Hippophae rhamnoides enzyme bacterial fertilizer, and preparation method and application thereof

Abstract

The application discloses seabuckthorn enzyme bacterial fertilizer, a preparation method and application thereof, and belongs to the technical field of microbial fertilizers. The seabuckthorn enzyme bacterial fertilizer is prepared by mixing seabuckthorn pomace fermentation liquor, ammonium salt, phosphate, trehalose and Bacillus subtilis in a mass ratio of 80:1.5-2.5:0.5-1.5:0.5-1.5:0.5-1.5, and then freezing and drying. The preparation method of the seabuckthorn pomace fermentation liquor comprises the following steps: after seabuckthorn pomace is soaked in water, the seabuckthorn pomace is crushed into homogenate; yeast, lactobacillus and white granulated sugar are added into the obtained homogenate, and then the mixture is uniformly mixed and sealed for fermentation; the adding amount of the yeast, the lactobacillus and the white granulated sugar is 0.05-0.1%, 0.2-0.3% and 5-10% of the mass of the homogenate, respectively. The application finds that the fermentation liquor of the seabuckthorn pomace after fermentation treatment and the Bacillus subtilis have a synergistic effect when they are used together, and the effect of the Bacillus subtilis can be significantly enhanced.

Description

TECHNICAL FIELD

[0001] The application relates to the technical field of ecological fertilizer, in particular to a sea buckthorn enzyme microbial fertilizer and a preparation method and application thereof. BACKGROUND

[0002] The microbial fertilizer, also known as biological fertilizer, biological fertilizer, bacterial fertilizer or inoculant, etc. The microbial fertilizer is "bacteria + fertilizer" in a figurative sense. On the one hand, the microbial fertilizer contains nutrients such as organic matter, nitrogen, phosphorus and potassium, which are necessary for crop growth. At the same time, the product also contains a large amount of beneficial microorganisms. The beneficial microorganisms reproduce in the soil and play a role in improving the soil and preventing diseases.

[0003] For example, the Chinese patent application with the publication number CN 112154759A discloses an environmentally friendly composite microbial fertilizer. The microbial fertilizer is composed of the following raw materials by weight: 5-10 parts of microbial liquid, 3-5 parts of plant regulator, 6-8 parts of Huangqin powder, 5-7 parts of Lianqiao powder, 4-6 parts of Gegen powder, 10-20 parts of earthworm powder, 2-3 parts of polyethylene oxide, 5-8 parts of soybean meal extract, 2-3 parts of potassium fulvic acid, 1-2 parts of amine ester, 2-5 parts of polybutene acid ammonium, 2-3 parts of brassinolide and 3-4 parts of chitosan oligosaccharide.

[0004] In recent years, agricultural product processing by-products have been used in the preparation of microbial fertilizers. For example, the Chinese patent application with the publication number CN 114634385A discloses a nitrogen-fixing microbial fertilizer using sea buckthorn pomace as a carrier. The preparation of Sediminibacillus sp. T-28 bacterial bodies is included. The sea buckthorn pomace is subjected to fixation, drying and crushing. The obtained sea buckthorn pomace powder is mixed with perlite, then sterilized to obtain a microbial fertilizer carrier. The Sediminibacillus sp. T-28 bacterial bodies are washed with a pH 8.0 TE buffer, then mixed with a pH 8.0 TE buffer to form a bacterial suspension. The microbial fertilizer carrier and the bacterial suspension are mixed and cultured at 30±1℃ for 3.5-4.5 days to obtain a nitrogen-fixing microbial fertilizer. In the application document, sea buckthorn pomace is directly used as a microbial fertilizer carrier. SUMMARY

[0005] Sea buckthorn fruit is rich in vitamins, amino acids and flavonoids. After the sea buckthorn fruit is squeezed, the by-product sea buckthorn pomace is obtained. The present application found that the fermentation liquid of sea buckthorn pomace and Bacillus subtilis can significantly enhance the effect of Bacillus subtilis when they are used together. They have a synergistic effect. Therefore, the present application proposes an invention application of a sea buckthorn enzyme microbial fertilizer and a preparation method and application thereof.

[0006] A sea-buckthorn enzyme bacterial fertilizer is obtained by uniformly mixing sea-buckthorn pomace fermentation liquor, ammonium salt, phosphate, trehalose and Bacillus subtilis in a mass ratio of 80:1.5-2.5:0.5-1.5:0.5-1.5:0.5-1.5 and then freeze-drying; the preparation method of the sea-buckthorn pomace fermentation liquor comprises the following steps:

[0007] The sea-buckthorn pomace is soaked with water and then crushed into homogenate;

[0008] Yeast, lactobacillus and white granulated sugar are added to the obtained homogenate and mixed uniformly, and then the mixture is sealed for fermentation;

[0009] The adding amount of the yeast, lactobacillus and white granulated sugar is respectively 0.05-0.1%, 0.2-0.3% and 5-10% of the mass of the homogenate.

[0010] Optionally, the viable count of the lactobacillus is generally required to be more than 100 million per gram, the yeast is preferably high-sugar-tolerant dry yeast, and the viable count of the Bacillus subtilis is generally required to be more than 40 billion per gram, which can be obtained by purchase.

[0011] Optionally, in the process of preparing the homogenate, the mixing mass ratio of the sea-buckthorn pomace to water is 1:0.8-1.2.

[0012] Optionally, the fermentation condition is 37°C; and the fermentation is carried out in a constant-temperature incubator for 50-65 hours.

[0013] Optionally, the adding amount of the yeast, lactobacillus and white granulated sugar is respectively 0.09%, 0.25% and 9% of the mass of the homogenate.

[0014] Optionally, the ammonium salt comprises carbamide and ammonium sulfate in a mass ratio of 1:2; and the phosphate comprises sodium dihydrogen phosphate and disodium hydrogen phosphate in a mass ratio of 1:1.

[0015] Optionally, the sea-buckthorn pomace fermentation liquor, ammonium salt, phosphate, trehalose and Bacillus subtilis are mixed in a mass ratio of 80:1.5-2:0.5-1:0.8-1.2:0.8-1.2.

[0016] The application further provides a preparation method of the sea-buckthorn enzyme bacterial fertilizer, comprising the following steps:

[0017] (1) preparing sea-buckthorn pomace fermentation liquor: sea-buckthorn pomace is soaked with water and then crushed into homogenate; yeast, lactobacillus and white granulated sugar are added to the obtained homogenate and mixed uniformly, and then the mixture is sealed for fermentation; the adding amount of the yeast, lactobacillus and white granulated sugar is respectively 0.05-0.1%, 0.2-0.3% and 5-10% of the mass of the homogenate;

[0018] (2) the sea-buckthorn pomace fermentation liquor, ammonium salt, phosphate, trehalose and Bacillus subtilis are mixed uniformly in a mass ratio of 80:1.5-2.5:0.5-1.5:0.5-1.5:0.5-1.5 and then freeze-dried to obtain the sea-buckthorn enzyme microbial fertilizer.

[0019] The application also provides application of the sea-buckthorn enzyme microbial fertilizer in soil improvement or fruit and vegetable planting.

[0020] Optionally, the sea-buckthorn enzyme microbial fertilizer is diluted at a dilution ratio of at least 1:500 and then irrigated to the root of the fruit and vegetable.

[0021] Optionally, the fruit and vegetable is Chinese flowering cabbage.

[0022] In the components, the sea-buckthorn pomace fermentation liquor has a significant synergistic effect on Bacillus subtilis; the phosphate is used to buffer the fertilizer solution and provide phosphorus; and the trehalose has a protective effect on the activity of the microbial fertilizer during the preservation process.

[0023] Compared with the prior art, the application has at least one of the following beneficial effects:

[0024] (1) The dry weight of Chinese flowering cabbage is negatively affected when the sea-buckthorn enzyme is used alone, and the dry weight of Chinese flowering cabbage is only slightly increased when Bacillus subtilis is used alone, but the dry weight of Chinese flowering cabbage is significantly increased when the sea-buckthorn enzyme and Bacillus subtilis are used together, which indicates that the sea-buckthorn enzyme and Bacillus subtilis have a synergistic effect on increasing the dry weight of Chinese flowering cabbage.

[0025] (2) The nitrogen content of Chinese flowering cabbage is negatively affected when the sea-buckthorn enzyme is used alone, and the nitrogen content of Chinese flowering cabbage is only slightly increased when Bacillus subtilis is used alone, but the nitrogen content of Chinese flowering cabbage is significantly increased when the sea-buckthorn enzyme and Bacillus subtilis are used together, which indicates that the sea-buckthorn enzyme and Bacillus subtilis have a synergistic effect on increasing the nitrogen content of Chinese flowering cabbage.

[0026] (3) The phosphorus and potassium contents are also significantly increased when the sea-buckthorn enzyme and Bacillus subtilis are used together.

[0027] (4) The sea-buckthorn enzyme and Bacillus subtilis are coupled and then applied to significantly improve the soil nutrients. DETAILED DESCRIPTION

[0028] The technical solutions of the application will be described clearly and completely below with reference to the embodiments of the application. Obviously, the described embodiments are only some of the embodiments of the application, but not all the embodiments. Based on the embodiments in the application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the application.

[0029] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description herein is for describing particular embodiments only and is not intended to be limiting of the application.

[0030] The sea buckthorn pomace used in the following examples is sea buckthorn pomace obtained after sea buckthorn juice is squeezed, and the water content thereof is approximately within 1-10%.

[0031] The lactobacillus and yeast used in the following examples are commercially available products, the effective viable count of the lactobacillus is more than 100 million per gram, and the yeast is preferably high-sugar-tolerant dry yeast; specifically, the lactobacillus can be Lactobacillus paracasei from Xi'an Juxing Original Biotechnology Co., Ltd., and the yeast can be general dry yeast from Angel Yeast Co., Ltd.

[0032] The bacillus subtilis used in the following examples is commercially available product, the effective viable count of the bacillus subtilis is more than 40 billion per gram, and specifically, the bacillus subtilis can be that from Hubei Qiming Bioengineering Co., Ltd.

[0033] Example 1 Preparation of sea buckthorn enzyme microbial fertilizer

[0034] Take 100 g of sea buckthorn pomace, add 100 g of sterile water to soak, then put it into a tissue crusher, crush at 12000 rpm to make a homogenate, add the homogenate into a fermentation tank which has been sterilized at high temperature, add 0.18 g of yeast, 0.50 g of lactobacillus (effective viable count: 100 million per gram) and 18 g of white sugar, and uniformly stir and seal.

[0035] Put the fermentation tank into a 37℃ constant temperature incubator to ferment for 60 hours, take 80 ml of supernatant after fermentation, add 1.5 g of carbamide, 0.3 g of ammonium sulfate, 0.3 g of sodium dihydrogen phosphate, 0.3 g of disodium hydrogen phosphate and 1 g of trehalose, and 1 g of bacillus subtilis (effective viable count: 40 billion per gram), and uniformly stir, then put it into a refrigerator at -80℃ for 2 days, then put it into a freeze dryer to freeze dry, and pack it with an aluminum foil bag (recommended to be stored in cold storage) to obtain a sea buckthorn enzyme microbial fertilizer.

[0036] Take one bag of the above microbial fertilizer, add 5-10 L of water, dissolve, and use it to irrigate fruits and vegetables.

[0037] Example 2 Application test of sea buckthorn enzyme microbial fertilizer

[0038] Group S (without microorganisms): take 100 g of sea buckthorn pomace, add 100 g of sterile water to soak, then put it into a tissue crusher, crush at 12000 rpm to make a homogenate, add the homogenate into a fermentation tank which has been sterilized at high temperature, add 0.18 g of yeast, 0.50 g of lactobacillus and 18 g of white sugar, and uniformly stir and seal.

[0039] Put the fermentation tank in a 37°C constant temperature incubator for 60 hours of fermentation. After the fermentation is completed, take 80 milliliters of supernatant, add 1.5 grams of carbamide, 0.3 grams of ammonium sulfate, 0.3 grams of sodium dihydrogen phosphate, 0.3 grams of disodium hydrogen phosphate, and 1 gram of trehalose, stir evenly, put in a -80°C refrigerator for 2 days, then put into a freeze dryer, freeze-dry, and pack into 1 bag (recommended to be stored in a refrigerator).

[0040] Take 1 bag of the above microbial fertilizer, add 5L-10L water, dissolve, and use it to irrigate the roots of fruits and vegetables.

[0041] S+M group (fermentation broth + bacteria): Take 100g of sea buckthorn pomace, add 100 grams of sterile water to soak, then put it into a tissue crusher, crush at 12000 rpm to make a homogenate, add the homogenate to a fermentation tank that has been sterilized at high temperature, add 0.18g of yeast, 0.50g of lactobacillus, and 18g of white sugar, stir evenly, and seal.

[0042] Put the fermentation tank in a 37°C constant temperature incubator for 60 hours of fermentation. After the fermentation is completed, take 80 milliliters of supernatant, add 1.5 grams of carbamide, 0.3 grams of ammonium sulfate, 0.3 grams of sodium dihydrogen phosphate, 0.3 grams of disodium hydrogen phosphate, 1 gram of trehalose, and 1 gram of bacillus subtilis, stir evenly, put in a -80°C refrigerator for 2 days, then put into a freeze dryer, freeze-dry, and pack into 1 bag (recommended to be stored in a refrigerator).

[0043] Take 1 bag of the above microbial fertilizer, add 5L-10L water, dissolve, and use it to irrigate the roots of fruits and vegetables. It is appropriate to irrigate the soil around the roots of fruits and vegetables.

[0044] M group (does not contain fermentation broth): 1 gram of bacillus subtilis is added to 5L-10L water, dissolved, and used to irrigate the roots of fruits and vegetables.

[0045] CK: pure water.

[0046] When used as a fertilizer for fruit and vegetable planting, take 1 bag of the above microbial fertilizer, dilute it with 5L of water for the S group, S+M group, M group, and CK group.

[0047] Take 1.8 kg of air-dried soil into a flowerpot, adjust its moisture content to 60% of the field water holding capacity, and carry out soil pre-cultivation. After the soil components are stable, sow 18 good, plump, and basically the same size of small cabbage seeds in each pot, and leave 7 small cabbages with good growth and basically the same growth conditions after germination. Apply 0.14 g of urea to each pot. During the growth of the small cabbages, irrigate them with the above-mentioned ferment or compound microbial fertilizer every 7 days, apply 45 mL to each pot, and irrigate the CK group with 45 mL of deionized water. At the same time, in addition to applying fertilizer once a week, irrigate each group with 20 mL of water every day to ensure the normal growth of the small cabbages, and record the growth of the small cabbages in each group. Harvest the small cabbages after 30 days of growth.

[0048] Dry the collected soil samples to determine the indicators. After the small cabbages mature, collect their aboveground parts for drying and indicator determination.

[0049] The determination of soil organic matter, alkali-hydrolyzable nitrogen content, available phosphorus content, and available potassium content is carried out according to the following relevant standards: "Soil Testing Part 6: Determination of Soil Organic Matter" (NY / T1121.6-2006), "Determination of Soil Alkali-hydrolyzable Nitrogen" (DB51 / T 1875-2014), "Determination of Available Phosphorus in Soil by Sodium Bicarbonate Extraction-Molybdenum Antimony Antispectrophotometric Method" (HJ704-2014), and "Determination of Soil Available Potassium and Slow-acting Potassium Content" (NY / T 889-2004).

[0050] The determination of small cabbage dry weight, nitrogen content, phosphorus content, and total potassium content is carried out by drying method and "Determination of Nitrogen, Phosphorus, and Potassium in Plants" (NY / T 2017-2011).

[0051] The test results are as follows:

[0052] Effect of microbial fertilizer on soil:

[0053] (1) Effect on soil organic matter

[0054] Table 1 Effect of different fertilizer treatments on soil organic matter

[0055]

[0056] (2) Effect on soil alkali-hydrolyzable nitrogen content

[0057] Table 2 Effect of different fertilizer treatments on soil alkali-hydrolyzable nitrogen content

[0058]

[0059] (3) Effect on soil available phosphorus content

[0060] Table 3 Effect of different fertilizer treatments on soil available phosphorus content

[0061]

[0062] (4) Effect on the available potassium content of soil

[0063] Table 4 Effect of different fertilization treatments on the available potassium content of soil

[0064]

[0065]

[0066] As can be seen from the results of Tables 1-4, the fertilization treatments of the three fertilizers except the CK group all improved the content of each index to some extent, but the improvement effect of the sea buckthorn enzyme and bacillus subtilis fertilizer (S+M) group on each index of soil was obviously better than that of the sea buckthorn enzyme group (S) or the bacillus subtilis fertilizer group (M) alone. The improvement effect of the sea buckthorn enzyme and bacillus subtilis fertilizer on soil nutrients was obviously higher than that of any one of them applied alone, and there was a synergistic effect between them.

[0067] Effect of microbial fertilizer on pakchoi

[0068] (1) Effect on the yield of pakchoi

[0069] Table 5 Effect of microbial fertilizer on the dry weight of pakchoi

[0070]

[0071] As can be seen from the results of Table 5, the application of S sea buckthorn enzyme fertilizer alone had a negative effect on the dry weight of pakchoi, the application of M bacillus subtilis fertilizer alone could improve the dry weight of pakchoi, but the improvement effect was limited, only about 30%, while the application of S sea buckthorn enzyme and M bacillus subtilis together could increase the dry weight of pakchoi by nearly 100%, which indicated that there was a synergistic effect between S sea buckthorn enzyme and M bacillus subtilis in improving the dry weight of pakchoi.

[0072] (2) Effect on the nitrogen content of pakchoi

[0073] Table 6 Effect of different fertilization treatments on the nitrogen content of pakchoi

[0074]

[0075] As can be seen from the results of Table 6, the application of S sea buckthorn enzyme fertilizer alone had a negative effect on the nitrogen content of pakchoi, the application of M bacillus subtilis fertilizer alone could improve the dry weight of pakchoi, but the improvement effect was limited, only about 22%, while the application of S sea buckthorn enzyme and M bacillus subtilis together could increase the dry weight of pakchoi by more than 60%, which indicated that there was a synergistic effect between S sea buckthorn enzyme and M bacillus subtilis in improving the nitrogen content of pakchoi.

[0076] (3) Effect on phosphorus content of Chinese flowering cabbage

[0077] Table 7 Effect of different fertilization treatments on phosphorus content of Chinese flowering cabbage

[0078]

[0079] (4) Effect on total potassium content of Chinese flowering cabbage

[0080] Table 8 Effect of different fertilization treatments on total potassium content of Chinese flowering cabbage

[0081]

[0082] From the above results of Tables 5-8, it can be seen that:

[0083] The application of S sea buckthorn enzyme fertilizer alone showed a negative effect on the dry weight and nitrogen content of Chinese flowering cabbage, but when combined with M Bacillus subtilis fertilizer, it could significantly increase the yield and nitrogen content of Chinese flowering cabbage, and also had a significant effect on the phosphorus and potassium contents.

[0084] The above examples only express several embodiments of the present application, and the description is more specific and detailed, but it should not be understood as limiting the scope of the patent. It should be noted that for ordinary skilled persons in the art, without departing from the concept of the present application, several modifications and improvements can be made, which are all within the scope of the present application. Therefore, the protection scope of the present patent should be subject to the appended claims.

Claims

1. The application of sea buckthorn enzyme microbial fertilizer in soil improvement or bok choy cultivation, characterized in that, The seabuckthorn enzyme fertilizer is prepared by mixing seabuckthorn fruit residue fermentation liquid, ammonium salt, phosphate, trehalose and Bacillus subtilis in a mass ratio of 80:1.5~2.5:0.5~1.5:0.5~1.5:0.5~1.5 and then freeze-drying. In the application of the bok choy planting, the sea buckthorn enzyme fertilizer increases the dry weight of the bok choy and improves its nitrogen content; The preparation method of the sea buckthorn pomace fermentation liquid includes: Sea buckthorn fruit residue is soaked in water at a mixing mass ratio of 1:0.8~1.2 and then crushed into a homogeneous slurry; Add yeast, lactobacillus and white sugar to the obtained homogenate and mix well. Then ferment in a constant temperature incubator at 37°C for 50-65 hours and seal for fermentation. The amounts of yeast, lactobacillus, and white sugar added are 0.05-0.1%, 0.2-0.3%, and 5-10% of the mass of the homogenate, respectively.

2. The application according to claim 1, characterized in that, In the application of the bok choy cultivation, the sea buckthorn enzyme fertilizer is diluted at a ratio of at least 1:500 and then applied to the roots of the bok choy.

3. The application according to claim 1, characterized in that, The amounts of yeast, lactobacillus, and granulated sugar added are 0.09%, 0.25%, and 9% of the mass of the homogenate, respectively.

4. The application according to claim 1, characterized in that, The ammonium salt comprises carbamide and ammonium sulfate in a mass ratio of 1:2; the phosphate salt comprises sodium dihydrogen phosphate and disodium hydrogen phosphate in a mass ratio of 1:

1.

5. A seabuckthorn enzyme fertilizer, characterized in that, It is obtained by mixing sea buckthorn fruit pomace fermentation broth, ammonium salt, phosphate, trehalose and Bacillus subtilis in a mass ratio of 80:1.5~2.5:0.5~1.5:0.5~1.5:0.5~1.5 and then freeze-drying. The preparation method of the sea buckthorn pomace fermentation liquid includes: Sea buckthorn fruit residue is soaked in water at a mixing mass ratio of 1:0.8~1.2 and then crushed into a homogeneous slurry; Add yeast, lactobacillus and white sugar to the obtained homogenate and mix well. Then ferment in a constant temperature incubator at 37°C for 50-65 hours and seal for fermentation. The amounts of yeast, lactobacillus, and white sugar added are 0.05-0.1%, 0.2-0.3%, and 5-10% of the mass of the homogenate, respectively.

6. The seabuckthorn enzyme fertilizer according to claim 5, characterized in that, The amounts of yeast, lactobacillus, and granulated sugar added are 0.09%, 0.25%, and 9% of the mass of the homogenate, respectively.

7. The seabuckthorn enzyme fertilizer according to claim 5, characterized in that, The ammonium salt comprises carbamide and ammonium sulfate in a mass ratio of 1:2; the phosphate salt comprises sodium dihydrogen phosphate and disodium hydrogen phosphate in a mass ratio of 1:1.

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