Black fungus chaff extracting solution as well as preparation method and application thereof

Abstract

The invention discloses a black fungus chaff extracting solution as well as a preparation method and application thereof, and belongs to the technical field of mushroom chaff reutilization. The preparation method of the black fungus chaff extracting solution 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 later use; adding the black fungus chaff powder into a citric acid solution, placing in a high-pressure container, and performing pressurized soaking extraction to obtain a solid-liquid mixture for later use; and centrifuging the solid-liquid mixture, and taking supernate to obtain the product. According to the preparation method, effective components in the black fungus chaff can be better and more sufficiently extracted, the content of soluble solids in the black fungus chaff extracting solution is increased, the black fungus chaff extracting solution is applied to planting of malva verticillata, gynura bicolor and mint for the first time, the germination rate and the vigor of seeds can be remarkably increased when the seeds are soaked, and the yield of the seeds is increased. Growth of different plants is adjusted, and a good growth effect is achieved.

Description

Technical Field

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

[0002] The resource development and utilization of mushroom residues is a bottleneck problem 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 voted by industry insiders; among them, the efficient recycling technology of edible fungi residues ranked first among the top ten technical issues with 84.0% of the votes.

[0003] my country is the world's largest producer of black fungus (Auricularia auricula), accounting for more than 90% of the world's output. The scale of the black fungus industry currently ranks second in the country, and it also has a large planting area in Hebei Province. The output of black fungus bran is huge, and it is rich in a variety of nutrients and physiologically active substances, such as protein, cellulose, and polysaccharides. It has a high recycling value. During its large-scale cultivation, a large amount of waste bran is generated. Due to the lack of large-scale treatment methods, most of the waste black fungus bran is randomly dumped in the fields, and a small amount is used for incineration, which will cause ecological problems such as agricultural non-point source pollution and air pollution and waste of resources, which is not in line with the concept of green development.

[0004] Although mushroom husk has many uses, its utilization rate is not high in actual production, and it has not yet been scaled up and promoted. The traditional methods for extracting mushroom husk are hot water extraction and ultrasonic assisted extraction, but the extraction time is long and the extraction efficiency is low.

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

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

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

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

[0009] (1) selecting fresh black fungus bran without mildew, naturally air-drying, crushing, and sieving to obtain black fungus bran powder;

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

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

[0012] The invention has the beneficial effects that, after the mushroom bran is crushed, reacted with an acid solvent and pressurized, more contents are released and the extraction efficiency is high. Therefore, the preparation method of the invention can better and more fully extract the effective components in the black fungus mushroom bran, and improve the soluble solid content in the black fungus mushroom bran extraction liquid. Compared with the mushroom bran, the contents of crude protein, crude fat, polysaccharide, calcium, phosphorus, potassium, total flavonoids and other components in the extraction liquid are all improved.

[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 further technical solution is as follows: as the solid-liquid ratio increases, the soluble solid content tends to increase first and then decrease. The reason is that the larger the volume of the leaching solution, the more complete the contact between the solvent and the bran, and more contents are dissolved in the same time. When the solid-liquid ratio is higher than the optimal ratio, the soluble solid content decreases. If the solid-liquid ratio is too large, more time and energy will be consumed. If the solid-liquid ratio is too large, the concentration of the solvent in the leaching solution decreases, resulting in a decrease in the extraction efficiency.

[0016] The solvent penetrates the cell membrane of the raw material, dissolves the soluble substances, forms a concentration difference between the inside and outside of the cell, and permeates the cell membrane to achieve the purpose of extraction. Increasing the concentration of the medium can promote the release of the extract, because a higher concentration 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, and accelerates the transfer of solute molecules from the solid phase to the liquid phase, thereby improving the leaching efficiency.

[0019] The main principle of the influence of pressurization time on the extraction effect is that increasing the 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 bran extract liquid prepared by the method.

[0022] The invention also provides an application of the black fungus bran extract in plant planting.

[0023] The beneficial effects of the present invention are as follows: the present invention applies the black fungus bran extract liquid to plant planting, the black fungus bran extract liquid can significantly improve the germination rate and seed vitality of the seeds when soaking plant seeds, and spraying the bran extract liquid with different concentrations can regulate the growth of different plants and achieve a good growth effect.

[0024] Mushroom bran is the solid culture medium left after the cultivation of edible fungi. During the cultivation process of edible fungi, a large amount of nutrients and active substances will remain in the bran, including mycelium, cellulose degraded by microorganisms, crude protein, crude fat, secondary metabolites of mycelium and minerals, etc., which has a high resource utilization value. Mushroom bran contains a variety of amino acids, which can be absorbed and utilized by plants to promote plant growth and development, enhance plant resistance, improve soil, increase crop yields, reduce the toxic effects of heavy metal ions on plants, and improve 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 back vegetable or mint.

[0027] The beneficial effects of adopting the above-mentioned further technical scheme are as follows: the present invention applies the black fungus bran extract to the cultivation of winter vegetable, purple-backed vegetable and mint for the first time. The black fungus bran extract can significantly improve the germination rate and seed vitality of winter vegetable, purple-backed vegetable 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 vegetable, purple-backed vegetable and mint, achieving a good growth effect.

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

[0029] (1) Wash the seeds of winter melon, purple melon or mint with distilled water for later use;

[0030] (2) diluting the black fungus bran extract with water to obtain a dilution of the black fungus bran extract for standby use;

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

[0032] (4) planting the cultured winter melon seeds, purple back vegetable seeds or mint seeds, and spraying the leaves with the diluted solution of the black fungus bran extract when the winter melon seeds, purple back vegetable or mint grow two leaves.

[0033] Further, in step (3), the soaking time is 24 hours, the concentration of the dilution of the black fungus bran extract is 20%-100%, 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, and the culture days are 7 days;

[0034] In step (4), when the winter vegetable grows two leaves, the black fungus bran extract with a concentration of 20%-100% is sprayed on the leaves, 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 winter vegetable grows two leaves, the black fungus bran extract with a concentration of 60% is sprayed on the leaves, or when the purple back vegetable 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 solid-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 the pressurized pressure in Test Examples 13-15 on the soluble solids content of the black fungus bran extract.

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

[0042] Figure 5It is the response surface and contour diagram between factors AB.

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

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

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

[0046] Fig. 9 Response surface and contour diagram between factors BD.

[0047] Fig.10 Response surface and contour plot between factors CD. DETAILED DESCRIPTION

[0048] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0049] Example 1

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

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

[0052] (2) adding 5 g of black fungus chaff powder to a citric acid solution and placing the mixture 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 is 1:30 g / mL, the concentration of the citric acid solution is 0.25 g / L, the pressurized pressure is 0.1 MPa, and the pressurization time is 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 bran extract liquid comprises the following steps:

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

[0058] (2) adding 5 g of black fungus chaff powder to a citric acid solution and placing the mixture 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 is 1:30 g / mL, the concentration of the citric acid solution is 0.25 g / L, the pressurized pressure is 0.1 MPa, and the pressurization time is 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 a black fungus bran extract.

[0060] Test Example 2

[0061] Compared with Experimental Example 1, except that the solid-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 solid-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 solid-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 solid-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 solid-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 solid-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 bran extract liquid comprises the following steps:

[0074] (1) selecting fresh and mildew-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) adding 5 g of black fungus chaff powder to a citric acid solution and placing the mixture 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 is 1:30 g / mL, the concentration of the citric acid solution is 0.25 g / L, the pressurized pressure is 0.1 MPa, and the pressurization time is 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 a black fungus bran extract.

[0077] Test Example 9

[0078] Compared with Experimental Example 8, except that the pressurization time is 10 minutes, other steps and parameters are exactly the same as those of Experimental Example 8.

[0079] Test Example 10

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

[0081] Test Example 11

[0082] Compared with Experimental Example 8, except that the pressurization time is 25 minutes, other steps and parameters are exactly the same as those of Experimental Example 8.

[0083] Test Example 12

[0084] Compared with Experimental Example 8, except that the pressurization time is 30 minutes, other steps and parameters are exactly the same as those of Experimental Example 8.

[0085] Test Example 13

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

[0087] (1) selecting fresh and mildew-free black fungus bran of the fourth batch, naturally air-drying, crushing, and passing through an 80-mesh sieve to obtain black fungus bran powder;

[0088] (2) adding 5 g of black fungus chaff powder to a citric acid solution and placing the mixture 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 is 1:30 g / mL, the concentration of the citric acid solution is 0.25 g / L, the pressurized pressure is 0.1 MPa, and the pressurization time is 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 a 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 those of 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 bran extract liquid comprises the following steps:

[0096] (1) selecting fresh black fungus chaff from the fifth batch without mildew, naturally air-drying, crushing, and passing through an 80-mesh sieve to obtain black fungus chaff powder;

[0097] (2) adding 5 g of black fungus chaff powder to a citric acid solution and placing the mixture 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 is 1:30 g / mL, the concentration of the citric acid solution is 0.25 g / L, the pressurized pressure is 0.1 MPa, and the pressurization time is 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 a 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 bran extract liquid comprises the following steps:

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

[0110] (2) adding 5 g of black fungus chaff powder to distilled water and placing it 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 distilled water is 1:30 g / mL, the pressurized pressure is 0.1 MPa, and the pressurization time is 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 solid 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 bran extract liquid comprises the following steps:

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

[0116] (2) adding 5 g of black fungus chaff powder to a NaHCO3 solution and placing it in a high-pressure steam sterilizer for pressurized soaking extraction to obtain a solid-liquid mixture, wherein the solid-liquid ratio of the black fungus chaff powder to the NaHCO3 solution is 1:35 g / mL, the concentration of the NaHCO3 solution is 8 g / L, the pressurized pressure is 0.1 MPa, and the pressurization time is 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 is used to prepare black fungus bran extract, comprising the following steps:

[0121] (1) selecting fresh black fungus chaff from the fifth batch without mildew, naturally air-drying, crushing, and passing through an 80-mesh sieve to obtain black fungus chaff powder;

[0122] (2) Add citric acid solution to 5 g of black fungus bran powder and place it in a constant temperature water bath for extraction to obtain a solid-liquid mixture. The solid-liquid ratio of black fungus bran 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 of the two are not completely the same, and the process parameters are not completely 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 solid content of this comparative example in the hot water extraction method is the highest.

[0125] Comparative Example 4

[0126] The ultrasonic-assisted extraction method is used to prepare black fungus bran extract, which comprises the following steps:

[0127] (1) selecting fresh black fungus chaff from the fifth batch without mildew, naturally air-drying, crushing, and passing through an 80-mesh sieve to obtain black fungus chaff powder;

[0128] (2) Add 5 g of black fungus bran powder to a citric acid solution and place it in an ultrasonic cleaner for extraction to obtain a solid-liquid mixture. The solid-liquid ratio of the black fungus bran powder to the citric acid solution is 1:30 g / mL, the concentration of the 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 of the two are not exactly the same, and the process parameters are not exactly the same. The influencing factors of the ultrasonic-assisted extraction method include the solid-liquid ratio, ultrasonic time, ultrasonic temperature, and extraction medium. After testing, the soluble solid content of this comparative example in the ultrasonic-assisted extraction method is the highest.

[0131] Determination of soluble solid content: Use a handheld refractometer to determine the soluble solid content of the bran extract. The operation method is as follows: First, calibrate to 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 them on the detection prism, close the cover to avoid bubbles. Hold the refractometer and align the light-inlet plate with a bright light source, observe through the eyepiece, turn the handwheel to adjust the eyepiece, and make the white-blue dividing line in the field of view clear. The scale value of the dividing line is the soluble solid 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 condition of weakly acidic aqueous solution, with the increase of solid-liquid ratio, the soluble solid content first increases and then decreases, and the maximum is 2.49% when the solid-liquid ratio is 1:35.

[0134] (2) Effect of pressing 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 is the largest at 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 bran extract liquid comprises the following steps:

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

[0143] (2) adding 5 g of black fungus bran powder to a citric acid solution and placing 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 a black fungus bran extract.

[0145] After the best response area is determined by single factor test, the response surface method is used to further optimize the interaction between the influencing factors and determine the optimal process route. At the same time, after optimization by Box-Behnken design experiment, the optimized process route is verified to test its reliability and accuracy.

[0146] According to the results of the single factor test, A pressure, B material-liquid ratio, C pressurization time, and D acid concentration were selected as influencing factors, and Y soluble solid content was used as the response value. The design table of the four-factor three-level Box-Behnken test is shown in Table 1, and the test results are shown in Table 2.

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

[0148]

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

[0150]

[0151]

[0152] Box-Behnken test ANOVA

[0153] Design-Expert.V8.0.6.1 software was used to analyze and process the test data and perform regression analysis on the interaction between various 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 P value of the model is less than 0.0001, indicating that the regression equation is significant. The entire model has a good fit in the test 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 test 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 value of the four factors is less than 0.0001, indicating that they have a very significant impact on the response value. The P value of the four-factor interaction term also shows that the interaction between different influencing factors has a large effect. Therefore, this model can be used to predict the actual situation of the 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 in the figure, all response surface 3D graphs are open downward and have a highest point, indicating that the response value increases with the increase of each factor value, and then gradually decreases after reaching the maximum value. The optimal formula can be determined by the response surface graph within the experimental range. The contour graphs are all elliptical, indicating that the interaction between the two factors is significant. The interaction of factors AB has the greatest impact on the soluble solids content of bran, and its response surface 3D graph has the steepest slope.

[0161] Verification test results

[0162] The best process parameters obtained by software analysis are: pressure 0.1MPa, pressurization time 20.28min, material-liquid ratio 1:30.39, citric acid concentration 0.25g / L. At this time, the theoretical value of soluble solid content is 2.53%. According to actual operation, the parameters are adjusted to pressure 0.1MPa, pressurization time 20min, material-liquid ratio 1:30, citric acid concentration 0.25g / L. The experiment is repeated under this condition, and the soluble solid content is 2.51%. The relative error with the predicted value is 0.79%, which is within the allowable range, indicating that the model has a high degree of fit and the prediction is correct.

[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 the gallic acid standard curve was determined, 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 according to the curve.

[0174] The determination of indoleacetic acid (IAA), gibberellin A3 (GA3), and abscisic acid (ABA) content was based on the methods in the literature. Zhang Yongmei, Ma Huiling, Zhang Suoke, et al. Method for determining four endogenous hormones in Kentucky bluegrass by HPLC [J]. Acta Grasslanda 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 Example 1 product nutritional composition test and results

[0177]

[0178]

[0179] Table 5: Test results of functional components of products in Example 1

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

[0183] After testing, it was found that compared with the 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 chaff extract in planting winter vegetables comprises the following steps:

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

[0187] (2) diluting the black fungus bran extract of Example 1 with water to obtain a dilution of the black fungus bran extract, which is then used for standby use;

[0188] (3) soaking winter melon seeds in a 60% dilution of black fungus bran extract for 24 hours, rinsing with clean water several times after soaking, placing the seeds in a culture dish and then placing them in a constant temperature incubator at 25° C. for culturing to allow the seeds to germinate, keeping the bud bed moist during the culturing period, and culturing for 7 days;

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

[0190] Example 3

[0191] The application of black fungus chaff extract in planting 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 bran extract of Example 1 with water to obtain a dilution of the black fungus bran extract, which is then used for standby use;

[0194] (3) soaking purple-backed cabbage seeds in a diluted solution of 80% black fungus bran extract for 24 h, rinsing with clean water several times after soaking, placing the seeds in a culture dish and then placing them in a constant temperature incubator at 25° C. for culturing to allow the seeds to germinate, keeping the bud bed moist during the culturing period, and culturing for 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 bran extract of Example 1 with water to obtain a dilution of the black fungus bran extract, which is then used for standby use;

[0200] (3) soaking mint seeds in a diluted solution of 80% black fungus bran extract for 24 hours, rinsing with clean water several times after soaking, placing the seeds in a culture dish and then placing them in a constant temperature incubator at 25° C. for culturing 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, and starting to spray the leaves with 80% black fungus bran extract when the mint seeds grow two leaves, with a spraying amount of 10-15 mL / plant, once every two days for a total of 30 days.

[0202] 1. Seed soaking test of winter cabbage, purple cabbage and mint

[0203] The seeds of winter vegetable, purple back vegetable and mint were repeatedly washed with distilled water for standby use. Five treatments were set 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 for 24 hours, with three groups of parallel treatments. 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, with clean water treatment as a control. The culture dish was placed in a 25°C constant temperature climate incubator 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 on germination rate and seed vitality of winter cabbage, purple 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 with the extract of fungus bran had a promoting effect on the germination rate and seed vitality of wintercress, purple-backed cabbage and mint. Among them, the 60% concentration of the extract of fungus bran had the best effect on the germination of wintercress, with a germination rate of 95.3%, an increase of 12.4% over the control, and a seed vitality of 95.4%, an increase of 36.1% over the control. The 80% concentration of the extract of fungus bran had the best effect on the germination of purple-backed cabbage, with a germination rate of 94.2%, an increase of 8.0% over the control, and a seed vitality of 88.4%, an increase of 19.0% over the control. The 80% concentration of the extract of fungus bran had the best effect on the germination of mint, with a germination rate of 95.3%, an increase of 10.1% over the control, and a seed vitality of 96.1%, an increase of 27.3% over the control. The extract of black fungus bran could significantly improve the germination rate and seed vitality of wintercress, purple-backed cabbage and mint seeds.

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

[0211] In the spraying test, all the cultivated seeds of Examples 2-4 were planted in the ground. Example 1 The chaff extract was set to 5 concentration gradients, with the chaff extract as the mother liquor gradient dilution, respectively 20%, 40%, 60%, 80%, 100% concentration, and the spraying test was started when the winter vegetable, purple back vegetable and mint grew two leaves. The leaves were sprayed evenly, and clear water was used as the control. Three groups were parallel for each treatment, and the spraying was performed once every two days, and the spraying amount 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, the soil was repeatedly washed off with clean water, and 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 for each treatment, and the widest part of the leaves was measured with a vernier caliper to take 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 soil was dried in an oven to a constant weight, and then weighed to obtain the aboveground biomass.

[0217] (2) Physiological indicators

[0218] Before the test, the leaves need to be carefully rinsed with clean water repeatedly to avoid the residue of the bran extract affecting the test results. The soluble protein content was determined by Coomassie Brilliant Blue staining. The soluble sugar content was determined by anthrone colorimetry. The chlorophyll content was determined by 95% ethanol grinding-centrifugation method. The malondialdehyde (MDA) content was determined by thiobarbituric acid method. The superoxide dismutase (SOD) activity was determined by pyrogallol autooxidation method. The peroxidase (POD) activity was determined by guaiacol method.

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

[0220] Table 7 Effects of spraying treatment on early growth indicators of wintercress, purple back vegetable 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 were blank controls.

[0224] Table 8 Effects of spraying treatment on early physiological indicators of wintercress, purple back vegetable and mint

[0225]

[0226]

[0227]

[0228] Table 9 Effects of spraying treatment on early physiological parameters of wintercress, purple back vegetable 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] The effects of different spraying concentrations on the plant height, leaf width, aboveground biomass, chlorophyll a, chlorophyll b, malondialdehyde, SOD, POD, soluble sugar content and other indicators of wintercress, purple back vegetable, and mint were comprehensively studied. The results showed that the 60% concentration of the bran extract had the most significant effect on the germination and early growth of wintercress, and the 80% concentration of the bran extract had the most significant effect on the germination and early growth of purple back vegetable and mint. Therefore, spraying different concentrations of bran extract can regulate the growth of different plants and achieve good growth effects.

[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 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 will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for preparing a black fungus bran extract, characterized in that: The following steps are involved: (1) selecting fresh black fungus bran without mildew, naturally air-drying, crushing, and sieving to obtain black fungus bran powder; (2) adding black fungus bran powder to a citric acid solution, placing the solution in a high-pressure container and pressurizing and soaking the solution to obtain a solid-liquid mixture; (3) centrifuging the solid-liquid mixture, taking the supernatant, and obtaining the black fungus bran extraction liquid.

2. The method for preparing a black fungus bran extract according to claim 1, wherein: In step (1), the sieving is through a 60-80 mesh sieve.

3. The method for preparing a black fungus bran extract according to claim 1, wherein: In step (2), the solid-liquid ratio of the black fungus bran powder to the citric acid solution is 1:30 g / mL, and the concentration of the citric acid solution is 0.25 g / L.

4. The method for preparing a black fungus bran extract according to claim 1, wherein: In step (2), the pressurization pressure is 0.1 MPa and the pressurization time is 20 min.

5. The method for preparing a black fungus bran extract according to claim 1, wherein: In step (3), the centrifugal speed is 4000 r / min and the centrifugal time is 10 min.

6. A black fungus bran extract prepared by the method according to any one of claims 1 to 5.

7. Use of the black fungus bran extract according to claim 6 in plant cultivation.

8. The application of black fungus bran extract in plant planting according to claim 7, characterized in that: The plant is winter melon, purple-backed vegetable or mint.

9. The application of black fungus bran extract in plant planting according to claim 8, characterized in that: The following steps are involved: (1) Wash the seeds of winter melon, purple melon or mint with distilled water for later use; (2) diluting the black fungus bran extract according to claim 8 with water to obtain a dilution of the black fungus bran extract for standby use; (3) soaking winter melon seeds, purple back melon seeds or mint seeds in the diluted solution of black fungus bran extract, rinsing with clean water several times after soaking, placing the seeds in a culture dish and then placing them in a constant temperature incubator for culturing to allow the seeds to germinate, and keeping the bud bed moist during the culturing period; (4) planting the cultured winter melon seeds, purple back vegetable seeds or mint seeds, and spraying the leaves with the diluted solution of the black fungus bran extract when the winter melon seeds, purple back vegetable or mint grow two leaves.

10. The application of black fungus bran extract in plant planting according to claim 9, 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%, 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, and the culture days are 7 days; In step (4), when the winter vegetable grows two leaves, the black fungus bran extract with a concentration of 20%-100% is sprayed on the leaves, once every two days, for a total of 30 days.

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