阅读说明：本技术 聚-γ-谷氨酸作为肥料增效剂在农作物中的应用 (Application of poly-gamma-glutamic acid as fertilizer synergist in crops ) 是由 彭伟 曾庆茁 李强 宋巍 于 2021-08-31 设计创作，主要内容包括：本发明属于肥料增效剂领域,具体公开了聚-γ-谷氨酸作为肥料增效剂在农作物中的应用,包括在农作物栽种前整地时向土壤施聚-γ-谷氨酸,和在农作物生长期对植株地上部喷施聚-γ-谷氨酸；所述土施聚-γ-谷氨酸的施用量为每公顷1-3kg,并与常规肥料混合均匀后一起撒施；所述作物生长期,每公顷作物喷施聚-γ-谷氨酸2.5-7.5kg。本发明公开的聚-γ-谷氨酸作为肥料增效剂,施肥量小,增产效果好,营养全面、成分明确可控,制备方法简单,可大规模工业化制备,适合用于谷物,果蔬类常见作物的栽种。(The invention belongs to the field of fertilizer synergists, and particularly discloses application of poly-gamma-glutamic acid as a fertilizer synergist in crops, which comprises the steps of applying the poly-gamma-glutamic acid to soil during soil preparation before crop planting, and spraying the poly-gamma-glutamic acid on the overground part of a plant in the growth period of the crops; the application amount of the poly-gamma-glutamic acid applied to the soil is 1-3kg per hectare, and the poly-gamma-glutamic acid applied to the soil and the conventional fertilizer are uniformly mixed and then are applied together; in the growth period of the crops, 2.5-7.5kg of poly-gamma-glutamic acid is sprayed on each hectare of the crops. The poly-gamma-glutamic acid disclosed by the invention is used as a fertilizer synergist, has the advantages of small fertilizing amount, good yield increasing effect, comprehensive nutrition, definite and controllable components, simple preparation method, large-scale industrial preparation and suitability for planting common crops such as grains, fruits and vegetables.)

1. The application of the poly-gamma-glutamic acid as a fertilizer synergist in crops is characterized by comprising the steps of applying the poly-gamma-glutamic acid to soil when preparing soil before the crops are planted, and spraying the poly-gamma-glutamic acid on the overground part of a plant in the growth period of the crops;

the application amount of the poly-gamma-glutamic acid applied to the soil is 1-3kg per hectare, and the poly-gamma-glutamic acid applied to the soil and the conventional fertilizer are uniformly mixed and then are applied together;

in the growth period of the crops, 2.5-7.5kg of poly-gamma-glutamic acid is sprayed on each hectare of the crops.

2. The use of poly-gamma-glutamic acid as a fertilizer synergist in crops as claimed in claim 1, which is a high molecular polymer formed from D-glutamic acid (D-Glu) and L-glutamic acid (L-Glu) via α -amino group and γ -carboxyl group in the form of peptide bond, and has an average molecular weight of 1000-3000 kD.

3. The application of poly-gamma-glutamic acid as a fertilizer synergist in crops of claim 1, wherein the poly-gamma-glutamic acid is prepared into a compound preparation, and the compound preparation consists of the following components in parts by weight:

4. the application of the poly-gamma-glutamic acid as a fertilizer synergist in crops is characterized in that the compound fertilizer consists of the following raw materials in parts by weight:

5. the application of the poly-gamma-glutamic acid as a fertilizer synergist in crops as claimed in claim 3, wherein the organic fertilizer comprises the following raw materials in parts by weight:

6. the application of the poly-gamma-glutamic acid as a fertilizer synergist in crops as claimed in claim 3, wherein the composite probiotic granule is in a dry powder state and comprises the following components in percentage by weight:

50% of bacillus subtilis, 25% of azospirillum, 15% of rhizobium, 5% of streptomyces griseus and 5% of yeast.

7. The application of the poly-gamma-glutamic acid as a fertilizer synergist in crops as claimed in claim 3, wherein the traditional Chinese medicine deinsectization particles comprise the following raw materials in parts by weight:

8. the use of poly-gamma-glutamic acid as a fertilizer synergist in crops of claim 3, wherein the preparation method of the yeast fermentation broth dry powder particle comprises the following steps:

(1) preparation of culture medium and recovery of yeast

Dissolving 20g of glucose, 20g of peptone, 10g of yeast extract and 20g of agar with distilled water, fixing the volume to 1000mL, sterilizing for 20-30 min at 121 ℃ by using an autoclave, adjusting the pH to 5.0-5.5 to prepare a culture medium, taking out the saccharomyces cerevisiae from a refrigerator for resuscitation, and culturing on the culture medium for 24h to obtain reactivated saccharomyces cerevisiae;

(2) basic liquid medium and gradient medium preparation

The preparation method of the basic liquid culture medium comprises the following steps: sequentially adding 20g of glucose, 20g of peptone and 10g of yeast extract into 1000ml of distilled water, uniformly stirring, and adjusting the pH value to 5.0-5.2 for later use;

the preparation method of the gradient culture medium comprises the following steps: respectively adding a boron source and a selenium source into the basic liquid culture medium to obtain a zinc culture medium with boron contents of 10mg/L, 20mg/L, 40mg/L, 80mg/L, 160mg/L, 320mg/L and 640mg/L and a selenium culture medium with selenium contents of 2mg/L, 4mg/L, 8mg/L, 16mg/L, 32mg/L, 64mg/L and 128mg/L respectively, wherein the boron culture medium and the basic liquid culture medium with boron content of 0mg/L form a boron gradient culture medium, and the selenium culture medium and the basic liquid culture medium with selenium content of 0mg/L form a selenium gradient culture medium for later use;

(3) gradient culture of yeast

Culturing the saccharomyces cerevisiae obtained by the treatment of the step (1) on the boron gradient culture medium prepared in the step (2) for 36 hours at the culture temperature of 28 ℃, and after the culture is finished, culturing the saccharomyces cerevisiae on the selenium gradient culture medium prepared in the step (2) for 24 hours at the culture temperature of 30 ℃;

(4) preparation of seed culture Medium

Obtaining fresh saccharification waste liquid in the beer production process, centrifuging for 15min at the rotating speed of 5000r/min, then adding 5.5% of cane sugar, 0.5% of urea, 0.25% of salt and 0.02% of sodium sulfide according to the weight ratio, uniformly mixing, sterilizing for 15-20min at 121 ℃, and adjusting the pH value to 5.0-5.5 to prepare a seed culture medium;

(5) preparation of fermented seeds

Inoculating the saccharomyces cerevisiae treated in the step (3) into the seed culture medium prepared in the step (4) according to an inoculation proportion of 10%, and placing the inoculated culture medium in a constant-temperature incubator at 30 ℃ for culturing for 24h to obtain saccharomyces cerevisiae fermentation seeds;

(6) preparation of fermentation broth

The fermentation liquor is prepared by utilizing saccharification waste liquid in the beer production process, and the specific preparation method comprises the following steps:

preparing fermentation liquor by using saccharification waste liquid: taking fresh saccharification waste liquid in the beer production process, centrifuging for 15min at the rotating speed of 5000r/min, then sequentially adding 2.0% of cane sugar, 25% of corn flour, 0.5% of urea, 0.25% of salt and 0.02% of sodium sulfide according to the weight ratio, sterilizing for 15-20min at 121 ℃, adjusting the pH value to 5.0-5.5, and preparing fermentation liquor;

(7) microbial fermentation

Adding 25% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor prepared in the step (6) according to the weight, uniformly mixing, respectively inoculating the saccharomyces cerevisiae fermentation seeds prepared in the step (5) into the fermentation liquor according to the inoculation proportion of 5%, fermenting at the temperature of 30 ℃, adding 25% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor again after fermenting for 4 hours, uniformly stirring, continuously fermenting for 8 hours, adding 25% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor again, uniformly stirring, continuously fermenting for 12 hours, adding the rest 20% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor, uniformly stirring, and continuously fermenting for 24 hours to complete the fermentation process;

(8) centrifuging the yeast fermentation liquor, collecting solids, and performing low-temperature vacuum drying and granulation;

collecting solid, centrifuging the filtrate at 10000rpm for 10min to obtain solid precipitate, mixing the solid obtained by filtration and the solid precipitate obtained by centrifugation, and vacuum drying and granulating in vacuum drying granulator under 0.01Mpa at 35 deg.C.

9. The use of poly-gamma-glutamic acid as a fertilizer synergist in crops of claim 3, wherein the complex formulation is prepared by the steps of:

1) weighing various raw materials of the organic fertilizer, drying the raw materials in vacuum at low temperature, granulating the dried raw materials, and uniformly mixing the granulated raw materials with the compound fertilizer for later use;

2) weighing poly-gamma-glutamic acid for later use;

3) weighing composite probiotic particles and traditional Chinese medicine deinsectization particles, uniformly stirring and mixing at the temperature of below 40 ℃, adding the mixture obtained in the step 2), mixing, adding the mixture obtained in the step 1), fully mixing, adding yeast fermentation liquid dry powder particles, crushing, and sieving with a 200-mesh sieve to obtain composite preparation dry powder;

4) when in use, the dry powder of the compound preparation prepared in the step 3) is diluted with water by 1:100 for use.

10. Use of poly-gamma-glutamic acid as a fertilizer synergist in crops of claim 1, wherein the crops comprise vegetables, cereals, soybeans.

Technical Field

The invention belongs to the field of fertilizer synergists, and particularly discloses application of poly-gamma-glutamic acid serving as a fertilizer synergist in crops.

Background

Fertilizer synergist refers to an active substance for the purpose of increasing nutrient availability. The nutrient solution can increase the supply of crop nutrients by retaining nitrogen and activating phosphorus and potassium elements which are difficult to utilize in soil, and plays a certain role in regulating the physiological functions of plants. The fertilizer is usually added into conventional fertilizers, so that the application amount of the fertilizer can be properly reduced, and the utilization rate of the fertilizer is improved. The fertilizer synergist has various types, and can be functionally divided into a nitrification inhibitor, a urease inhibitor, a nutrient activator, a water-retaining agent and the like. The composition can be divided into organic activator, inorganic activator, biological activator or mixed activator. The fertilizer synergist has the functions of promoting nutrient effectiveness and improving nutrient utilization rate under certain conditions, but the function is limited, and the fertilizer synergist is combined with fertilizer application, so that the high yield and stable yield of crops cannot be achieved by the fertilizer synergist.

The polymerized amino acid can increase the absorption of plant to nutrition, save the fertilizer usage amount and improve the crop yield. A polymeric amino acid that has been used as a fertilizer synergist at present is polyaspartic acid (PAA). The Donlar company in the United states has first utilized chemical synthesis, i.e., thermal polymerization, to produce TPA (thermal polyaspartate), which is already industrialized and commercialized, and is applied to fertilizers. TPA is an active ingredient in a fertilizer accelerator product Amisorb, and because the TPA contains a plurality of free carboxyl groups, the TPA has higher affinity to nutrients in the fertilizer, can accelerate the formation of roots and increase tillering, and Amisorb is proved to be applicable to crops such as wheat, corn, soybean and the like, and has the effects of increasing the yield, saving the using amount of the fertilizer and improving the quality of the crops. However, the biodegradability of the accelerator is not yet satisfactory. Poly-gamma-glutamic acid (Poly-gamma-glutamic acid) is anionic polyamino acid generated by microbial fermentation in nature, and has the characteristics of water solubility, water absorbability, biodegradability, affinity to metal ions, no toxicity, frost resistance, environmental friendliness, resistance to protease and the like, so that the Poly-gamma-glutamic acid has wide application prospects in the fields of agriculture, food, medicines, cosmetics, fiber light chemical industry and the like. At present, the research on poly-gamma-glutamic acid as a fertilizer synergist in agriculture is very limited, and the poly-gamma-glutamic acid is worthy of further research.

Disclosure of Invention

In order to solve the problems, the invention discloses application of poly-gamma-glutamic acid as a fertilizer synergist in crops, and the poly-gamma-glutamic acid disclosed by the invention as the fertilizer synergist has the advantages of small fertilizing amount, good yield increasing effect, comprehensive nutrition, definite and controllable components, simple preparation method, large-scale industrial preparation and suitability for planting common crops such as grains, fruits and vegetables.

The technical scheme of the invention is as follows:

the application of poly-gamma-glutamic acid as a fertilizer synergist in crops comprises applying the poly-gamma-glutamic acid to soil when preparing soil before planting the crops, and spraying the poly-gamma-glutamic acid on the overground part of a plant in the growth period of the crops;

the application amount of the poly-gamma-glutamic acid applied to the soil is 1-3kg per hectare, and the poly-gamma-glutamic acid applied to the soil and the conventional fertilizer are uniformly mixed and then are applied together;

in the growth period of the crops, 2.5-7.5kg of poly-gamma-glutamic acid is sprayed on each hectare of the crops.

Further, the poly-gamma-glutamic acid is used as a fertilizer synergist in crops, is a high molecular polymer formed by peptide bonds of D-glutamic acid (D-Glu) and L-glutamic acid (L-Glu) through alpha-amino and gamma-carboxyl, and has an average molecular weight of 1000-3000 kD.

Further, the poly-gamma-glutamic acid is applied to crops as a fertilizer synergist, and is prepared into a compound preparation for use, wherein the compound preparation comprises the following components in parts by weight:

further, the poly-gamma-glutamic acid is applied to crops as a fertilizer synergist, and the compound fertilizer consists of the following raw materials in parts by weight:

further, the poly-gamma-glutamic acid is applied to crops as a fertilizer synergist, and the organic fertilizer is prepared from the following raw materials in parts by weight:

further, the poly-gamma-glutamic acid is applied to crops as a fertilizer synergist, and the composite probiotic granule is in a dry powder state and comprises the following components in percentage by weight:

50% of bacillus subtilis, 25% of azospirillum, 15% of rhizobium, 5% of streptomyces griseus and 5% of yeast.

Further, the application of the poly-gamma-glutamic acid as a fertilizer synergist in crops, the traditional Chinese medicine deinsectization granules comprise the following raw materials in parts by weight:

further, the application of the poly-gamma-glutamic acid as a fertilizer synergist in crops, and the preparation method of the yeast fermentation liquid dry powder particles comprises the following steps:

(1) preparation of culture medium and recovery of yeast

Dissolving 20g of glucose, 20g of peptone, 10g of yeast extract and 20g of agar with distilled water, fixing the volume to 1000mL, sterilizing for 20-30 min at 121 ℃ by using an autoclave, adjusting the pH to 5.0-5.5 to prepare a culture medium, taking out the saccharomyces cerevisiae from a refrigerator for resuscitation, and culturing on the culture medium for 24h to obtain reactivated saccharomyces cerevisiae;

(2) basic liquid medium and gradient medium preparation

The preparation method of the basic liquid culture medium comprises the following steps: sequentially adding 20g of glucose, 20g of peptone and 10g of yeast extract into 1000ml of distilled water, uniformly stirring, and adjusting the pH value to 5.0-5.2 for later use;

the preparation method of the gradient culture medium comprises the following steps: respectively adding a boron source and a selenium source into the basic liquid culture medium to obtain a zinc culture medium with boron contents of 10mg/L, 20mg/L, 40mg/L, 80mg/L, 160mg/L, 320mg/L and 640mg/L and a selenium culture medium with selenium contents of 2mg/L, 4mg/L, 8mg/L, 16mg/L, 32mg/L, 64mg/L and 128mg/L respectively, wherein the boron culture medium and the basic liquid culture medium with boron content of 0mg/L form a boron gradient culture medium, and the selenium culture medium and the basic liquid culture medium with selenium content of 0mg/L form a selenium gradient culture medium for later use;

(3) gradient culture of yeast

Culturing the saccharomyces cerevisiae obtained by the treatment of the step (1) on the boron gradient culture medium prepared in the step (2) for 36 hours at the culture temperature of 28 ℃, and after the culture is finished, culturing the saccharomyces cerevisiae on the selenium gradient culture medium prepared in the step (2) for 24 hours at the culture temperature of 30 ℃;

(4) preparation of seed culture Medium

Obtaining fresh saccharification waste liquid in the beer production process, centrifuging for 15min at the rotating speed of 5000r/min, then adding 5.5% of cane sugar, 0.5% of urea, 0.25% of salt and 0.02% of sodium sulfide according to the weight ratio, uniformly mixing, sterilizing for 15-20min at 121 ℃, and adjusting the pH value to 5.0-5.5 to prepare a seed culture medium;

(5) preparation of fermented seeds

Inoculating the saccharomyces cerevisiae treated in the step (3) into the seed culture medium prepared in the step (4) according to an inoculation proportion of 10%, and placing the inoculated culture medium in a constant-temperature incubator at 30 ℃ for culturing for 24h to obtain saccharomyces cerevisiae fermentation seeds;

(6) preparation of fermentation broth

The fermentation liquor is prepared by utilizing saccharification waste liquid in the beer production process, and the specific preparation method comprises the following steps:

preparing fermentation liquor by using saccharification waste liquid: taking fresh saccharification waste liquid in the beer production process, centrifuging for 15min at the rotating speed of 5000r/min, then sequentially adding 2.0% of cane sugar, 25% of corn flour, 0.5% of urea, 0.25% of salt and 0.02% of sodium sulfide according to the weight ratio, sterilizing for 15-20min at 121 ℃, adjusting the pH value to 5.0-5.5, and preparing fermentation liquor;

(7) microbial fermentation

Adding 25% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor prepared in the step (6) according to the weight, uniformly mixing, respectively inoculating the saccharomyces cerevisiae fermentation seeds prepared in the step (5) into the fermentation liquor according to the inoculation proportion of 5%, fermenting at the temperature of 30 ℃, adding 25% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor again after fermenting for 4 hours, uniformly stirring, continuously fermenting for 8 hours, adding 25% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor again, uniformly stirring, continuously fermenting for 12 hours, adding the rest 20% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor, uniformly stirring, and continuously fermenting for 24 hours to complete the fermentation process;

(8) centrifuging the yeast fermentation liquor, collecting solids, and performing low-temperature vacuum drying and granulation;

collecting solid, centrifuging the filtrate at 10000rpm for 10min to obtain solid precipitate, mixing the solid obtained by filtration and the solid precipitate obtained by centrifugation, and vacuum drying and granulating in vacuum drying granulator under 0.01Mpa at 35 deg.C.

Further, the poly-gamma-glutamic acid is applied to crops as a fertilizer synergist, and the compound preparation is prepared by the following steps:

1) weighing various raw materials of the organic fertilizer, drying the raw materials in vacuum at low temperature, granulating the dried raw materials, and uniformly mixing the granulated raw materials with the compound fertilizer for later use;

2) weighing poly-gamma-glutamic acid for later use;

3) weighing composite probiotic particles and traditional Chinese medicine deinsectization particles, uniformly stirring and mixing at the temperature of below 40 ℃, adding the mixture obtained in the step 2), mixing, adding the mixture obtained in the step 1), fully mixing, adding yeast fermentation liquid dry powder particles, crushing, and sieving with a 200-mesh sieve to obtain composite preparation dry powder;

4) when in use, the dry powder of the compound preparation prepared in the step 3) is diluted with water by 1:100 for use.

Furthermore, the poly-gamma-glutamic acid is applied to crops as a fertilizer synergist, and the crops comprise vegetables, grains and soybeans.

According to the technical scheme, the invention has the following beneficial effects:

the invention discloses application of poly-gamma-glutamic acid as a fertilizer synergist in crops, preferably, the poly-gamma-glutamic acid is prepared into a compound preparation for use, and the compound preparation carefully selects compound fertilizer, organic fertilizer, yeast fermentation broth dry powder particles, compound probiotic particles and traditional Chinese medicine deinsectization particles; after yeast fermentation, centrifuging the fermentation liquor, collecting solids, and performing low-temperature vacuum drying and granulation to obtain the yeast fermentation liquor dry powder particles; the compound fertilizer of the invention provides nitrogen source, phosphate and calcium salt which are necessary for the growth of crops, and meanwhile, the added organic fertilizer increases biological organic matters and trace elements for the growth of solanaceae plants, thus improving the comprehensive nutrition and disease resistance of crops; the yeast fermentation liquid dry powder particles contain rich protein, boron and selenium elements, can promote the root development of crops, and improve the oxidation resistance in plants, thereby improving the stress resistance and the anti-aging capability of the plants and ensuring the normal growth of the plants; the composite probiotic particles further enhance the fertility of the soil of the root system of the plant, create good microbial ecology, promote the generation and dilution of nitrogen elements and prevent the growth of harmful bacteria; the traditional Chinese medicine deinsectization granules have strong insect expelling smell and insect killing capability, can reduce the plant diseases and insect pests of roots, reduce the application of pesticides, and are green and pollution-free.

In conclusion, the poly-gamma-glutamic acid disclosed by the invention is applied to crops as a fertilizer synergist, has the advantages of small fertilizing amount, good yield increasing effect, comprehensive nutrition, definite and controllable components, simple preparation method and suitability for large-scale industrial preparation, and is suitable for planting common crops such as grains, fruits and vegetables.

Detailed Description

The invention will be further illustrated by the following specific examples, all of which are commercially available and are intended to be illustrative and not limiting.

Example 1

The application of the poly-gamma-glutamic acid as a fertilizer synergist in wheat planting is characterized by comprising the steps of applying the poly-gamma-glutamic acid to soil during soil preparation before the wheat is planted, and spraying the poly-gamma-glutamic acid on the overground part of a plant during the growth period of the wheat;

the application amount of the poly-gamma-glutamic acid applied to the soil is 1kg per hectare, and the poly-gamma-glutamic acid applied to the soil and the conventional fertilizer are uniformly mixed and then are applied together;

in the growth period of the crops, 2.5kg of poly-gamma-glutamic acid is sprayed on each hectare of the crops.

The poly-gamma-glutamic acid is a high molecular polymer formed by D type glutamic acid (D-Glu) and L type glutamic acid (L-Glu) in a peptide bond form through alpha-amino and gamma-carboxyl, and has an average molecular weight of 1000 kD.

The poly-gamma-glutamic acid is prepared into a compound preparation for use, and the compound preparation consists of the following components in parts by weight:

the compound fertilizer is composed of the following raw materials in parts by weight:

the organic fertilizer is prepared from the following raw materials in parts by weight:

the composite probiotic granule is in a dry powder state and comprises the following components in percentage by weight:

50% of bacillus subtilis, 25% of azospirillum, 15% of rhizobium, 5% of streptomyces griseus and 5% of yeast.

The traditional Chinese medicine deinsectization granules comprise the following raw materials in parts by weight:

the preparation method of the yeast fermentation liquid dry powder particles comprises the following steps:

(1) preparation of culture medium and recovery of yeast

Dissolving 20g of glucose, 20g of peptone, 10g of yeast extract and 20g of agar with distilled water, fixing the volume to 1000mL, sterilizing for 20-30 min at 121 ℃ by using an autoclave, adjusting the pH to 5.0-5.5 to prepare a culture medium, taking out the saccharomyces cerevisiae from a refrigerator for resuscitation, and culturing on the culture medium for 24h to obtain reactivated saccharomyces cerevisiae;

(2) basic liquid medium and gradient medium preparation

The preparation method of the basic liquid culture medium comprises the following steps: sequentially adding 20g of glucose, 20g of peptone and 10g of yeast extract into 1000ml of distilled water, uniformly stirring, and adjusting the pH value to 5.0-5.2 for later use;

the preparation method of the gradient culture medium comprises the following steps: respectively adding a boron source and a selenium source into the basic liquid culture medium to obtain a zinc culture medium with boron contents of 10mg/L, 20mg/L, 40mg/L, 80mg/L, 160mg/L, 320mg/L and 640mg/L and a selenium culture medium with selenium contents of 2mg/L, 4mg/L, 8mg/L, 16mg/L, 32mg/L, 64mg/L and 128mg/L respectively, wherein the boron culture medium and the basic liquid culture medium with boron content of 0mg/L form a boron gradient culture medium, and the selenium culture medium and the basic liquid culture medium with selenium content of 0mg/L form a selenium gradient culture medium for later use;

(3) gradient culture of yeast

Culturing the saccharomyces cerevisiae obtained by the treatment of the step (1) on the boron gradient culture medium prepared in the step (2) for 36 hours at the culture temperature of 28 ℃, and after the culture is finished, culturing the saccharomyces cerevisiae on the selenium gradient culture medium prepared in the step (2) for 24 hours at the culture temperature of 30 ℃;

(4) preparation of seed culture Medium

Obtaining fresh saccharification waste liquid in the beer production process, centrifuging for 15min at the rotating speed of 5000r/min, then adding 5.5% of cane sugar, 0.5% of urea, 0.25% of salt and 0.02% of sodium sulfide according to the weight ratio, uniformly mixing, sterilizing for 15-20min at 121 ℃, and adjusting the pH value to 5.0-5.5 to prepare a seed culture medium;

(5) preparation of fermented seeds

Inoculating the saccharomyces cerevisiae treated in the step (3) into the seed culture medium prepared in the step (4) according to an inoculation proportion of 10%, and placing the inoculated culture medium in a constant-temperature incubator at 30 ℃ for culturing for 24h to obtain saccharomyces cerevisiae fermentation seeds;

(6) preparation of fermentation broth

The fermentation liquor is prepared by utilizing saccharification waste liquid in the beer production process, and the specific preparation method comprises the following steps:

preparing fermentation liquor by using saccharification waste liquid: taking fresh saccharification waste liquid in the beer production process, centrifuging for 15min at the rotating speed of 5000r/min, then sequentially adding 2.0% of cane sugar, 25% of corn flour, 0.5% of urea, 0.25% of salt and 0.02% of sodium sulfide according to the weight ratio, sterilizing for 15-20min at 121 ℃, adjusting the pH value to 5.0-5.5, and preparing fermentation liquor;

(7) microbial fermentation

Adding 25% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor prepared in the step (6) according to the weight, uniformly mixing, respectively inoculating the saccharomyces cerevisiae fermentation seeds prepared in the step (5) into the fermentation liquor according to the inoculation proportion of 5%, fermenting at the temperature of 30 ℃, adding 25% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor again after fermenting for 4 hours, uniformly stirring, continuously fermenting for 8 hours, adding 25% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor again, uniformly stirring, continuously fermenting for 12 hours, adding the rest 20% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor, uniformly stirring, and continuously fermenting for 24 hours to complete the fermentation process;

(8) centrifuging the yeast fermentation liquor, collecting solids, and performing low-temperature vacuum drying and granulation;

collecting solid, centrifuging the filtrate at 10000rpm for 10min to obtain solid precipitate, mixing the solid obtained by filtration and the solid precipitate obtained by centrifugation, and vacuum drying and granulating in vacuum drying granulator under 0.01Mpa at 35 deg.C.

The compound preparation is prepared by the following steps:

1) weighing various raw materials of the organic fertilizer, drying the raw materials in vacuum at low temperature, granulating the dried raw materials, and uniformly mixing the granulated raw materials with the compound fertilizer for later use;

2) weighing poly-gamma-glutamic acid for later use;

3) weighing composite probiotic particles and traditional Chinese medicine deinsectization particles, uniformly stirring and mixing at the temperature of below 40 ℃, adding the mixture obtained in the step 2), mixing, adding the mixture obtained in the step 1), fully mixing, adding yeast fermentation liquid dry powder particles, crushing, and sieving with a 200-mesh sieve to obtain composite preparation dry powder;

4) when in use, the dry powder of the compound preparation prepared in the step 3) is diluted with water by 1:100 for use.

Example 2

The application of the poly-gamma-glutamic acid as a fertilizer synergist in wheat planting is characterized by comprising the steps of applying the poly-gamma-glutamic acid to soil during soil preparation before the wheat is planted, and spraying the poly-gamma-glutamic acid on the overground part of a plant during the growth period of the wheat;

the application amount of the poly-gamma-glutamic acid applied to the soil is 2kg per hectare, and the poly-gamma-glutamic acid applied to the soil and the conventional fertilizer are uniformly mixed and then are applied together;

in the growth period of the crops, 5kg of poly-gamma-glutamic acid is sprayed on each hectare of the crops.

The poly-gamma-glutamic acid is a high molecular polymer formed by D type glutamic acid (D-Glu) and L type glutamic acid (L-Glu) in a peptide bond form through alpha-amino and gamma-carboxyl, and has an average molecular weight of 2000 kD.

The poly-gamma-glutamic acid is prepared into a compound preparation for use, and the compound preparation consists of the following components in parts by weight:

the compound fertilizer is composed of the following raw materials in parts by weight:

the organic fertilizer is prepared from the following raw materials in parts by weight:

the composite probiotic granule is in a dry powder state and comprises the following components in percentage by weight:

50% of bacillus subtilis, 25% of azospirillum, 15% of rhizobium, 5% of streptomyces griseus and 5% of yeast.

The traditional Chinese medicine deinsectization granules comprise the following raw materials in parts by weight:

the preparation method of the yeast fermentation liquid dry powder particles comprises the following steps:

(1) preparation of culture medium and recovery of yeast

Dissolving 20g of glucose, 20g of peptone, 10g of yeast extract and 20g of agar with distilled water, fixing the volume to 1000mL, sterilizing for 20-30 min at 121 ℃ by using an autoclave, adjusting the pH to 5.0-5.5 to prepare a culture medium, taking out the saccharomyces cerevisiae from a refrigerator for resuscitation, and culturing on the culture medium for 24h to obtain reactivated saccharomyces cerevisiae;

(2) basic liquid medium and gradient medium preparation

The preparation method of the basic liquid culture medium comprises the following steps: sequentially adding 20g of glucose, 20g of peptone and 10g of yeast extract into 1000ml of distilled water, uniformly stirring, and adjusting the pH value to 5.0-5.2 for later use;

the preparation method of the gradient culture medium comprises the following steps: respectively adding a boron source and a selenium source into the basic liquid culture medium to obtain a zinc culture medium with boron contents of 10mg/L, 20mg/L, 40mg/L, 80mg/L, 160mg/L, 320mg/L and 640mg/L and a selenium culture medium with selenium contents of 2mg/L, 4mg/L, 8mg/L, 16mg/L, 32mg/L, 64mg/L and 128mg/L respectively, wherein the boron culture medium and the basic liquid culture medium with boron content of 0mg/L form a boron gradient culture medium, and the selenium culture medium and the basic liquid culture medium with selenium content of 0mg/L form a selenium gradient culture medium for later use;

(3) gradient culture of yeast

Culturing the saccharomyces cerevisiae obtained by the treatment of the step (1) on the boron gradient culture medium prepared in the step (2) for 36 hours at the culture temperature of 28 ℃, and after the culture is finished, culturing the saccharomyces cerevisiae on the selenium gradient culture medium prepared in the step (2) for 24 hours at the culture temperature of 30 ℃;

(4) preparation of seed culture Medium

Obtaining fresh saccharification waste liquid in the beer production process, centrifuging for 15min at the rotating speed of 5000r/min, then adding 5.5% of cane sugar, 0.5% of urea, 0.25% of salt and 0.02% of sodium sulfide according to the weight ratio, uniformly mixing, sterilizing for 15-20min at 121 ℃, and adjusting the pH value to 5.0-5.5 to prepare a seed culture medium;

(5) preparation of fermented seeds

Inoculating the saccharomyces cerevisiae treated in the step (3) into the seed culture medium prepared in the step (4) according to an inoculation proportion of 10%, and placing the inoculated culture medium in a constant-temperature incubator at 30 ℃ for culturing for 24h to obtain saccharomyces cerevisiae fermentation seeds;

(6) preparation of fermentation broth

The fermentation liquor is prepared by utilizing saccharification waste liquid in the beer production process, and the specific preparation method comprises the following steps:

preparing fermentation liquor by using saccharification waste liquid: taking fresh saccharification waste liquid in the beer production process, centrifuging for 15min at the rotating speed of 5000r/min, then sequentially adding 2.0% of cane sugar, 25% of corn flour, 0.5% of urea, 0.25% of salt and 0.02% of sodium sulfide according to the weight ratio, sterilizing for 15-20min at 121 ℃, adjusting the pH value to 5.0-5.5, and preparing fermentation liquor;

(7) microbial fermentation

Adding 25% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor prepared in the step (6) according to the weight, uniformly mixing, respectively inoculating the saccharomyces cerevisiae fermentation seeds prepared in the step (5) into the fermentation liquor according to the inoculation proportion of 5%, fermenting at the temperature of 30 ℃, adding 25% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor again after fermenting for 4 hours, uniformly stirring, continuously fermenting for 8 hours, adding 25% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor again, uniformly stirring, continuously fermenting for 12 hours, adding the rest 20% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor, uniformly stirring, and continuously fermenting for 24 hours to complete the fermentation process;

(8) centrifuging the yeast fermentation liquor, collecting solids, and performing low-temperature vacuum drying and granulation;

collecting solid, centrifuging the filtrate at 10000rpm for 10min to obtain solid precipitate, mixing the solid obtained by filtration and the solid precipitate obtained by centrifugation, and vacuum drying and granulating in vacuum drying granulator under 0.01Mpa at 35 deg.C.

The compound preparation is prepared by the following steps:

1) weighing various raw materials of the organic fertilizer, drying the raw materials in vacuum at low temperature, granulating the dried raw materials, and uniformly mixing the granulated raw materials with the compound fertilizer for later use;

2) weighing poly-gamma-glutamic acid for later use;

3) weighing composite probiotic particles and traditional Chinese medicine deinsectization particles, uniformly stirring and mixing at the temperature of below 40 ℃, adding the mixture obtained in the step 2), mixing, adding the mixture obtained in the step 1), fully mixing, adding yeast fermentation liquid dry powder particles, crushing, and sieving with a 200-mesh sieve to obtain composite preparation dry powder;

4) when in use, the dry powder of the compound preparation prepared in the step 3) is diluted with water by 1:100 for use.

Example 3

The application of the poly-gamma-glutamic acid as a fertilizer synergist in wheat planting is characterized by comprising the steps of applying the poly-gamma-glutamic acid to soil during soil preparation before the wheat is planted, and spraying the poly-gamma-glutamic acid on the overground part of a plant during the growth period of the wheat;

the application amount of the poly-gamma-glutamic acid applied to the soil is 3kg per hectare, and the poly-gamma-glutamic acid applied to the soil and the conventional fertilizer are uniformly mixed and then are applied together;

in the growth period of the crops, 7.5kg of poly-gamma-glutamic acid is sprayed on each hectare of the crops.

The poly-gamma-glutamic acid is a high molecular polymer formed by D-type glutamic acid (D-Glu) and L-type glutamic acid (L-Glu) in a peptide bond form through alpha-amino and gamma-carboxyl, and has an average molecular weight of 3000 kD.

The poly-gamma-glutamic acid is prepared into a compound preparation for use, and the compound preparation consists of the following components in parts by weight:

the compound fertilizer is composed of the following raw materials in parts by weight:

the organic fertilizer is prepared from the following raw materials in parts by weight:

the composite probiotic granule is in a dry powder state and comprises the following components in percentage by weight:

50% of bacillus subtilis, 25% of azospirillum, 15% of rhizobium, 5% of streptomyces griseus and 5% of yeast.

The traditional Chinese medicine deinsectization granules comprise the following raw materials in parts by weight:

the preparation method of the yeast fermentation liquid dry powder particles comprises the following steps:

(1) preparation of culture medium and recovery of yeast

Dissolving 20g of glucose, 20g of peptone, 10g of yeast extract and 20g of agar with distilled water, fixing the volume to 1000mL, sterilizing for 20-30 min at 121 ℃ by using an autoclave, adjusting the pH to 5.0-5.5 to prepare a culture medium, taking out the saccharomyces cerevisiae from a refrigerator for resuscitation, and culturing on the culture medium for 24h to obtain reactivated saccharomyces cerevisiae;

(2) basic liquid medium and gradient medium preparation

The preparation method of the basic liquid culture medium comprises the following steps: sequentially adding 20g of glucose, 20g of peptone and 10g of yeast extract into 1000ml of distilled water, uniformly stirring, and adjusting the pH value to 5.0-5.2 for later use;

the preparation method of the gradient culture medium comprises the following steps: respectively adding a boron source and a selenium source into the basic liquid culture medium to obtain a zinc culture medium with boron contents of 10mg/L, 20mg/L, 40mg/L, 80mg/L, 160mg/L, 320mg/L and 640mg/L and a selenium culture medium with selenium contents of 2mg/L, 4mg/L, 8mg/L, 16mg/L, 32mg/L, 64mg/L and 128mg/L respectively, wherein the boron culture medium and the basic liquid culture medium with boron content of 0mg/L form a boron gradient culture medium, and the selenium culture medium and the basic liquid culture medium with selenium content of 0mg/L form a selenium gradient culture medium for later use;

(3) gradient culture of yeast

Culturing the saccharomyces cerevisiae obtained by the treatment of the step (1) on the boron gradient culture medium prepared in the step (2) for 36 hours at the culture temperature of 28 ℃, and after the culture is finished, culturing the saccharomyces cerevisiae on the selenium gradient culture medium prepared in the step (2) for 24 hours at the culture temperature of 30 ℃;

(4) preparation of seed culture Medium

Obtaining fresh saccharification waste liquid in the beer production process, centrifuging for 15min at the rotating speed of 5000r/min, then adding 5.5% of cane sugar, 0.5% of urea, 0.25% of salt and 0.02% of sodium sulfide according to the weight ratio, uniformly mixing, sterilizing for 15-20min at 121 ℃, and adjusting the pH value to 5.0-5.5 to prepare a seed culture medium;

(5) preparation of fermented seeds

Inoculating the saccharomyces cerevisiae treated in the step (3) into the seed culture medium prepared in the step (4) according to an inoculation proportion of 10%, and placing the inoculated culture medium in a constant-temperature incubator at 30 ℃ for culturing for 24h to obtain saccharomyces cerevisiae fermentation seeds;

(6) preparation of fermentation broth

The fermentation liquor is prepared by utilizing saccharification waste liquid in the beer production process, and the specific preparation method comprises the following steps:

preparing fermentation liquor by using saccharification waste liquid: taking fresh saccharification waste liquid in the beer production process, centrifuging for 15min at the rotating speed of 5000r/min, then sequentially adding 2.0% of cane sugar, 25% of corn flour, 0.5% of urea, 0.25% of salt and 0.02% of sodium sulfide according to the weight ratio, sterilizing for 15-20min at 121 ℃, adjusting the pH value to 5.0-5.5, and preparing fermentation liquor;

(7) microbial fermentation

Adding 25% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor prepared in the step (6) according to the weight, uniformly mixing, respectively inoculating the saccharomyces cerevisiae fermentation seeds prepared in the step (5) into the fermentation liquor according to the inoculation proportion of 5%, fermenting at the temperature of 30 ℃, adding 25% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor again after fermenting for 4 hours, uniformly stirring, continuously fermenting for 8 hours, adding 25% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor again, uniformly stirring, continuously fermenting for 12 hours, adding the rest 20% of sodium tetraborate and sodium selenite to be fermented into the fermentation liquor, uniformly stirring, and continuously fermenting for 24 hours to complete the fermentation process;

(8) centrifuging the yeast fermentation liquor, collecting solids, and performing low-temperature vacuum drying and granulation;

collecting solid, centrifuging the filtrate at 10000rpm for 10min to obtain solid precipitate, mixing the solid obtained by filtration and the solid precipitate obtained by centrifugation, and vacuum drying and granulating in vacuum drying granulator under 0.01Mpa at 35 deg.C.

The compound preparation is prepared by the following steps:

1) weighing various raw materials of the organic fertilizer, drying the raw materials in vacuum at low temperature, granulating the dried raw materials, and uniformly mixing the granulated raw materials with the compound fertilizer for later use;

2) weighing poly-gamma-glutamic acid for later use;

3) weighing composite probiotic particles and traditional Chinese medicine deinsectization particles, uniformly stirring and mixing at the temperature of below 40 ℃, adding the mixture obtained in the step 2), mixing, adding the mixture obtained in the step 1), fully mixing, adding yeast fermentation liquid dry powder particles, crushing, and sieving with a 200-mesh sieve to obtain composite preparation dry powder;

4) when in use, the dry powder of the compound preparation prepared in the step 3) is diluted with water by 1:100 for use.

Comparative example 1

The application of the poly-gamma-glutamic acid as a fertilizer synergist in wheat is characterized by comprising the steps of applying the poly-gamma-glutamic acid to soil during soil preparation before wheat is planted, and spraying the poly-gamma-glutamic acid on the overground part of a plant during the growth period of the wheat; the application amount of the poly-gamma-glutamic acid applied to the soil is 2kg per hectare, and the poly-gamma-glutamic acid applied to the soil and the conventional fertilizer are uniformly mixed and then are applied together; in the growth period of the crops, 5kg of poly-gamma-glutamic acid is sprayed on each hectare of the crops;

the poly-gamma-glutamic acid is a high molecular polymer formed by D type glutamic acid (D-Glu) and L type glutamic acid (L-Glu) in a peptide bond form through alpha-amino and gamma-carboxyl, and has an average molecular weight of 2000 kD;

the poly-gamma-glutamic acid is prepared into a compound preparation for use, and the compound preparation consists of the following components in parts by weight:

i.e. not containing compound fertilizer, the other steps are the same as the example 2.

Comparative example 2

The application of the poly-gamma-glutamic acid as a fertilizer synergist in wheat is characterized by comprising the steps of applying the poly-gamma-glutamic acid to soil during soil preparation before wheat is planted, and spraying the poly-gamma-glutamic acid on the overground part of a plant during the growth period of the wheat; the application amount of the poly-gamma-glutamic acid applied to the soil is 2kg per hectare, and the poly-gamma-glutamic acid applied to the soil and the conventional fertilizer are uniformly mixed and then are applied together; in the growth period of the crops, 5kg of poly-gamma-glutamic acid is sprayed on each hectare of the crops;

the poly-gamma-glutamic acid is a high molecular polymer formed by D type glutamic acid (D-Glu) and L type glutamic acid (L-Glu) in a peptide bond form through alpha-amino and gamma-carboxyl, and has an average molecular weight of 2000 kD;

the poly-gamma-glutamic acid is prepared into a compound preparation for use, and the compound preparation consists of the following components in parts by weight:

i.e. containing no organic fertilizer, the rest is the same as example 2.

Comparative example 3

The application of the poly-gamma-glutamic acid as a fertilizer synergist in wheat is characterized by comprising the steps of applying the poly-gamma-glutamic acid to soil during soil preparation before wheat is planted, and spraying the poly-gamma-glutamic acid on the overground part of a plant during the growth period of the wheat; the application amount of the poly-gamma-glutamic acid applied to the soil is 3kg per hectare, and the poly-gamma-glutamic acid applied to the soil and the conventional fertilizer are uniformly mixed and then are applied together; in the growth period of the crops, 5kg of poly-gamma-glutamic acid is sprayed on each hectare of the crops;

the poly-gamma-glutamic acid is a high molecular polymer formed by D type glutamic acid (D-Glu) and L type glutamic acid (L-Glu) in a peptide bond form through alpha-amino and gamma-carboxyl, and has an average molecular weight of 2000 kD;

the poly-gamma-glutamic acid is prepared into a compound preparation for use, and the compound preparation consists of the following components in parts by weight:

i.e. without complex probiotic particles, otherwise as in example 2.

Comparative example 4

The application of the poly-gamma-glutamic acid as a fertilizer synergist in wheat is characterized by comprising the steps of applying the poly-gamma-glutamic acid to soil during soil preparation before wheat is planted, and spraying the poly-gamma-glutamic acid on the overground part of a plant during the growth period of the wheat; the application amount of the poly-gamma-glutamic acid applied to the soil is 2kg per hectare, and the poly-gamma-glutamic acid applied to the soil and the conventional fertilizer are uniformly mixed and then are applied together; in the growth period of the crops, 5kg of poly-gamma-glutamic acid is sprayed on each hectare of the crops;

the poly-gamma-glutamic acid is a high molecular polymer formed by D type glutamic acid (D-Glu) and L type glutamic acid (L-Glu) in a peptide bond form through alpha-amino and gamma-carboxyl, and has an average molecular weight of 2000 kD;

i.e. no Chinese medicinal deinsectization granules, the other steps are the same as the example 2.

Comparative example 5

The application of the poly-gamma-glutamic acid as a fertilizer synergist in wheat is characterized by comprising the steps of applying the poly-gamma-glutamic acid to soil during soil preparation before wheat is planted, and spraying the poly-gamma-glutamic acid on the overground part of a plant during the growth period of the wheat; the application amount of the poly-gamma-glutamic acid applied to the soil is 2kg per hectare, and the poly-gamma-glutamic acid applied to the soil and the conventional fertilizer are uniformly mixed and then are applied together; in the growth period of the crops, 5kg of poly-gamma-glutamic acid is sprayed on each hectare of the crops;

the poly-gamma-glutamic acid is a high molecular polymer formed by D type glutamic acid (D-Glu) and L type glutamic acid (L-Glu) in a peptide bond form through alpha-amino and gamma-carboxyl, and has an average molecular weight of 2000 kD;

i.e., no yeast broth dry powder particles, as in example 2.

Test example

The wheat grown by the methods of examples 1 to 3 and comparative examples 1 to 5 was subjected to comparative tests, and the wheat variety was west agriculture 511, and the results are shown in table 1.

Table 1 wheat growth.

	Mu yield (kg)	Protein content (%)	Thousand seed weight	Gibberella zeae grain of wheat%
					Example 1	586	15.2	45.3	1.3
Example 2	597	15.5	46.1	1.4
					Example 3	601	15.6	46.3	1.2
Comparative example 1	565	13.9	41.2	2.1
					Comparative example 2	571	14.1	42.3	2.3
Comparative example 3	575	14.2	42.6	3.0
					Comparative example 4	570	14.4	42.3	3.7
Comparative example 5	576	14.1	41.5	2.0

According to the test example, compared with the comparative example, the application of the poly-gamma-glutamic acid as the fertilizer synergist in the wheat has the advantages of high wheat yield per mu, high protein content, high thousand grain weight, low disease rate and good wheat quality.

The above are only preferred embodiments of the present invention, and the scope of the present invention should not be limited thereby, and all the equivalent changes and modifications made by the claims and the summary of the invention should be covered by the protection scope of the present patent application.