Application of 2, 3-butanediol in promoting seed germination
By using 2,3-butanediol as a germination agent, the problems of unstable seed germination and reduced stubborn seed vitality are solved, and the rapid, efficient and environmentally friendly results of seed germination are achieved.
Patent Information
- Application Number
- CN202510596855.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-05-09
- Publication Date
- 2025-07-29
AI Technical Summary
The prior art has unstable effects in promoting seed germination. The use of gibberellin causes seedlings to grow long and reduce the reversibility of seedlings, and the vitality of refractory seeds is reduced after storage, and there is a lack of effective germination promoters.
2,3-butanediol is used as a germination agent, with a concentration of 25-400μM, and it promotes seed germination with specific light and temperature conditions, and is suitable for normal and stubborn seeds.
It accelerates the germination of normal seeds, improves the vitality of stubborn seeds, broadens the scope of application, and is non-toxic and pollution-free, and is environmentally friendly.
Smart Images
Figure SMS_1 
Figure SMS_2 
Figure SMS_3
Abstract
Description
Technical Field
[0001] The present invention belongs to the field of bioengineering technology, and particularly relates to the application of 2,3-butanediol in promoting seed germination. Background Art
[0002] Seeds are the main carriers for plants to reproduce offspring and are also the most important production materials in agricultural production. Seed germination is the basis for seedling establishment and the start of the plant life cycle. The induction, maintenance, and release of seed dormancy are important physiological processes in seed plants. Seeds usually enter dormancy after maturity and initiate germination when the environment is suitable.
[0003] Current research shows that various exogenous and endogenous factors can affect seed germination. In traditional agricultural techniques, physical methods such as seed soaking or temperature regulation are generally used to break dormancy and then promote seed germination, but their effects are often unstable. Gibberellin, as a commonly used plant hormone for promoting seed germination, has a very significant effect on promoting seed germination. However, applying gibberellin during the germination period often leads to seedling spindling and reduces the stress tolerance of seedlings. Therefore, developing a new germination promoter that can replace gibberellin is of great significance for agricultural production.
[0004] Compared with orthodox seeds, recalcitrant seeds are not tolerant to low temperature and dehydration, are prone to losing viability, and have a short storage life. Therefore, developing reagents (i.e., germination promoters) that can effectively improve the viability and germination of recalcitrant seeds after storage is of great significance for the long-term preservation of recalcitrant seeds. Summary of the Invention
[0005] The object of the present invention is to provide the application of 2,3-butanediol in promoting seed germination.
[0006] The object of the present invention is achieved as follows: the application of 2,3-butanediol in promoting seed germination, and the application concentration of the 2,3-butanediol is 25 - 400 μM.
[0007] The method for promoting seed germination using 2,3-butanediol is as follows: 1) Sow the seeds into a germination / solid medium or solution with a 2,3-butanediol concentration of 100 - 200 μM / L. Arabidopsis thaliana and tobacco seeds need to be vernalized at 4°C for 3 days; 2) Then transfer the medium to a greenhouse with a temperature of 22 or 28°C, a photoperiod of 14 or 16 h of light and 10 or 8 h of darkness, and a light intensity of about 2500 - 3000 lx for cultivation.
[0008] The beneficial effects of the present invention are as follows: The present invention uses 2,3-butanediol as a germination promoter, which can not only accelerate the germination of orthodox seeds such as Arabidopsis thaliana, tobacco, and upland / lowland rice seeds, but also improve the vigor of recalcitrant seeds after storage. At the same time, it can also improve the germination of Arabidopsis thaliana, tobacco, upland / lowland rice, and Hopea exalata seeds under adversity, broadening the application scope of 2,3-butanediol. Compared with the commonly used germination promoter - gibberellin, 2,3-butanediol shows a comparable germination-promoting effect and has the advantages of being non-toxic, pollution-free, and environmentally friendly. It not only provides a new way to improve the emergence rate and emergence rate of crops, but also provides a new solution for enhancing the vigor of recalcitrant seeds after storage. Detailed implementation mode
[0009] The following further illustrates the present invention with reference to embodiments, but the present invention is not limited in any way. Any transformation or substitution based on the teachings of the present invention belongs to the protection scope of the present invention.
[0010] The present invention provides the application of 2,3-butanediol in promoting seed germination, and the application concentration of 2,3-butanediol is 25 - 400 μM.
[0011] The application concentration of 2,3-butanediol is 100 - 200 μM.
[0012] The seeds are Arabidopsis thaliana, tobacco, rice, upland rice or Hopea exalata seeds.
[0013] The Arabidopsis thaliana seeds are of the Columbia ecotype (Col-0), the tobacco is Honghuadajinyuan (HD-1), the rice is Nipponbare, and the upland rice is Luyin 46 (LY46).
[0014] The 2,3-butanediol can promote seed germination in high-salt and low-temperature environments.
[0015] The present invention also provides a method for promoting seed germination, which is realized according to the following steps: 1) Sow the seeds into a germination / solid medium or solution with a 2,3-butanediol concentration of 100 - 200 μM. Arabidopsis thaliana and tobacco seeds need to be vernalized at 4°C for 3 days; 2) Then transfer the medium to a greenhouse with a temperature of 22 or 28°C, a photoperiod of 14 or 16 h of light and 10 or 8 h of darkness, and a light intensity of 2500 - 3000 lx for cultivation.
[0016] The germination medium is 1 / 2 MS medium.
[0017] The formula of the 1 / 2 MS medium is: 2.2025 g / L MS medium, 10 g / L sucrose, 6.5 g / L agar.
[0018] The formula of the solid medium is: 6.5 g / L agar.
[0019] Example 1 Effects of Different Concentrations of 2,3-Butanediol on Seed Germination of Arabidopsis thaliana, Tobacco, Rice and Upland Rice Experimental method: Select mature and plump seeds of Arabidopsis thaliana, tobacco, rice and upland rice. After disinfecting with 20% sodium hypochlorite solution for 10 - 15 min, rinse with distilled water 4 - 6 times, and sow them in the germination medium or solution containing 25 μM, 50 μM, 100 μM, 200 μM, 400 μM 2,3-butanediol. Use 0 μM 2,3-butanediol as the control. According to the optimal growth conditions of the seeds, place them in a greenhouse at a temperature of 22 or 28 °C, with a 16 / 8 h light cycle or a 14 / 10 h light cycle, and a light intensity of 2500 - 3000 lx for germination. Count the germination rate of Arabidopsis thaliana seeds after 36 h and the cotyledon greening rate after 75 h. Count the germination rate of tobacco seeds after 4 d and the cotyledon greening rate after 8 d. Count the germination rate of rice seeds after 3 d. Count the germination rate of upland rice seeds after 3 d. The experiment was repeated three times.
[0020] Germination rate (%) = Number of germinated seeds / Number of tested seeds * 100% Cotyledon greening rate (%) = Number of seeds with cotyledons / Number of tested seeds * 100% The results of the effects of different concentrations of 2,3-butanediol on seed germination of Arabidopsis thaliana, tobacco, rice and upland rice are shown in Table 1.
[0021] Table 1 Results of the Effects of Different Concentrations of 2,3-Butanediol on Seed Germination of Arabidopsis thaliana, Tobacco, Rice and Upland Rice Note: The data in the table are compared between the same columns of the same species; the values are mean ± standard deviation; different lowercase letters in the same column represent that their corresponding means are significantly different at the p < 0.05 level. The same below.
[0022] As can be seen from Table 1, the seeds of 4 plants all showed an increase in germination rate after being treated with 2,3-butanediol. For Arabidopsis seeds, the optimal concentration of 2,3-butanediol for promoting germination was 200 μM. Under this optimal condition, the germination promotion efficiency of 2,3-butanediol on Arabidopsis seeds was 143.72%, and the cotyledon emergence promotion efficiency was 162.88%. For tobacco seeds, the optimal concentration of 2,3-butanediol for promoting germination was 200 μM. Under this optimal condition, the germination promotion efficiency of 2,3-butanediol on tobacco seeds was 124.45%, and the cotyledon emergence promotion efficiency was 307.47%. For rice seeds, the optimal concentration of 2,3-butanediol for promoting germination was 100 μM. Under this optimal condition, the germination promotion efficiency of 2,3-butanediol on rice seeds was 86.44%. For upland rice seeds, the optimal concentration of 2,3-butanediol for promoting germination was 200 μM. Under this optimal condition, the germination promotion efficiency of 2,3-butanediol on upland rice seeds was 99.96%. It shows that 2,3-butanediol can significantly increase the germination rate of Arabidopsis, tobacco, rice and upland rice seeds, and has the best germination promotion effect when the concentration of 2,3-butanediol is 100-200 μM.
[0023] Example 2 Effect of 2,3-butanediol on the viability of Hopea exalata seeds after storage Select freshly picked Hopea exalata seeds, rinse them with distilled water 4-6 times, and sow them in a solid medium containing 0 μM 2,3-butanediol. Place them in a greenhouse at a temperature of 28 °C, a light cycle of 14 / 10 h, and a light intensity of 2500-3000 lx for germination. The germination rate of Hopea exalata seeds was counted after 16 days, and the experiment was repeated three times, with 25 seeds in each repetition.
[0024] Select Hopea exalata seeds stored in the dark at 22 °C for 3 weeks, rinse them with distilled water 4-6 times, and sow them in a solid medium containing 200 μM 2,3-butanediol. Using 0 μM 2,3-butanediol as a control, place them in a greenhouse at a temperature of 28 °C, a light cycle of 14 / 10 h, and a light intensity of 2500-3000 lx for germination. The germination rate of Hopea exalata seeds was counted after 18 days, and the experiment was repeated three times, with 20 seeds in each repetition.
[0025] Table 2 Results of the effect of 2,3-butanediol on the viability of Hopea exalata seeds after storage From the data in Table 2, it can be seen that freshly harvested Hopea exalata seeds could almost completely germinate at 16 days, while the germination rate of Hopea exalata seeds after storage for 3 weeks decreased significantly. After treatment with 2,3-butanediol, the germination rate of stored Hopea exalata seeds increased by 66.68%, indicating that 2,3-butanediol can improve the viability of stored Hopea exalata seeds to promote their germination.
[0026] Example 3 Effect of 2,3-butanediol on the germination of Arabidopsis, tobacco and rice seeds under high-salt environment Experimental method: Select mature and plump Arabidopsis thaliana, tobacco, and rice seeds. After disinfecting them with sodium hypochlorite solution at a concentration of 20% for 10 - 15 min, rinse them with distilled water 4 - 6 times and then sow them. Sow Arabidopsis thaliana seeds in a germination medium containing 125 mM NaCl and 100 μM 2,3 - butanediol, sow tobacco seeds in a germination medium containing 125 mM NaCl and 200 μM 2,3 - butanediol, and sow rice seeds in a solution containing 100 mM NaCl and 50 μM 2,3 - butanediol. According to the optimal growth conditions of the seeds, place them in a greenhouse at a temperature of 22 or 28 °C, with a 16 / 8 - hour light - dark cycle or a 14 / 10 - hour light - dark cycle, and a light intensity of 2500 - 3000 lx to germinate. Count the germination rate of Arabidopsis thaliana seeds after 3 days and the cotyledon greening rate after 5 days. Count the germination rate of tobacco seeds after 5 days and the cotyledon greening rate after 9 days. Count the germination rate of rice seeds after 4 days, and repeat the experiment three times. The specific method is the same as that in Example 1.
[0027] The results of the effect of 2,3 - butanediol on the germination of Arabidopsis thaliana, tobacco, and rice seeds under high - salt conditions are shown in Table 3.
[0028] Table 3 Results of the effect of 2,3 - butanediol on the germination of Arabidopsis thaliana, tobacco, and rice seeds under high - salt conditions As can be seen from Table 3, compared with the control group, the germination of seeds of the three plants was inhibited under high - salt conditions. After adding a certain concentration of 2,3 - butanediol to the experimental group, the germination rate or cotyledon greening rate of the seeds of the three plants was significantly increased. Compared with the experimental group treated with NaCl, after adding 2,3 - butanediol, the germination rate of Arabidopsis thaliana seeds increased by 69.19%, the cotyledon greening rate of Arabidopsis thaliana increased by 181.05%, the germination rate of tobacco seeds increased by 57.75%, the cotyledon greening rate of tobacco increased by 123.81%, and the germination rate of rice seeds increased by 66.67%.
[0029] Example 4 Effect of 2,3 - butanediol on the germination of Arabidopsis thaliana, tobacco, rice, and Hopea exalata seeds under low - temperature conditions Experimental method: Select mature and plump Arabidopsis thaliana, tobacco, and rice seeds, and select Hopea exalata seeds after storage for 3 weeks. After disinfecting Arabidopsis thaliana, tobacco, and rice seeds with a 20% sodium hypochlorite solution for 10 - 15 min, rinse them with distilled water 4 - 6 times and then sow. Rinse Hopea exalata seeds with distilled water 4 - 6 times and then sow. Sow the seeds in a germination / solid medium or solution containing 200 μM 2,3 - butanediol. According to the optimal growth conditions of the seeds, place the control group in a greenhouse at a temperature of 22 or 28°C, with a 16 / 8 h photoperiod or a 14 / 10 h photoperiod, and a light intensity of 2500 - 3000 lx for germination. Place the experimental group in an incubator at a temperature of 12 or 15°C, with the same photoperiod and light intensity as the control group for germination. Count the germination rate of Arabidopsis thaliana seeds after 4 days and the cotyledon greening rate after 8 days. Count the germination rate of tobacco seeds after 7 days and the cotyledon greening rate after 13 days. Count the germination rate of rice seeds after 8 days, and repeat the experiment three times. Count the germination rate of Hopea exalata seeds after 18 days, and repeat the experiment three times. The specific method is the same as that in Example 1.
[0030] The results of the effect of 2,3 - butanediol on the germination of Arabidopsis thaliana, tobacco, and rice seeds under low - temperature conditions are shown in Table 4.
[0031] Table 4 Results of the effect of 2,3 - butanediol on the germination of Arabidopsis thaliana, tobacco, and rice seeds under low - temperature conditions As can be seen from Table 4, compared with the control group, the germination of the seeds of the three plants was inhibited under low - temperature conditions. After adding a certain concentration of 2,3 - butanediol to the experimental group, the germination rate or cotyledon greening rate of the seeds of the three plants was significantly increased. Compared with the control group under cold treatment, after adding 2,3 - butanediol, the germination rate of Arabidopsis thaliana seeds increased by 74.23%, the cotyledon greening rate of Arabidopsis thaliana increased by 51.27%, the germination rate of tobacco seeds increased by 67.77%, the cotyledon greening rate of tobacco increased by 33.32%, and the germination rate of rice seeds increased by 27.27%.
[0032] Example 5 Effect of different concentrations of 2,3 - butanediol and gibberellin on the germination of Arabidopsis thaliana seeds Experimental method: Select mature and plump Arabidopsis thaliana seeds. After disinfecting them with a 20% sodium hypochlorite solution for 10 min, rinse them with distilled water 4 - 6 times and then sow. Sow them in a germination medium containing 100 μM and 200 μM 2,3 - butanediol and containing 5 μM and 10 μM gibberellin (GA3). After sowing, place them in a greenhouse at a temperature of 22°C, with a 16 h light and 8 h dark photoperiod, and a light intensity of 2500 - 3000 lx, and count the germination rate after 36 h and the cotyledon greening rate after 75 h. The specific method is the same as that in Example 1.
[0033] The results of the effects of 2,3-butanediol and gibberellin at different concentrations on the germination of Arabidopsis thaliana seeds are shown in Table 5.
[0034] Table 5 Results of the effects of 2,3-butanediol and gibberellin at different concentrations on the germination of Arabidopsis thaliana seeds As can be seen from Table 5, compared with the control group, gibberellin at 5 μM and 10 μM and 2,3-butanediol at 100 μM and 200 μM can all significantly promote the germination of Arabidopsis thaliana seeds. Compared with 5 μM gibberellin, the treatment with 100 μM 2,3-butanediol has the same effect on promoting germination and cotyledon greening. Compared with 10 μM gibberellin, the treatment with 200 μM 2,3-butanediol has the same effect on promoting germination and cotyledon greening. Therefore, 2,3-butanediol has the potential to replace gibberellin to promote seed germination in agricultural applications.
[0035] The embodiments of the present application have been described above. The above description is exemplary and not exhaustive, and is not limited to the disclosed embodiments. Many modifications and variations are obvious to those of ordinary skill in the art in the technical field without departing from the scope and spirit of the described embodiments. The choice of terms used herein is intended to best explain the principles of the embodiments, practical applications, or improvements to the technologies in the market, or to enable other ordinary skill in the art in the technical field to understand the embodiments disclosed herein.
Claims
1. Use of 1,2,3 - butanetriol in promoting seed germination, characterized in that, The application concentration of the 2,3-butanediol is 25 - 400 μM.
2. Use of 2,3-butanediol according to claim 1 in promoting seed germination, characterized in that, The application concentration of the 2,3-butanediol is 100 - 200 μM.
3. The application of 2,3-butanediol according to claim 1 in promoting seed germination, characterized in that, The seeds are Arabidopsis thaliana, tobacco, rice, upland rice, and Hopea exalata seeds.
4. The application of 2,3-butanediol according to claim 1 in promoting seed germination, characterized in that, The 2,3-butanediol can promote the germination of seeds under high-salt and low-temperature environments.
5. A method for promoting seed germination, characterized in that, It is realized according to the following steps: 1) Sow the seeds into a germination / solid medium or solution with a 2,3-butanediol concentration of 100 - 200 μM; 2) Then transfer the medium to a greenhouse with a temperature of 22 or 28 °C, a photoperiod of 14 or 16 h of light and 10 or 8 h of darkness for cultivation, and the light intensity is 2500 - 3000 lx.
6. The method for promoting seed germination according to claim 5, wherein The germination medium is 1 / 2 MS medium.
7. The method for promoting seed germination according to claim 6, wherein The formula of the 1 / 2 MS medium is: 2.2025 g / L MS medium, 10 g / L sucrose, 6.5 g / L agar.