Stress-resistant agent for alfalfa planting and use method of stress-resistant agent

Abstract

According to the stress-resistant agent for alfalfa planting and the use method of the stress-resistant agent, the synergistic effect of betaine, glucose and monopotassium phosphate is utilized, and a seed soaking or foliage spraying mode is adopted, so that on one hand, expression of a series of stress-resistant related genes can be induced, ROS generated by stress excitation can be eliminated, and the stress resistance of alfalfa can be improved; the damage of reactive oxygen free radicals and the like to plasma membranes is reduced, the activity of antioxidant enzymes is enhanced, the accumulation of osmotic regulation substances is enhanced, and the chlorophyll synthesis of alfalfa leaves is remarkably improved, so that the stress resistance of plants is enhanced, and the damage of adversity stress to alfalfa plants is relieved. The stress-resistant agent is simple in component, free of pungent smell and good in stress-resistant effect, has the characteristics of no environmental pollution, zero emission and no residue, and can obviously promote germination of alfalfa seeds, regulate growth of alfalfa seedlings and improve stress resistance of the alfalfa seedlings.

Description

Technical Field

[0001] The present application relates to the field of stress-resistant planting of leguminous plants, and in particular to a stress-resistant agent for alfalfa planting and a method for using the same. Background Art

[0002] Forage is an important basic support for the development of the grass-livestock industry. With the improvement of residents' consumption level, the demand for dietary structure of Chinese residents has changed. The proportion of meat, eggs and milk consumption in residents' food has gradually increased. The growth of people's demand for dairy products has led to the vigorous development of the alfalfa industry. The increase in alfalfa production capacity can, on the one hand, alleviate the problem of insufficient high-quality forage in my country; on the other hand, it can also improve economic benefits while achieving ecological governance of "locking wind and sand, controlling salinization, improving ecology, and beautifying the environment."

[0003] However, the climate and environment in the north of my country, where alfalfa bases are concentrated, are harsh and the land is barren, so the growth of forage grass often faces severe adverse stress. Biological stress caused by vegetation degradation, as well as abiotic stress caused by drought, salinity, cold damage, heavy metal toxins, etc., will seriously affect the growth of alfalfa. Among them, drought, salinity, and extreme temperatures are the main factors that affect the distribution of alfalfa planting and limit its yield and safety.

[0004] Given the adverse growth environment of leguminous forage, enhancing its stress resistance is the key to meeting the growing demand for high-quality forage in my country's animal husbandry. Therefore, exploring the response of alfalfa to salinity stress and low temperatures in winter and spring and updating mitigation measures to enhance alfalfa tolerance are of great significance to ensuring the safe production of high-quality forage in my country. Summary of the invention

[0005] In view of the deficiencies of the prior art, the present application provides an alfalfa planting stress-resistant agent and a method of using the same. The present application uses betaine, glucose and potassium dihydrogen phosphate to prepare a highly effective stress-resistant agent to improve the synthesis efficiency of chlorophyll in alfalfa and resist the damage of ROS and free radicals to alfalfa during adverse stress. The present application specifically adopts the following technical solutions.

[0006] Firstly, to achieve the above purpose, a stress resistance agent for alfalfa planting is proposed, which comprises: 0.5-0.7 parts by weight of betaine; 0.2-0.3 parts by weight of potassium dihydrogen phosphate; and 0.5-0.7 parts by weight of glucose dissolved in distilled water.

[0007] Optionally, the stress-resistant agent for alfalfa planting as described above further comprises 0.5 parts by weight of a non-ionic surfactant.

[0008] Optionally, the stress-resistant agent for alfalfa planting as described above, wherein each kilogram of the stress-resistant agent comprises: 6g betaine, 2g potassium dihydrogen phosphate, 5g glucose, and the balance is water.

[0009] Optionally, the stress-resistant agent for alfalfa planting as described above, wherein per kilogram of the stress-resistant agent, 6g of betaine, 2.5g of potassium dihydrogen phosphate, 6g of glucose, and 5ml of non-ionic surfactant are dissolved.

[0010] Optionally, in any of the above-described stress-resistant agents for alfalfa cultivation, the non-ionic surfactant is Tween 20.

[0011] Optionally, the stress-resistant agent for alfalfa cultivation as described above is prepared according to the following steps: first, betaine, potassium dihydrogen phosphate and glucose are added to 500 ml of distilled water to fully dissolve them; then, water is added to the dissolved mixture to make the volume 1000 ml.

[0012] The stress-resistant agent for alfalfa planting as described above, wherein the pH value of the stress-resistant agent is between 5.5 and 6.

[0013] At the same time, to achieve the above-mentioned purpose, the present application also provides a method for using the stress-resistant agent for alfalfa planting as described above, the steps of which include: using the stress-resistant agent to soak alfalfa seeds 3-5 days before sowing; spraying the stress-resistant agent on the alfalfa leaf surface in the morning and evening of the 6th to 8th day after the alfalfa turns green in spring, when the stomata of the leaves are open; and / or spraying the stress-resistant agent on the alfalfa leaf surface once a day in the morning and evening of 6-8 days before wintering, when the stomata of the alfalfa leaves are open; and / or spraying the stress-resistant agent on the alfalfa leaf surface in the morning and evening of 6-8 days after the alfalfa turns green, when the stomata of the alfalfa leaves are open; and / or spraying the alfalfa plants in the evening on sunny days 2-5 days before the arrival of the winter cold wave.

[0014] Optionally, in any of the above methods for using the stress-resistant agent for alfalfa cultivation, when soaking alfalfa seeds, the seeds need to be soaked for 24 hours and then dried in the shade.

[0015] Optionally, in the method of using any of the stress-resistant agents for alfalfa cultivation as described above, when spraying alfalfa leaves, the method is to spray until water droplets of the anti-cold protective agent hang on the leaves, or spray at a dosage of 750 kg per hectare. Beneficial Effects

[0016] The stress-resistant agent for alfalfa planting provided by the present application utilizes the synergistic effect between betaine, glucose and potassium dihydrogen phosphate. Through seed soaking or foliar spraying, on the one hand, it can induce the expression of a series of stress-resistant related genes, reduce the damage of reactive oxygen free radicals to the plasma membrane, remove the ROS stimulated by adversity, enhance the activity of antioxidant enzymes and enhance the accumulation of osmotic regulating substances, significantly improve the synthesis of chlorophyll in alfalfa leaves, thereby enhancing the stress resistance of the plant and alleviating the damage of adversity stress to alfalfa plants. The stress-resistant agent of the present application is weakly acidic, can effectively reduce the irritating smell of the preparation and reduce the requirements for transportation and storage conditions. The stress-resistant agent of the present application has simple ingredients, no irritating smell, good stress-resistant effect, and has the characteristics of pollution-free environment, zero emission and no residue, which can significantly promote the germination of alfalfa seeds, regulate the growth of alfalfa seedlings, and improve the stress resistance of seedlings.

[0017] The innovation of the present invention is that it uses relatively simple chemical components and a relatively small amount to enable alfalfa to grow and resist stress while achieving high yield. In addition, the components of the present invention are environmentally friendly and pollution-free, with a simple ratio and easy operation. The reason why the present application can achieve such an anti-stress effect with a small amount of simple components is that: First, betaine is an important osmotic protectant, mainly in plant chloroplasts and cytoplasm. Exogenous betaine can enhance the chlorophyll content of leaves, photosynthesis capacity, antioxidant system activity and antioxidant accumulation in plant cells under low temperature stress, stabilize plant cell membranes and maintain plant stress resistance. This application utilizes the characteristics of exogenous betaine that is easily absorbed and transported to the entire plant body through plant leaves and roots, and is not easily metabolized for a long time. On the one hand, it ensures that betaine achieves a certain relief and protection effect in the plant body. On the other hand, it can promote the electron transfer rate of the COX pathway through the synergy of betaine and potassium dihydrogen phosphate, thereby promoting mitochondrial ATP content, increasing the activity of mitochondrial cytochrome cell oxidase, and achieving stress resistance. The reason is that when plants suffer from adverse stress, the accumulation of betaine in the plant body can help plant cells maintain a stable osmotic pressure. It gradually accumulates in the chloroplasts and cytoplasm of the plant to reduce the osmotic potential of the cells and protect the cells from damage by adverse environments. Under salt stress, it can also increase the activity of antioxidant enzymes to varying degrees, reduce the damage of salt stress to the protective enzyme system, reduce the damage of active oxygen free radicals to the plasma membrane and the aggravation of membrane lipid peroxidation, protect the cell plasma membrane to maintain stability and integrity, and thus maintain the normal physiological functions and metabolic activities of the organism.

[0018] Secondly, glucose is the main energy source for plants and also an important raw material for synthesizing other organic substances. If the absolute amount of glucose in the plant is insufficient, the immunity and resistance of the plant will be reduced, and it will be vulnerable to the attack of pests and diseases. Under normal circumstances, plants convert carbon dioxide and water into glucose through photosynthesis and release oxygen. Various biological and abiotic stresses (such as high salt, drought and pathogen infection, etc.) will lead to energy deficit in plants and inhibit plant growth or even death, so maintaining energy balance in the body is crucial for plants to resist adversity. The present application promotes balanced photosynthesis, respiration and fatty acid β-oxidation processes through exogenous soluble sugars, while enhancing the redox process of cells, and can also use the glucose inside the plant to synthesize other organic substances that are also crucial for the growth and development of plants, such as proteins, fats and nucleic acids. Sugar, as a signal molecule, regulates the content of intracellular carbohydrates, the activity of metabolism-related enzymes, and controls gene expression. During salt stress, the present application can increase the content of glucose and fructose in salt-sensitive rice by exogenous glucose, and improve the salt tolerance of salt-sensitive rice. Analysis of Arabidopsis gene transcription levels showed that glucose can induce the expression of a large number of stress response genes. In addition, the expression levels of various sugar metabolism enzymes in Arabidopsis were significantly increased under abiotic stress, indicating that sugar signals play an important regulatory role in plant response to stress.

[0019] Third, the potassium dihydrogen phosphate in the ratio of this application is a high-concentration binary compound fertilizer containing phosphorus and potassium elements. It has a high nutritional value and a low salt index, and will not cause burns to seeds, seedlings and roots. The application uses potassium dihydrogen phosphate, which not only promotes the absorption of nitrogen and phosphorus by crops, increases grain weight and yield, but also can be used in conjunction with betaine and glucose to enhance crop stress resistance and promote the healing of damaged crop tissues. Potassium dihydrogen phosphate is safe and non-toxic, 100% plant nutrition, and is used for foliar fertilizer spraying. It has a high absorption and utilization rate, can supplement the nutrient deficiency of crops during critical periods, and can also regulate crop growth, make crop metabolism vigorous, and achieve the effect of increasing quality and yield. In particular, the potassium element in potassium dihydrogen phosphate, as the highest cation in plant cells, can pass through the biofilm at high speed, as a key auxiliary factor for most enzymes, and promote the activation of enzyme activity in the antioxidant enzyme system; phosphorus compounds and nucleic acids can participate in the differentiation of flower buds. The application can increase the number of inflorescences by synchronously applying phosphorus fertilizers, ensure that the differentiation of flower buds under adversity is not inhibited, and maintain plant activity.

[0020] In addition, the stress-resistant agent of the present application can also provide a slightly acidic solvent environment through potassium dihydrogen phosphate at the same time, and can directly dissolve each solvent. The components of the stress-resistant agent of the present application are simple, the concentration is low, and the toxicity is low. The surface tension of the aqueous solution can be further reduced by the high surface activity of non-ionic surfactants such as Tween, so that the number of micelle aggregation of the stress-resistant agent is increased, and strong solubilization is guaranteed. In addition, since non-ionic surfactants such as Tween are not charged in the solution and will not bind to proteins, the low toxicity can further guarantee the biological effect of the above-mentioned preparation. Therefore, the preparation provided by the present application can achieve a non-irritating odor. It soaks seeds with stress-resistant agent before sowing alfalfa; and / or sprays the above-mentioned stress-resistant agent on the alfalfa leaf surface when the stomata of the alfalfa leaves are opened in the morning and evening 6-8 days before the alfalfa overwintering; and / or sprays the above-mentioned stress-resistant agent on the alfalfa leaf surface when the stomata of the alfalfa leaves are opened in the morning and evening 6-8 days after the alfalfa turns green. Alternatively, 2-5 days before the arrival of the winter cold wave weather, choose a sunny evening to spray the stress-resistant agent on the alfalfa plants.

[0021] The seed soaking of the stress-resistant agent of the present application can improve the stress resistance of alfalfa under saline-alkali stress, increase the chlorophyll content of alfalfa leaves, enhance various antioxidant properties, reduce the accumulation of active oxygen, and contribute to the establishment of alfalfa; the spraying of the stress-resistant agent can enhance the growth performance of alfalfa in a low-temperature wintering environment, and can stabilize the quality of alfalfa while increasing the yield of alfalfa, thereby improving production efficiency.

[0022] Other features and advantages of the present application will be set forth in the following description, and in part will become apparent from the description, or may be understood by practicing the present application. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] The accompanying drawings are used to provide a further understanding of the present application and constitute a part of the specification. Together with the embodiments of the present application, they are used to explain the present application and do not constitute a limitation of the present application. In the accompanying drawings: Figure 1 It is a comparison diagram of the stress-resistant agent of the present application and the control group plants; DETAILED DESCRIPTION

[0024] In order to make the purpose and technical solution of the embodiment of the present application clearer, the technical solution of the embodiment of the present application will be clearly and completely described in conjunction with the drawings of the embodiment of the present application. Obviously, the described embodiment is a part of the embodiment of the present application, not all of the embodiments. Based on the described embodiment of the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of this application.

[0025] It will be understood by those skilled in the art that, unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as generally understood by those skilled in the art to which this application belongs. It should also be understood that terms such as those defined in common dictionaries should be understood to have meanings consistent with the meanings in the context of the prior art, and will not be interpreted with idealized or overly formal meanings unless defined as herein.

[0026] The meaning of "and / or" described in this application means that the situations where each exists alone or both exist at the same time are included.

[0027] When seeds are under stress, on the one hand, the activities of various enzymes on the biomembrane will be affected, thus causing abnormal changes in cell physiology and biochemistry, which will limit the ATP produced by the COX respiratory pathway in the respiratory electron transfer on the inner membrane of the mitochondria during seed germination. ATP provides the ability for seed germination and is the key to seed germination. Under adverse stress, the electron transfer rate of the COX pathway is significantly suppressed, which limits ATP synthesis and affects seed germination.

[0028] Taking into account the fact that under adversity, COX respiration, which promotes ATP synthesis during seed germination, contributes more, this application uses betaine as an important osmotic protectant, and utilizes its characteristics of being easily absorbed through plant leaves and root parts and transported to the entire plant body, stabilizing the structure and function of biological macromolecules, stabilizing key enzymes of the triacetic acid cycle, photosystems, terminal oxidases and peripheral polypeptides and other physiological functions under osmotic stress. Through its synergy with potassium dihydrogen phosphate, the activity of enzymes in alfalfa cells is increased, thereby increasing the COX respiration electron transfer rate under adversity and the proportion of COX in total respiration. Under the premise that the respiration of germinating seeds in adversity is limited, this application uses betaine to regulate electron transfer, and regulates the distribution of electrons to the COX respiratory pathway that produces ATP, thereby promoting the production of more ATP in the cell to supply seed germination, increasing the activity of mitochondrial cytochrome cell oxidases, and delaying the damage to plants caused by the decline in ATP content in adversity. At the same time, glucose is also used as an important raw material for synthesizing other organic substances, and glucose is used as a signal molecule to regulate the content of intracellular carbohydrates, the activity of metabolism-related enzymes, and the characteristics of controlling the expression of related genes. The expression of various related antioxidant enzyme genes is increased by supplementing exogenous glucose, the stability of the biofilm is enhanced, and the biomass accumulation is improved. In addition, the present application also simultaneously utilizes the synergistic effect between potassium dihydrogen phosphate and glucose, and while providing sufficient nutrition to alfalfa plants, the enzyme activity in the antioxidant enzyme system is further activated through the synergy of phosphorus, potassium and betaine, so that the plants can resist adverse stress more quickly. Example 1

[0029] This embodiment provides an anti-adverse agent, which is composed of the following components with a total weight of 1 kg: 5g betaine, (weight 6‰) 2g potassium dihydrogen phosphate, (weight 2‰) 5g glucose, (weight 5‰) The balance is water.

[0030] The above-mentioned alfalfa stress resistance agent can be prepared according to the following steps: The first step is to add 5 g of glucose, 2 g of potassium dihydrogen phosphate, 6 g of betaine, and 5 ml of Tween 20 solubilizer into 500 ml of distilled water to fully dissolve them to obtain a first mixed solution; In the second step, water is added to the first mixed solution obtained in the first step to make the volume to 1000 ml.

[0031] For alfalfa crops, the above stress resistance agents can be used specifically in any of the following scenarios: Before sowing alfalfa, soak the seeds with stress-resistant agent; and / or spray the alfalfa leaves with the above stress-resistant agent in the morning and evening when the stomata of the alfalfa leaves are open 6-8 days before the alfalfa overwintering; and / or spray the alfalfa leaves with the above stress-resistant agent in the morning and evening when the stomata of the alfalfa leaves are open 6-8 days after the alfalfa turns green. Optionally, the method of use as described above further includes: spraying the alfalfa plants with the stress-resistant agent in the evening on a sunny day 2-5 days before the arrival of the winter cold wave.

[0032] When using anti-adverse agents: Each time you soak alfalfa seeds, you need to soak them for 24 hours and use them after drying in the shade. Each time the alfalfa plants are sprayed, the anti-cold protective agent is sprayed until water drops hang on the leaves, or sprayed at a dosage of 750 kg per hectare.

[0033] By comparing the growth conditions of alfalfa in the same environment without using stress-resistant agents, it can be seen that in a saline-alkali environment, soaking seeds with the stress-resistant agent of the present application can increase the aboveground dry weight of alfalfa by 24.14% and the antioxidant enzyme activity by 14.6%~102% compared with not using stress-resistant agents; in a low temperature environment, spraying the stress-resistant agent of the present application can increase the alfalfa plant height by 5.9%, the leaf antioxidant enzyme activity by 30%~91%, and the alfalfa hay yield by 16.1% compared with not using stress-resistant agents.

[0034] By comparing the growth conditions of alfalfa using a single dose of betaine (5 g / kg) of the same concentration in the same environment, it can be seen that the stress-resistant agent seed soaking of the present application can increase the aboveground dry weight of alfalfa by 13.65% and the antioxidant enzyme activity by 10.5%~88.2% compared with the single dose of betaine of the same concentration; in a saline-alkali environment, the stress-resistant agent spraying of the present application can increase the alfalfa plant height by 3.6% and the alfalfa hay yield by 9.5% compared with the single dose of betaine of the same concentration.

[0035] By comparing the growth conditions of alfalfa using a single dose of glucose (5 g / kg) of the same concentration in the same environment, it can be seen that in a saline-alkali environment, soaking seeds with the stress-resistant agent of the present application can increase the aboveground dry weight of alfalfa by 9.6% and the antioxidant enzyme activity by 9.8% to 66.4% compared with a single dose of glucose of the same concentration; in a low temperature environment, spraying the stress-resistant agent of the present application can increase the plant height of alfalfa by 4.2% and the alfalfa hay yield by 8.7% compared with a single dose of glucose of the same concentration.

[0036] By comparing the growth conditions of alfalfa using a single dose of potassium dihydrogen phosphate (2 g / kg) of the same concentration in the same environment, it can be seen that in a saline-alkali environment, the stress-resistant agent seed soaking of the present application can increase the aboveground dry weight of alfalfa by 7.5% and the antioxidant enzyme activity by 11.6%~85.4% compared with a single dose of potassium dihydrogen phosphate of the same concentration; in a low temperature environment, the stress-resistant agent spraying of the present application can increase the alfalfa plant height by 4.0% and the alfalfa hay yield by 7.8% compared with a single dose of potassium dihydrogen phosphate of the same concentration. Example 2

[0037] This embodiment provides an anti-adverse agent, which is composed of the following components with a total weight of 1 kg: 6g betaine 3g potassium dihydrogen phosphate 6g glucose 5ml Tween 20 solubilizer, The balance is water.

[0038] Optionally, the alfalfa stress resistance agent as described above, wherein the preparation steps are as follows: The first step is to add 6 g of glucose, 3 g of potassium dihydrogen phosphate, 6 g of betaine, and 5 ml of Tween 20 solubilizer into 500 ml of distilled water to fully dissolve them to obtain a first mixed solution; In the second step, water is added to the first mixed solution obtained in the first step to make the volume to 1000 ml.

[0039] Alternatively, for alfalfa crops, the above-mentioned stress resistance agents can also be used specifically in the following scenarios: Before sowing alfalfa, the seeds are soaked with stress-resistant agents; and / or the stress-resistant agents are sprayed on the alfalfa leaves in the morning and evening when the stomata of the alfalfa leaves are open 6-8 days before the alfalfa overwintering.

[0040] By comparing the growth conditions of alfalfa using lower concentrations of the same agent (2g betaine + 1.5g potassium dihydrogen phosphate + 4g glucose) in the same environment, it can be seen that in a saline-alkali environment, the stress-resistant agent seed soaking of the present application can increase the aboveground dry weight of alfalfa by 8.12% and the antioxidant enzyme activity by 6.83-54.14% compared with a low-concentration preparation of the same component; in a low-temperature environment, the stress-resistant agent spraying of the present application can increase the alfalfa plant height by 2.7% and the alfalfa hay yield by 6.5% compared with a low-concentration preparation of the same component.

[0041] By comparing the growth conditions of alfalfa using higher concentrations of the same agent (8g betaine + 4g potassium dihydrogen phosphate + 8g glucose) in the same environment, it can be seen that in a saline-alkali environment, the stress-resistant agent seed soaking of the present application can increase the aboveground dry weight of alfalfa by 7.39% and the antioxidant enzyme activity by 8.65-49.27% ​​compared with a high-concentration preparation of the same component; in a low temperature environment, the stress-resistant agent spraying of the present application can increase the hay yield of alfalfa plants by 7.5% compared with a high-concentration preparation of the same component.

[0042] By comparing the growth conditions of alfalfa using a similar effect agent 1 (0.6μl methyl jasmonate + 3g potassium dihydrogen phosphate + 6g glucose) in the same environment, it can be seen that in a saline-alkali environment, the stress-resistant agent seed soaking of the present application can increase the aboveground dry weight of alfalfa by 10.29% and the antioxidant enzyme activity by 3.56~75.19% compared with the similar effect agent 1; in a low temperature environment, the stress-resistant agent spraying of the present application can increase the alfalfa plant height by 2.6% and the alfalfa hay yield by 7.9% compared with the similar effect agent 1.

[0043] By comparing the growth conditions of alfalfa using similar effect agent 2 (0.6g betaine + 3g potassium nitrate and phosphate + 6g glucose) in the same environment, it can be seen that in a saline-alkali environment, the stress-resistant agent seed soaking of the present application can increase the aboveground dry weight of alfalfa by 7.28% and the antioxidant enzyme activity by 5.81~60.24% compared with the similar effect agent 2; in a low temperature environment, the stress-resistant agent spraying of the present application can increase the alfalfa plant height by 5.3% and the alfalfa hay yield by 5.9% compared with the similar effect agent 2.

[0044] By comparing the growth conditions of alfalfa using similar effect agent 3 (0.6g betaine + 3g potassium dihydrogen phosphate + 6g sucrose) in the same environment, it can be seen that in a saline-alkali environment, the stress-resistant agent seed soaking of the present application can increase the aboveground dry weight of alfalfa by 5.13% and the antioxidant enzyme activity by 1.95~50.87% compared with the similar effect agent 3; in a low temperature environment, the stress-resistant agent spraying of the present application can increase the alfalfa plant height by 4.1% and the alfalfa hay yield by 6.3% compared with the similar effect agent 3.

[0045] By comparing the growth conditions of alfalfa using similar effect agent 4 (0.6μl methyl jasmonate + 3g potassium nitrate and 6g sucrose) in the same environment, it can be seen that in a saline-alkali environment, the stress-resistant agent seed soaking of the present application can increase the aboveground dry weight of alfalfa by 12.35% and the antioxidant enzyme activity by 15.84~99.12% compared with the similar effect agent 4; in a low temperature environment, the stress-resistant agent spraying of the present application can increase the alfalfa plant height by 9.5% and the alfalfa hay yield by 8.9% compared with the similar effect agent 4. Example 3

[0046] This embodiment provides an anti-adverse agent, which is composed of the following components with a total weight of 1 kg: 7g betaine, 2.5g potassium dihydrogen phosphate, 7g glucose, 5ml Tween 20 solubilizer, The following test was conducted to compare the protective effect of the stress-resistant agent obtained by diluting the above compound agent on alfalfa: 1) Experimental Example 1: Effects of seed soaking with stress-resistant agents on the growth of alfalfa seedlings under saline-alkali stress The test variety was WL525. The coated seeds were washed to remove the coating. The alfalfa seeds were rinsed with tap water for several times, disinfected with 0.1% hypochlorous acid solution for 10 minutes, rinsed with distilled water 3 times, and then disinfected with 75% ethanol for 5 minutes and rinsed with distilled water 3 times to control the moisture. The seeds were then soaked for 24 hours, treated with stress-resistant soaking agents and blank controls. After the soaking, the seeds were dried in the shade for later use. 20 seeds of uniform size and full grains were selected for each treatment. Each treatment had 4 replicates and were evenly placed in a culture dish (9 cm in diameter) with two layers of filter paper. 10 ml of a composite salt-alkali mixture (40 mmol / l) was added and cultured in a constant temperature incubator at 25°C. The weight was weighed regularly every day, and the salt-alkali solution was added to the initial weight. The germination was considered when the radicle length reached 1 / 2 of the seed length. The experiment ended on the 7th day, and various physiological growth indicators were measured. The phenotypic growth status of alfalfa after salt-alkali treatment was compared. Figure 1 As shown: The left side of the figure is the group treated with stress-resistant agent soaking seeds, and the right side is the group treated with saline-alkali without soaking seeds.

[0047] The comparison of alfalfa growth status with the control is shown in Table 1, and the data are the average values ​​of 4 repeated experiments.

[0048] from Figure 1 It can be seen that: Alfalfa seeds were soaked with stress-resistant agents and then subjected to saline-alkali stress. After 10 days of saline-alkali stress, the saline-alkali control group had obvious short plants, short roots, and wilted leaves, while the stress-resistant agent soaked seeds had well-developed roots, taller plants, and better growth. Table 1 Comparison of germination status of alfalfa seedlings under stress resistance agent soaking and control conditions

[0049] Studies have shown that the germination potential, germination rate, germination index, and vitality index of alfalfa seeds are significantly reduced under saline-alkali stress. As can be seen from Table 1, compared with the saline-alkali control, the germination potential, germination rate, germination index, and vitality index of alfalfa seeds treated with the stress-resistant agent of the present invention are 102.0%, 15.6%, and 14.6% higher, respectively. It can be seen that soaking seeds with stress-resistant agents can improve the germination ability of alfalfa seeds under saline-alkali stress.

[0050] Table 2 Comparison of the growth of alfalfa seedlings under stress resistance agent soaking and control conditions

[0051] Salt-alkali stress can significantly reduce the number of alfalfa roots, root length, seedling length, aboveground dry weight, and leaf SPAD value. As can be seen from Table 2, under salt-alkali stress, the number of alfalfa roots, root length, seedling length, aboveground dry weight, and leaf SPAD value of the alfalfa seedlings treated with stress-resistant agents were significantly higher than those of the saline-alkali control, which were 52.75%, 43.17%, 25.82%, 24.14%, and 21.27% higher, respectively. It can be seen that stress-resistant agent soaking can effectively improve the salt-alkali tolerance of alfalfa seedlings.

[0052] Table 3 Comparison of antioxidant system activity of alfalfa seedlings under stress-resistant agent soaking and control conditions

[0053] The antioxidant system of alfalfa is activated under adversity. Salt-alkali stress significantly increases the superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) of alfalfa seedlings. The accumulation of active oxygen significantly increases the content of harmful substance malondialdehyde (MDA). As shown in Table 3, under salt-alkali stress, compared with the salt-alkali control, the activities of SOD, CAT, and POD of alfalfa seedlings soaked with stress-resistant agents increased by 102.0%, 15.6%, and 14.6%, respectively, and the content of MDA decreased by 44.5%. This shows that soaking with stress-resistant agents can enhance the antioxidant enzyme activity of alfalfa seedlings.

[0054] 2) Experimental Example 2: Effect of stress resistance agent spraying on the wintering and greening of alfalfa under low temperature stress in saline-alkali soil The field spraying test was conducted in the saline-alkali land of the Yellow River beach area from 2023 to 2024. The test material was WL525, which was mowed in October 2023 and waited for spring to return to green. From December 15 to 17, 2023, the temperature during the wintering period of alfalfa was below zero, and the lowest temperature reached -12°C. The spraying treatment was from December 9 to 10, 2023, spraying twice a day, with the same amount of water spraying as the control. Three days after the spraying, various physiological indicators of alfalfa leaves in the test area were measured. The yield and nutritional quality of alfalfa were measured at the budding stage (April 12) the following year.

[0055] Table 4 Comparison of antioxidant system activity of alfalfa leaves under stress resistance spraying and control conditions

[0056] As shown in Table 4, under low temperature stress, compared with the low temperature control, the SOD, CAT, and POD activities of alfalfa leaves sprayed with stress-resistant agents increased by 91.1%, 53.9%, and 34.7%, respectively, and the MDA content decreased by 30.1%. This indicates that the spraying of stress-resistant agents can enhance the antioxidant activity of alfalfa under low temperature stress during the wintering period, reduce its MDA content, and delay plant senescence.

[0057] Table 5 Comparison of agronomic traits and grass yield of alfalfa under stress resistance spraying and control conditions

[0058] Studies have shown that low temperature stress can significantly reduce the chlorophyll content of alfalfa, thereby reducing photosynthetic efficiency, reducing the growth rate of plants, resulting in reduced plant height and grass yield. As shown in Table 5, under low temperature stress, compared with the low temperature control, the plant height, leaf SPAD, and branch number of alfalfa sprayed with stress-resistant agents increased by 5.9%, 5.4%, and 6.6%, respectively, and the fresh grass yield and hay yield of alfalfa sprayed with stress-resistant agents increased by 14.1% and 16.1%, respectively. This shows that stress-resistant agent spraying can increase alfalfa grass yield under low temperature stress during the wintering period of alfalfa.

[0059] Table 6 Comparison of alfalfa yield and quality under stress resistance agent spraying and control conditions

[0060] As shown in Table 6, under low temperature stress, the crude protein content and relative feeding value of the anti-stress agent spraying did not change significantly compared with the low temperature control, while the crude fat content increased by 6.0%, the crude ash content decreased by 12.1%, and the relative feeding quality increased by 4.2%. It can be seen that under low temperature stress, the anti-stress agent spraying can significantly increase the alfalfa hay yield without reducing the hay quality.

[0061] The above is only an implementation method of the present application, and its description is relatively specific and detailed, but it cannot be understood as limiting the scope of the patent of the present application. It should be pointed out that for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present application, which all belong to the protection scope of the present application.

Claims

1. An anti-stress agent for alfalfa planting, characterized in that: Included dissolved in distilled water: 0.5-0.7 parts by weight of betaine; 0.2-0.3 parts by weight of potassium dihydrogen phosphate; 0.5-0.7 parts by weight of glucose.

2. The stress-resistant agent for alfalfa planting according to claim 1, characterized in that: Also included is 0.5 parts by weight of a nonionic surfactant.

3. The stress-resistant agent for alfalfa planting according to claim 1, characterized in that: Each kilogram of the anti-adverse agent includes: 6g betaine, 2g potassium dihydrogen phosphate, 5g glucose, and the balance is water.

4. The stress-resistant agent for alfalfa planting according to claim 1, characterized in that: Each kilogram of the anti-adverse agent contains: 6g betaine, 2.5g potassium dihydrogen phosphate, 6g glucose, and 5ml non-ionic surfactant.

5. The stress-resistant agent for alfalfa planting according to any one of claims 1 to 4, characterized in that: The non-ionic surfactant was Tween 20.

6. The stress-resistant agent for alfalfa planting according to any one of claims 1 to 5, characterized in that: Follow the steps below to configure: First, add betaine, potassium dihydrogen phosphate, and glucose into 500 ml of distilled water and fully dissolve them; Then, water was added to the mixed solution obtained by dissolution to make the volume up to 1000 ml.

7. The stress-resistant agent for alfalfa planting according to any one of claims 1 to 6, characterized in that: The pH value of the stress-resistant agent is between 5.5 and 6.

8. A method for using the stress-resistant agent for alfalfa planting according to any one of claims 1 to 7, characterized in that the steps include: Using the stress-resistant agent to soak alfalfa seeds 3-5 days before sowing; and / or spraying the stress-resistant agent on the alfalfa leaves in the morning and evening on the 6th to 8th day after the alfalfa turns green in spring, when the stomata of the leaves are open; and / or spraying the stress-resistant agent on the alfalfa leaves once a day on the 6th to 8th day before the alfalfa overwintering, when the stomata of the leaves are open; And / or spray alfalfa plants on a sunny evening 2-5 days before the onset of winter cold weather.

9. The method for using the stress-resistant agent for alfalfa planting according to claim 8, characterized in that: When soaking alfalfa seeds, soak them for 24 hours and then dry them in the shade.

10. The method for using the stress-resistant agent for alfalfa planting according to claim 8, characterized in that: When spraying alfalfa leaves, spray until the anti-cold protective agent drops onto the leaves, or spray at a dosage of 750 kg per hectare.

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