A nutritional adjuvant for increasing sugarcane yield and its preparation method

Abstract

This invention discloses a nutrient adjuvant for increasing sugarcane yield and its preparation method, comprising the following steps: Making Enzyme 1: Collect protein casing waste, add a pre-set fermenting agent to the protein casing waste and ferment for 6-10 hours, sterilize, cool and filter; the filtrate is Enzyme 1. Making Enzyme 2: Take Imperata cylindrica root, dilute with water and pulp, separate the pulp and residue, take the pulp, add PBS mother liquor to the pulp and then inoculate with bacterial powder, seal and ferment for 40-52 hours to obtain Enzyme 2. Making Enzyme 3: Take Mesona chinensis and alcohol solution, shake repeatedly for 5-8 hours, filter, dilute the filtrate 10-20 times, then inoculate with bacterial solution, maintaining an aeration rate of 0.08 m³ / h. 3 / (m 3 ·min)-0.15m 3 / (m 3 The mixture is fermented aerobically for 4-6 days (min), filtered, and the pH of the filtrate is adjusted to 7.0-8.0 to obtain enzyme 3. Enzyme 1, enzyme 2, and enzyme 3 are mixed to prepare a nutrient adjuvant for increasing sugarcane yield. Compared with the prior art, this invention prepares a nutrient adjuvant containing high levels of acid-soluble proteins, flavonoids, organic acids and their salts, and effective active bacteria. This nutrient adjuvant promotes the development of sugarcane roots, increases sugarcane leaf area, improves the rate of photosynthesis, and increases sugarcane yield; it also improves soil degradation and nutrient imbalance caused by continuous sugarcane cropping, and restores soil ecology and fertility.

Description

Technical Field

[0001] This invention relates to the field of enzyme processing technology, and in particular to a nutritional adjuvant for increasing sugarcane yield and its preparation method. Background Technology

[0002] The sugarcane industry is one of the important pillar industries of Guangxi's economy, and the most crucial link in the sugarcane industry is sugarcane production. The entire sugarcane production process includes land preparation, planting, ridging, fertilization, hilling, and harvesting. Currently, the commonly used sugarcane planting method is ratooning, where after the sugarcane stalks of the previous planting cycle are harvested, the sugarcane buds at the base of the stalks in the ground sprout and grow into new plants under suitable environmental conditions. Due to the characteristics of sugarcane cultivation, and the poor yield of other crops after turning over land previously planted with sugarcane, farmers often choose to continuously crop sugarcane on the same plot. However, continuous cropping of sugarcane has led to a gradual decline in sugarcane yield per acre year by year.

[0003] When sugarcane growth is poor, farmers commonly use chemical fertilizers such as nitrogen, phosphorus, and potassium fertilizers to increase yield per acre. While the application of chemical fertilizers can increase sugarcane yield to some extent, it also depletes soil organic matter, exacerbates nutrient imbalance, worsens soil physical and chemical properties, and reduces soil biodiversity and content. The pursuit of yield through fertilizer application exacerbates soil degradation, which in turn leads to further decline in sugarcane yield. This yield reduction then prompts the application of even more fertilizer, creating a vicious cycle.

[0004] Agricultural enzymes are products made by fermenting agricultural biological resources. They combine the soil physicochemical properties improvement of organic fertilizers with the rapid yield and quality improvement of chemical fertilizers. However, existing agricultural enzymes are mainly obtained by anaerobic fermentation of brown sugar, fruits and vegetables and water in a specific ratio. The fermentation time is long and the nitrogen source content is low. At present, no enzymes have been found to be made from animal protein waste and plants.

[0005] Studies have found that the processing of bovine collagen casings generates a large amount of waste, including bovine collagen and bovine keratin. This waste comprises waste hides, bovine hair, and casings, collectively referred to as protein casing waste. A large-scale artificial casing processing plant generates hundreds of tons of protein casing waste daily, a massive amount that is difficult to treat and degrade using environmental protection facilities. Therefore, the utilization of protein casing waste is an urgent problem to be solved. Summary of the Invention

[0006] This invention provides a nutrient adjuvant for increasing sugarcane yield, aiming to overcome the problem that the excessive application of chemical fertilizers to increase yield in continuously cropped sugarcane exacerbates the decline in organic matter and soil degradation, thereby further reducing sugarcane yield.

[0007] The present invention also provides a method for preparing a nutrient adjuvant for increasing sugarcane yield, which aims to solve the problem that protein casing waste is difficult to treat and degrade through environmental protection facilities, and to overcome the deficiency of not utilizing animal protein waste and special plant resources to prepare enzymes containing high levels of acid-soluble proteins, flavonoids, small molecule organic acids and effective active bacteria, so as to improve the yield-increasing effect of the nutrient adjuvant on sugarcane.

[0008] To achieve the above objectives, the present invention provides a method for preparing a nutrient adjuvant for increasing sugarcane yield, the method comprising:

[0009] S1, Making Enzyme 1: Collect protein casing waste, add preset fermentation agent for 6-10 h, sterilize, cool and filter, the filtrate is Enzyme 1;

[0010] S2, making enzyme 2: take Imperata cylindrica root, add water and pulp, separate the pulp and residue, take the pulp liquid, add PBS mother liquor to the pulp liquid and then inoculate with bacterial powder, seal and ferment for 40-52 h to obtain enzyme 2.

[0011] S3, making enzyme 3: Take grass jelly and alcohol solution, shake back and forth for 5-8 hours, filter and dilute the filtrate 10-20 times, then inoculate with bacterial solution, maintain the aeration rate of 0.08 m³ / (m³·min)-0.15 m³ / (m³·min), aerobic fermentation for 4-6 days, filter and adjust the pH of the filtrate to 7-8.0 to obtain enzyme 3;

[0012] S4 is a nutrient adjuvant for increasing sugarcane yield, made by mixing enzyme 1, enzyme 2 and enzyme 3.

[0013] Preferably, in the above technical solution, in the step of fermentation treatment with a preset fermenting agent, the preset fermenting agent is one or more of Bacillus subtilis dry powder, Bacillus amyloliquefaciens dry powder, alkaline protease, and neutral protease.

[0014] Preferably, in the above technical solution, the preset fermentation agent is 0.5-1‰ Bacillus subtilis dry powder, 0.5-1‰ Bacillus amyloliquefaciens dry powder, 1-2‰ alkaline protease and 1-2‰ neutral protease.

[0015] Preferably, in the above technical solution, in step S1, the specific method for preparing enzyme 1 is as follows: collect protein casing waste, add quicklime to the protein casing waste, heat to boiling and maintain for 2-4 h, when cooled to 45-55℃, add a preset fermentation agent for fermentation treatment for 6-10 h, sterilize, cool and filter, and the filtrate is enzyme 1.

[0016] Preferably, in the above technical solution, the bacterial powder is 0.5-1‰ Lactobacillus plantarum dry powder and 0.5-1‰ Pediococcus spp. dry powder.

[0017] Preferably, in the above technical solution, in step S3, the bacterial solution is a 2-3% pasteurized acetic acid bacteria solution.

[0018] Preferably, in the above technical solution, the pasteurized acetic acid bacteria solution is obtained by inoculating pasteurized acetic acid bacteria into an acetic acid bacteria culture medium. The acetic acid bacteria culture medium is prepared by mixing 10-15g glucose, 10-15g yeast extract, 8-10g calcium carbonate, 2.5-5g sodium acetate, 1.5-3g calcium lactate and 1000-120mL PBS, sterilizing at 115-121℃ for 30-40min, cooling and then adding 50-60mL alcohol and mixing evenly.

[0019] Preferably, in the above technical solution, the aerobic fermentation refers to maintaining an oxygen flow rate of 0.08 m³ / (m³·min)-0.15 m³ / (m³·min) during the fermentation process.

[0020] Preferably, in the above technical solution, in step S4, the nutritional aid is prepared by mixing 20-60 parts of enzyme 1, 10-40 parts of enzyme 2 and 10-40 parts of enzyme 3.

[0021] A nutrient adjuvant for increasing sugarcane yield, prepared by the method described above.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] The nutritional adjuvant of the present invention promotes sugarcane yield increase in the following ways: (1) Bacillus subtilis, Bacillus amyloliquefaciens, alkaline protease and neutral protease are used to promote the degradation of protein casing waste into acid-soluble protein. The acid-soluble protein, together with flavonoids and organic acids and their salts, promotes the development of sugarcane roots, increases sugarcane leaf area and improves photosynthesis rate, thereby increasing sugarcane yield; (2) Before the fermentation of protein casing waste, quicklime and high-temperature boiling are used to neutralize the acidity of protein casing waste and promote the denaturation of protein casing waste, providing a more suitable fermentation environment for fermentation agents and enzymes, thereby increasing the fermentation speed; (3) Fermentation of Imperata cylindrica and Mesona chinensis with microbial agents increases the flavonoid content and effective active bacteria content and their metabolites in the nutritional adjuvant. The synergistic effect of acid-soluble protein is conducive to the formation of a stable microbial community in the soil, increasing soil biomass and promoting soil structure improvement. Detailed Implementation

[0024] The technical solutions in the embodiments of this invention are clearly and completely described. Obviously, the described embodiments are only a part of the embodiments of this invention, and not all of them. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention. Example 1

[0025] A method for preparing a nutrient adjuvant for increasing sugarcane yield, the method comprising:

[0026] S1, Making Enzyme 1: Collect protein casing waste, add 2% quicklime to the protein casing waste, heat to boiling and maintain for 4 hours to obtain fermentation liquid. When the fermentation liquid is cooled to 45°C, add 0.5‰ Bacillus subtilis dry powder, 0.5‰ Bacillus amyloliquefaciens dry powder, 1‰ alkaline protease, and 1‰ neutral protease to the fermentation liquid. After stirring, ferment for 8 hours. After fermentation, raise the temperature to 100°C and maintain for 30 minutes. Filter, and the filtrate is Enzyme 1.

[0027] S2, Making Enzyme 2: Take 10 parts of Imperata cylindrica root and 90 parts of water to make a pulp, separate the pulp and residue, take the pulp liquid, add 1‰ PBS mother liquor to the pulp liquid, stir evenly, add 0.5‰ Lactobacillus plantarum dry powder and 0.5‰ Pediococcus spp. dry powder and stir evenly, and anaerobic ferment at 36℃ for 48 h to obtain enzyme 2.

[0028] Preparation of PBS stock solution: Weigh 34.0 g of potassium dihydrogen phosphate and dissolve it in 500 mL of distilled water. Adjust the pH to 7.2 with approximately 175 mL of 1 mol / L sodium hydroxide solution. Dilute to 1000 mL with distilled water and store in a refrigerator. Dilute the stock solution 1000 times before use.

[0029] S3, making enzyme 3: Take 10 parts of grass jelly and 100 parts of 60% alcohol solution, shake repeatedly for 8 hours, filter and take the filtrate, then dilute it 20 times, inoculate with 2% Pasteurella acetic acid bacteria solution and stir well, maintain the oxygenation rate of 0.08 m³ / (m³·min)-0.15 m³ / (m³·min), aerobic fermentation for 6 days, filter after fermentation, adjust the pH of the filtrate to 8.0 with 1 mol / L potassium hydroxide solution to obtain enzyme 3;

[0030] Pasteurella acetic acid bacteria culture was obtained by inoculating Pasteurella acetic acid bacteria into acetic acid bacteria culture medium. The preparation method of acetic acid bacteria culture medium is as follows: 10 g glucose, 10 g yeast extract, 8 g calcium carbonate, 2.5 g sodium acetate, 1.5 g calcium lactate and 1000 mL PBS are sterilized at 115℃ for 30 min, and after cooling, 50 mL of alcohol is added and mixed evenly.

[0031] S4, 60 parts of enzyme 1, 40 parts of enzyme 2 and 40 parts of enzyme 3 are mixed to prepare a nutritional adjuvant for increasing sugarcane yield. Example 2

[0032] A method for preparing a nutrient adjuvant for increasing sugarcane yield, the method comprising:

[0033] S1, Making Enzyme 1: Collect protein casing waste, add 2% quicklime to the protein casing waste, heat to boiling and maintain for 2 hours to obtain fermentation liquid. When the fermentation liquid is cooled to 55°C, add 1‰ Bacillus subtilis dry powder, 1‰ Bacillus amyloliquefaciens dry powder, 2‰ alkaline protease, and 2‰ neutral protease to the fermentation liquid. After stirring, ferment for 10 hours. After fermentation, raise the temperature to 100°C and maintain for 40 minutes. Filter, and the filtrate is Enzyme 1.

[0034] S2, making enzyme 2: Take 10 parts of Imperata cylindrica root and 90 parts of water to make a pulp, separate the pulp and residue, take the pulp liquid, add 1‰ PBS mother liquor to the pulp liquid, stir evenly, add 1‰ Lactobacillus plantarum dry powder and 1‰ Pediococcus dry powder and stir evenly, and anaerobic ferment at 36℃ for 48h to obtain enzyme 2.

[0035] Preparation of PBS stock solution: Weigh 34.0 g of potassium dihydrogen phosphate and dissolve it in 500 mL of distilled water. Adjust the pH to 7.2 with approximately 175 mL of 1 mol / L sodium hydroxide solution. Dilute to 1000 mL with distilled water and store in a refrigerator. Dilute the stock solution 1000 times before use.

[0036] S3, making enzyme 3: Take 10 parts of grass jelly and 100 parts of 60% alcohol solution, shake repeatedly for 8 hours, filter and take the filtrate, then dilute it 10-20 times, inoculate with 3% Pasteurella acetic acid bacteria solution and stir well, maintain the oxygenation rate of 0.08 m³ / (m³·min)-0.15 m³ / (m³·min), and aerobic ferment for 5 days. After fermentation, adjust the pH of the fermentation broth to 7.0 with 1mol / L potassium hydroxide solution to obtain enzyme 3;

[0037] Pasteurella acetic acid bacteria culture was obtained by inoculating Pasteurella acetic acid bacteria into acetic acid bacteria culture medium. The preparation method of acetic acid bacteria culture medium is as follows: 10 g glucose, 10 g yeast extract, 8 g calcium carbonate, 2.5 g sodium acetate, 1.5 g calcium lactate and 1000 mL PBS are sterilized at 115℃ for 30 min, and after cooling, 50 mL of alcohol is added and mixed evenly.

[0038] S4, 20 parts of enzyme 1, 10 parts of enzyme 2 and 40 parts of enzyme 3 are mixed to prepare a nutritional adjuvant for increasing sugarcane yield. Example 3

[0039] A method for preparing a nutrient adjuvant for increasing sugarcane yield, the method comprising:

[0040] S1, Making Enzyme 1: Collect protein casing waste, crush it, add 2% quicklime to the protein casing waste, heat to boiling and maintain for 4 hours to obtain fermentation liquid. When the fermentation liquid is cooled to 55°C, add 0.5‰ Bacillus subtilis dry powder, 0.5‰ Bacillus amyloliquefaciens dry powder, 1‰ alkaline protease, and 1‰ neutral protease to the fermentation liquid. After stirring, ferment for 10 hours. After fermentation, raise the temperature to 100°C and maintain for 40 minutes. Filter, and the filtrate is Enzyme 1.

[0041] S2, Making Enzyme 2: Take 10 parts of Imperata cylindrica root and 90 parts of water to make a pulp, separate the pulp and residue, take the pulp liquid, add 1‰ PBS mother liquor to the pulp liquid, stir evenly, add 0.5‰ Lactobacillus plantarum dry powder and 0.5‰ Pediococcus spp. dry powder and stir evenly, and anaerobic ferment at 36℃ for 48 h to obtain enzyme 2.

[0042] Preparation of PBS stock solution: Weigh 34.0 g of potassium dihydrogen phosphate and dissolve it in 500 mL of distilled water. Adjust the pH to 7.2 with approximately 175 mL of 1 mol / L sodium hydroxide solution. Dilute to 1000 mL with distilled water and store in a refrigerator. Dilute the stock solution 1000 times before use.

[0043] S3, making enzyme 3: Take 10 parts of grass jelly and 100 parts of 60% alcohol solution, shake repeatedly for 5 hours, filter and take the filtrate, then dilute it 10-20 times, inoculate with 2% Pasteurella acetic acid bacteria solution and stir well, maintain the oxygenation rate of 0.08 m³ / (m³·min)-0.15 m³ / (m³·min), and aerobic ferment for 6 days. After fermentation, adjust the pH of the fermentation broth to 8.0 with 1 mol / L potassium hydroxide solution to obtain enzyme 3;

[0044] Pasteurella acetic acid bacteria culture was obtained by inoculating Pasteurella acetic acid bacteria into acetic acid bacteria culture medium. The preparation method of acetic acid bacteria culture medium is as follows: 10 g glucose, 10 g yeast extract, 8 g calcium carbonate, 2.5 g sodium acetate, 1.5 g calcium lactate and 1000 mL PBS, sterilized at 115℃ for 30 min, cooled and mixed with 50 mL alcohol.

[0045] S4, mix 20 parts of enzyme 1, 20 parts of enzyme 2 and 30 parts of enzyme 3 to prepare a nutritional adjuvant for increasing sugarcane yield. Example 4

[0046] A method for preparing a nutrient adjuvant for increasing sugarcane yield, the method comprising:

[0047] S1, Making Enzyme 1: Collect protein casing waste, crush it, add 2% quicklime to the protein casing waste, heat to boiling and maintain for 2 hours to obtain fermentation liquid. When the fermentation liquid is cooled to 45°C, add 1‰ Bacillus subtilis dry powder, 1‰ Bacillus amyloliquefaciens dry powder, 2‰ alkaline protease, and 2‰ neutral protease to the fermentation liquid. After stirring, ferment for 10 hours. After fermentation, raise the temperature to 100°C and maintain for 40 minutes. Filter, and the filtrate is Enzyme 1.

[0048] S2, Making Enzyme 2: Take 10 parts of Imperata cylindrica root and 90 parts of water to make a pulp, separate the pulp and residue, take the pulp liquid, add 1‰ PBS mother liquor to the pulp liquid, stir evenly, add 0.5‰ Lactobacillus plantarum dry powder and 0.5‰ Pediococcus spp. dry powder and stir evenly, and anaerobic ferment at 36℃ for 48 h to obtain enzyme 2.

[0049] Preparation of PBS stock solution: Weigh 34.0 g of potassium dihydrogen phosphate and dissolve it in 500 mL of distilled water. Adjust the pH to 7.2 with approximately 175 mL of 1 mol / L sodium hydroxide solution. Dilute to 1000 mL with distilled water and store in a refrigerator. Dilute the stock solution 1000 times before use.

[0050] S3, making enzyme 3: Take 10 parts of grass jelly and 100 parts of 60% alcohol solution, shake repeatedly for 8 hours, filter and take the filtrate, then dilute it 10-20 times, inoculate with 3% Pasteurella acetic acid bacteria solution and stir well, maintain the oxygenation rate of 0.08 m³ / (m³·min)-0.15 m³ / (m³·min), and aerobic ferment for 6 days. After fermentation, adjust the pH of the fermentation broth to 8.0 with 1mol / L potassium hydroxide solution to obtain enzyme 3;

[0051] Pasteurella acetic acid bacteria culture was obtained by inoculating Pasteurella acetic acid bacteria into acetic acid bacteria culture medium. The preparation method of acetic acid bacteria culture medium is as follows: 10 g glucose, 10 g yeast extract, 8 g calcium carbonate, 2.5 g sodium acetate, 1.5 g calcium lactate and 1000 mL PBS are sterilized at 115℃ for 30 min, and after cooling, 50 mL of alcohol is added and mixed evenly.

[0052] S4, 40 parts of enzyme 1, 20 parts of enzyme 2 and 20 parts of enzyme 3 are mixed to prepare a nutritional adjuvant for increasing sugarcane yield.

[0053] Comparative Example 1

[0054] To make enzyme 1: Collect protein casing waste, add 0.5‰ Bacillus subtilis dry powder, 0.5‰ Bacillus amyloliquefaciens dry powder, 1‰ alkaline protease, and 1‰ neutral protease to the protein casing waste, stir well, and ferment for 12 hours. After fermentation, raise the temperature to 100℃ and maintain it for 30 minutes. Filter, and the filtrate is enzyme 1.

[0055] Comparative Example 2

[0056] To make enzyme 1: Collect protein casing waste, add 1% quicklime to the protein casing waste, heat to boiling and maintain for 2-4 hours to obtain fermentation liquid. When the fermentation liquid is cooled to 45-55℃, add 1‰ alkaline protease and 1‰ neutral protease to the fermentation liquid, stir well and ferment for 8 hours. After fermentation, raise the temperature to 100℃ and maintain for 30 minutes. Filter, and the filtrate is enzyme 1.

[0057] Comparative Example 3

[0058] To make enzyme 1: Collect protein casing waste, add 2% quicklime to the protein casing waste, heat to boiling and maintain for 2-4 hours to obtain fermentation liquid. When the fermentation liquid is cooled to 45-55℃, add 0.5‰ Bacillus subtilis dry powder and 0.5‰ Bacillus amyloliquefaciens dry powder to the fermentation liquid, stir well and ferment for 8 hours. After fermentation, raise the temperature to 100℃ and maintain for 30 minutes. Filter, and the filtrate is enzyme 1.

[0059] The acid-soluble protein content of Examples 1 and Comparative Examples 1-3 after fermentation for 6h, 8h, 10h, and 12h is shown in Table 1 below.

[0060] Table 1. Changes in the content of acid-soluble protein in Enzyme 1 of Examples 1 and Comparative Examples 1-3 with prolonged fermentation time.

[0061] Group 6h 8h 10h 12h Example 1 20.6 25.8 25.8 25.9 Comparative Example 1 1.5 1.9 2.7 3.5 Comparative Example 2 6.7 7.6 9.4 12.2 Comparative Example 3 7.3 9.1 12.9 15.6

[0062] As shown in Table 1, under the same fermentation time, compared with Example 1, Comparative Examples 1, 2, and 3 had higher contents of acid-soluble proteins, indicating that the fermentation process conditions of the present invention are conducive to the degradation of protein casing waste into acid-soluble proteins by enzymes or microbial agents. The principle may be that the protein casing waste is weakly acidic and alkaline in quicklime solution. Under alkaline heating conditions, the protein carboxyl groups in the protein casing waste undergo an alkaline hydrolysis reaction, forming smaller compounds. Simultaneously, this alters the protein's spatial conformation, increasing the contact area and contact sites between bacteria and enzymes, thus accelerating the decomposition rate of the protein casing waste. Furthermore, increasing the pH value of the protein casing waste eliminates the need for additional alkaline pH adjustment, reducing fermentation steps and costs. The combined use of Bacillus subtilis powder, Bacillus amyloliquefaciens powder, alkaline protease, and neutral protease enables synergistic degradation by bacteria and enzymes, resulting in even faster degradation of the protein casing waste.

[0063] After fermentation for 10 hours in Comparative Examples 1-3, Enzyme 1 was obtained. The nutritional components of Enzyme 1 are as follows:

[0064] Group Acid-soluble protein (g / L) Organic acids and their salts (mg / L) Effective viable bacteria count (CFU / mL) pH Example 1 25.8 0.6 ＜10 7.5-8.5 Comparative Example 1 2.7 0.4 ＜10 5.5-6.5 Comparative Example 2 9.4 0.7 ＜10 6.5-7.5 Comparative Example 3 12.9 0.6 ＜10 7.5-8.5

[0065] As can be seen from the table above, the organic acids and their salts in Example 1, Comparative Example 1, Comparative Example 2 and Comparative Example 3 are almost zero because the protein casing waste contains almost no sugars; the pH values ​​of Comparative Example 1 and Comparative Example 2 are lower than those in Example 1 and Comparative Example 3 because the percentage of quicklime added is different.

[0066] After fermenting comparative examples 1-3 for 10 hours, enzyme 1 was obtained. This enzyme was then mixed with enzyme 2 and enzyme 3 prepared by the following method:

[0067] To make enzyme 2: Take 10 parts of Imperata cylindrica root and 90 parts of water and slurry them. Separate the pulp and residue, take the pulp, add 1‰ PBS mother liquor to the pulp, stir evenly, add 0.5‰ Lactobacillus plantarum dry powder and 0.5‰ Pediococcus spp. dry powder and stir evenly. Perform anaerobic fermentation at 36℃ for 48 h to obtain enzyme 2.

[0068] Preparation of PBS stock solution: Weigh 34.0 g of potassium dihydrogen phosphate and dissolve it in 500 mL of distilled water. Adjust the pH to 7.2 with approximately 175 mL of 1 mol / L sodium hydroxide solution. Dilute to 1000 mL with distilled water and store in a refrigerator.

[0069] The preparation method of enzyme 3 is as follows:

[0070] To prepare enzyme 3: Take 10 parts of grass jelly and 100 parts of 60% alcohol solution, shake repeatedly for 5-8 hours, filter and collect the filtrate, then dilute it 10-20 times, inoculate with 2% pasteurized acetic acid bacteria solution and stir well. Maintain an aeration rate of 0.08 m³ / (m³·min)-0.15 m³ / (m³·min), and ferment aerobically for 6 days. After fermentation, filter and adjust the pH of the filtrate to 8.0 with 1 mol / L potassium hydroxide solution to obtain enzyme 3.

[0071] Pasteurella acetic acid bacteria culture was obtained by inoculating Pasteurella acetic acid bacteria into acetic acid bacteria culture medium. The preparation method of acetic acid bacteria culture medium is as follows: 10 g glucose, 10 g yeast extract, 8 g calcium carbonate, 2.5 g sodium acetate, 1.5 g calcium lactate and 1000 mL PBS are sterilized at 115℃ for 30 min, and after cooling, 50 mL of alcohol is added and mixed evenly.

[0072] 60 parts of enzyme 1 prepared in Comparative Example 1, 40 parts of enzyme 2 and 40 parts of enzyme 3 were mixed to prepare a nutrient adjuvant for increasing sugarcane yield.

[0073] 60 parts of enzyme 1 prepared in Comparative Example 2, 40 parts of enzyme 2 and 40 parts of enzyme 3 were mixed to prepare a nutrient adjuvant for increasing sugarcane yield.

[0074] 60 parts of enzyme 1 prepared in Comparative Example 3, 40 parts of enzyme 2, and 40 parts of enzyme 3 were mixed to prepare a nutrient adjuvant for increasing sugarcane yield.

[0075] Comparative Example 4

[0076] To prepare enzyme 2: Take 10 parts of Imperata cylindrica root and add 90 parts of water to make a pulp. Separate the pulp from the residue and take the pulp liquid. Add 1‰ PBS mother liquor to the pulp liquid and stir evenly. Add 0.5‰ Lactobacillus plantarum dry powder and 0.5‰ Bacillus amyloliquefaciens dry powder and stir evenly. Perform anaerobic fermentation at 36℃ for 48 h to obtain enzyme 2.

[0077] Comparative Example 5

[0078] To make enzyme 3: Take 10 parts of grass jelly and 100 parts of 60% alcohol solution, shake repeatedly for 5-8 hours, filter and collect the filtrate, then dilute it 10-20 times, inoculate with 2% pasteurized acetic acid bacteria solution and stir well, maintain an aeration rate of 0.01 m³ / (m³·min)-0.05 m³ / (m³·min), and aerobic ferment for 6 days. After fermentation, filter to obtain enzyme 3.

[0079] Pasteurella acetic acid bacteria culture was obtained by inoculating Pasteurella acetic acid bacteria into acetic acid bacteria culture medium. The preparation method of acetic acid bacteria culture medium is as follows: 10 g glucose, 10 g yeast extract, 8 g calcium carbonate, 2.5 g sodium acetate, 1.5 g calcium lactate, 1000 mL PBS, sterilized at 115℃ for 30 min, and 50 mL alcohol was added after cooling.

[0080] Comparative Example 6

[0081] S1, collect protein casing waste, add 2% quicklime to the protein casing waste, heat to boiling and maintain for 2-4 hours to obtain fermentation liquid;

[0082] S2, take 10 parts Imperata cylindrica root and 10 parts Mesona chinensis, add 100 parts water and pulverize, separate the pulp and residue, and take the pulp liquid;

[0083] S3, mix the liquid to be fermented with the slurry, add 0.5‰ Bacillus subtilis dry powder, 0.5‰ Bacillus amyloliquefaciens dry powder, 1‰ alkaline protease, 1‰ neutral protease, 0.5‰ Lactobacillus plantarum dry powder and 0.5‰ Pediococcus dry powder, stir well and ferment for 8 days. After fermentation, raise the temperature to 100℃ and maintain for 30 min, then filter. The filtrate is a nutrient adjuvant for increasing sugarcane yield.

[0084] The nutritional components of Examples 1, 2, 3, Comparative Examples 1, 2, 3, 4, 5, and 6 were tested according to industry standard QB / T5323-2018. The nutritional components are shown in Table 2 below.

[0085] Table 2. Nutritional composition of the nutritional supplements in each specific embodiment.

[0086] Group Acid-soluble protein (g / L) Flavonoids (mg / L) Organic acids and their salts (mg / L) Effective viable bacteria count (CFU / mL) pH Example 1 25.6 3.6 825 <![CDATA[1×10 7 ]]> 5.5-8.5 Example 2 25.8 4.4 837 <![CDATA[1×10 7 ]]> 5.5-8.5 Example 3 24.4 3.8 822 <![CDATA[1×10 7 ]]> 5.5-8.5 Example 4 25.8 3.6 823 <![CDATA[1×10 7 ]]> 5.5-8.5 Comparative Example 1 2.7 3.7 823 <![CDATA[1×10 7 ]]> 4.5-6.5 Comparative Example 2 9.4 3.6 821 <![CDATA[1×10 7 ]]> 5.0-7.5 Comparative Example 3 12.9 3.8 825 <![CDATA[1×10 7 ]]> 5.5-8.5 Comparative Example 4 0 1.8 362 <![CDATA[1×10 7 ]]> 3 Comparative Example 5 0 1.4 416 <![CDATA[1×10 6 ]]> 3 Comparative Example 6 16.3 3.2 749 ＜10 5.5-8.5

[0087] As can be seen from the table above, compared with Example 1, the flavonoid content of Comparative Examples 1-3 is not significantly different, indicating that the flavonoids mainly come from enzymes 2 and 3. Enzymes 2 and 3, obtained using the same fermentation method and conditions as Example 1, may have slight variations due to the activity of the microbial agent or the condition of the raw materials, but the changes are not significant. The organic acids and their salts have changed considerably. For example, the organic acids and their salts in Example 1, Comparative Examples 1, 2, and 3 increased from zero to over 800, because enzymes 1 and 3 contain more organic acids and their salts. Compared with Comparative Example 6, Example 1 fermented the protein casing waste, Imperata cylindrica root, and Mesona chinensis separately and then mixed them. Each enzyme was fermented more completely by the microbial agent or enzyme agent, resulting in more acid-soluble proteins, flavonoids, organic acids, and effective viable bacteria.

[0088] The heavy metal content of Examples 1, 2, 3, Comparative Examples 1, 2, 3, 4, 5, and 6 was tested according to industry standard QB / T5323-2018, and the heavy metal content is shown in Table 3 below. The heavy metal testing included total arsenic and its compounds (As), total cadmium and its compounds (Cd), total lead and its compounds (Pb), total chromium and its compounds (Cr), and total mercury and its compounds (Hg).

[0089] Table 3. Heavy metal content of nutritional supplements in various specific embodiments

[0090] Group Hg (mg / kg) As (mg / kg) Cd (mg / kg) Pb (mg / kg) Cr (mg / kg) Example 1 0.01 0.002 0.3 0.4 0.7 Example 2 0.01 0.003 0.3 0.4 0.8 Example 3 0.01 0.002 0.4 0.4 0.7 Example 4 0.01 0.002 0.4 0.5 0.8 Comparative Example 1 0.01 0.002 0.4 0.5 0.8 Comparative Example 2 0.02 0.002 0.4 0.4 0.8 Comparative Example 3 0.01 0.004 0.5 0.4 0.8 Comparative Example 4 0.01 0.003 0.4 0.6 0.9 Comparative Example 5 0.01 0.002 0.4 0.4 1.0 Comparative Example 6 0.02 0.003 0.5 0.6 0.8

[0091] The nutrient adjuvants prepared in Examples 1-3 and Comparative Examples 1-6 were applied to the sugarcane fields of the corresponding experimental plots. The application method was as follows: 1 kg / mu was applied 4 times per sugarcane planting cycle, with a total application rate of 4 kg / mu. The adjuvants were diluted 200 times with water before spraying or drip irrigation. The sugarcane yield of each experimental plot at harvest time is shown in Table 4 below.

[0092] Table 4 Sugarcane yield after application of nutrient adjuvants

[0093] Group Yield (kg / mu) Example 1 6560 Example 2 6810 Example 3 6550 Example 4 6680 Comparative Example 1 4780 Comparative Example 2 5120 Comparative Example 3 5360 Comparative Example 4 4130 Comparative Example 5 4270 Comparative Example 6 5730

[0094] After harvesting, the soil nutrient status was compared between the nutrient adjuvant prepared according to this invention and the plots that were conventionally fertilized with compound fertilizer. The results are shown in Table 5 below.

[0095] Table 5. Soil nutrient status of soil blocks treated with nutrient adjuvants and fertilizers

[0096] Yield (kg / mu) Organic matter (g / kg) <![CDATA[Bacteria (1×10 6 CFU / g)]]> Nutritional supplements 6560 31.9 139.67 Compound fertilizer 5970 20.7 86.55

[0097] This invention can be implemented in various ways and is not limited to the embodiments described. Those skilled in the art will understand that the invention can be implemented in other specific ways without changing the technical concept or essential features. Therefore, it should be understood that the embodiments described above are exemplary and not intended to limit the invention.

Claims

1. A method for preparing a nutrient adjuvant for increasing sugarcane yield, characterized in that, include: S1, Making Enzyme 1: Collect protein casing waste, add quicklime to the protein casing waste, heat to boiling and maintain for 2-4 hours. When cooled to 45-55℃, add a preset fermentation agent and ferment for 6-10 hours. Sterilize, cool and filter. The filtrate is Enzyme 1. The preset fermentation agent is 0.5-1‰ Bacillus subtilis dry powder, 0.5-1‰ Bacillus amyloliquefaciens dry powder, 1-2‰ alkaline protease and 1-2‰ neutral protease. S2, Making Enzyme 2: Take Imperata cylindrica root, add water and slurry, separate the pulp and residue, take the slurry, add PBS mother liquor to the slurry and then inoculate with bacterial powder, seal and ferment for 40-52 h to obtain enzyme 2, wherein the bacterial powder is 0.5-1‰ Lactobacillus plantarum dry powder and 0.5-1‰ Pediococcus spp. dry powder. S3, making enzyme 3: Take grass jelly and alcohol solution, shake back and forth for 5-8 hours, filter and dilute the filtrate 10-20 times, then inoculate with bacterial solution, aerobic fermentation for 4-6 days, filter and adjust the pH of the filtrate to 7.0-8.0 to obtain enzyme 3, wherein the bacterial solution is 2-3% pasteurized acetic acid bacteria solution; S4 is a nutrient adjuvant for increasing sugarcane yield, made by mixing 20-60 parts of enzyme 1, 10-40 parts of enzyme 2 and 10-40 parts of enzyme 3.

2. The method for preparing the nutrient adjuvant for increasing sugarcane yield as described in claim 1, characterized in that, The pasteurized acetic acid bacteria solution was obtained by inoculating pasteurized acetic acid bacteria into an acetic acid bacteria culture medium. The acetic acid bacteria culture medium was prepared by mixing 10-15g glucose, 10-15g yeast extract, 8-10g calcium carbonate, 2.5-5g sodium acetate, 1.5-3g calcium lactate and 1000mL PBS, sterilizing at 115-121℃ for 30-40min, cooling and then adding 50-60mL of alcohol and mixing thoroughly.

3. The method for preparing the nutrient adjuvant for increasing sugarcane yield as described in claim 1, characterized in that, The aerobic fermentation refers to maintaining an oxygen flow rate of 0.08 m³ / (m³·min) to 0.15 m³ / (m³·min) during the fermentation process.

4. A nutrient adjuvant for increasing sugarcane yield, prepared by any one of the preparation methods described in claims 1-3.