CN115997765A - Spermidine-containing plant germination accelerator and application thereof in promoting germination of plant seeds - Google Patents
Spermidine-containing plant germination accelerator and application thereof in promoting germination of plant seeds Download PDFInfo
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- CN115997765A CN115997765A CN202211507163.XA CN202211507163A CN115997765A CN 115997765 A CN115997765 A CN 115997765A CN 202211507163 A CN202211507163 A CN 202211507163A CN 115997765 A CN115997765 A CN 115997765A
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- ATHGHQPFGPMSJY-UHFFFAOYSA-N spermidine Chemical compound NCCCCNCCCN ATHGHQPFGPMSJY-UHFFFAOYSA-N 0.000 title claims abstract description 91
- 230000035784 germination Effects 0.000 title claims abstract description 85
- 229940063673 spermidine Drugs 0.000 title claims abstract description 45
- 230000001737 promoting effect Effects 0.000 title claims abstract description 16
- 241000196324 Embryophyta Species 0.000 claims abstract description 58
- 235000017587 Medicago sativa ssp. sativa Nutrition 0.000 claims abstract description 45
- 241000219823 Medicago Species 0.000 claims abstract description 44
- 230000015784 hyperosmotic salinity response Effects 0.000 claims abstract description 9
- 238000000034 method Methods 0.000 claims description 26
- 150000003839 salts Chemical class 0.000 abstract description 11
- 238000012360 testing method Methods 0.000 abstract description 10
- 230000007226 seed germination Effects 0.000 abstract description 7
- 230000004071 biological effect Effects 0.000 abstract description 3
- 230000002708 enhancing effect Effects 0.000 abstract description 3
- 238000002360 preparation method Methods 0.000 abstract description 2
- 238000011282 treatment Methods 0.000 description 22
- 230000035882 stress Effects 0.000 description 19
- 238000002791 soaking Methods 0.000 description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 14
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical group [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 12
- 230000000694 effects Effects 0.000 description 8
- 239000008367 deionised water Substances 0.000 description 7
- 229910021641 deionized water Inorganic materials 0.000 description 7
- 239000003795 chemical substances by application Substances 0.000 description 6
- 239000011780 sodium chloride Substances 0.000 description 6
- 239000005708 Sodium hypochlorite Substances 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- 238000004659 sterilization and disinfection Methods 0.000 description 4
- 238000010998 test method Methods 0.000 description 4
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000005648 plant growth regulator Substances 0.000 description 3
- 229920000768 polyamine Polymers 0.000 description 3
- 239000008223 sterile water Substances 0.000 description 3
- 241000238631 Hexapoda Species 0.000 description 2
- 241000607479 Yersinia pestis Species 0.000 description 2
- 230000001133 acceleration Effects 0.000 description 2
- 238000000540 analysis of variance Methods 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000007619 statistical method Methods 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 240000004658 Medicago sativa Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000010485 coping Effects 0.000 description 1
- 239000006184 cosolvent Substances 0.000 description 1
- 230000000003 effect on germination Effects 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000003630 growth substance Substances 0.000 description 1
- 150000002611 lead compounds Chemical class 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
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- 238000003786 synthesis reaction Methods 0.000 description 1
- 231100000027 toxicology Toxicity 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
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Abstract
The invention discloses a plant germination accelerating agent containing spermidine and application thereof in promoting plant seed germination. The invention also discloses application of the spermidine-containing plant germination accelerating agent in improving the stress tolerance of plant seeds. The biological activity test shows that the plant germination accelerating agent containing spermidine has the capability of promoting the germination of alfalfa seeds and enhancing the salt stress resistance of the alfalfa seeds, and the spermidine can effectively promote the germination of the alfalfa seeds and enhance the salt tolerance of the alfalfa seeds when being applied at a low concentration (0.1 mmol/L). Meanwhile, spermidine has the advantages of easy preparation and low cost, and has high agricultural application and popularization values.
Description
Technical Field
The invention belongs to the field of plant growth regulators, and particularly relates to a plant germination accelerator containing spermidine and application thereof in promoting germination of plant seeds, in particular to a plant germination accelerator containing spermidine and application thereof in promoting germination of alfalfa seeds.
Background
The shortage of high-quality pasture is one of the main limiting factors for developing pasture and animal husbandry in China. Alfalfa is used as an important pasture crop, and in recent years, although tension of pasture demand is relieved by introducing foreign high-quality germplasm resources, the problems of low germination rate, weak seedling period, poor stress resistance and the like of alfalfa seeds still exist, and a large amount of alfalfa seeds need to be imported every year to meet planting demands.
The plant growth regulator is used as an important branch of pesticides, is widely used in crop field management, and has important effects in promoting seed germination and enhancing plant stress resistance. The development of the novel plant growth regulator is a complex work which needs to consume manpower and material resources, and the development process mainly comprises the main processes of functional discovery and chemical synthesis, activity evaluation, action mechanism research, toxicology evaluation, registration and the like of lead compounds.
Polyamines (PAs) are chemical substances widely existing in organisms and are a class of aliphatic nitrogenous bases with biological activity. As an important growth regulator of higher plants, polyamine participates in physiological processes such as growth and development, maturation and aging, stress resistance and the like of the plants.
Disclosure of Invention
The invention aims to solve the technical problem of how to promote the germination of plant seeds and improve the stress tolerance of the plant seeds.
In order to solve the technical problems, the invention firstly provides new application of spermidine or spermidine solution.
The present invention provides the use of spermidine or a spermidine solution in any of the following 1) -4):
1) Promoting germination of plant seeds;
2) Preparing a product for promoting germination of plant seeds;
3) Improving the stress tolerance of plant seeds;
4) And preparing a product for improving the stress tolerance of plant seeds.
In the above application, the promotion of germination of plant seeds is embodied as improvement of germination rate and/or germination index of plant seeds.
In the above application, the stress tolerance may specifically be salt tolerance.
Further, the salt tolerance is sodium chloride tolerance.
Still further, the sodium chloride concentration was 200mmol/L.
In the above application, the product may be a plant germination accelerator.
In order to solve the technical problems, the invention also provides a plant germination accelerating agent.
The plant germination accelerating agent provided by the invention comprises spermidine or spermidine solution.
In practical application, the product or the plant germination accelerating agent can also contain other components such as synergists, cosolvents and the like according to practical needs.
In order to solve the technical problems, the invention also provides a method for promoting the germination of plant seeds or improving the stress tolerance of the plant seeds.
The method for promoting germination of plant seeds or improving stress tolerance of plant seeds provided by the invention comprises the step of treating plant seeds with spermidine or spermidine solution.
In the method, the promotion of plant seed germination is embodied as improvement of the germination rate and/or germination index of plant seeds.
In the above method, the stress tolerance may specifically be salt tolerance.
Further, the salt tolerance is sodium chloride tolerance.
Still further, the sodium chloride concentration was 200mmol/L.
In the above method, the treatment mode may be seed soaking.
Further, the seed soaking time can be 12-24 hours, and specifically can be 24 hours.
In the method, the seed disinfection step is further included before the treatment.
Further, the seed disinfection method can be carried out according to the following steps: the seeds were sterilized with sodium hypochlorite solution (e.g., 0.3% sodium hypochlorite solution) for 5min, then rinsed with sterile water and the surface of the seeds was blotted with filter paper.
The method further comprises the step of accelerating germination of the treated seeds.
Further, the germination accelerating method can be carried out according to the following steps: placing the treated seeds into culture dishes (100 seeds in each culture dish) paved with filter paper, dripping deionized water to keep the filter paper moist, and then illuminating the culture dishes at the temperature of 25 ℃ for 12 h/d; germination was performed in the dark at 12h/d and 20 ℃.
In any of the above applications or plant germination accelerating agents or methods, the concentration of the spermidine solution may be 0.05-0.2mmol/L or 0.05-0.1mmol/L or 0.1-0.2mmol/L or 0.05mmol/L or 0.1mmol/L or 0.2mmol/L, preferably 0.1mmol/L.
Further, the solvent of the spermidine solution is deionized water.
In any of the above applications or plant germination accelerating agents or methods, the spermidine has a structural formula shown in formula I,
in any of the above applications or plant germination accelerating agents or methods, the plant may be alfalfa; the alfalfa may be alfalfa; the variety of the alfalfa can be in particular Zhongjing No. 4.
The invention provides a plant germination accelerator with spermidine as an active ingredient, which is verified by a related biological activity test, and the plant germination accelerator has the capability of promoting germination of alfalfa seeds and enhancing salt stress resistance of the alfalfa seeds, and the spermidine can be applied at a low concentration (0.1 mmol/L) to effectively promote germination of the alfalfa seeds and enhance salt tolerance of the alfalfa seeds. Meanwhile, spermidine has the advantages of easy preparation and low cost, and has high agricultural application and popularization values.
Drawings
FIG. 1 is a graph showing the effect of spermidine on the germination of alfalfa seeds.
FIG. 2 shows the effect of spermidine on salt stress resistance of alfalfa seeds.
Detailed Description
The following detailed description of the invention is provided in connection with the accompanying drawings that are presented to illustrate the invention and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the invention in any way.
The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are carried out according to techniques or conditions described in the literature in the field or according to the product specifications. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified. The quantitative experiments in the following examples were performed in triplicate unless otherwise specified.
Spermidine in the examples below is the product of Meryer/Michael under the product number 124-20-9|M04069-5G and has the formula I.
EXAMPLE 1 promotion of alfalfa seed germination by spermidine
1. Test site and materials
The germination test is carried out on full alfalfa (alfalfa No. 4) seeds with consistent sizes and no plant diseases and insect pests in the artificial climatic chamber of the plant protection building of Beijing Chinese university.
2. Test method
1. Seed disinfection
Sterilizing all seeds with 0.3% sodium hypochlorite solution for 5min, washing with sterile water, and sucking water on the surface of the seeds with filter paper to obtain sterilized seeds.
2. Seed treatment
The disinfected seeds are randomly divided into two groups, and an experimental group and a control group are set according to the different treatment methods of alfalfa seeds, wherein each group of treatment methods are as follows:
experimental group (Spd): alfalfa seeds were treated by soaking in 0.1mmol/L spermidine solution (deionized water as solvent).
Control group (CK): soaking alfalfa seeds in deionized water.
3. Accelerating germination
After 24h of seed soaking treatment, the seeds after seed soaking treatment are placed into culture dishes paved with filter paper for germination, 100 seeds are placed into each culture dish, deionized water is dripped to keep the filter paper moist, and the culture dishes are placed into a climatic chamber for germination. The climatic chamber conditions are that the illumination is 12h/d and the temperature is 25 ℃; dark for 12h/d,20 ℃. The germination of the seeds is counted every 12 hours (germination is based on the same length of the radicle of the seeds as the length of the seeds). The observation was continued for 10 days, and the observation was ended on the 10 th day and the seedling length of 10 seeds was randomly determined.
4. Statistical analysis of data
To investigate the effect of spermidine on alfalfa seed germination, the measured seed germination-related data were counted and analyzed after 10 days of experimental treatment. According to the measurement standard in the national seed test procedure, the germination rate and the germination index of the seeds are calculated, and the specific calculation formula is as follows:
germination percentage (%) =number of germinated seeds/number of tested seeds×100%.
Germination Index (GI) = Σg t /D t (G t D is the germination number at t days t Days for germination up to t days).
All experimental data were analyzed using SPSS 25.0. The analysis method selects ANOVA for differential analysis, and when P is less than 0.05, the difference is obvious.
3. Test results
The results are shown in Table 1 and FIG. 1, and the test results show that: compared with the control group, the germination rate of alfalfa seeds after seed soaking treatment in the Spd group is remarkably improved (figure 1 a); the germination index of alfalfa seeds after seed soaking treatment of the Spd group is also remarkably improved (figure 1 b), which shows that the seed soaking treatment of the Spd has remarkable promotion effect on germination of alfalfa seeds. Therefore, spermidine has the function of promoting germination of alfalfa seeds.
Table 1: influence of spermidine seed soaking on germination rate and germination index of alfalfa seeds
| Treatment of | Germination percentage (%) | Germination index |
| CK | 83.33±3.09b | 55±0.53b |
| Spd | 92.33±3.30a | 74.47±5.21a |
Note that: the different letters in the same column represent significant differences between treatments (P < 0.05)
Example 2 Effect of spermidine on salt stress resistance of alfalfa seeds
1. Test site and materials
The germination test is carried out on full alfalfa (alfalfa No. 4) seeds with consistent sizes and no plant diseases and insect pests in the artificial climatic chamber of the plant protection building of Beijing Chinese university.
2. Test method
1. Seed disinfection
Sterilizing all seeds with 0.3% sodium hypochlorite solution for 5min, washing with sterile water, and sucking water on the surface of the seeds with filter paper to obtain sterilized seeds.
2. Seed treatment
The disinfected seeds are randomly divided into two groups, and an experimental group and a control group are set according to the different treatment methods of alfalfa seeds, wherein each group of treatment methods are as follows:
experimental group (Spd): alfalfa seeds were treated by soaking in 0.1mmol/L spermidine solution (deionized water as solvent).
Control group (CK): soaking alfalfa seeds in deionized water.
3. Salt stress resistance experiment
After 24h of seed soaking treatment, the seeds after seed soaking treatment are placed in culture dishes paved with filter paper for germination acceleration, 100 seeds are placed in each culture dish, then 4mL of NaCl solution (solvent is water) with the concentration of 200mmol/L is dripped into each culture dish, the weight of each culture dish is weighed, and the culture dishes are placed in a climatic chamber for germination acceleration. The climatic chamber conditions are that the illumination is 12h/d and the temperature is 25 ℃; dark for 12h/d,20 ℃. Every 12 hours, the germination condition of the seeds is observed and counted (the germination is based on the condition that the length of radicle of the seeds is the same as that of the seeds), and the water in each culture dish is supplemented by a weighing method in the next day so as to ensure that the salt concentration is consistent. The observation was continued for 10 days, and the observation was ended on the 10 th day and the seedling length of 10 seeds was randomly determined.
4. Statistical analysis of data
To investigate the effect of spermidine on salt stress resistance of alfalfa seeds, the measured seed germination-related data were counted and analyzed 10 days after the experimental treatment. According to the measurement standard in the national seed test procedure, the germination rate and the germination index of the seeds are calculated, and the specific calculation formula is as follows:
germination percentage (%) =number of germinated seeds/number of tested seeds×100%.
Germination Index (GI) = Σg t /D t (G t D is the germination number at t days t Days for germination up to t days).
All experimental data were analyzed using SPSS 25.0. The analysis method selects ANOVA for differential analysis, and when P is less than 0.05, the difference is obvious.
3. Test results
The results are shown in Table 2 and FIG. 2, and the test results show that: compared with the control group, the seed soaking treatment of the Spd group obviously improves the germination rate (figure 2 a) and the germination index (figure 2 b) of the alfalfa seeds under the salt stress, which proves that the Spd has the capability of improving the salt stress coping capacity of the alfalfa seeds, and can improve the tolerance of the alfalfa seeds under the salt stress. Therefore, spermidine has the function of improving the salt tolerance of alfalfa seeds.
Table 2: effect of spermidine on salt stress resistance of alfalfa seeds
| Treatment of | Germination percentage (%) | Germination index |
| NaCl | 78.67±5.44b | 37.83±3.30b |
| Spd | 91.33±3.30a | 46.34±2.36a |
Note that: the different letters in the same column represent significant differences between treatments (P < 0.05)
The present invention is described in detail above. It will be apparent to those skilled in the art that the present invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with respect to specific embodiments, it will be appreciated that the invention may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. The application of some of the basic features may be done in accordance with the scope of the claims that follow.
Claims (10)
1. Use of spermidine or a spermidine solution in any of the following 1) -4):
1) Promoting germination of plant seeds;
2) Preparing a product for promoting germination of plant seeds;
3) Improving the stress tolerance of plant seeds;
4) And preparing a product for improving the stress tolerance of plant seeds.
2. The use according to claim 1, characterized in that: the promotion of germination of plant seeds is reflected in an increase in the germination rate and/or germination index of plant seeds.
3. The use according to claim 1, characterized in that: the stress tolerance is salt tolerance.
4. A plant germination agent comprising spermidine or a spermidine solution.
5. A method of promoting germination of plant seeds comprising the step of treating the plant seeds with spermidine or a spermidine solution.
6. The method according to claim 5, wherein: the promotion of germination of plant seeds is reflected in an increase in the germination rate and/or germination index of plant seeds.
7. A method for improving stress tolerance of a plant seed comprising the step of treating the plant seed with spermidine or a spermidine solution.
8. The method according to claim 7, wherein: the stress tolerance is salt tolerance.
9. A use according to any one of claims 1 to 3 or a plant germination agent according to claim 4 or a method according to any one of claims 5 to 8, characterized in that: the concentration of the spermidine solution is 0.1mmol/L.
10. A use according to any one of claims 1 to 3 or a plant germination agent according to claim 4 or a method according to any one of claims 5 to 8, characterized in that: the plant is alfalfa; or, the alfalfa is alfalfa.
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