Medicago sativa seed germinating agent as well as preparation method and application thereof

Abstract

The invention provides an alfalfa seed germinator as well as a preparation method and application thereof, and relates to the technical field of plant breeding. The seed germination agent disclosed by the invention is prepared from 5 to 15 mL of Ectoine, 1 to 10 mL of betaine and 0.5 to 3 mL of glycerol. Through scientific proportioning of the raw materials, the drought and saline-alkali stress resistance of the alfalfa seeds can be remarkably improved, and the germination rate and radicle and germ lengths of the alfalfa seeds in drought and saline-alkali environments are increased. The seed germinator is simple in formula and preparation process, low in cost, convenient to use and environment-friendly, and has wide market space and application prospect.

Description

Technical Field

[0001] The present invention relates to the technical field of plant breeding, in particular to an alfalfa seed germination agent and a preparation method and application thereof. Background Art

[0002] Soil salinization is one of the important issues affecting agricultural production and ecosystem balance worldwide, especially in arid and semi-arid saline-alkali areas. Salt and drought stress have a serious impact on seed germination, growth and development, and yield of crops.

[0003] Alfalfa (Medicago sativa L.) is a high-protein, adaptable, and high-quality forage grass with diverse ecological functions and high nutritional value. Its drought- and cold-tolerant nature, along with a certain degree of salt- and alkali-tolerant properties, makes it crucial for addressing saline-alkali soil issues and a common plant used in saline-alkali experiments. Furthermore, alfalfa offers significant economic benefits and environmental benefits, making it widely used in animal husbandry. However, with increasing soil salinization and drought, the germination and growth of alfalfa under drought stress conditions are limited. Therefore, improving alfalfa's drought tolerance, increasing its germination rate, and improving its growth in drought-prone environments are crucial for ecological restoration and forage production.

[0004] Ectoine, a natural protective substance synthesized by extreme microorganisms, has been proven to reduce cell damage under various stress conditions. In particular, it can enhance the resistance of proteins to radiation and high salt concentration oxidative damage in extreme environments, thereby enhancing the survival ability of plants in adversity.

[0005] In view of this, the present invention provides an alfalfa seed germination agent, its preparation method, and its use. The seed germination agent of the present invention can significantly improve the ability of alfalfa seeds to resist drought and saline-alkali stress, increasing their germination rate and the length of their radicle and plumule in drought and saline-alkali environments. Furthermore, the seed germination agent of the present invention has a simple formulation and preparation process, is low-cost, easy to use, and environmentally friendly, and has important application value in ecological restoration and forage production. Summary of the Invention

[0006] In order to overcome the deficiencies of the above-mentioned prior art, the purpose of the present invention is to provide an agent for promoting the germination of alfalfa seeds and a preparation method thereof, which has a simple formula, is suitable for large-scale production, can promote the germination and production application of alfalfa seeds, can increase the germination rate of seeds, and improve seed resistance.

[0007] An agent for promoting germination of alfalfa seeds is prepared from the following raw materials: 5-15 mL of 1-10 mmol / L ectoine, 1-10 mL of 5-15 mmol / L betaine, and 0.5-3 mL of 3-7 mmol / L glycerol.

[0008] The second aspect of the present invention is to provide a method for preparing a seed germination agent, which specifically comprises the following steps:

[0009] S1. Prepare 1-10 mmol / L of ectoine, 5-15 mmol / L of betaine, and 3-7 mmol / L of glycerol at molar concentrations.

[0010] S2. Add 1-10 mL of the Ectoine solution, 1-10 mL of the betaine solution, and 0.5-3 mL of the glycerol solution prepared in S1 into the beaker in sequence, and stir for 1-5 minutes until mixed to obtain a seed germination agent.

[0011] The third aspect of the present invention is to provide a seed germination agent for use in alfalfa planting.

[0012] Preferably, the seed germination agent of the present invention is used for planting alfalfa in arid and semi-arid saline-alkali areas.

[0013] Compared with the prior art, the present invention has the following beneficial technical effects:

[0014] The invention significantly improves the ability of alfalfa seeds to resist drought and saline-alkali stress by scientifically proportioning natural bioactive ingredients such as seed germination agent Ectoine, betaine and glycerol, thereby increasing the germination rate and the length of radicle and plumule of alfalfa seeds under drought and saline-alkali conditions.

[0015] The seed germination agent of the present invention has simple formula and preparation process, low cost, convenient use, and is environmentally friendly. It is particularly suitable for alfalfa planting in arid and semi-arid areas and has broad market space and application prospects.

[0016] Reference numerals

[0017] Figure 1 This is a diagram of some alfalfa seed germination experiments of the present invention. DETAILED DESCRIPTION

[0018] A first aspect of the present invention is to provide a seed germination agent, which comprises 5-15 mL of 1-10 mmol / L ectoine, 1-10 mL of 5-15 mmol / L betaine, and 0.5-3 mL of 3-7 mmol / L glycerol.

[0019] Ectoine, a natural protective substance synthesized by extreme microorganisms, has been proven to reduce cell damage under various stress conditions. In particular, it can enhance the resistance of proteins to radiation and high salt concentration oxidative damage in extreme environments, thereby enhancing the survival ability of plants in adversity.

[0020] The seed germination agent of the present invention uses ectoine and betaine as its main ingredients, leveraging their synergistic effect to effectively promote the germination of alfalfa seeds under drought and saline-alkali stress, significantly increasing the seed germination rate and enhancing the stress resistance of seedlings. Ectoine improves the hydration of the lipid membrane surface by balancing the osmotic pressure inside and outside the cells, enhancing the fluidization of the lipid membrane, and maintaining the stability of the molecular structure of proteins or nucleic acids, thereby enhancing the stress resistance of alfalfa. Betaine can reduce the impact of adverse environments on alfalfa, improving its ability to adapt to adverse conditions.

[0021] The second aspect of the present invention is to provide a method for preparing a seed germination agent, which specifically comprises the following steps:

[0022] S1. Prepare 1-10 mmol / L of ectoine, 5-15 mmol / L of betaine, and 3-7 mmol / L of glycerol at molar concentrations.

[0023] S2. Add 1-10 mL of the Ectoine solution, 1-10 mL of the betaine solution, and 0.5-3 mL of the glycerol solution prepared in S1 into the beaker in sequence, and stir for 1-5 minutes until mixed to obtain a seed germination agent.

[0024] The preparation process employed in the present invention is simple, easy to operate, low-cost, and environmentally friendly and safe. The entire preparation process requires only three steps and is carried out entirely at room temperature, avoiding damage to active ingredients such as ectoine and betaine by high temperatures. A step-by-step dissolution method is employed to ensure full dissolution and uniform distribution of each component. The present invention uses distilled water as a solvent, is free of organic solvent pollution, and is environmentally friendly. Through precise raw material ratios and a preparation process, the stability and reproducibility of product quality are ensured. Furthermore, the preparation method is simple to operate, does not require complex equipment or high-temperature and high-pressure conditions, and has easily controllable process parameters. The production cost is low, and it is convenient for industrialized mass production, laying a good technical foundation for the industrial application of alfalfa seed germination agents.

[0025] The third aspect of the present invention is to provide a seed germination agent for use in alfalfa planting.

[0026] The seed germination agent of the present invention is preferably used for planting alfalfa in arid and semi-arid saline-alkali areas to increase the germination rate of alfalfa seeds and the length of the radicle and plumule.

[0027] The following will clearly and completely describe the technical solutions of the present invention in conjunction with the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0028] Unless otherwise specified, all experiments were repeated three times and the results were expressed as the mean value.

[0029] Example 1 A seed germination agent, comprising 5 mL of 5 mmol / L ectoine, 10 mL of 5 mmol / L betaine, and 0.5 mL of 3 mmol / L glycerol;

[0030] The preparation method of the seed germination agent is as follows:

[0031] S1. Prepare 5 mmol / L ectoine solution, 5 mmol / L betaine solution, and 3 mmol / L glycerol solution at molar concentrations.

[0032] S2. Add the Ectoine solution, betaine solution, and glycerol solution prepared in S1 into the beaker in sequence, and stir for 5 minutes until mixed to obtain a seed germination agent.

[0033] Example 2 A seed germination agent is prepared, comprising 5 mL of 10 mmol / L ectoine, 10 mL of 5 mmol / L betaine, and 0.5 mL of 3 mmol / L glycerol.

[0034] The preparation method of the seed germination agent is the same as that of Example 1 except that the concentration of the solution prepared in step S1 is different.

[0035] Example 3 A seed germination agent is prepared, comprising 5 mL of 5 mmol / L ectoine, 10 mL of 10 mmol / L betaine, and 0.5 mL of 3 mmol / L glycerol.

[0036] The preparation method of the seed germination agent is the same as that of Example 1 except that the concentration of the prepared solution is different.

[0037] Test Case

[0038] 200 healthy and plump alfalfa seeds were selected, disinfected by soaking in 75% alcohol for 5 min, then rinsed with clean water and dried. Eight layers of gauze were spread flat in a petri dish as a germination bed, with 40 seeds placed in each petri dish and the petri dishes were numbered. Drought stress was simulated with 15% PEG-6000 solution, and salt stress was simulated with 80 mmol / L NaCl solution.

[0039] Group 1 (drought group): the exogenous Ectoine concentration was set to 0 mmol / L, i.e., 15% PEG 10 mL was used as the solvent;

[0040] Group 2 (saline-alkali group): the exogenous ectoine concentration was set to 0 mmol / L, that is, 8 mL of distilled water containing 80 mmol / L NaCl (2 mL) was used as the solvent;

[0041] Group 3-1: 80 mmol / L NaCl (2 mL) + 5 mmol / L Ectoine (8 mL) + 3 mmol / L glycerol (0.5 mL);

[0042] Group 3-2: 80 mmol / L NaCl (2 mL) + 5 mmol / L betaine (8 mL) + 3 mmol / L glycerol (0.5 mL);

[0043] Group 3-3: 80 mmol / L NaCl (2 mL) + 5 mmol / L ectoine (6 mL) + 5 mmol / L betaine (2 mL) + 3 mmol / L glycerol (0.5 mL);

[0044] Group 4-1: 15% PEG-6000 (10 mL) + 5 mmol / L Ectoine (5 mL) + 3 mmol / L glycerol (0.5 mL);

[0045] Group 4-2: 15% PEG-6000 (10 mL) + 5 mmol / L betaine (5 mL) + 3 mmol / L glycerol (0.5 mL);

[0046] Group 4-3: 15% PEG-6000 (10 mL) + 5 mmol / L ectoine (4 mL) + 10 mmol / L betaine (1 mL) + 3 mmol / L glycerol (0.5 mL);

[0047] Group 5 (control group): distilled water.

[0048] The preparation methods of the above germination agents were the same as those in Example 1, and the differences in ingredients were appropriately adjusted according to the conditions of each group.

[0049] Five replicates were set up in each group to observe the effect of drought stress on alfalfa seed germination. The number of seeds germinated was recorded every day, and the germination rate was calculated on the 10th day after seed germination. The radicle length and plumule length were measured with a vernier caliper. The results are shown in Table 1.

[0050] Table 1 Germination rate, radicle length and plumule length of alfalfa

[0051] As shown in Table 1, in the germination process of the present invention, the specific ratio of Ectoine and betaine can synergistically improve seed vigor and significantly increase the germination rate, radicle length and plumule length of alfalfa seeds under drought stress.

[0052] The above-described embodiments merely illustrate several implementations of the present invention, and while their descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patent. It should be noted that a person skilled in the art would be able to make numerous variations and improvements without departing from the spirit of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the patent for this invention shall be determined by the appended claims.

Claims

1. A seed germination agent, characterized in that Includes Ectoine 5-15mL, Betaine 1-10mL and Glycerol 0.5-3mL.

2. A seed germination agent according to claim 1, characterized in that, The concentration of Ectoine in the seed germination agent is 1-10 mmol / L.

3. A seed germination agent according to claim 1, characterized in that, The concentration of betaine in the seed germination agent is 5-15 mmol / L.

4. A seed germination agent according to claim 1, characterized in that, The concentration of glycerol in the seed germination agent is 3-7 mol / L.

5. The method for preparing the seed germination agent according to any one of claims 1 to 4, characterized in that: The following steps are involved: S1. Prepare 1-10 mmol / L of ectoine, 5-15 mmol / L of betaine, and 3-7 mmol / L of glycerol at molar concentrations. S2. Add 1-10 mL of the Ectoine solution, 1-10 mL of the betaine solution, and 0.5-3 mL of the glycerol solution prepared in S1 into the beaker in sequence, and stir for 1-5 minutes until mixed to obtain a seed germination agent.

6. Application of a seed germination agent in alfalfa planting, characterized in that: The seed germination agent is the seed germination agent according to any one of claims 1 to 4 or the seed germination agent prepared according to the method according to claim 5.

7. The use according to claim 6, characterized in that The seed germination agent is used for planting alfalfa in arid, semi-arid and saline-alkali areas.

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