A microbial preparation suitable for saline-alkali soil, a preparation method and application thereof

Abstract

The present application belongs to the technical field of microbial preparation, and particularly relates to a microbial preparation suitable for saline-alkali soil, a preparation method and application thereof. The microbial preparation comprises, in terms of weight parts, 4-25 parts of Bacillus subtilis, 4-30 parts of Saccharomyces cerevisiae, 40-85 parts of a synergistic carrier and 5-20 parts of a protective carrier. The synergistic carrier comprises one or more than two selected from the group of yeast metabolites, mineral humic acid and amino acid powder. The protective carrier comprises one or more than two selected from the group of malt dextrin, trehalose, corn starch and attapulgite. The present application combines Bacillus subtilis and Saccharomyces cerevisiae, and combines the synergistic carrier and the protective carrier, so that the obtained microbial preparation has excellent effects of improving saline-alkali soil, promoting crop growth and improving the salt-alkali resistance of crops. The preparation process of the microbial preparation is simple and easy to operate, the quality is stable, the cost is low, and the microbial preparation is more suitable for industrial production.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of microbial preparation, and particularly relates to a microbial preparation suitable for saline-alkali soil, a preparation method and application thereof. BACKGROUND

[0002] The saline-alkali soil is a general term for saline soil and alkaline soil. Soil salinization refers to a geological process and phenomenon that the content of salt in the soil surface increases continuously under the action of nature and human beings, and exceeds a certain limit. The saline-alkali soil affects the absorption of nutrients by crops, reduces the availability of soil nutrients, and deteriorates the physical and biological properties of the soil. Studies have shown that a high content of salt in the soil can seriously affect the yield of crops.

[0003] The current biological improvement technology for saline-alkali soil uses the life activities of microorganisms to increase the content of nitrogen, available phosphorus and available potassium in the soil, convert some substances in the soil that cannot be directly utilized by plants into nutrients that can be absorbed and utilized by plants, inhibit plant pathogenic bacteria by regulating the material level of the crops, improve the soil fertility, and reduce the salt content and pH of the saline-alkali soil, so as to improve the soil conditions of the saline-alkali soil. At present, some strains with growth promotion and disease resistance have been made into microbial fertilizers or microbial pesticides, but they are mainly applied to conventional soil, and the application in saline-alkali soil is not ideal. Therefore, it is urgent to develop a microbial preparation suitable for saline-alkali soil and having excellent performance. SUMMARY

[0004] The technical problem solved by the present application is that the effect of the microbial fertilizer in the prior art is not ideal in the application in saline-alkali soil.

[0005] In view of the above technical problems, the present application provides a microbial preparation suitable for saline-alkali soil, a preparation method and application thereof.

[0006] Specifically, the present application provides the following technical solutions:

[0007] In a first aspect, the present application provides a microbial preparation suitable for saline-alkali soil, which comprises, by weight: 4-25 parts of Bacillus subtilis, 4-30 parts of Saccharomyces cerevisiae, 40-85 parts of a synergistic carrier, and 5-20 parts of a protective carrier; wherein the synergistic carrier comprises one or two or more selected from the group of substances consisting of yeast metabolites, mineral humic acid and amino acid powder; and the protective carrier comprises one or two or more selected from the group of substances consisting of malt dextrin, trehalose, corn starch and attapulgite.

[0008] Preferably, the microbial preparation comprises 4-20 parts of Bacillus subtilis, more preferably 4-13 parts of Bacillus subtilis, and more preferably 4-6 parts of Bacillus subtilis, by weight. And / or

[0009] Preferably, the microbial preparation comprises 4-20 parts by weight of Saccharomyces cerevisiae; more preferably, the microbial preparation comprises 4-6 parts by weight of Saccharomyces cerevisiae. And / or

[0010] Preferably, the microbial preparation comprises 60-85 parts by weight of the synergistic carrier; more preferably, the microbial preparation comprises 75-85 parts by weight of the synergistic carrier; still more preferably, the microbial preparation comprises 78-82 parts by weight of the synergistic carrier. And / or

[0011] Preferably, the microbial preparation comprises 5-15 parts by weight of the protective carrier; more preferably, the microbial preparation comprises 5-12 parts by weight of the protective carrier; still more preferably, the microbial preparation comprises 8-11 parts by weight of the protective carrier. And / or

[0012] Preferably, the Bacillus subtilis is Bacillus subtilis subsp. subtilis tc-4 (Bacillus subtilis subsp. subtilis tc-4), which was preserved in the China Center for Type Culture Collection (CCTCC) on April 17, 2023, and the preservation number is CCTCC NO: M 2023541. And / or

[0013] Preferably, the Saccharomyces cerevisiae is Saccharomyces cerevisiae Gaojiuyao YY (Saccharomyces cerevisiae Gaojiuyao YY), which was preserved in the China Center for Type Culture Collection (CCTCC) on February 1, 2021, and the preservation number is CCTCC NO: M 2021171; or the Saccharomyces cerevisiae is Saccharomyces cerevisiae d8.8, which was preserved in the China Center for Type Culture Collection (CCTCC) on March 29, 2017, and the preservation number is CCTCC NO: M 2017148; more preferably, the Saccharomyces cerevisiae is Saccharomyces cerevisiae Gaojiuyao YY (Saccharomyces cerevisiae Gaojiuyao YY), which was preserved in the China Center for Type Culture Collection (CCTCC) on February 1, 2021, and the preservation number is CCTCC NO: M 2021171.

[0014] And / or

[0015] Preferably, the synergistic carrier comprises a yeast metabolite; more preferably, the yeast metabolite is a metabolite produced by fermentation of molasses after extraction of yeast; further preferably, the molasses comprises sugar beet molasses; and / or, more preferably, the content of fulvic acid in the yeast metabolite is 15% to 30% by dry weight of the yeast metabolite; further preferably, the content of betaine in the yeast metabolite is 6% to 15% by dry weight of the yeast metabolite.

[0016] Preferably, the protective carrier comprises one or more than two selected from the group consisting of trehalose, corn starch and attapulgite; more preferably, the protective carrier comprises corn starch and / or attapulgite.

[0017] Preferably, the effective viable cell number of Bacillus subtilis in the microbial preparation is 4 billion to 25 billion CFU / g; more preferably, the effective viable cell number of Bacillus subtilis in the microbial preparation is 4 billion to 20 billion CFU / g.

[0018] Preferably, the effective viable cell number of Saccharomyces cerevisiae in the microbial preparation is 0.8 billion to 6 billion CFU / g; more preferably, the effective viable cell number of Saccharomyces cerevisiae in the microbial preparation is 0.8 billion to 4 billion CFU / g.

[0019] and / or

[0020] Preferably, the Bacillus subtilis is Bacillus subtilis powder, and the preparation method comprises the following steps:

[0021] Step 1: activation of Bacillus subtilis;

[0022] Step 2: inoculating the activated Bacillus subtilis into a culture medium for culture to obtain a seed liquid; and

[0023] Step 3: inoculating the seed liquid into a fermentation culture medium for culture to obtain a fermentation liquid, and drying the fermentation liquid to obtain Bacillus subtilis powder;

[0024] More preferably, in Step 1, the culture medium used for activation comprises 0.5% to 1.5% of proteose peptone, 0.1% to 0.5% of beef extract, 0.3% to 0.7% of sodium chloride and 1.5% to 2% of agar by mass of the culture medium, and the rest is water; further preferably, the culture medium used for activation comprises 1% of proteose peptone, 0.3% of beef extract, 0.5% of sodium chloride and 1.5% to 2% of agar, and the rest is water; and / or

[0025] More preferably, in Step 1, the culture temperature is 35 to 39°C; and / or, the culture time is 21 to 27 h; and / or

[0026] Further preferably, the Bacillus subtilis after cultivation is further activated;

[0027] More preferably, the culture medium used for activation comprises 0.5% to 1.5% of proteose peptone, 0.1% to 0.5% of beef extract, 0.3% to 0.7% of sodium chloride and 1.5% to 2% of agar, with the rest being water, based on the mass of the culture medium used for activation; most preferably, the culture medium used for activation comprises 1% of proteose peptone, 0.3% of beef extract, 0.5% of sodium chloride and 1.5% to 2% of agar, with the rest being water; and / or

[0028] Preferably, the temperature of the further cultivation is 35 to 39°C; and / or, the time of the cultivation is 21 to 27h; and / or,

[0029] More preferably, in step 2, the culture medium comprises 0.5% to 1.5% of proteose peptone, 0.1% to 0.5% of beef powder, 0.3% to 0.7% of sodium chloride and 0.05% to 0.15% of glucose, with the rest being water, based on the total mass of the culture medium; more preferably, the culture medium comprises 1% of proteose peptone, 0.3% of beef powder, 0.5% of sodium chloride and 0.1% of glucose, with the rest being water; and / or, the temperature of the cultivation is 35 to 39°C; and / or, the time of the cultivation is 10 to 15h; and / or, the rotation speed is 100 to 200rpm; and / or

[0030] More preferably, in step 3, the culture medium comprises 3% to 5% of corn powder, 3% to 5% of soybean meal powder, 0.05% to 0.15% of magnesium sulfate and 0.05% to 0.15% of calcium chloride, with the rest being water, based on the total mass of the culture medium; more preferably, the culture medium comprises 4% of corn powder, 4% of soybean meal powder, 0.1% of magnesium sulfate and 0.1% of calcium chloride, with the rest being water; and / or, the temperature of the cultivation is 35 to 39°C; and / or, the time of the cultivation is 15 to 20h; and / or, the ventilation amount is 20 to 50L / min; and / or, the tank pressure is 0.2 to 0.5atmospheres; and / or preferably, the Saccharomyces cerevisiae is a Saccharomyces cerevisiae powder, and the preparation method thereof comprises the following steps:

[0031] Step 1: activation of Saccharomyces cerevisiae;

[0032] Step 2: the activated Saccharomyces cerevisiae is inoculated into a culture medium for cultivation to obtain a seed liquid;

[0033] Step 3: the seed liquid is inoculated into a fermentation culture medium for cultivation to obtain a fermentation liquid, and the Saccharomyces cerevisiae powder is obtained after separation and drying;

[0034] More preferably, in step 1, the culture medium used for activation comprises 0.5% to 1.5% peptone, 1.5% to 2.5% glucose, 0.1% to 1% yeast extract powder and 1.1 to 1.7% agar, with the rest being water, based on the total mass of the culture medium used for activation; more preferably, the culture medium used for activation comprises 1% peptone, 2% glucose, 0.5% yeast extract powder and 1.4% agar, with the rest being water; and / or, the temperature of the culture is 28 to 32℃; and / or, the time of the culture is 21 to 27h; and / or,

[0035] More preferably, in step 2, the activated Saccharomyces cerevisiae is inoculated into the culture medium used for the first-stage seed liquid to obtain the first-stage seed liquid; then, the first-stage seed liquid is inoculated into the culture medium used for the second-stage seed liquid to obtain the second-stage seed liquid (also referred to as the seed liquid); more preferably, the culture conditions of the first-stage seed liquid are as follows: the temperature of the culture is 28 to 32℃; and / or, the time of the culture is 12 to 18h; and / or, more preferably, the composition of the culture medium used for the first-stage seed liquid is as follows: 90 to 110g of sucrose, 18 to 22g of yeast extract powder, 0.5 to 1.5g of magnesium sulfate and 0.5 to 1.5g of zinc sulfate per liter of water; more preferably, the composition of the culture medium used for the first-stage seed liquid is as follows: 100g of sucrose, 20g of yeast extract powder, 1g of magnesium sulfate and 1g of zinc sulfate per liter of water; and / or, more preferably, the culture conditions of the second-stage seed liquid are as follows: the inoculation amount of the first-stage seed liquid is 1% to 3%, based on the volume of the culture medium; and / or, the temperature of the culture is 28 to 32℃; and / or, the time of the culture is 12 to 18h; and / or, the culture medium used for the second-stage seed liquid comprises sugar beet molasses; and / or,

[0036] More preferably, in step 3, the inoculation amount of the seed liquid is 1% to 3%, based on the volume of the culture medium; and / or, the culture medium comprises sugar beet molasses; and / or, the temperature of the culture is 28 to 32℃; and / or, the time of the culture is 12 to 18h.

[0037] In a second aspect, the present application provides a preparation method of a microbial preparation suitable for saline-alkali soil, comprising the following steps:

[0038] The synergistic carrier, Bacillus subtilis, Saccharomyces cerevisiae and protective carrier are mixed to obtain the microbial preparation.

[0039] Preferably, the microbial preparation suitable for saline-alkali soil is prepared by the above-mentioned preparation method of the microbial preparation suitable for saline-alkali soil.

[0040] In a third aspect, the present application provides the use of the microbial preparation suitable for saline-alkali soil in improving saline-alkali soil.

[0041] Preferably, the application amount of the microbial preparation suitable for saline-alkali soil is 10 to 20kg / hm2 ; and / or

[0042] Preferably, the microbial preparation is used by drip irrigation or flushing, and is suitable for the seedling growth period of crops. And / or

[0043] Preferably, the improvement of saline-alkali soil includes improving soil fertility, reducing soil alkalinity, and / or reducing soil conductivity.

[0044] In a fourth aspect, the present application provides a microbial preparation suitable for saline-alkali soil for promoting the growth of crops and / or improving the salt-alkali tolerance of crops.

[0045] Preferably, the crops are legume crops. And / or

[0046] Preferably, the application amount of the microbial preparation suitable for saline-alkali soil is 10-20 kg / hm 2 . And / or

[0047] Preferably, the microbial preparation is used by drip irrigation or flushing, and is suitable for the seedling growth period of crops.

[0048] In a fifth aspect, the present application provides a composition for improving saline-alkali soil, which includes the above-mentioned microbial preparation suitable for saline-alkali soil.

[0049] Preferably, the improvement of saline-alkali soil includes improving soil fertility, reducing soil alkalinity, and / or reducing soil conductivity.

[0050] In a sixth aspect, the present application provides a composition for promoting the growth of crops and / or improving the salt-alkali tolerance of crops, which includes the above-mentioned microbial preparation suitable for saline-alkali soil.

[0051] Advantages of the present application

[0052] (1) The present application uses Bacillus subtilis and Saccharomyces cerevisiae in cooperation, and combines with a synergistic carrier and a protective carrier, so that the obtained microbial preparation has excellent effects of improving saline-alkali soil, specifically in improving soil fertility, reducing soil alkalinity, and reducing soil conductivity.

[0053] (2) The microbial preparation provided by the present application also has the effects of promoting the growth of crops and improving the salt-alkali tolerance of crops. It is especially suitable for legume crops and has more excellent effects in legume crops.

[0054] (3) In the present application, especially the microbial preparation obtained by combining Bacillus subtilis subsp. subtilis tc-4 and Saccharomyces cerevisiae high alcohol yeast YY with a protective carrier and a synergistic carrier, and by regulating the content ratio of the four components, the synergistic effect between the components makes it have more excellent effects of improving saline-alkali soil, and also has more excellent effects of promoting growth and resisting stress on crops.

[0055] (4) The microbial preparation provided by the present application is safe for crops and the environment, reduces the input of chemical substances, and reduces environmental pressure.

[0056] (5) The preparation process of the microbial preparation provided by the present application is simple and easy to operate, and the cost is relatively low, which is more suitable for industrialized production.

[0057] Strain preservation information

[0058] In the present application, Bacillus subtilis subsp. subtilis tc-4 (Bacillus subtilis subsp. subtilis tc-4) was preserved in China Center for Type Culture Collection (CCTCC) on April 17, 2023, with the preservation number CCTCC NO: M 2023541 and the preservation address China, Wuhan, Wuhan University, postal code 430072, telephone (027) 68754052. The Bacillus subtilis subsp. subtilis tc-4 is disclosed in Chinese Patent No. 202311624023.5.

[0059] In the present application, Saccharomyces cerevisiae high alcohol yeast YY (Saccharomyces cerevisiae high alcohol yeast YY) was preserved in China Center for Type Culture Collection (CCTCC) on February 1, 2021, with the preservation number CCTCC NO: M2021171 and the preservation address China, Wuhan, Wuhan University, postal code 430072, telephone (027) 68754052. The Saccharomyces cerevisiae high alcohol yeast YY is disclosed in Chinese Patent No. 202110342874.5.

[0060] In the present application, Saccharomyces cerevisiae d8.8 (Saccharomyces cerevisiae d8.8) was preserved in China Center for Type Culture Collection (CCTCC) on March 29, 2017, with the preservation number CCTCC NO: M2017148 and the preservation address China, Wuhan, Wuhan University, postal code 430072, telephone (027) 68754052. The Saccharomyces cerevisiae d8.8 is disclosed in Chinese Patent No. 201710519421.9. BRIEF DESCRIPTION OF DRAWINGS

[0061] Figure 1 Figure of growth promotion effect evaluation of the microbial preparation prepared for the example.

[0062] Figure 2 Figure of stress resistance effect evaluation of the microbial preparation prepared for the example. DETAILED DESCRIPTION

[0063] As described above, the present application aims to provide a microbial preparation suitable for saline-alkali soil, and a preparation method and application thereof.

[0064] In the present application, the first object is to provide a microbial preparation suitable for saline-alkali soil, comprising, in parts by weight: 4-25 parts of Bacillus subtilis, 4-30 parts of Saccharomyces cerevisiae, 40-85 parts of a synergistic carrier, and 5-20 parts of a protective carrier; wherein the synergistic carrier comprises one or more than two selected from the group of substances consisting of yeast metabolites, humic acid from mineral sources, and amino acid powder; and the protective carrier comprises one or more than two selected from the group of substances consisting of malt dextrin, trehalose, corn starch, and attapulgite.

[0065] Preferably, the microbial preparation comprises 4-25 parts of Bacillus subtilis, for example, it can comprise 4-22 parts, 4-21 parts, 4-15 parts, or 4-6 parts of Bacillus subtilis. The Bacillus subtilis is preferably Bacillus subtilis subsp. subtilis tc-4. For example, in some embodiments, the microbial preparation comprises 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 parts, or comprises Bacillus subtilis within a numerical range constituted by any two of the above-mentioned specific numerical values as endpoints, preferably Bacillus subtilis powder, more preferably Bacillus subtilis subsp. subtilis tc-4 powder (which is a powder prepared from Bacillus subtilis subsp. subtilis tc-4); and / or,

[0066] Preferably, the microbial preparation comprises 4-30 parts of Saccharomyces cerevisiae, for example, it can comprise 4-25 parts, 4-22 parts, 4-21 parts, 4-15 parts, or 4-6 parts of Saccharomyces cerevisiae. The Saccharomyces cerevisiae is preferably Saccharomyces cerevisiae high alcohol yeast YY. For example, in some embodiments, the microbial preparation comprises 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 parts, or comprises Saccharomyces cerevisiae within a numerical range constituted by any two of the above-mentioned specific numerical values as endpoints, preferably Saccharomyces cerevisiae powder, more preferably Saccharomyces cerevisiae high alcohol yeast YY powder (which is a powder prepared from Saccharomyces cerevisiae high alcohol yeast YY); and / or,

[0067] Preferably, the microbial preparation includes 40-85 parts of a synergistic carrier, for example, it can include 50-85 parts, 55-85 parts, 70-85 parts, 75-85 parts, or 77-83 parts of a synergistic carrier. The synergistic carrier includes one or more than two selected from the group consisting of yeast metabolites, humic acid of mineral origin, and amino acid powder; preferably, the synergistic carrier is yeast metabolites. For example, in some embodiments, the microbial preparation includes 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, or 85 parts, or a number within a range defined by any two of the above-mentioned specific numbers as endpoints, of a synergistic carrier, preferably yeast metabolites; and / or,

[0068] Preferably, the microbial preparation includes 5-20 parts of a protective carrier, for example, it can include 5-18 parts, 5-16 parts, 5-13 parts, 5-11 parts, or 7-13 parts of a protective carrier. The protective carrier includes one or more than two selected from the group consisting of malt dextrin, trehalose, corn starch, and attapulgite; preferably, the protective carrier is corn starch and / or attapulgite. For example, in some embodiments, the microbial preparation includes 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 parts, or a number within a range defined by any two of the above-mentioned specific numbers as endpoints, of a protective carrier, preferably corn starch and / or attapulgite.

[0069] In the present application, the synergistic carrier refers to a carrier that can promote the function of the microbial agent, to some extent, reduce the addition amount of the microbial agent, and synergistically act with other components. The synergistic carrier includes one or more than two of yeast metabolites, humic acid of mineral origin, and amino acid powder; preferably, the synergistic carrier is yeast metabolites. The yeast metabolites are metabolites extracted from yeast after fermentation using molasses as raw material; preferably, the molasses includes sugar beet molasses. The most preferred yeast metabolites are because the yeast metabolites contain betaine and other stress resistance factors, and also contain fulvic acid and other soil conservation factor components, which synergistically improve the soil environment, enhance plant resistance, and increase yield and quality. More preferably, the yeast metabolites contain 15%-30% fulvic acid and / or 6%-15% betaine.

[0070] In the present application, the protective carrier refers to a kind of carrier that can improve the survival rate of live bacteria in microbial preparation, and prolong the shelf life of bacterial agent. In some specific embodiments, the protective carrier is selected from one or more of malt dextrin, trehalose, corn starch and attapulgite. Malt dextrin, trehalose, corn starch and / or attapulgite are selected as the synergistic carrier because they can effectively improve the alkalinity of the soil, optimize the physicochemical properties of the soil, and reduce the loss of soil moisture. Preferably, the protective carrier is corn starch and / or attapulgite because it is inexpensive and has a more significant protective effect on live bacteria in microbial preparations.

[0071] In the present application, the Bacillus subtilis contains dormant spores, which can help the bacteria to survive in adverse environments, produce growth-promoting substances IAA, promote plant growth, produce stress-resistant substances, and enhance the resistance of plants to adverse environments. The Saccharomyces cerevisiae contains a large amount of trehalose, glucan, mannose oligosaccharide, growth-promoting substance IAA, stress-resistant substance salicylic acid and jasmonic acid, etc., which enhance the stress resistance of the yeast itself and the applied plants. Bacillus subtilis and Saccharomyces cerevisiae, both of which have strong stress resistance, can survive in saline-alkali environments, produce growth-promoting and stress-resistant substances, and the synergistic effect of the two bacteria enhances the growth performance of plants in saline-alkali environments and produces polysaccharide substances to improve soil structure.

[0072] In the present application, the microbial preparation obtained by combining Bacillus subtilis subsp. subtilis tc-4 and Saccharomyces cerevisiae high-yeast YY with a protective carrier and a synergistic carrier, and by adjusting the content ratio of the four components, has excellent effects of improving saline-alkali soil, and has excellent growth-promoting and stress-resistant effects on crops. The content of each component and / or the type of component are within the preferred range, and the effects are better. In a more preferred range, the improvement of saline-alkali soil and the growth-promoting and stress-resistant effects on crops can be further improved. Preferably, the microbial preparation of the present application is particularly suitable for legume crops and has more excellent effects when applied to legume crops.

[0073] The second object of the present application is to provide a preparation method of a microbial preparation, which comprises the following steps: sequentially adding a synergistic carrier, Bacillus subtilis powder, Saccharomyces cerevisiae powder and a protective carrier, and mixing in a mixing bin for 5-8 min to obtain a microbial preparation.

[0074] Preferably, the preparation method of the Bacillus subtilis powder comprises the following steps:

[0075] Step 1: inoculate Bacillus subtilis in a culture medium including 0.5% to 1.5% of proteose peptone, 0.1% to 0.5% of beef extract, 0.3% to 0.7% of sodium chloride, 1.5% to 2% of agar and water, and incubate at a temperature of 35 to 39℃ for 21 to 27 hours; inoculate the obtained single colony in a culture medium including 0.5% to 1.5% of proteose peptone, 0.1% to 0.5% of beef extract, 0.3% to 0.7% of sodium chloride, 1.5% to 2% of agar and water, and continue to incubate at a temperature of 35 to 39℃ for 21 to 27 hours to obtain activated Bacillus subtilis;

[0076] Step 2: inoculate the single colony of the activated Bacillus subtilis in a culture medium including 0.5% to 1.5% of proteose peptone, 0.1% to 0.5% of beef extract, 0.3% to 0.7% of sodium chloride, 0.05% to 0.15% of glucose and water, and incubate at a temperature of 35 to 39℃ and a rotation speed of 100 to 200 rpm for 10 to 15 hours to obtain a seed liquid;

[0077] Step 3: inoculate the seed liquid in a culture medium including 3% to 5% of corn powder, 3% to 5% of soybean meal powder, 0.05% to 0.15% of magnesium sulfate, 0.05% to 0.15% of calcium chloride and water, and incubate at a temperature of 35 to 39℃, an air flow of 20 to 50 L / min and a tank pressure of 0.2 to 0.5 atm for 15 to 20 hours to obtain a fermentation liquid. The fermentation liquid is subjected to spray drying to obtain Bacillus subtilis powder.

[0078] And / or, preferably, the preparation method of the Saccharomyces cerevisiae powder comprises the following steps:

[0079] Step 1: inoculate Saccharomyces cerevisiae in a culture medium including 0.5% to 1.5% of peptone, 1.5% to 2.5% of glucose, 0.1% to 1% of yeast extract powder, 1.1% to 1.7% of agar and water, and incubate at a temperature of 28 to 32℃ for 21 to 27 hours;

[0080] Step 2: inoculate the activated Saccharomyces cerevisiae in a culture medium including 90 to 110 g of sucrose, 18 to 22 g of yeast extract powder, 0.5 to 1.5 g of magnesium sulfate and 0.5 to 1.5 g of zinc sulfate per liter of water to obtain a primary seed liquid, and incubate at a temperature of 28 to 32℃ for 12 to 18 hours; then, inoculate the primary seed liquid into a secondary seed liquid culture medium containing sugar beet molasses at a volume percentage of 1% to 3% to obtain a secondary seed liquid (also referred to as a seed liquid), and incubate at a temperature of 28 to 32℃ for 12 to 18 hours;

[0081] Step 3: the seed liquid is inoculated into the fermentation medium containing the beet molasses at a volume percentage of 1-3% of the volume of the medium, and cultured at a temperature of 28-32°C for 12-18 hours to obtain a fermentation liquid, which is separated and dried by drum extrusion to obtain the yeast powder.

[0082] In the present application, a third object is to provide a microbial preparation suitable for saline-alkali soil for improving saline-alkali soil, promoting crop growth and / or improving the salt-tolerance of legume crops. The microbial preparation is used by drip irrigation or flush application, and is suitable for the seedling growth period of crops, with an application amount of 10-20 kg / hm 2 .

[0083] The various reagents / instruments used in the examples and comparative examples of the present application are all conventional commercially available products, unless otherwise specified. The experimental materials used in the present application are shown in the following table:

[0084] Table 1 Experimental materials and suppliers

[0085]

[0086] In the present application, the NA plate is a purchased nutrient agar NA, which includes 1% proteose peptone, 0.3% beef extract, 0.5% sodium chloride, 1.5% agar, and the rest is water, in terms of mass percentage. The NB medium is a purchased nutrient broth NB, which includes 1% proteose peptone, 0.3% beef extract, 0.5% sodium chloride, 0.1% glucose, and the rest is water, in terms of mass percentage. The YPD plate is a purchased YPD agar (yeast extract peptone dextrose agar), which includes 1% peptone, 2% glucose, 0.5% yeast extract, and 1.4% agar, and the rest is water, in terms of mass percentage.

[0087] The source of the beet molasses in the present application is not limited in any way, and any commercially available or self-made beet molasses can be used in the present application. Preferably, the present inventors have found that as long as the commercially available or self-made beet molasses has a betaine content of ≥3 wt%, it can be used in the present application; more preferably, the beet molasses has a betaine content of 3 wt%-8 wt%. In some specific embodiments, the beet molasses is from COFCO Sugar.

[0088] In the present application, the yeast metabolites include yeast polysaccharide (calculated as β-glucan + mannoglucan) ≥2.5%, fulvic acid ≥15%, betaine ≥6%, medium elements (Ca+Mg) ≥1%, total nutrients (N, P2O5 and K2O) ≥10%, organic matter content ≥15%, and water-insoluble substances ≤5%.

[0089] In some embodiments, the present application does not make any limitation on the source of the yeast metabolite, and any commercially available or self-made yeast metabolite can be used in the present application. Specifically, the present inventors have found that as long as the commercially available or self-made yeast metabolite has betaine content ≥ 6% and fulvic acid content ≥ 15%, it can be used in the present application.

[0090] Preferably, the yeast metabolite comprises 24%-26% of crude protein, 50%-55% of crude ash, 3%-5% of total nitrogen, 15%-28% of organic matter, 15%-30% of fulvic acid, and / or 6%-15% of betaine, based on the weight of the yeast metabolite.

[0091] More preferably, the yeast metabolite further comprises 4%-5.5% of calcium, 0.1%-1% of magnesium, 7%-8% of sodium, 1%-2% of β-glucan, 2%-3% of mannans, 2%-3% of humic acid, 13-15% of potassium, 0.1%-1% of phosphorus, 3%-4% of sulfur, 15-18% of chloride ion, and / or 9%-13% of hydrolyzed amino acid.

[0092] In the present application, the detected components of the yeast metabolite purchased from Ili Fubang New Agricultural Co., Ltd. are shown in Table 2 below:

[0093] Table 2 Detected components of the yeast metabolite

[0094]

[0095]

[0096]

[0097] In the present application, the detection method of mannans comprises the following steps:

[0098] Sample treatment: accurately weigh 400 mg (accurate to 0.1 mg) of the sample into a 20 mL heat-resistant glass screw cap test tube, add 6.0 mL of hydrochloric acid (37%), carefully cap the test tube and mix with a vortex mixer to obtain a uniform suspension. Place the test tube in a 30°C water bath for 45 min, and mix once every 15 min with a vortex mixer. Then quantitatively transfer the suspension into a 200 mL Durham bottle, wash the test tube with about 100 mL-120 mL of water several times, and pour the washing liquid into the Durham bottle. Place the Durham bottle in an autoclave and treat at 121°C for 60 min. After cooling, adjust the pH of the solution to 6-7 with sodium hydroxide solution, and then dilute to 200 mL. Filter with a 0.45 micron pore size cellulose acetate membrane for use.

[0099] Chromatographic conditions: pure water was used as mobile phase, the flow rate was 0.5 mL / min, the column temperature was 80 ℃, and then the sample was injected after the baseline of the instrument was stable.

[0100] Preparation of standard curve: 1, 2, 3, 4, 5 mL of mannose standard solution (1 g / L, 0.2500 g of mannose was weighed and diluted to 250 mL with pure water) was taken into a 10 mL volumetric flask, and diluted to the mark with high-purity water to obtain a standard series of 100, 200, 300, 400, and 500 mg / L of mannose. Under the above chromatographic conditions, 20 μL of the sample was accurately injected to obtain the regression equation between the chromatographic peak area and the mass concentration of the standard substance, and the standard curve was drawn.

[0101] Determination of samples and controls: under the same chromatographic conditions, the treated sample was injected into the chromatograph, and the retention time and peak area of each chromatographic peak were recorded. The retention time of the sugar standard chromatographic peak was used for qualitative analysis, and the peak area of the sugar standard chromatographic peak was used for quantitative analysis.

[0102] Result calculation: the content of mannose was calculated according to the following formula:

[0103]

[0104] In the formula:

[0105] X - the content of mannose in the sample, %;

[0106] A - the content of mannose in the sample solution, mg / L, which was obtained by checking the sample solution on the standard curve according to the peak area of the sample solution;

[0107] m - the mass of the sample, g;

[0108] 0.2 - the volume of the sample control after treatment and dilution, L;

[0109] 0.9 - the coefficient for converting mannose to mannose.

[0110] In the present application, the total nitrogen content of the yeast extract powder is ≥9.0%, the amino nitrogen content is ≥3.0%, the ash content is ≤15%, the sodium chloride content is ≤2%, and the moisture content is ≤6%. In some specific embodiments, the present application does not make any limitation on the source of the yeast extract powder, and any commercially available or self-made yeast extract powder can be used in the present application. Specifically, the present inventors have found that as long as the total nitrogen content of the commercially available or self-made yeast extract powder is ≥9.0% and / or the amino nitrogen content is ≥3.0%, it can be used in the present application. Preferably, the total nitrogen content of the yeast extract powder is 9% to 15%, and / or the amino nitrogen content is 3% to 5%.

[0111] Preferably, the yeast extract includes 41,000 to 42,000 mg / kg of trace elements based on the weight of the yeast extract. More preferably, the trace elements include 32,760 to 32,770 mg / kg of potassium, 5,836 to 5,846 mg / kg of sodium, 53 to 53 mg / kg of calcium, and 2,029 to 2,039 mg / kg of magnesium. Still more preferably, the trace elements include 32,767.33 mg / kg of potassium, 5,841.92 mg / kg of sodium, 53.71 mg / kg of calcium, and 2,034.06 mg / kg of magnesium.

[0112] and / or, preferably, the yeast extract powder comprises vitamins in an amount of 5,000-6,000 ppm by weight of the yeast extract powder; more preferably, the yeast extract powder comprises vitamin Bl in an amount of 2-3 ppm (detection method: GB 5009.84-2016 National Food Safety Standard Determination of Vitamin Bl in Foods-Method 1-High Performance Liquid Chromatography), vitamin B2 in an amount of 38-44 ppm (detection method: GB 5009.85-2016 National Food Safety Standard Determination of Vitamin B2 in Foods-Method 1-High Performance Liquid Chromatography), vitamin B5 in an amount of 78-84 ppm (detection method: GB 5009.210-2023 National Food Safety Standard Determination of Pantothenic Acid in Foods-Method 3-Microbiological Method), vitamin B6 in an amount of 10-13 ppm (detection method: GB 5009.154-2023 National Food Safety Standard Determination of Vitamin B6 in Foods-Method 3-High Performance Liquid Chromatography-Fluorescence Detection Method), vitamin B7 in an amount of 4-7 ppm (detection method: GB 5009.259-2023 National Food Safety Standard Determination of Biotin in Foods-Method 2-Microbiological Method), vitamin B9 in an amount of 23-27 ppm (detection method: GB 5009.211-2022 National Food Safety Standard Determination of Folic Acid in Foods), vitamin B12 in an amount of 0.15-0.25 pg / 100 g (detection method: GB 5009.285-2022 National Food Safety Standard Determination of Vitamin B12 in Foods-Method 1-Liquid Chromatography Method), choline in an amount of 3,010-3,025 ppm (detection method: GB 5413.20-2022 National Food Safety Standard Determination of Choline in Infant and Baby Foods and Dairy Products-Method 3-Liquid Chromatography-Tandem Mass Spectrometry Method), inositol in an amount of 2,010-2,020 ppm (detection method: GB 5009.270-2023 National Food Safety Standard Determination of Inositol in Foods-Method 1-Gas Chromatography Method), and / or nicotinic acid in an amount of 310-325 ppm (detection method: GB 5009.89-2023 National Food Safety Standard Determination of Nicotinic Acid and Nicotinamide in Foods-Method 1-High Performance Liquid Chromatography Method). Still more preferably, the yeast extract powder comprises vitamin Bl in an amount of 2.6 ppm, vitamin B2 in an amount of 41.6 ppm, vitamin B5 in an amount of 81.3 ppm, vitamin B6 in an amount of 11.5 ppm, vitamin B7 in an amount of 5.79 ppm, vitamin B9 in an amount of 25.1 ppm, vitamin B12 in an amount of 0.21 pg / 100 g, choline in an amount of 3,017 ppm, inositol in an amount of 2,016 ppm, and / or nicotinic acid in an amount of 318 ppm.

[0113] And / or, preferably, the yeast extract includes 57% to 61% of hydrolyzed amino acids by weight of the yeast extract. More preferably, the yeast extract includes 59.16% of hydrolyzed amino acids.

[0114] In the present application, the total available nutrient content in the soil is the total content of hydrolyzable nitrogen, available phosphorus and available potassium, wherein the determination of hydrolyzable nitrogen content refers to the determination of hydrolyzable nitrogen in the determination of forest soil nitrogen of the People's Republic of China Forestry Industry Standard LY / T1228-2015; the determination of available phosphorus content refers to the determination of available phosphorus in soil testing Part 7 of the People's Republic of China Agricultural Industry Standard NY / T1121.7-2014; the determination of available potassium content refers to the determination of soil available potassium and slow available potassium content of the People's Republic of China Agricultural Industry Standard NY / T889-2004. The determination of organic matter content in soil refers to the determination of soil organic matter of the People's Republic of China Agricultural Industry Standard NY / T 1121.6-2006 Soil Testing Part 6. The soil conductivity refers to the determination of soil conductivity by electrode method of the People's Republic of China National Environmental Protection Standard HJ 802-2017. The soluble salt concentration in soil is tested by conductivity meter. The soil pH refers to the determination of soil pH of the People's Republic of China Agricultural Industry Standard NY / T 1121.2-2006 Soil Testing Part 2.

[0115] In order to better understand the technical solutions of the present application, the technical solutions of the present application will be described in detail below in combination with specific embodiments.

[0116] Example 1

[0117] (1) A formula of a microbial preparation suitable for saline-alkali soil

[0118] A microbial preparation suitable for saline-alkali soil, which includes 80 parts of yeast metabolites, 5 parts of Bacillus subtilis powder, 5 parts of Saccharomyces cerevisiae powder and 10 parts of corn starch by weight. Among them, the content of fulvic acid in yeast metabolites is 26.12%, and the content of betaine is 11.9%.

[0119] Among them, the Bacillus subtilis in the Bacillus subtilis powder is Bacillus subtilis subsp. subtilis tc-4 (Bacillus subtilis subsp. subtilis tc-4), which was preserved in the China Center for Type Culture Collection (CCTCC) on April 17, 2023, with the preservation number CCTCC NO: M 2023541 and the preservation address China, Wuhan, Wuhan University, postal code: 430072; telephone: (027) 68754052. The Bacillus subtilis subsp. subtilis tc-4 is disclosed in Chinese Patent No. 202311624023.5.

[0120] The preparation method of the Bacillus subtilis powder comprises the following steps:

[0121] Step 1: Using an inoculation loop to dip the Bacillus subtilis subsp. subtilis tc-4 bacterial liquid glycerol-preserved at a temperature of -80℃, performing streak culture on a NA plate (i.e. a purchased nutrient agar NA), and culturing at a temperature of 37℃ for 24h to obtain a single colony. The single colony is picked and further streaked on a NA plate, and cultured at a temperature of 37℃ for 24h to obtain activated Bacillus subtilis tc-4.

[0122] Step 2: An inoculation loop is used to pick an entire single colony, and inoculated into a flask containing 200mL of NB culture medium, and placed in a shaking bed for shaking culture at a temperature of 37℃ for 12h at a speed of 160r / min to obtain a seed liquid.

[0123] Step 3: 15L of liquid culture medium is first loaded into a 30L fermentation tank, and the preparation method of the liquid culture medium comprises: adding 4% corn powder, 4% soybean meal powder, 0.1% magnesium sulfate and 0.1% calcium chloride based on the total mass of the culture medium, and then adding tap water to 15L, adjusting the pH to 7, and high-temperature high-pressure sterilization at a temperature of 121℃ for 30min. Then, the 200mL seed liquid obtained in step 2 is all transferred into the 30L fermentation tank for fermentation culture, and the culture conditions are: air volume 20-50L / min, tank pressure 0.2-0.5atm, and ventilation fermentation at a temperature of 37℃ for 17-19h. Finally, the fermentation liquid is subjected to spray drying to obtain the Bacillus subtilis powder.

[0124] The Saccharomyces cerevisiae in the Saccharomyces cerevisiae powder is Saccharomyces cerevisiae high alcohol yeast YY, which was preserved in the China Center for Type Culture Collection (CCTCC) on February 1, 2021, with a preservation number of CCTCC NO: M 2021171 and a preservation address of China, Wuhan, Wuhan University, postal code: 430072, telephone: (027) 68754052. The Saccharomyces cerevisiae high alcohol yeast YY is disclosed in Chinese Patent No. 202110342874.5.

[0125] The preparation method of the Saccharomyces cerevisiae powder comprises the following steps:

[0126] Step 1: Using an inoculation loop to dip the Saccharomyces cerevisiae high alcohol yeast YY (i.e. Saccharomyces cerevisiae high alcohol yeast YY) bacterial liquid glycerol-preserved at a temperature of -80℃, performing plate streaking on a YPD agar culture medium, and culturing at a temperature of 30℃ for 24h to obtain a single colony.

[0127] Step 2: inoculate a complete single colony into the medium used for the first-stage seed liquid and culture at a temperature of 30℃ for 15h to obtain the first-stage seed liquid of the yeast. The preparation method of the medium used for the first-stage seed liquid comprises: dissolving 100g of sucrose, 20g of yeast extract powder, 1g of magnesium sulfate and 1g of zinc sulfate in 1L of water, and adjusting the pH to 4.8 with concentrated sulfuric acid.

[0128] The first-stage seed liquid of the yeast is inoculated into the medium used for the second-stage seed liquid at an inoculation amount of 1% to 3% by volume of the medium, and cultured at a temperature of 30℃ for 15h. The culture liquid is separated by a separator, and the precipitate is washed with water to obtain the seed liquid of the yeast (also referred to as the second-stage seed liquid). The preparation method of the medium used for the second-stage seed liquid comprises: taking 8L of sugar beet molasses and adjusting the pH to 5.5 with concentrated sulfuric acid.

[0129] Step 3: the seed liquid of the yeast is inoculated into a fermentation tank at an inoculation amount of 1% to 3% by volume of the medium, and fermented at a temperature of 30℃ for 15h to obtain the yeast liquid. The preparation method of the medium in the fermentation tank comprises the following steps: taking sugar beet molasses and adjusting the pH to 5.5 with concentrated sulfuric acid. The yeast liquid is separated by a separator, the precipitate is washed with water, and then drum extrusion drying is performed to obtain the yeast powder.

[0130] (2) A preparation method of a microbial preparation suitable for saline-alkali soil

[0131] A preparation method of a microbial preparation suitable for saline-alkali soil, comprising the following steps:

[0132] The above-mentioned yeast metabolites, Bacillus subtilis powder, Saccharomyces cerevisiae powder and corn starch are put into a mixing bin and mixed for 5 to 8min to obtain the microbial preparation.

[0133] Example 2

[0134] (1) A formula of a microbial preparation suitable for saline-alkali soil

[0135] A microbial preparation suitable for saline-alkali soil, comprising 40 parts of yeast metabolites, 20 parts of Bacillus subtilis powder, 13 parts of Saccharomyces cerevisiae powder, 5 parts of corn starch and 10 parts of attapulgite by weight. The content of fulvic acid in the yeast metabolites is 26.12%, and the content of betaine is 11.9%.

[0136] The Bacillus subtilis in the Bacillus subtilis powder is Bacillus subtilis subsp. subtilis tc-4. The preparation method of the Bacillus subtilis powder is the same as the preparation method of the Bacillus subtilis powder in Embodiment 1.

[0137] The Saccharomyces cerevisiae in the Saccharomyces cerevisiae powder is Saccharomyces cerevisiae high alcohol yeast YY. The preparation method of the Saccharomyces cerevisiae powder is the same as the preparation method of the Saccharomyces cerevisiae powder in Embodiment 1.

[0138] (2) A preparation method of a microbial agent suitable for saline-alkali soil

[0139] The preparation method of the microbial agent suitable for saline-alkali soil comprises the following steps:

[0140] The above yeast metabolite, Bacillus subtilis powder, Saccharomyces cerevisiae powder, corn starch and attapulgite are put into a mixing bin, and mixed for 5-8 minutes to obtain the microbial agent.

[0141] Embodiment 3

[0142] (1) A formula of a microbial agent suitable for saline-alkali soil

[0143] The microbial agent suitable for saline-alkali soil comprises 60 parts of yeast metabolite, 13 parts of Bacillus subtilis powder, 20 parts of Saccharomyces cerevisiae powder and 5 parts of corn starch by weight. The content of fulvic acid in the yeast metabolite is 26.12%, and the content of betaine is 11.9%.

[0144] The Bacillus subtilis in the Bacillus subtilis powder is Bacillus subtilis subsp. subtilis tc-4. The preparation method of the Bacillus subtilis powder is the same as the preparation method of the Bacillus subtilis powder in Embodiment 1.

[0145] The Saccharomyces cerevisiae in the Saccharomyces cerevisiae powder is Saccharomyces cerevisiae high alcohol yeast YY. The preparation method of the Saccharomyces cerevisiae powder is the same as the preparation method of the Saccharomyces cerevisiae powder in Embodiment 1.

[0146] (2) A preparation method of a microbial agent suitable for saline-alkali soil

[0147] The preparation method of the microbial agent suitable for saline-alkali soil comprises the following steps:

[0148] The above yeast metabolite, Bacillus subtilis powder, Saccharomyces cerevisiae powder and corn starch are put into a mixing bin and mixed for 5-8 minutes to obtain the microbial preparation.

[0149] Example 4

[0150] (1) A formula of a microbial preparation suitable for saline-alkali soil

[0151] A microbial preparation suitable for saline-alkali soil, which comprises 80 parts of yeast metabolite, 25 parts of Bacillus subtilis powder, 30 parts of Saccharomyces cerevisiae powder and 10 parts of corn starch by weight. The content of fulvic acid in the yeast metabolite is 26.12%, and the content of betaine is 11.9%.

[0152] The Bacillus subtilis in the Bacillus subtilis powder is Bacillus subtilis subsp. subtilis tc-4 (Bacillus subtilis subsp. subtilis tc-4). The preparation method of the Bacillus subtilis powder is the same as that of the Bacillus subtilis powder in Example 1.

[0153] The Saccharomyces cerevisiae in the Saccharomyces cerevisiae powder is Saccharomyces cerevisiae high alcohol yeast YY (Saccharomyces cerevisiae high alcohol yeast YY). The preparation method of the Saccharomyces cerevisiae powder is the same as that of the Saccharomyces cerevisiae powder in Example 1.

[0154] (2) A preparation method of a microbial preparation suitable for saline-alkali soil

[0155] The preparation method of a microbial preparation suitable for saline-alkali soil comprises the following steps:

[0156] The above yeast metabolite, Bacillus subtilis powder, Saccharomyces cerevisiae powder and corn starch are put into a mixing bin and mixed for 5-8 minutes to obtain the microbial preparation.

[0157] Example 5

[0158] (1) A formula of a microbial preparation suitable for saline-alkali soil

[0159] A microbial preparation suitable for saline-alkali soil, which comprises 80 parts of yeast metabolite, 25 parts of Bacillus subtilis powder, 30 parts of Saccharomyces cerevisiae powder and 10 parts of corn starch by weight. The content of fulvic acid in the yeast metabolite is 26.12%, and the content of betaine is 11.9%.

[0160] The Bacillus subtilis in the Bacillus subtilis powder is Bacillus subtilis subsp. subtilis tc-4. The preparation method of the Bacillus subtilis powder is the same as the preparation method of the Bacillus subtilis powder in Embodiment 1.

[0161] The Saccharomyces cerevisiae in the Saccharomyces cerevisiae powder is Saccharomyces cerevisiae high alcohol yeast YY. The preparation method of the Saccharomyces cerevisiae powder is the same as the preparation method of the Saccharomyces cerevisiae powder in Embodiment 1.

[0162] (1) A formula of a microbial preparation suitable for saline-alkali soil

[0163] A preparation method of a microbial preparation suitable for saline-alkali soil, comprising the following steps:

[0164] The yeast metabolite, the Bacillus subtilis powder, the Saccharomyces cerevisiae powder and the corn starch are put into the mixing bin, mixed for 5-8 minutes, and the microbial preparation is obtained.

[0165] Embodiment 6

[0166] Compared with Embodiment 1, the Bacillus subtilis powder prepared from Bacillus subtilis subsp. subtilis tc-4 is replaced by a commercially available Bacillus subtilis powder (powder, product name: Bacillus subtilis, selling company: Shandong Blue Bio-technology Co., Ltd.). The specific process is as follows:

[0167] (1) A formula of a microbial preparation suitable for saline-alkali soil

[0168] A microbial preparation suitable for saline-alkali soil, comprising 80 parts of yeast metabolite, 5 parts of Bacillus subtilis powder (commercially available), 5 parts of Saccharomyces cerevisiae powder and 10 parts of corn starch by weight. The content of fulvic acid in the yeast metabolite is 26.12%, and the content of betaine is 11.9%.

[0169] The Bacillus subtilis powder is a commercially available Bacillus subtilis powder, and the selling company is Shandong Blue Bio-technology Co., Ltd.

[0170] The Saccharomyces cerevisiae in the Saccharomyces cerevisiae powder is Saccharomyces cerevisiae high alcohol yeast YY. The preparation method of the Saccharomyces cerevisiae powder is the same as the preparation method of the Saccharomyces cerevisiae powder in Embodiment 1.

[0171] (2) A preparation method of a microbial preparation suitable for saline-alkali soil

[0172] A preparation method of a microbial preparation suitable for saline-alkali soil, comprising the following steps:

[0173] The above yeast metabolites, Bacillus subtilis powder (commercially available), Saccharomyces cerevisiae powder and corn starch are put into a mixing bin and mixed for 5-8 min to obtain the microbial preparation.

[0174] Example 7

[0175] Compared with Example 4, Saccharomyces cerevisiae high alcohol yeast YY (Saccharomyces cerevisiae high alcohol yeast YY) in the Saccharomyces cerevisiae powder is replaced by Saccharomyces cerevisiae d8.8 (Saccharomyces cerevisiae d8.8), which was preserved in the China Center for Type Culture Collection (CCTCC) on March 29, 2017, with the preservation number of CCTCC NO: M2017148 and the preservation address of Wuhan University, Wuhan, China, with the postal code of 430072 and the telephone number of (027) 68754052. The Saccharomyces cerevisiae d8.8 is disclosed in Chinese Patent No. 201710519421.9. The details are as follows:

[0176] (1) A formula of a microbial preparation suitable for saline-alkali soil

[0177] A microbial preparation suitable for saline-alkali soil, comprising 80 parts of yeast metabolites, 25 parts of Bacillus subtilis powder, 30 parts of Saccharomyces cerevisiae powder and 10 parts of corn starch by weight. Among them, the content of fulvic acid in the yeast metabolites is 26.12%, and the content of betaine is 11.9%.

[0178] Among them, the Bacillus subtilis in the Bacillus subtilis powder is Bacillus subtilis subsp. subtilis tc-4 (Bacillus subtilis subsp. subtilis tc-4). The preparation method of the Bacillus subtilis powder is the same as the preparation method of the Bacillus subtilis powder in Example 1.

[0179] Among them, the Saccharomyces cerevisiae in the Saccharomyces cerevisiae powder is Saccharomyces cerevisiae d8.8 (Saccharomyces cerevisiae d8.8).

[0180] The preparation method of the Saccharomyces cerevisiae powder comprises the following steps:

[0181] Step 1: Using an inoculation loop to dip the Saccharomyces cerevisiae d8.8 bacterial solution preserved in glycerol at a temperature of -80℃, performing streaking on YPD agar, and obtaining single colonies after 24h culture at a temperature of 30℃.

[0182] Step 2: inoculate a complete single colony into the medium used for the first-stage seed liquid and culture at a temperature of 30℃ for 15h to obtain the first-stage seed liquid of the yeast. The preparation method of the medium used for the first-stage seed liquid comprises: dissolving 100g of sucrose, 20g of yeast extract powder, 1g of magnesium sulfate and 1g of zinc sulfate in 1L of water, and adjusting the pH to 4.8 with concentrated sulfuric acid.

[0183] The first-stage seed liquid of the yeast is inoculated into the medium used for the second-stage seed liquid at an inoculation amount of 1% to 3% by volume of the medium, and cultured at a temperature of 30℃ for 15h. The culture liquid is separated by a separator, and the precipitate is washed with water to obtain the seed liquid of the yeast (also referred to as the second-stage seed liquid). The preparation method of the medium used for the second-stage seed liquid comprises: taking 8L of sugar beet molasses and adjusting the pH to 5.5 with concentrated sulfuric acid.

[0184] Step 3: the seed liquid of the yeast is inoculated into a fermentation tank at an inoculation amount of 1% to 3% by volume of the medium, and fermented at a temperature of 30℃ for 15h to obtain the yeast liquid. The preparation method of the medium in the fermentation tank comprises the following steps: taking sugar beet molasses and adjusting the pH to 5.5 with concentrated sulfuric acid. The yeast liquid is separated by a separator, the precipitate is washed with water, and then drum extrusion drying is performed to obtain the yeast powder.

[0185] (2) A preparation method of a microbial preparation suitable for saline-alkali soil

[0186] A preparation method of a microbial preparation suitable for saline-alkali soil, comprising the following steps:

[0187] The above-mentioned yeast metabolites, Bacillus subtilis powder, Saccharomyces cerevisiae powder (strain: Saccharomyces cerevisiae d8.8) and corn starch are put into a mixing bin and mixed for 5 to 8min to obtain the microbial preparation.

[0188] Application Example 1: Stability Experiment

[0189] The evaluation of stability comprises the following steps: the microbial preparation prepared in the example is stored at room temperature indoors, and is stored in a woven bag and an inner film bag from inside to outside. The data of 12 months of storage are determined by sampling and testing once a month. The effective viable cell count is determined by referring to the plate counting method in the Agricultural Industry Standard of the People's Republic of China NY / T 2321-2013 Microbial Fertilizer Product Test Procedure, the survival rate and the coefficient of variation (i.e. CV value) of one year of storage are calculated, and the calculation method is as follows:

[0190]

[0191] Wherein, X is survival rate, unit is %, A is the effective viable count of the microbial preparation prepared to be stored for 12 months, unit is CFU / g; B is the initial effective viable count of the microbial preparation prepared (the initial effective viable count refers to the microbial preparation with storage time of 0 month), unit is CFU / g.

[0192]

[0193] Wherein, CV is coefficient of variation, unit is %, SD is standard deviation of the effective viable count for 12 months; Mean is the monthly average value of the effective viable count for 12 months.

[0194] Table 3 stability index results of the microbial preparation prepared in the examples

[0195]

[0196] From the above table 3, in the microbial preparation prepared in the examples 1-7, the survival rate of Bacillus subtilis reaches more than 75%, and the CV value is within 15%; the survival rate of Saccharomyces cerevisiae reaches more than 59%, and the CV value is within 15.1%. Among them, in the microbial preparation prepared in the examples 4-7, the survival rate of Bacillus subtilis is 75.0%-79.9%, the CV value is 10.9%-13.7%, the survival rate of Saccharomyces cerevisiae is 59.5%-72.9%, and the CV value is 11.8%-15.1%, which shows that the stability of the microbial preparation prepared in the examples 4-7 is good. In the microbial preparation prepared in the examples 1-3, the survival rate of Bacillus subtilis is 85.0%-88.3%, the CV value is 8.9%-9.6% (less than 10%), the survival rate of Saccharomyces cerevisiae is 73.6%-84.0%, and the CV value is 9.3%-9.8% (less than 10%), the effective viable count data of Bacillus subtilis and Saccharomyces cerevisiae fluctuate less, which shows that the stability of the microbial preparation obtained is better. The above data shows that the raw material components and their contents are in the preferred range, which can achieve more excellent technical effects.

[0197] Application example 2: fertility experiment

[0198] Experimental sample: the microbial preparation sample prepared in the examples 1-7.

[0199] Experimental method: the alkaline nutrients of the test plot in Xilicun, Zhanhua district, Binzhou city of Shandong province are classified, which belongs to middle and lower level, and the soil is severely alkaline and slightly salinized. The planted crop is soybean, and the variety is Zhonghuang 13.

[0200] The test plot is divided into eight small plots with the same size, and the area of each small plot is 30m 2, the base fertilizer is conventional fertilization (compound fertilizer and organic fertilizer). In the early stage of soybean planting (July-August), the seven experimental samples prepared in Examples 1-7 are applied at a rate of 1000 g / mu, and are applied twice by root irrigation. After the soybean is harvested, soil samples are taken for physicochemical property analysis.

[0201] Table 4: Result data of soil saline-alkali physicochemical properties

[0202]

[0203] In Table 4, CK represents the control group, i.e., only conventional fertilization without adding the microbial preparation of the application, and the rest of the conditions are the same as the examples.

[0204] As can be seen from Table 4 above, compared with CK, after adding the microbial preparation of the application, the total nutrient and organic matter content are increased, and the conductivity, soluble salt content and pH value are all decreased, which shows that the microbial preparation of the application has the effect of improving soil fertility and improving soil alkalinity. Among them, in Examples 4-7, the total nutrient content is increased by 5.90%-10.69%, the organic matter content is increased by 2.06%-7.94%, the conductivity is decreased by 0.61%-2.03%, the soluble salt content is decreased by 3.22%-9.68%, and the pH value is decreased by 2.76%-6.28%. The above data show that the microbial preparation prepared in Examples 4-7 has the effect of improving soil fertility and improving saline-alkali soil. In Examples 1-3, the total nutrient content is increased by 16.01%-21.30%, the organic matter content is increased by 8.35%-12.37%, the conductivity is decreased by 1.09%-22.29%, the soluble salt content is decreased by 6.45%-15.81%, and the pH value is decreased by 7.72%-10.80%. The above data show that the microbial preparation prepared in Examples 1-3 has the effect of improving soil fertility and improving soil alkalinity. The above data show that the raw material components and their contents are within the preferred range, and better technical effects can be achieved.

[0205] Application Example 3: Growth Promotion Experiment

[0206] Experimental samples: microbial preparation samples prepared in Examples 1-7.

[0207] Experimental method: potting method is adopted, and root irrigation treatment is carried out when the soybean sprouts 2 cm. The control group CK is treated with water, and the seven treatment groups are treated with the microbial preparation samples prepared in Example 1, respectively. Three repetitions are set for each treatment group. 2 g of the microbial preparation experimental sample prepared in Example is weighed, diluted 500 times, and 200 mL of the experimental sample solution is taken for each pot for root irrigation. The treatment is repeated after 7 days, and the relevant growth data is counted after 14 days. The specific data are shown in Table 5 and Figure 1 .

[0208] Table 5: Result data of soybean growth promotion experiment

[0209] Item Soybean stem length / cm CK 9.23±0.64 Example 1 13.13±0.95 Example 2 12.05±1.12 Example 3 12.53±1.08 Example 4 11.53±0.62 Example 5 10.40±0.9 Example 6 9.60±1.12 Example 7 10.00±1.05

[0210] Figure 1 Table 5: Result data of soybean stem length after applying microbial preparation prepared by examples 1-7. Figure 1 As shown in Table 5, after applying microbial preparation prepared by examples of the present application, the stem length of soybean is increased. The stem length of soybean in examples 1-3 is 12.05-13.13 cm, which is increased by 30.55%-42.25% compared with the blank control group CK, and is greater than 30%; the stem length of soybean in examples 4-7 is 9.60-11.53 cm, which is increased by 4.00%-24.92% compared with the blank control group CK. The above data show that the growth promotion effect of examples 1-3 on soybean stem length is more significant than that of examples 4-7; this indicates that the raw material components and their contents in the preferred range can achieve more excellent technical effects.

[0211] Application Example 4: Stress Resistance Experiment

[0212] Experimental sample: microbial preparation samples prepared by examples 1-7.

[0213] Experimental method: potting method was used, and root irrigation experiment was carried out at the soybean seedling stage. Two control groups were set, which were water control group CK1 and salt water control group CK2. Seven treatment groups were treated with microbial preparation samples prepared by examples 1-7, and three replicates were set for each treatment group. Except for the water control group CK1, the salt water control group CK2 and examples 1-7 were treated with 200 mL of 0.6% salt water once at 2 cm of soybean emergence. 2 g of microbial preparation experimental sample prepared by examples was weighed, diluted 500 times, and 200 mL of experimental sample solution was taken for each pot for root irrigation. The treatment was repeated after 7 days, and the growth data was counted after 14 days. The specific data are shown in Table 6 and Figure 2

[0214] Table 6: Result data of soybean stress resistance experiment

[0215] Item Soybean stem length / cm CK1 8.25±0.50 CK2 4.07±0.76 Example 1 9.60±0.49 Example 2 8.28±0.22 Example 3 8.58±0.48 Example 4 7.70±0.24 Example 5 8.00±0.26 Example 6 7.70±0.79 Example 7 7.60±0.17

[0216] Figure 2 Table 6: Result data of soybean stem length after applying microbial preparation prepared by examples 1-7 in simulated saline-alkali soil. As shown in Table 6, after applying microbial preparation prepared by examples of the present application, the stem length of soybean is increased. The stem length of soybean in examples 1-3 is 12.05-13.13 cm, which is increased by 30.55%-42.25% compared with the blank control group CK, and is greater than 30%; the stem length of soybean in examples 4-7 is 9.60-11.53 cm, which is increased by 4.00%-24.92% compared with the blank control group CK. The above data show that the growth promotion effect of examples 1-3 on soybean stem length is more significant than that of examples 4-7; this indicates that the raw material components and their contents in the preferred range can achieve more excellent technical effects. Figure 2 ​As shown in Table 6, after applying the microbial preparation prepared by the embodiments of the present application, the stem length of the soybean growing in saline-alkali soil is improved. Among them, the stem length of the soybean in the example 1-3 group is 8.28-9.60 cm, which is increased by 103.44%-135.87% compared with the salt water control group CK2, and the growth is consistent with the water control group CK1, and the example 1 group is better than the water control group CK1. The stem length of the soybean in the example 4-7 group is 7.60-8.00 cm, which is increased by 86.73%-96.56% compared with the salt water control group CK2, and the growth is equivalent to the water control group CK1. The above data shows that the growth-promoting effect of the example 1-3 group on the soybean growing in saline-alkali soil is more significant, and the stress resistance effect is more excellent than that of the example 4-7 group; this shows that the raw material components and their contents in the preferred range can achieve more excellent technical effects.

[0217] Although the specific embodiments of the present application are described above, they are not intended to limit the scope of protection of the present application. Various modifications or variations made by those skilled in the art on the basis of the technical solutions of the present application without creative labor are still within the scope of protection of the present application.

Claims

1. A microbial preparation suitable for saline-alkali soil, characterized in that, By weight, it includes: 4-25 parts Bacillus subtilis, 4-30 parts Saccharomyces cerevisiae, 40-85 parts synergistic carrier and 5-20 parts protective carrier; Wherein, the enhancing carrier is yeast metabolite; the protective carrier is corn starch and / or attapulgite; and the Bacillus subtilis is Bacillus subtilis subsp. tc-4 (…). Bacillus subtilis subsp subtilis tc-4), with accession number CCTCC NO: M 2023541; the brewing yeast strain is brewing yeast high-alcohol yeast YY ( Saccharomyces cerevisiae The yeast strain is *Saccharomyces cerevisiae* YY, with accession number CCTCC NO: M 2021171; or the yeast strain is *Saccharomyces cerevisiae* d8.8 (…). Saccharomyces cerevisiae d8.8), with accession number CCTCC NO: M2017148.

2. The microbial preparation suitable for saline-alkali soil according to claim 1, wherein, The microbial preparation comprises, by weight, 4-20 parts of Bacillus subtilis, and / or 4-20 parts of Saccharomyces cerevisiae, and / or 60-85 parts of synergistic carrier, and / or 5-15 parts of protective carrier.

3. The microbial preparation suitable for saline-alkali soil according to claim 1, wherein, The microbial preparation comprises, by weight, 4-13 parts of Bacillus subtilis, and / or 4-6 parts of Saccharomyces cerevisiae, and / or 75-85 parts of synergistic carrier, and / or 5-12 parts of protective carrier.

4. The microbial preparation suitable for saline-alkali soil according to claim 1, wherein, The microbial preparation comprises, by weight, 4-6 parts of Bacillus subtilis, and / or 78-82 parts of synergistic carrier and 8-11 parts of protective carrier.

5. The microbial preparation suitable for saline-alkali soil according to claim 1, wherein, The yeast metabolites are the metabolic products of yeast extracted from molasses through fermentation.

6. The microbial preparation suitable for saline-alkali soil according to claim 2, wherein, The yeast metabolites are the metabolic products of yeast extracted from molasses through fermentation.

7. The microbial preparation suitable for saline-alkali soil according to claim 3, wherein, The yeast metabolites are the metabolic products of yeast extracted from molasses through fermentation.

8. The microbial preparation suitable for saline-alkali soil according to claim 4, wherein, The yeast metabolites are the metabolic products of yeast extracted from molasses through fermentation.

9. The microbial preparation suitable for saline-alkali soil according to claim 5, wherein, The molasses includes beet molasses.

10. The microbial preparation suitable for saline-alkali soil according to claim 6, wherein, The molasses includes beet molasses.

11. The microbial preparation suitable for saline-alkali soil according to claim 7, wherein, The molasses includes beet molasses.

12. The microbial preparation suitable for saline-alkali soil according to claim 8, wherein, The molasses includes beet molasses.

13. The microbial preparation suitable for saline-alkali soil according to claim 1, wherein, The yeast metabolites contain 15% to 30% fulvic acid by dry weight, and / or the yeast metabolites contain 6% to 15% betaine.

14. The microbial preparation suitable for saline-alkali soil according to claim 2, wherein, The yeast metabolites contain 15% to 30% fulvic acid by dry weight, and / or the yeast metabolites contain 6% to 15% betaine.

15. The microbial preparation suitable for saline-alkali soil according to claim 3, wherein, The yeast metabolites contain 15% to 30% fulvic acid by dry weight, and / or the yeast metabolites contain 6% to 15% betaine.

16. The microbial preparation suitable for saline-alkali soil according to claim 4, wherein, The yeast metabolites contain 15% to 30% fulvic acid by dry weight, and / or the yeast metabolites contain 6% to 15% betaine.

17. The microbial preparation suitable for saline-alkali soil according to claim 5, wherein, The yeast metabolites contain 15% to 30% fulvic acid by dry weight, and / or the yeast metabolites contain 6% to 15% betaine.

18. The microbial preparation suitable for saline-alkali soil according to claim 6, wherein, The yeast metabolites contain 15% to 30% fulvic acid by dry weight, and / or the yeast metabolites contain 6% to 15% betaine.

19. The microbial preparation suitable for saline-alkali soil according to claim 7, wherein, The yeast metabolites contain 15% to 30% fulvic acid by dry weight, and / or the yeast metabolites contain 6% to 15% betaine.

20. The microbial preparation suitable for saline-alkali soil according to claim 8, wherein, The yeast metabolites contain 15% to 30% fulvic acid by dry weight, and / or the yeast metabolites contain 6% to 15% betaine.

21. The microbial preparation suitable for saline-alkali soil according to claim 9, wherein, The yeast metabolites contain 15% to 30% fulvic acid by dry weight, and / or the yeast metabolites contain 6% to 15% betaine.

22. The microbial preparation suitable for saline-alkali soil according to claim 10, wherein, The yeast metabolites contain 15% to 30% fulvic acid by dry weight, and / or the yeast metabolites contain 6% to 15% betaine.

23. The microbial preparation suitable for saline-alkali soil according to claim 11, wherein, The yeast metabolites contain 15% to 30% fulvic acid by dry weight, and / or the yeast metabolites contain 6% to 15% betaine.

24. The microbial preparation suitable for saline-alkali soil according to claim 12, wherein, The yeast metabolites contain 15% to 30% fulvic acid by dry weight, and / or the yeast metabolites contain 6% to 15% betaine.

25. The microbial preparation suitable for saline-alkali soil according to any one of claims 1-24, wherein, The effective viable count of Bacillus subtilis in the microbial preparation is 4 billion to 25 billion CFU / g, and / or the effective viable count of Saccharomyces cerevisiae in the microbial preparation is 800 million to 6 billion CFU / g.

26. The microbial preparation suitable for saline-alkali soil according to any one of claims 1-24, wherein, The effective viable count of Bacillus subtilis in the microbial preparation is 4 billion to 20 billion CFU / g, and / or the effective viable count of Saccharomyces cerevisiae in the microbial preparation is 800 million to 4 billion CFU / g.

27. The microbial preparation suitable for saline-alkali soil according to any one of claims 1-24, wherein, The Bacillus subtilis is Bacillus subtilis powder, and its preparation method includes the following steps: Step 1: Activation of Bacillus subtilis; Step 2: Inoculate the activated Bacillus subtilis into the culture medium for cultivation to obtain seed culture; and Step 3: Inoculate the seed liquid into the fermentation medium for cultivation to obtain fermentation broth, and then dry it to obtain Bacillus subtilis powder.

28. The microbial preparation suitable for saline-alkali soil according to claim 25, wherein, The Bacillus subtilis is Bacillus subtilis powder, and its preparation method includes the following steps: Step 1: Activation of Bacillus subtilis; Step 2: Inoculate the activated Bacillus subtilis into the culture medium for cultivation to obtain seed culture; and Step 3: Inoculate the seed liquid into the fermentation medium for cultivation to obtain fermentation broth, and then dry it to obtain Bacillus subtilis powder.

29. The microbial preparation suitable for saline-alkali soil according to claim 27, wherein, In step 1, the activation medium consists of 0.5%-1.5% peptone, 0.1%-0.5% beef extract, 0.3%-0.7% sodium chloride, and 1.5%-2% agar, with the remainder being water, based on the mass of the medium used for activation.

30. The microbial preparation suitable for saline-alkali soil according to claim 28, wherein, In step 1, the activation medium consists of 0.5%-1.5% peptone, 0.1%-0.5% beef extract, 0.3%-0.7% sodium chloride, and 1.5%-2% agar, with the remainder being water, based on the mass of the medium used for activation.

31. The microbial preparation suitable for saline-alkali soil according to claim 27, wherein, In step 1, the temperature during activation is 35~39℃, and / or the activation time is 21~27h.

32. The microbial preparation suitable for saline-alkali soil according to claim 28, wherein, In step 1, the temperature during activation is 35~39℃, and / or the activation time is 21~27h.

33. The microbial preparation suitable for saline-alkali soil according to claim 27, wherein, In step 1, the cultured Bacillus subtilis is further activated.

34. The microbial preparation suitable for saline-alkali soil according to claim 28, wherein, In step 1, the cultured Bacillus subtilis is further activated.

35. The microbial preparation suitable for saline-alkali soil according to claim 33, wherein, The culture medium used for further activation comprises, by weight, 0.5% to 1.5% peptone, 0.1% to 0.5% beef extract, 0.3% to 0.7% sodium chloride, and 1.5% to 2% agar, with the remainder being water.

36. The microbial preparation suitable for saline-alkali soil according to claim 34, wherein, The culture medium used for further activation comprises, by weight, 0.5% to 1.5% peptone, 0.1% to 0.5% beef extract, 0.3% to 0.7% sodium chloride, and 1.5% to 2% agar, with the remainder being water.

37. The microbial preparation suitable for saline-alkali soil according to claim 33, wherein, The temperature for the further activation culture is 35~39℃, and / or the time for the further activation culture is 21~27h.

38. The microbial preparation suitable for saline-alkali soil according to claim 34, wherein, The temperature for the further activation culture is 35~39℃, and / or the time for the further activation culture is 21~27h.

39. The microbial preparation suitable for saline-alkali soil according to claim 27, wherein, In step 2, the culture medium, by total mass, includes 0.5%~1.5% peptone, 0.1%~0.5% beef meal, 0.3%~0.7% sodium chloride and 0.05%~0.15% glucose, with the remainder being water.

40. The microbial preparation suitable for saline-alkali soil according to claim 28, wherein, In step 2, the culture medium, by total mass, includes 0.5%~1.5% peptone, 0.1%~0.5% beef meal, 0.3%~0.7% sodium chloride and 0.05%~0.15% glucose, with the remainder being water.

41. The microbial preparation suitable for saline-alkali soil according to claim 27, wherein, In step 2, the culture temperature is 35~39℃, and / or the culture time is 10~15h, and / or the rotation speed during culture is 100~200rpm.

42. The microbial preparation suitable for saline-alkali soil according to claim 28, wherein, In step 2, the culture temperature is 35~39℃, and / or the culture time is 10~15h, and / or the rotation speed during culture is 100~200rpm.

43. The microbial preparation suitable for saline-alkali soil according to claim 27, wherein, In step 3, the culture medium, by total mass, includes 3%~5% corn flour, 3%~5% soybean meal, 0.05%~0.15% magnesium sulfate and 0.05%~0.15% calcium chloride, with the remainder being water.

44. The microbial preparation suitable for saline-alkali soil according to claim 28, wherein, In step 3, the culture medium, by total mass, includes 3%~5% corn flour, 3%~5% soybean meal, 0.05%~0.15% magnesium sulfate and 0.05%~0.15% calcium chloride, with the remainder being water.

45. The microbial preparation suitable for saline-alkali soil according to claim 27, wherein, In step 3, the fermentation medium is cultured at a temperature of 35~39℃, and / or the fermentation medium is cultured for 15~20h.

46. ​​The microbial preparation suitable for saline-alkali soil according to claim 28, wherein, In step 3, the fermentation medium is cultured at a temperature of 35~39℃, and / or the fermentation medium is cultured for 15~20h.

47. The microbial preparation suitable for saline-alkali soil according to claim 27, wherein, In step 3, the aeration rate of the fermentation medium is 20~50L / min, and / or the pressure of the fermentation medium is 0.2~0.5 standard atmospheres.

48. The microbial preparation suitable for saline-alkali soil according to claim 28, wherein, In step 3, the aeration rate of the fermentation medium is 20~50L / min, and / or the pressure of the fermentation medium is 0.2~0.5 standard atmospheres.

49. The microbial preparation suitable for saline-alkali soil according to any one of claims 1-24, wherein, The brewer's yeast is brewer's yeast powder, and its preparation method includes the following steps: Step 1: Activation of brewer's yeast; Step 2: Inoculate the activated Saccharomyces cerevisiae into the culture medium for cultivation to obtain seed culture; and Step 3: Inoculate the seed liquid into the fermentation medium for culture to obtain fermentation broth. After separation and drying, obtain the brewing yeast powder.

50. The microbial preparation suitable for saline-alkali soil according to claim 25, wherein, The brewer's yeast is brewer's yeast powder, and its preparation method includes the following steps: Step 1: Activation of brewer's yeast; Step 2: Inoculate the activated Saccharomyces cerevisiae into the culture medium for cultivation to obtain seed culture; and Step 3: Inoculate the seed liquid into the fermentation medium for culture to obtain fermentation broth. After separation and drying, obtain the brewing yeast powder.

51. The microbial preparation suitable for saline-alkali soil according to claim 49, wherein, In step 1, the activation medium consists of 0.5% to 1.5% peptone, 1.5% to 2.5% glucose, 0.1% to 1% yeast extract, and 1.1% to 1.7% agar, with the remainder being water.

52. The microbial preparation suitable for saline-alkali soil according to claim 50, wherein, In step 1, the activation medium consists of 0.5% to 1.5% peptone, 1.5% to 2.5% glucose, 0.1% to 1% yeast extract, and 1.1% to 1.7% agar, with the remainder being water.

53. The microbial preparation suitable for saline-alkali soil according to claim 49, wherein, In step 1, the activation culture temperature is 28~32℃, and / or the activation culture time is 21~27h.

54. The microbial preparation suitable for saline-alkali soil according to claim 50, wherein, In step 1, the activation culture temperature is 28~32℃, and / or the activation culture time is 21~27h.

55. The microbial preparation suitable for saline-alkali soil according to claim 49, wherein, In step 2, the activated Saccharomyces cerevisiae are inoculated into the culture medium used for the primary seed culture and cultured to obtain the primary seed culture. Then, the primary seed culture is inoculated into the culture medium used for the secondary seed culture and cultured to obtain the secondary seed culture.

56. The microbial preparation suitable for saline-alkali soil according to claim 50, wherein, In step 2, the activated Saccharomyces cerevisiae are inoculated into the culture medium used for the primary seed culture and cultured to obtain the primary seed culture. Then, the primary seed culture is inoculated into the culture medium used for the secondary seed culture and cultured to obtain the secondary seed culture.

57. The microbial preparation suitable for saline-alkali soil according to claim 55, wherein, The culture conditions for the primary seed solution are: a culture temperature of 28~32℃ and / or a culture time of 12~18h.

58. The microbial preparation suitable for saline-alkali soil according to claim 56, wherein, The culture conditions for the primary seed solution are: a culture temperature of 28~32℃ and / or a culture time of 12~18h.

59. The microbial preparation suitable for saline-alkali soil according to claim 55, wherein, The culture medium used for the primary seed culture consists of 90-110g sucrose, 18-22g yeast extract, 0.5-1.5g magnesium sulfate, and 0.5-1.5g zinc sulfate per liter of water.

60. The microbial preparation suitable for saline-alkali soil according to claim 56, wherein, The culture medium used for the primary seed culture consists of 90-110g sucrose, 18-22g yeast extract, 0.5-1.5g magnesium sulfate, and 0.5-1.5g zinc sulfate per liter of water.

61. The microbial preparation suitable for saline-alkali soil according to claim 55, wherein, The inoculation amount of the primary seed culture is 1% to 3% based on the volume of the culture medium.

62. The microbial preparation suitable for saline-alkali soil according to claim 56, wherein, The inoculation amount of the primary seed culture is 1% to 3% based on the volume of the culture medium.

63. The microbial preparation suitable for saline-alkali soil according to claim 55, wherein, The temperature at which the primary seed culture is inoculated into the culture medium for the secondary seed culture is 28-32°C, and / or the inoculation time of the primary seed culture into the culture medium for the secondary seed culture is 12-18 hours.

64. The microbial preparation suitable for saline-alkali soil according to claim 56, wherein, The temperature at which the primary seed culture is inoculated into the culture medium for the secondary seed culture is 28-32°C, and / or the inoculation time of the primary seed culture into the culture medium for the secondary seed culture is 12-18 hours.

65. The microbial preparation suitable for saline-alkali soil according to claim 55, wherein, The culture medium used for the secondary seed liquid includes beet molasses.

66. The microbial preparation suitable for saline-alkali soil according to claim 56, wherein, The culture medium used for the secondary seed liquid includes beet molasses.

67. The microbial preparation suitable for saline-alkali soil according to claim 49, wherein, In step 3, the inoculation amount of the seed solution is 1% to 3% based on the volume of the culture medium.

68. The microbial preparation suitable for saline-alkali soil according to claim 50, wherein, In step 3, the inoculation amount of the seed solution is 1% to 3% based on the volume of the culture medium.

69. The microbial preparation suitable for saline-alkali soil according to claim 49, wherein, In step 3, the fermentation medium includes beet molasses.

70. The microbial preparation suitable for saline-alkali soil according to claim 50, wherein, In step 3, the fermentation medium includes beet molasses.

71. The microbial preparation suitable for saline-alkali soil according to claim 49, wherein, In step 3, the temperature at which the seed culture is inoculated into the fermentation medium is 28~32℃, and / or the time for inoculating the seed culture into the fermentation medium is 12~18h.

72. The microbial preparation suitable for saline-alkali soil according to claim 50, wherein, In step 3, the temperature at which the seed culture is inoculated into the fermentation medium is 28~32℃, and / or the time for inoculating the seed culture into the fermentation medium is 12~18h.

73. A method for preparing a microbial agent suitable for saline-alkali soil according to any one of claims 1-72, characterized in that, Includes the following steps: A microbial preparation is obtained by mixing an enhancing carrier, Bacillus subtilis, Saccharomyces cerevisiae, and a protective carrier, wherein the Bacillus subtilis is Bacillus subtilis subsp. tc-4. Bacillus subtilis subsp subtilis tc-4), with accession number CCTCC NO: M 2023541; the brewing yeast strain is brewing yeast high-alcohol yeast YY ( Saccharomyces cerevisiae The yeast strain is *Saccharomyces cerevisiae* YY, with accession number CCTCC NO: M 2021171; or the yeast strain is *Saccharomyces cerevisiae* d8.8 (…). Saccharomyces cerevisiae d8.8), with accession number CCTCC NO: M2017148.

74. A microbial preparation suitable for saline-alkali soil, characterized in that, It is prepared by the method for preparing microbial agents suitable for saline-alkali soil as described in claim 73.

75. The application of a microbial preparation suitable for saline-alkali soil in the improvement of saline-alkali soil, characterized in that, The microbial preparation suitable for saline-alkali soil is the microbial preparation suitable for saline-alkali soil according to any one of claims 1-72 or the microbial preparation suitable for saline-alkali soil obtained by the preparation method according to claim 73.

76. The application of the microbial preparation for saline-alkali soil according to claim 75 in the improvement of saline-alkali soil, wherein the improvement of saline-alkali soil includes improving soil fertility, reducing soil alkalinity and / or reducing soil electrical conductivity.

77. The application according to claim 75 in improving saline-alkali soil, wherein, The application rate of the microbial agent suitable for saline-alkali soil is 10-20 kg / hm. 2 .

78. The application of the microbial preparation suitable for saline-alkali soil according to claim 76 in the improvement of saline-alkali soil, wherein, The application rate of the microbial agent suitable for saline-alkali soil is 10-20 kg / hm. 2 .

79. The application of the microbial preparation suitable for saline-alkali soil according to any one of claims 75-78 in the improvement of saline-alkali soil, wherein the microbial preparation is used by drip irrigation or fertigation and is suitable for the seedling growth period of crops.

80. The application of a microbial preparation suitable for saline-alkali soil in promoting crop growth and / or enhancing crop salt and alkali tolerance, characterized in that, The microbial preparation suitable for saline-alkali soil is any one of the microbial preparations suitable for saline-alkali soil according to claims 1-72 or the microbial preparation suitable for saline-alkali soil prepared by the method described in claim 73.

81. The application of the microbial preparation suitable for saline-alkali soil according to claim 80 in promoting crop growth and / or enhancing crop salt tolerance, wherein the crop is a legume.

82. The application of the microbial preparation suitable for saline-alkali soil according to claim 81 in promoting crop growth and / or enhancing crop salt and alkali tolerance, wherein the legume crop is soybean.

83. The application according to any one of claims 80-82 in promoting crop growth and / or enhancing crop salt and alkali tolerance, wherein, The application rate of the microbial agent suitable for saline-alkali soil is 10-20 kg / hm. 2 .

84. The application according to any one of claims 80-82 in promoting crop growth and / or enhancing crop salt and alkali tolerance, wherein, The microbial preparation is used for drip irrigation or fertigation and is suitable for the seedling stage of crops.

85. The application according to claim 83 in promoting crop growth and / or enhancing crop salt and alkali tolerance, wherein, The microbial preparation is used for drip irrigation or fertigation and is suitable for the seedling stage of crops.

86. A composition for improving saline-alkali soil, characterized in that, The composition for improving saline-alkali soil includes the microbial preparation suitable for saline-alkali soil as described in any one of claims 1-72 or the microbial preparation suitable for saline-alkali soil prepared by the method described in claim 73.

87. The composition according to claim 86, wherein, The improvement of saline-alkali soil includes enhancing soil fertility, reducing soil alkalinity and / or reducing soil electrical conductivity.

88. A composition for promoting crop growth and / or enhancing crop salt and alkali tolerance, characterized in that, The composition for crop growth and / or enhancing crop salt tolerance includes the microbial preparation suitable for saline-alkali soil as described in any one of claims 1-72 or the microbial preparation suitable for saline-alkali soil prepared by the method described in claim 73.

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