicate significant differences (P<0.05).

[0223] CK1-CK3 served as blank controls.

[0224] Table 8 Effects of spraying treatment on early physiological indicators of wintercress, purple-backed cabbage and mint

[0225]

[0226]

[0227]

[0228] Table 9 Effects of spraying treatments on early physiological indicators of wintercress, purple-backed cabbage and mint (continued)

[0229] Note: Data in the same column but different rows without the same lowercase letter indicate significant differences (P<0.05). CK1-CK3 are blank controls.

[0230] A comprehensive study of the effects of different spray concentrations on plant height, leaf width, aboveground biomass, chlorophyll a, chlorophyll b, malondialdehyde, SOD, POD, and soluble sugar content of wintergreen, violet, and mint revealed that a 60% concentration of the fungus bran extract had the most significant effect on the germination and early growth of wintergreen, while an 80% concentration had the most significant effects on the germination and early growth of violet and mint. Therefore, spraying different concentrations of fungus bran extract can regulate the growth of different plants and achieve optimal growth outcomes.

[0231] The disclosed embodiments are described to enable one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be readily apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention is not limited to the embodiments shown herein but is to be construed in the widest possible manner consistent with the principles and novel features disclosed herein.

Claims

1. An application of a black fungus chaff extract in plant planting, characterized in that: The plant is winter melon, purple back melon or mint; The preparation method of the black fungus chaff extract comprises the following steps: (1) Select fresh and mold-free black fungus chaff, air-dry it, crush it, and sieve it to obtain black fungus chaff powder; (2) Adding black fungus husk powder to a citric acid solution and placing the solution in a high-pressure container for pressurized soaking and extraction to obtain a solid-liquid mixture; (3) centrifuging the solid-liquid mixture and taking the supernatant to obtain the black fungus bran extract; In step (1), the sieving is through a 60-80 mesh sieve; In step (2), the material-liquid ratio of the black fungus chaff powder and the citric acid solution is 1:30 g / mL, and the concentration of the citric acid solution is 0.25 g / L; In step (2), the pressurization pressure is 0.1 MPa and the pressurization time is 20 min; In step (3), the centrifugal speed is 4000 r / min and the centrifugal time is 10 min.

2. according to the application of black fungus chaff extracting liquid in plant planting of claim 1, it is characterized in that, The following steps are involved: (1) Wash the wintercress, purple-backed cabbage or mint seeds with distilled water and set aside; (2) diluting the black fungus husk extract with water to obtain a dilution of the black fungus husk extract for later use; (3) Soak the seeds of wintergreen cabbage, purple cabbage or mint in the diluted solution of the black fungus husk extract. Rinse the seeds with clean water several times after soaking. Place the seeds in a culture dish and then place them in a constant temperature incubator for culturing to allow the seeds to germinate. Keep the bud bed moist during the culturing period. (4) Plant the cultivated winter melon seeds, purple-backed cabbage seeds or mint seeds, and spray the leaves with the diluted solution of the black fungus bran extract when the winter melon, purple-backed cabbage or mint grow two leaves.

3. according to the application of black fungus chaff extracting liquid in plant planting described in claim 2, it is characterized in that, In step (3), the soaking time is 24 hours, the concentration of the dilution of the black fungus bran extract is 20%-100%, and the seeds are placed in a culture dish and cultured in a constant temperature incubator at 20-25°C to allow the seeds to germinate for 7 days; In step (4), when the wintercress grows two leaves, the black fungus husk extract with a concentration of 20%-100% is sprayed on the leaves, and the spraying is done once every two days for a total of 30 days.

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