Promotion of exogenous choline glutarate on mung bean seed germination

Abstract

The present invention belongs to the technical field of seed germination, and specifically relates to the application of exogenous choline glutarate in the germination of mung bean seeds. Choline glutarate is prepared according to the following steps: at room temperature, an aqueous solution of choline hydroxide is mixed with glutaric acid, and then reacted on a shaker for 48 - 72 h at a shaker speed of 180 - 220 r / min. After the reaction is completed, the water is removed to obtain choline glutarate. The present invention uses self-synthesized choline glutarate as a seed soaking agent, and choline glutarate can significantly promote the germination of mung bean seeds under normal conditions and PEG-simulated drought stress, and improve the germination quality.

Description

Technical Field

[0001] The present invention belongs to the technical field of seed germination, specifically, it is about the promoting effect of exogenous choline glutarate on the germination of mung bean seeds. Background Art

[0002] Mung bean is one of the main miscellaneous grain crops in China, with good edible and nutritional value. In the northwest region of China, including Inner Mongolia, Shanxi, northern Shaanxi and other places, are all advantageous areas for mung bean production. The above areas are widely distributed in the junction area of the Loess Plateau and the desert steppe, belonging to the temperate monsoon climate, with rain and heat in the same season. The rainfall in spring is scarce, and mung beans are usually planted around the middle and late May. The scarce rainfall and drought in the soil can easily lead to difficulties in the emergence of mung bean seeds, a decline in sowing quality, and seriously restrict the improvement of mung bean yield in related areas.

[0003] At present, the domestic research on the germination technology of mung bean seeds is relatively scarce. The commonly used method in the industry is still the germination acceleration technology of mung bean seeds. The main method is as follows: Fill a clean container with warm water at 50 - 55°C, and the water volume is 5 times the volume of the seeds. Immerse the seeds in the warm water and stir constantly. Stop stirring when the water temperature drops to 30°C, and then soak for 8 - 12 hours to make the seeds absorb enough water. Then take out the seeds, drain the water, wrap them with gauze, and then wrap them with a wet gunny bag piece or towel, and place them in a basin for germination acceleration. This method is called warm water soaking, which is the most commonly used germination acceleration method at present. This method is simple and convenient, but there are also many problems: The emergence of seeds is slow, the germination potential is weak, the emergence is not uniform, the phenomenon of large and small seedlings is serious. If the seeds are dormant, the emergence speed is slower, and the germination rate is low. The germination of mung bean seeds is slow and not uniform, and it is easy to naturally age during storage, resulting in a decrease in vitality.

[0004] Taking appropriate methods to soak seeds before sowing crops is an effective means to improve the germination quality of seeds. At present, there is still a lack of practical soaking reagents before sowing in mung bean production. The research on a few reported compounds, such as salicylic acid and other technologies, is still not comprehensive enough. Especially, there is a lack of soaking reagents that can promote the germination of mung bean seeds under drought stress. Therefore, it is very necessary to find a soaking agent that can improve the germination of mung bean seeds under normal conditions and drought stress. Summary of the Invention

[0005] In view of the above deficiencies of the prior art, the purpose of the present invention is to provide the promoting effect of exogenous choline glutarate on the germination of mung bean seeds. The present invention uses self-synthesized choline glutarate as the soaking agent, and choline glutarate can significantly promote the germination of mung bean seeds under normal conditions and PEG-simulated drought stress, and improve the germination quality.

[0006] To solve the above technical problems, the present invention adopts the following technical solutions:

[0007] Application of exogenous choline glutarate in mung bean seed germination, wherein the structure of choline glutarate is shown in formula (I):

[0008]

[0009] Preferably, the choline glutarate is prepared according to the following steps:

[0010] At room temperature, an aqueous solution of choline hydroxide is mixed with glutaric acid, and then reacted on a shaker for 48 - 72 h, and then the water is removed to obtain choline glutarate.

[0011] Preferably, the molar ratio of choline hydroxide to glutaric acid is 1:1.

[0012] Preferably, the mixing method is: under stirring conditions, glutaric acid is added dropwise to the aqueous solution of choline hydroxide and mixed.

[0013] Preferably, the rotation speed of the shaker is 180 - 220 r / min.

[0014] Preferably, the soaking concentration of the choline glutarate is 100 - 300 mg / L.

[0015] Preferably, the soaking concentration of the choline glutarate is 200 mg / L.

[0016] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0017] 1. Choline glutarate is a novel choline compound, and there has been no relevant report on its promoting effect on plant growth before. In this application, using self - synthesized choline glutarate as the material, it is found that soaking seeds with this compound can significantly promote the germination of mung bean seeds under normal conditions and simulated drought stress, and improve the germination quality. Aiming at the problems such as difficult emergence, weak seedling growth, and drought during the germination period that may occur during the sowing process of crops such as mung beans, the present invention aims to improve the sowing quality by soaking seeds with choline glutarate.

[0018] 2. The mechanism of action of choline glutarate as a seed - soaking agent on mung beans is as follows: Through our research, the promoting effect of choline glutarate seed - soaking on the germination of mung bean seeds under normal conditions and simulated drought stress may have the following two mechanisms of action:

[0019] (1) Choline glutarate seed - soaking has an obvious promoting effect on the growth of roots after seed germination. Especially under drought conditions, developed roots are beneficial to strong seedlings of plants, enhancing water absorption capacity and improving drought resistance;

[0020] (2) Choline glutarate seed - soaking increases the antioxidant enzyme activity of mung bean seedlings and reduces the content of malondialdehyde, indicating that it helps to alleviate oxidative damage during seed germination and enhance antioxidant capacity.

[0021] 3. In this application, different compounds were screened and different seed soaking concentrations were used. It was found that soaking mung bean seeds with 200 mg / L choline glutarate could significantly improve the germination rate of mung bean seeds and the growth conditions of roots and buds after germination under normal conditions and drought stress. This method is easy to operate, and the reagents used are all common reagents that are easily available. The synthesis and preparation costs are low. The raw materials and the product choline glutarate used in the synthesis process are all green compounds with extremely low toxicity, and no by-products are generated during the synthesis process, featuring environmental friendliness. This discovery fills the technical gap in the reagent for soaking mung bean seeds before sowing, especially in improving the germination quality of mung bean seeds under drought stress. Therefore, the application of the technology of the present invention can provide support for effectively improving the emergence quality of mung beans in the excellent mung bean growing areas in the northwest of China. Description of the Drawings

[0022] Figure 1 It is a physical diagram of choline glutarate prepared in Example 1 of the present invention;

[0023] Figure 2 It is a hydrogen spectrum diagram of choline glutarate prepared in Example 1 of the present invention;

[0024] Figure 3 It is a screening experiment diagram of soaking mung bean seeds with different concentrations of choline glutarate of the present invention;

[0025] Figure 4 It is a physical control diagram of soaking mung beans with different samples of the present invention. Detailed Embodiments

[0026] The following describes the detailed embodiments of the present invention in detail, but it should be understood that the protection scope of the present invention is not limited by the detailed embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts belong to the protection scope of the present invention. The experimental methods described in the embodiments of the present invention are all conventional methods unless otherwise specified.

[0027] Example 1

[0028] The preparation method of choline glutarate includes the following steps:

[0029] Choline glutarate is synthesized using aqueous choline hydroxide solution and glutaric acid. The volume ratio of choline hydroxide to water in the aqueous choline hydroxide solution is 46:100. The synthesis method is mainly the neutralization method, and the specific steps are as follows:

[0030] At room temperature, an aqueous solution of choline hydroxide and glutaric acid are mixed in a molar ratio of 1:1. The mixing method is to gradually add small amounts drop by drop. The glutaric acid is added dropwise to the aqueous solution of choline hydroxide, and the whole process is carried out on a magnetic stirrer. Then, it is reacted in a shaker for 48 h at a shaker temperature of 18 °C and a rotation speed of 200 r / min. After the reaction is completed, the water is removed using a rotary evaporator at 55 °C to obtain choline glutarate.

[0031] Example 2

[0032] A preparation method of choline glutarate, comprising the following steps:

[0033] Choline glutarate is synthesized using two compounds, an aqueous solution of choline hydroxide and glutaric acid. The volume ratio of choline hydroxide to water in the aqueous solution of choline hydroxide is 40:100. The synthesis method is mainly the neutralization method, and the specific steps are as follows:

[0034] At room temperature, an aqueous solution of choline hydroxide and glutaric acid are mixed in a molar ratio of 1:1. The mixing method is to gradually add small amounts drop by drop. The glutaric acid is added dropwise to the aqueous solution of choline hydroxide, and the whole process is carried out on a magnetic stirrer. Then, it is reacted in a shaker for 72 h at a shaker temperature of 25 °C and a rotation speed of 220 r / min. After the reaction is completed, the water is removed using a rotary evaporator at 55 °C to obtain choline glutarate.

[0035] Example 3

[0036] A preparation method of choline glutarate, comprising the following steps:

[0037] Choline glutarate is synthesized using two compounds, an aqueous solution of choline hydroxide and glutaric acid. The volume ratio of choline hydroxide to water in the aqueous solution of choline hydroxide is 50:100. The synthesis method is mainly the neutralization method, and the specific steps are as follows:

[0038] At room temperature, an aqueous solution of choline hydroxide and glutaric acid are mixed in a molar ratio of 1:1. The mixing method is to gradually add small amounts drop by drop. The glutaric acid is added dropwise to the aqueous solution of choline hydroxide, and the whole process is carried out on a magnetic stirrer. Then, it is reacted in a shaker for 60 h at a shaker temperature of 35 °C and a rotation speed of 180 r / min. After the reaction is completed, the water is removed using a rotary evaporator at 55 °C to obtain choline glutarate.

[0039] 1. Structure analysis

[0040] In Examples 1 - 3 of the present invention, choline glutarate with high purity was prepared, and the yield of the obtained choline glutarate was greater than 95%. It is a liquid at room temperature and is light yellow to light brown ( Figure 1 ). The structure of the choline glutarate prepared in Example 1 was identified by nuclear magnetic resonance spectroscopy ( Figure 2) From the integral ratio of hydrogen atoms in the hydrogen spectrum, it can be seen that the purity of the synthesized choline glutarate is greater than 95%, verifying the correctness of the synthesized structure.

[0041] Table 1 Identification results of nuclear magnetic resonance spectroscopy of choline glutarate

[0042]

[0043] 2. Screening experiment on the soaking concentration of choline glutarate:

[0044] The optimal soaking concentration of choline glutarate was determined through a concentration gradient screening experiment. The different concentration ranges used were: 0 mg / L, 50 mg / L, 100 mg / L, 200 mg / L, 300 mg / L. The results are as Figure 3 shown. It can be seen that soaking mung bean seeds with choline glutarate at different concentrations can significantly increase the fresh weight of the roots of mung beans. Among them, the increase in the fresh weight of the roots is the highest at the soaking concentration of 200 mg / L. Therefore, 200 mg / L was determined as the optimal soaking concentration.

[0045] 3. Analysis of the effect of soaking mung bean seeds with choline glutarate on the germination of mung beans under normal conditions and simulated drought stress

[0046] (1) Research method:

[0047] Two treatment groups were set up, a control group (soaking seeds with distilled water treatment) and an experimental group (soaking seeds with choline glutarate). Among them, the soaking concentration of choline glutarate was 200 mg / L. The differences between the experimental group and the control group were compared to analyze the effect of soaking mung bean seeds with choline glutarate on the germination of mung beans. Using "Jilv 13" mung beans as materials, the seeds in the experimental group were soaked in a choline glutarate (200 mg / L) solution for 12 h, and the seeds in the control group were soaked in distilled water for 12 h. The soaked seeds were placed in a plastic germination box to germinate (germination box specifications: length × width × height = 19 cm × 13 cm × 13 cm). Filter paper was laid at the bottom of the germination box. For seed germination under normal conditions, 40 mL of distilled water was added to each germination box to wet the filter paper and then germinate. For seed germination under simulated drought stress conditions, 40 mL of PEG solution (13%) was added to each germination box to wet the filter paper and then germinate.

[0048] Both the experimental group and the control group were set up with 4 replicates. Each replicate was to place 30 mung bean seeds in a germination box. The germination situation was investigated regularly every day, and samples were taken after 7 days. For each replicate of each treatment, 3 plants with consistent growth were selected to measure their growth indexes and retain the fresh samples of roots and buds. The activities of two antioxidant enzymes were measured with the retained fresh samples: superoxide dismutase (SOD) and peroxidase (POD) respectively. At the same time, the index representing the degree of oxidative damage was measured: the content of malondialdehyde (MDA).

[0049] (2) Research results:

[0050] 3.1. Effects of choline glutarate seed soaking on mung bean seed germination under normal conditions

[0051] As shown in Table 2, under normal germination conditions, soaking mung bean seeds with 200 mg / L choline glutarate significantly promoted the growth of both roots and shoots after germination, especially the promotion effect on root growth was more obvious. Compared with the control, the root length of mung beans soaked with choline glutarate increased significantly by 26.3%, and the root dry weight and shoot dry weight increased by 17% and 16.6% respectively. It can be seen that choline glutarate has an obvious promoting effect on mung bean seed germination under normal conditions.

[0052] Table 2 Effects of choline glutarate seed soaking on mung bean seed germination under normal conditions

[0053]

[0054] Note: The data in the table are presented in the form of mean ± standard deviation (n = 4), and "*" indicates a significant difference between the experimental group and the control group (p < 0.05).

[0055] 3.2. Effects of choline glutarate seed soaking on mung bean seed germination under drought stress

[0056] As shown in Table 3, in this application, the drought stress condition was simulated by using 13% polyethylene glycol (PEG6000). This concentration of polyethylene glycol solution is usually used to simulate moderate drought stress. Under the condition of PEG-simulated drought stress, compared with the control, soaking seeds with choline glutarate significantly increased the root length of mung beans after germination, with an increase amplitude of 26.1%. Compared with the control, the root dry weight and shoot dry weight increased by 27.1% and 6.8% respectively. It can be seen that choline glutarate also has an obvious promoting effect on mung bean seed germination under drought stress, and the effect on the root system is more obvious. Therefore, the above research results show that soaking seeds with choline glutarate can significantly promote the growth of mung bean seed roots under drought stress, which is beneficial to improving the growth status of seedlings after seed germination and enhancing the drought resistance of mung beans.

[0057] Table 3 Effects of choline glutarate seed soaking on mung bean seed germination under simulated drought stress conditions

[0058]

[0059] Note: The data in the table are presented in the form of mean ± standard deviation (n = 4), and "*" indicates a significant difference between the experimental group and the control group (p < 0.05);

[0060] The physical diagram is as Figure 4 shown, which is consistent with the conclusions obtained in Table 2 and Table 3.

[0061] 3.3 Effects of choline glutarate seed soaking on antioxidant enzyme activities (SOD, POD) and malondialdehyde (MDA) content in roots and buds of mung bean seeds under drought stress

[0062] As shown in Table 4, under the condition of PEG-simulated drought stress, compared with the control, seed soaking with choline glutarate significantly increased the antioxidant enzyme POD activity in the roots and buds of mung bean seeds after germination. This indicates that seed soaking with choline glutarate helps to enhance the antioxidant function of mung bean seeds, which is beneficial for them to resist drought stress. The content of malondialdehyde is an important indicator reflecting the degree of cytoplasmic membrane peroxidation, and an increase in its content usually represents that plants may suffer oxidative damage, and drought stress is also an important cause of plant oxidative damage. As shown in Table 4, compared with the control, seed soaking with choline glutarate significantly reduced the MDA content in the roots and buds of mung bean under drought stress, indicating that seed soaking with choline glutarate helps to alleviate the damage of drought stress to the germination of mung bean seeds and improve their germination status.

[0063] Table 4 Effects of choline glutarate seed soaking on antioxidant enzyme activities (SOD, POD) and malondialdehyde (MDA) content in roots and buds of mung bean seeds under simulated drought stress

[0064]

[0065]

[0066] Note: The data in the table are presented in the form of mean ± standard deviation (n = 4), and "*" indicates a significant difference between the experimental group and the control group (p < 0.05).

[0067] The above research results show that soaking seeds with exogenous choline glutarate can effectively improve the germination status of mung bean seeds under normal conditions and simulation, promote the growth of the roots of germinated seeds, and improve the drought resistance during germination.

[0068] Those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalent technologies, the present invention also intends to include these modifications and variations.

Claims

1. Application of exogenous choline glutarate in mung bean seed germination, characterized in that, The structure of the choline glutarate is shown in formula (I): （I）； The seed soaking concentration of the choline glutarate is 100 - 300 mg / L.

2. Use of exogenous choline glutarate in mung bean seed germination according to claim 1, characterized in that, The choline glutarate is prepared according to the following steps: At room temperature, an aqueous solution of choline hydroxide is mixed with glutaric acid, and then after reacting on a shaker for 48 - 72 h, the water is removed to obtain choline glutarate.

3. The application of exogenous choline glutarate in mung bean seed germination according to claim 1, characterized in that, The mixing method is as follows: under stirring conditions, glutaric acid is added dropwise to the aqueous solution of choline hydroxide and mixed.

4. The application of exogenous choline glutarate in mung bean seed germination according to claim 1, characterized in that, The rotation speed of the shaker is 180 - 220 r / min.

5. Use of exogenous choline glutarate in mung bean seed germination according to claim 1, characterized in that, The seed soaking concentration of the choline glutarate is 200 mg / L.

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