A microbial selenium protein preparation, its preparation method, usage and application

Abstract

This invention belongs to the field of turfgrass and forage grass maintenance technology, specifically relating to a microbial selenium protein preparation for alleviating drought stress in perennial ryegrass, its preparation method, application method, and application. The microbial selenium protein preparation comprises the following raw materials in specific weight proportions: peptone, beef extract, sucrose, salt, selenium-enriched mushroom powder, organosilicon, calcium amino acids, potassium humate, betaine, salicylic acid, and Bacillus natto. This microbial selenium protein preparation aims to solve the limitations of traditional drought-resistant methods and the poor effectiveness of single-component preparations. When using this microbial selenium protein preparation, it is first diluted and then applied as a root drench to ryegrass seedlings multiple times. The selenium protein preparation of this invention, through the synergistic effect of multiple components, can significantly improve the drought resistance of ryegrass. Furthermore, the preparation process of this protein preparation is simple, environmentally friendly, and efficient, suitable for ryegrass propagation and maintenance, and has broad application prospects.

Description

Technical Field

[0001] This invention belongs to the field of turfgrass and forage grass maintenance technology, specifically relating to a microbial selenium protein preparation for alleviating drought stress in perennial ryegrass, as well as its preparation method, usage method and application. Background Technology

[0002] Perennial ryegrass ( Perennial ryegrass Perennial ryegrass (L.) is a high-quality turfgrass and forage grass, characterized by strong tillering ability, good palatability, and excellent greening effect, and is widely planted in temperate and subtropical regions worldwide. However, with the intensification of global climate change and the increasing prominence of water scarcity, drought stress has become one of the main abiotic stress factors restricting the growth and development of perennial ryegrass. Drought stress disrupts the physiological metabolic balance of perennial ryegrass, leading to reduced plant height, decreased biomass, and reduced relative water content in leaves. It also causes cell membrane damage, increasing malondialdehyde content and relative conductivity, accelerating chlorophyll degradation, and inhibiting photosynthesis, which in severe cases can even lead to plant wilting and death. Especially under severe drought conditions, even if perennial ryegrass can adapt to the environment by adjusting its root growth strategy, its growth vigor and stress resistance will still decrease significantly, causing huge losses to lawn maintenance and livestock production.

[0003] Currently, traditional methods to alleviate drought stress in perennial ryegrass mainly include water-saving irrigation, breeding drought-resistant varieties, and applying chemical drought-resistant agents. However, water-saving irrigation is limited by region and facilities, drought-resistant breeding has a long cycle and high cost, and chemical drought-resistant agents are prone to causing soil pollution and ecological damage, making it difficult to meet the needs of efficient, environmentally friendly and large-scale application. Summary of the Invention

[0004] Based on the above technical background, the main objective of this invention is to provide a microbial selenium protein preparation for alleviating drought stress in perennial ryegrass, as well as its preparation method, usage method and application, so as to overcome the shortcomings of the prior art.

[0005] Based on the above background technology, the field has gradually focused on green and efficient biological regulation methods, among which the application of components such as Bacillus natto, selenium, potassium humate, and salicylic acid in plant drought resistance has received widespread attention. Bacillus natto, as a beneficial microorganism, can enhance plant stress resistance by secreting growth-promoting metabolites, improving the soil microecological environment, and enhancing the absorption capacity of plant roots. Selenium, as a trace element required for plant growth and development, can participate in the construction of the plant's antioxidant system. Selenium-containing metabolites generated through microbial transformation can effectively regulate plant osmotic balance and protect cell membrane integrity. Potassium humate, as a natural organic matter, can not only improve soil structure and promote root development, but also regulate plant stomatal conductance, reduce water transpiration, and enhance plant photosynthesis and osmotic regulation capacity. Salicylic acid is an important plant signaling molecule that can induce the activation of the plant's antioxidant system, regulate stomatal movement, and promote the accumulation of osmotic regulatory substances, thereby reducing oxidative damage caused by drought stress. Existing research has confirmed that these components, when applied individually, can improve plant drought resistance to a certain extent. However, the mechanisms of action of a single component are relatively simple and cannot comprehensively alleviate the combined effects of drought stress on plants.

[0006] Due to the limitations of single-component formulations, synergistic regulation by multiple beneficial factors has become a new direction in plant drought resistance research. Its advantage lies in the fact that each component can complement and enhance each other through different pathways, forming a comprehensive drought protection system with a more significant stress relief effect than a single component. However, achieving multi-factor synergistic regulation presents several technical challenges: First, the concentration ratio of each component is crucial; excessively high or low concentrations not only fail to exert a synergistic effect but may also produce antagonistic effects, impacting plant growth. Second, how to integrate different types of components, such as microorganisms, trace elements, and plant growth regulators, through reasonable processes to achieve efficient functioning of each component while ensuring the stability and safety of the formulation remains an unsolved problem in the field.

[0007] To achieve the aforementioned objectives, the technical solution adopted by this invention includes:

[0008] The first aspect of this invention is to provide a microbial selenium protein preparation for alleviating drought stress in perennial ryegrass, the microbial selenium protein preparation comprising the following raw materials in parts by weight: 30-50 parts by weight of peptone, 20-30 parts by weight of beef extract, 50-70 parts by weight of sucrose, 15-25 parts by weight of salt, 5-40 parts by weight of selenium-enriched mushroom powder (selenium content is 2‰), 5-15 parts by weight of organosilicon (ethoxylated trisiloxane content is 10%), 15-25 parts by weight of calcium amino acid (calcium content is 10%), 10-90 parts by weight of potassium humate (humic acid content is 50%), 15-25 parts by weight of betalain (total betalain pigment content is 3%), 1-20 parts by weight of salicylic acid (o-hydroxybenzoic acid content is 99%), and 180-220 parts by weight of Bacillus natto.

[0009] Preferably, the microbial selenium protein preparation comprises the following raw materials in parts by weight: 35-45 parts by weight of peptone, 22-28 parts by weight of beef extract, 55-65 parts by weight of sucrose, 17-22 parts by weight of salt, 10-30 parts by weight of selenium-enriched mushroom powder (content 2‰), 7-12 parts by weight of organosilicon (content 10%), 18-22 parts by weight of calcium amino acids (content 10%), 20-50 parts by weight of potassium humate (content 50%), 17-22 parts by weight of betalain (content 3%), 5-20 parts by weight of salicylic acid (content 99%), and 190-210 parts by weight of Bacillus natto.

[0010] More preferably, the microbial selenium protein preparation comprises the following raw materials in parts by weight: 40 parts by weight of peptone, 24 parts by weight of beef extract, 60 parts by weight of sucrose, 20 parts by weight of salt, 20 parts by weight of selenium-enriched mushroom powder (content 2‰), 10 parts by weight of organosilicon (content 10%), 20 parts by weight of calcium amino acids (content 10%), 30 parts by weight of potassium humate (content 50%), 20 parts by weight of betalain (content 3%), 10 parts by weight of salicylic acid (content 99%), and 200 parts by weight of Bacillus natto.

[0011] The functions and effects of each raw material in the microbial selenium protein preparation are as follows: Peptone: As a core nitrogen source, it can provide high-quality protein and amino acids for the growth and reproduction of Bacillus natto, ensuring the metabolic activity of microorganisms. At the same time, it can be absorbed by the roots of ryegrass, supplementing the nitrogen needed for plant growth and enhancing the plant's resistance to stress.

[0012] Beef extract: Rich in protein, polypeptides and minerals, it works synergistically with peptone to form a complex nutrient matrix, optimizing the nutrient structure of the culture medium. It can provide sufficient carbon and nitrogen sources for Bacillus natto, promote its secretion, and promote growth metabolites.

[0013] Sucrose: As a rapidly available carbon source, it provides energy for the aerobic fermentation of Bacillus natto, accelerating the proliferation and metabolism of the strain; on the other hand, it can regulate the osmotic pressure of the preparation and improve its stability. At the same time, after being absorbed by ryegrass, it can be converted into soluble sugar, enhancing the plant's osmotic regulation capacity and alleviating drought stress.

[0014] Salt: It can maintain the osmotic pressure balance of the protein preparation system, provide the necessary sodium element for microbial growth, regulate the permeability of ryegrass cell membranes, help plants maintain water balance, and enhance drought resistance.

[0015] Selenium-enriched mushroom powder: The core selenium source carrier contains organic selenium that is easily converted by microorganisms into selenium-containing metabolites (such as selenoproteins and selenium polysaccharides), which can activate the antioxidant system of ryegrass, protect cell membrane integrity, and reduce oxidative damage caused by drought; at the same time, it provides nutrition for Bacillus natto and helps promote its function.

[0016] Organosilicon: As a wetting and penetrating agent, it can reduce the surface tension of protein preparations, enhance the adhesion and penetration ability of diluted protein preparations in ryegrass roots and soil, and promote the absorption of nutrients by roots; at the same time, it can form a protective film on the plant surface to reduce water transpiration and loss.

[0017] Amino acid calcium: It can provide directly absorbable amino acid chelated calcium. Calcium can stabilize the cell membrane structure of ryegrass and reduce cell membrane leakage under drought stress. As an organic nutrient, amino acids can promote root development and improve the plant's absorption efficiency of water and nutrients.

[0018] Potassium humate: a natural organic matter that can improve soil structure, loosen soil, promote the expansion of ryegrass roots, regulate the stomatal conductance of plants, and reduce water transpiration; at the same time, it can chelate nutrients in the soil, improve nutrient utilization, enhance the plant's photosynthesis and osmotic regulation capabilities, and synergistically alleviate drought stress.

[0019] Betalains: As a natural antioxidant, it can eliminate reactive oxygen free radicals produced by plants under drought stress and reduce oxidative damage; at the same time, it has a certain water retention capacity, which can help maintain the relative water content of leaves, delay plant wilting, and improve the stability of the formulation and extend the shelf life.

[0020] Salicylic acid: a key plant signaling molecule that can induce the activation of the antioxidant enzyme system in ryegrass, promote the accumulation of osmotic regulators (soluble sugars, proline), regulate stomatal movement, reduce water consumption, alleviate membrane damage caused by drought, and enhance the plant's stress response.

[0021] Bacillus natto: a core functional microorganism that can secrete growth hormones, antibacterial substances and extracellular polysaccharides, improve the soil microecological environment, and inhibit the reproduction of harmful microorganisms; at the same time, it enhances the vitality of ryegrass roots, promotes the absorption of water and nutrients by the roots, and works with components such as selenium and potassium humate to build a comprehensive drought resistance and protection system.

[0022] The second aspect of this invention is to provide a method for preparing a microbial selenium protein preparation for alleviating drought stress in perennial ryegrass as described in the first aspect of this invention, the preparation method comprising the following steps: Step 1: Add water to peptone, beef extract, sucrose, salt, and selenium-enriched mushroom powder, bring to a boil, autoclave, and cool to obtain a culture medium; Step 2: Add organosilicon, calcium amino acids, potassium humate, betaine, salicylic acid and Bacillus natto to the culture medium and carry out aerobic fermentation to obtain microbial selenium protein preparation.

[0023] The steps described above are described in detail below.

[0024] In step 1, the conditions for autoclaving are: autoclaving at 120–125°C for 15–25 minutes.

[0025] Preferably, the conditions for autoclaving are: autoclaving at 121°C for 20 minutes.

[0026] After autoclaving, the culture medium is cooled to 25–35°C, preferably to 30°C, to obtain the culture medium.

[0027] In step 2, the conditions for aerobic fermentation are: 35-40℃, aerobic fermentation for 4-10 days.

[0028] Preferably, the conditions for aerobic fermentation are: aerobic fermentation at 37°C for 7 days.

[0029] The third aspect of the present invention is to provide an application of the microbial selenium protein preparation for alleviating drought stress in perennial ryegrass as described in the first aspect of the present invention, which can be used to propagate ryegrass and improve the drought resistance of perennial ryegrass.

[0030] A fourth aspect of the present invention is to provide a method of using the microbial selenium protein preparation for alleviating drought stress in perennial ryegrass as described in the first aspect of the present invention, the method comprising the following steps: Dilute the microbial selenium protein preparation with water 90 to 120 times to obtain a diluted solution of the biological selenium protein preparation. Then, apply the diluted solution to the roots of ryegrass seedlings, applying it 2 to 5 times in total, once every 2 to 5 days. The amount of diluted solution used is 40 to 60 ml.

[0031] Preferably, the method of use includes the following steps: Dilute the microbial selenium protein preparation with water 100 times to obtain a diluted solution of the biological selenium protein preparation. Then, apply the diluted solution to the roots of ryegrass seedlings for a total of 3 root irrigations, once every 3 days. The amount of diluted solution used is 50 ml.

[0032] The beneficial effects of this invention are as follows: (1) The biological selenium protein preparation of the present invention breaks through the limitations of single components and organically integrates Bacillus natto, selenium, potassium humate, salicylic acid and other components. Each component can complement each other and exert synergistic effects through different pathways: Bacillus natto can improve soil microecology, selenium-enriched mushroom powder can be converted into selenium-containing antioxidant products, potassium humate can optimize water use, and salicylic acid can activate plant stress resistance signals. The above components work together to form a comprehensive drought resistance and protection system, which can significantly improve the stress relief effect compared with a single component.

[0033] (2) The biological selenium protein preparation must be diluted with water before application. After application, it can increase the relative water content of ryegrass leaves and the plant height of ryegrass under drought stress, increase soluble sugar, reduce malondialdehyde content and relative conductivity, effectively protect cell membrane integrity, and enhance photosynthesis and osmotic regulation.

[0034] (3) The preparation process of this biological selenium protein preparation only requires two core operations, including high pressure sterilization and aerobic fermentation. The conditions are mild and easy to control, and the raw materials are easy to obtain. When using it, it only needs to be diluted and then applied to the roots. The operation is simple and there are no geographical or facility restrictions. Compared with water-saving irrigation, drought-resistant breeding and chemical drought-resistant agents, the biological selenium protein preparation and its application method described in this invention are more environmentally friendly and low-cost. It can be applied on a large scale to lawn maintenance and ryegrass planting in animal husbandry.

[0035] (4) The raw materials used in this biological selenium protein preparation are all natural organic matter, beneficial microorganisms and trace elements. No harmful chemical substances are added. After application, it will not cause soil pollution or ecological damage. It not only ensures the quality of ryegrass, but also meets the needs of green agricultural development, taking into account both economic and ecological value. Detailed Implementation

[0036] The present invention will now be described in detail, and its features and advantages will become clearer and more apparent from these descriptions.

[0037] Example The present invention is further illustrated below with specific examples. These embodiments are merely illustrative and not intended to limit the scope of the invention. All raw materials used in the embodiments of the present invention are commercially available.

[0038] Example 1 A microbial selenium protein preparation for alleviating drought stress in perennial ryegrass, the microbial selenium protein preparation being prepared from the following raw materials by weight: 40 g peptone, 24 g beef extract, 60 g sucrose, 20 g salt, 20 g selenium-enriched mushroom powder (content 2‰), 10 g organosilicon (content 10%), 20 g calcium amino acids (content 10%), 30 g potassium humate (content 50%), 20 g betalain (content 3%), 10 g salicylic acid (content 99%), and 200 g Bacillus natto.

[0039] A method for preparing a microbial selenium protein preparation to alleviate drought stress in perennial ryegrass, the preparation method comprising the following steps: Weigh each raw material according to the above-mentioned weight and set aside. Add 1000 mL of water to peptone, beef extract, sucrose, salt, and selenium-enriched mushroom powder, bring to a boil, autoclave at 121℃ for 20 min, and then cool to 38℃ to obtain the culture medium.

[0040] Organosilicon, calcium amino acids, potassium humate, betaine, and salicylic acid were added sequentially to the culture medium, followed by the addition of Bacillus natto. Fermentation was carried out at 37°C under aerobic conditions for one week to obtain the microbial selenium protein preparation.

[0041] Example 2 The microbial selenium protein preparation was prepared in a manner similar to that in Example 1, except that the amount of selenium-enriched mushroom powder (content 2‰) added was 5 grams.

[0042] Example 3 The preparation of the microbial selenium protein preparation was carried out in a manner similar to that in Example 1, except that the amount of selenium-enriched mushroom powder (content 2‰) added was 10 grams.

[0043] Example 4 The microbial selenium protein preparation was prepared in a manner similar to that in Example 1, except that the amount of selenium-enriched mushroom powder (content 2‰) added was 40 grams.

[0044] Example 5 The microbial selenium protein preparation was prepared in a manner similar to that in Example 1, except that the amount of potassium humate (content 50%) added was 10 grams.

[0045] Example 6 The microbial selenium protein preparation was prepared in a manner similar to that in Example 1, except that the amount of potassium humate (content 50%) added was 90 grams.

[0046] Example 7 The microbial selenium protein preparation was prepared in a manner similar to that in Example 1, except that the amount of salicylic acid (99% content) added was 1 gram.

[0047] Example 8 The microbial selenium protein preparation was prepared in a manner similar to that in Example 1, except that the amount of salicylic acid (99% content) added was 5 grams.

[0048] Example 9 The microbial selenium protein preparation was prepared in a manner similar to that in Example 1, except that the amount of salicylic acid (99% content) added was 20 grams.

[0049] Example 10 The method of using the microbial selenium protein preparation includes the following steps: Dilute the above-mentioned microbial selenium protein preparation 100 times with water before application. A root irrigation experiment was conducted on perennial ryegrass seedlings (Excellence variety), irrigating once every 3 days for a total of 3 times, using 50 ml of diluted solution. The control group was irrigated with an equal volume of deionized water. After three treatments, irrigation was stopped, and simulated natural drought treatment began. After 12 days of natural drought stress treatment, samples were taken and relevant indicators were measured. The results are shown in Table 1.

[0050] Experimental Example Propagation of perennial ryegrass: a. Select plump, uniformly sized perennial ryegrass seeds, disinfect them with 75% alcohol for about 1 minute, disinfect them with 1% sodium hypochlorite solution for 30 minutes, then rinse them three times with distilled water, and place the seeds in a petri dish with filter paper to germinate.

[0051] b. After seed germination, seedlings of uniform growth were planted in a substrate using 4-inch pots (10 cm × 10 cm × 9 cm). The substrate consisted of vermiculite and perlite in a 1:1 ratio, with 9 seedlings planted per pot. The materials were then cultured in an artificial climate chamber under the following conditions: 18 h light / 6 h dark (25℃ / 20℃). After 3 weeks of growth, the seedlings were used for subsequent experiments.

[0052] Experiment Example 1: Test on the effect of alleviating drought stress in perennial ryegrass The microbial selenium protein preparations obtained in Examples 1-4 were tested. The testing procedure was as follows: the microbial selenium protein preparations obtained in Examples 1-4 were diluted 100 times with water to obtain diluted protein preparation solutions. Root irrigation experiments were conducted on perennial ryegrass seedlings (Excellence) using these diluted solutions, irrigating once every 3 days for a total of 3 times, with each irrigation being 50 mL. The control group was irrigated with an equal volume of deionized water. After three treatments, irrigation was stopped, and simulated natural drought treatment began. After 12 days of natural drought stress treatment, samples were taken and relevant indicators were measured. The test results are shown in Table 1.

[0053] Table 1. Effects of different concentrations of selenium-enriched mushroom powder on the mitigation of drought stress in perennial ryegrass by microbial selenoprotein preparations.

[0054] Selenium-enriched mushroom powder can serve as a nutrient source and antioxidant for microorganisms, promoting the growth and metabolism of functional strains. It can also be transformed by microorganisms to generate selenium-containing metabolites, thereby improving drought resistance by regulating plant osmotic balance and protecting cell membrane integrity.

[0055] As shown in Table 1, adding 20 grams of selenium-enriched mushroom powder to 1000 mL of liquid culture medium and fermenting the resulting microbial selenium protein preparation significantly increased the plant height, relative leaf water content, soluble sugar and chlorophyll content of perennial ryegrass, while reducing the malondialdehyde content. This indicates that the amount of selenium-enriched mushroom powder added has a significant impact on the effect of the protein preparation on the drought resistance of ryegrass. Fermentation broth obtained by adding an appropriate concentration of selenium-enriched mushroom powder to the fermentation system can significantly improve the drought resistance of perennial ryegrass.

[0056] Experiment Example 2: Improvement Effect Test The microbial selenium protein preparations obtained in Examples 1, 5, and 6 were tested. The testing procedure was as follows: the microbial selenium protein preparations obtained in Examples 1, 5, and 6 were diluted 100 times with water to obtain diluted protein preparation solutions. Root irrigation experiments were conducted on perennial ryegrass seedlings (Excellence) using this diluted solution, irrigating once every 3 days for a total of 3 times, with each irrigation being 50 mL. The control group was irrigated with an equal volume of deionized water. After three treatments, irrigation was stopped, and simulated natural drought treatment began. After 12 days of natural drought stress treatment, samples were taken and relevant indicators were measured. The test results are shown in Table 2.

[0057] Table 2. Effects of different concentrations of potassium humate on the mitigation of drought stress in perennial ryegrass by microbial selenoprotein preparations.

[0058] Potassium humate itself has the effect of promoting plant growth, improving soil structure and enhancing stress resistance. This invention uses it as a component of microbial fermentation substrate, which can further improve the biological activity of fermentation liquid.

[0059] As shown in Table 2, adding an appropriate concentration of potassium humate to the microbial selenium protein preparation used in this invention can improve the drought resistance of perennial ryegrass. Compared with the control, the microbial selenium protein preparation obtained by adding 30 g of potassium humate (50% content) to 1000 mL of culture medium significantly increased the plant height, relative leaf water content, soluble sugar and chlorophyll content of perennial ryegrass, and decreased the malondialdehyde content. This indicates that the microbial selenium protein preparation obtained by adding an appropriate concentration of potassium humate to the fermentation system can improve the drought resistance of perennial ryegrass by improving plant water status, enhancing photosynthesis and osmotic regulation capacity, and reducing membrane damage.

[0060] Experiment Example 3: Improvement Effect Test The microbial selenium protein preparations obtained in Examples 1 and 7-9 were tested. The testing procedure was as follows: the microbial selenium protein preparations obtained in Examples 1, 5, and 6 were diluted 100 times with water to obtain diluted protein preparation solutions. Root irrigation experiments were conducted on perennial ryegrass seedlings (Excellence) using this diluted solution, irrigating once every 3 days for a total of 3 times, with each irrigation being 50 mL. The control group was irrigated with an equal volume of deionized water. After three treatments, irrigation was stopped, and simulated natural drought treatment began. After 12 days of natural drought stress treatment, samples were taken and relevant indicators were measured. The test results are shown in Table 3.

[0061] Table 3. Effects of salicylic acid content in protein formulations on the mitigation of drought stress in perennial ryegrass by microbial selenoprotein formulations.

[0062] Salicylic acid, as an important plant signaling molecule, can enhance plant stress resistance through inducing antioxidant systems, regulating stomatal movement, and promoting the accumulation of osmotic regulators. In this invention, adding 10 grams of salicylic acid to 1000 mL of a microbial selenoprotein preparation significantly increased the plant height, relative leaf water content, soluble sugar and chlorophyll content of perennial ryegrass, while decreasing malondialdehyde (MDA) content. These results indicate that an appropriate concentration of salicylic acid can effectively enhance the drought resistance of perennial ryegrass. This demonstrates that a microbial selenoprotein preparation obtained by adding an appropriate concentration of salicylic acid can significantly improve the drought resistance of perennial ryegrass.

[0063] Experiment Example 4 To investigate the effects of different dilution ratios of microbial selenoprotein preparations on their ability to alleviate drought stress in perennial ryegrass, we conducted a gradient experiment with different dilution ratios of microbial selenoprotein preparations: The microbial selenoprotein preparation obtained in Example 1 was diluted with water at different ratios (0, 50, 100, 200) for application. A root irrigation experiment was conducted on perennial ryegrass (Excellence), with irrigation every 3 days for a total of 3 times, each irrigation being 50 mL. The control group (group ck in Table 4) was irrigated with an equal volume of deionized water. After three treatments, irrigation was stopped, and simulated natural drought treatment began. After 12 days of natural drought stress treatment, samples were taken and relevant indicators were measured. The test results are shown in Table 4.

[0064] Table 4. Effects of different dilution ratios of microbial selenoprotein preparations on alleviating drought stress in perennial ryegrass.

[0065] Table 4 shows that diluting the microbial selenoprotein preparation prepared in this invention to different ratios and then applying it can improve the drought resistance of perennial ryegrass, exhibiting a significant concentration effect. The 100-fold dilution treatment showed the best effect, significantly increasing the plant height, relative leaf water content, soluble sugar and chlorophyll content of perennial ryegrass, while reducing malondialdehyde content. These results indicate that the 100-fold dilution of the microbial selenoprotein preparation can effectively alleviate the inhibitory effect of drought stress on the growth and physiological metabolism of perennial ryegrass by improving plant water status, enhancing photosynthesis and osmotic regulation, and reducing membrane damage.

[0066] The present invention has been described in detail above with reference to specific embodiments and exemplary examples; however, these descriptions should not be construed as limiting the present invention. Those skilled in the art will understand that various equivalent substitutions, modifications, or improvements can be made to the technical solutions and embodiments of the present invention without departing from the spirit and scope of the invention, and all such modifications and improvements fall within the scope of the present invention. The scope of protection of the present invention is defined by the appended claims.

Claims

1. A microbial selenium protein preparation for alleviating drought stress in perennial ryegrass, characterized in that, The microbial selenium protein preparation comprises the following raw materials in parts by weight: 30-50 parts peptone, 20-30 parts beef extract, 50-70 parts sucrose, 15-25 parts salt, 5-40 parts selenium-enriched mushroom powder, 5-15 parts organosilicon, 15-25 parts calcium amino acids, 10-90 parts potassium humate, 15-25 parts betaine, 1-20 parts salicylic acid, and 180-220 parts Bacillus natto.

2. The microbial selenium protein preparation for alleviating drought stress in perennial ryegrass according to claim 1, characterized in that, The microbial selenium protein preparation comprises the following raw materials in parts by weight: 35-45 parts peptone, 22-28 parts beef extract, 55-65 parts sucrose, 17-22 parts salt, 10-30 parts selenium-enriched mushroom powder, 7-12 parts organosilicon, 18-22 parts calcium amino acids, 20-50 parts potassium humate, 17-22 parts betaine, 5-20 parts salicylic acid, and 190-210 parts Bacillus natto.

3. The microbial selenium protein preparation for alleviating drought stress in perennial ryegrass according to claim 1, characterized in that, The microbial selenium protein preparation comprises the following raw materials in parts by weight: 40 parts by weight of peptone, 24 parts by weight of beef extract, 60 parts by weight of sucrose, 20 parts by weight of salt, 20 parts by weight of selenium-enriched mushroom powder, 10 parts by weight of organosilicon, 20 parts by weight of calcium amino acids, 30 parts by weight of potassium humate, 20 parts by weight of betaine, 10 parts by weight of salicylic acid, and 200 parts by weight of Bacillus natto.

4. A method for preparing a microbial selenium protein preparation for alleviating drought stress in perennial ryegrass as described in any one of claims 1 to 3, characterized in that, The preparation method includes the following steps: Step 1: Add water to peptone, beef extract, sucrose, salt, and selenium-enriched mushroom powder, bring to a boil, autoclave, and cool to obtain a culture medium; Step 2: Add organosilicon, calcium amino acids, potassium humate, betaine, salicylic acid and Bacillus natto to the culture medium and carry out aerobic fermentation to obtain microbial selenium protein preparation.

5. The preparation method according to claim 4, characterized in that, In step 1, The conditions for autoclaving are: autoclaving at 120–125°C for 15–25 minutes.

6. The preparation method according to claim 4, characterized in that, In step 2, The conditions for aerobic fermentation are: 35–40°C, aerobic fermentation for 4–10 days.

7. The application of a microbial selenium protein preparation for alleviating drought stress in perennial ryegrass as described in any one of claims 1 to 3, characterized in that, The microbial selenium protein preparation can be used to propagate ryegrass and improve the drought resistance of perennial ryegrass.

8. A method of using a microbial selenium protein preparation for alleviating drought stress in perennial ryegrass as described in any one of claims 1 to 3, characterized in that, The method of use includes the following steps: Dilute the microbial selenium protein preparation with water 90 to 120 times to obtain a diluted solution of the biological selenium protein preparation. Then, apply the diluted solution to the roots of ryegrass seedlings for root irrigation, for a total of 2 to 5 times, once every 2 to 5 days. The amount of diluted solution used is 40 to 60 mL.

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