CN113115605A - Method for promoting spinach seed germination by using carbon dots - Google Patents
Method for promoting spinach seed germination by using carbon dots Download PDFInfo
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- CN113115605A CN113115605A CN202110548990.2A CN202110548990A CN113115605A CN 113115605 A CN113115605 A CN 113115605A CN 202110548990 A CN202110548990 A CN 202110548990A CN 113115605 A CN113115605 A CN 113115605A
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Abstract
The invention discloses a method for promoting spinach seed germination by using carbon dots, and belongs to the technical field of agriculture. The method for promoting spinach seed germination by using the carbon dots comprises the steps of preparing the carbon dots into a carbon dot aqueous solution, applying the carbon dot aqueous solution to spinach seeds, and performing light-proof culture; wherein the carbon dots are spherical carbon nano-particles, the size is 2-9nm, and the element molar ratio of C, O is as follows: 1-6: 1; the concentration of the carbon dot aqueous solution is 1-50 mg/L. After spinach seeds are cultured for 10 days by the method, the germination rate reaches more than 63%, and compared with a method of solution culture without carbon spots, the method provided by the invention increases the fresh weight of the spinach seeds by more than 25% and increases the dry weight by more than 46.7%; the overground part is increased by more than 40.5 percent, and the root part is increased by more than 67.2 percent; the expression level of aquaporin gene is increased by 393.5% or more, and the water content of the seedling is increased by 25.9% or more.
Description
Technical Field
The invention relates to a method for promoting spinach seed germination by using carbon dots, and belongs to the technical field of agriculture.
Background
The germination of seeds is a very critical stage during the plant growth cycle and is also a stage that is very sensitive to abiotic environmental factors. Whether a plant seedling can grow rapidly and robustly is closely related to the germination capacity of the seed. Many scholars now have developed their own ideas for techniques to promote seed germination. The Castanea mollissima test finds that gibberellin and naphthylacetic acid have certain effects on improving the germination rate and the germination potential of celery seeds; the Wangwang printed article indicates that the seed of the leaf beet is soaked and treated by hydrogen peroxide, boric acid and monopotassium phosphate with certain concentration, so that the germination rate, the germination vigor and the germination index of the seed can be improved; the aged and dazzling soldiers find that seeds can be effectively promoted to germinate by soaking the seeds in 20% liquid detergent for 24 hours at the seed soaking temperature of 30 ℃ and the germination accelerating temperature of 25 ℃.
Spinach is an important worldwide green leaf vegetable, and has low germination rate and irregular emergence after field sowing due to the reasons of inconsistent seed maturity, strong dormancy habit, thick peel, germination inhibiting substances contained in the spinach and the like, and particularly has obvious reduction of the germination rate of old seeds in old years. Before sowing, spinach seeds are usually subjected to a germination treatment in order to increase the level of germination of the spinach seeds. The spinach germination rate can be improved to different degrees by husking, high-temperature treatment, seed soaking in warm soup, seed soaking in low temperature, ultraviolet radiation, ultrasonic waves and seed coat breaking by rubbing. Researches on spinach seed germination by treatment with various chemical reagents, such as choline chloride, hydrogen peroxide, polyethylene glycol (PEG100), potassium nitrate, and boric acid seed soaking have also been reported. But has the problem of low germination rate.
Therefore, how to promote the spinach seed germination and improve the germination rate is a technical problem which needs to be solved urgently at present.
Carbon Dots (CDs), as a new class of Carbon-based fluorescent ENMs, are widely used in the fields of bio-imaging, drug delivery, catalysis, optoelectronic devices, etc. due to their excellent properties, such as wide light absorption, adjustable fluorescence, high luminous efficiency, photobleaching resistance, high chemical stability, good biocompatibility, low toxicity, etc. In recent years, CDs have been shown to promote crop growth, improve plant photosynthesis, and enhance plant resistance and nitrogen fixation. However, there is no document disclosing whether it can stably and efficiently promote the germination of crop seeds.
Disclosure of Invention
[ problem ] to
The conventionally adopted spinach seed germination method has the problem of low germination rate.
[ solution ]
In order to solve the problems, the carbon dots are applied to the spinach seed germination stage, and the water absorption efficiency is increased through the action of the aquaporin, so that the seed germination is promoted.
The first purpose of the invention is to provide a method for promoting spinach seed germination by using carbon dots, which is to prepare the carbon dots into a carbon dot aqueous solution, apply the carbon dot aqueous solution to spinach seeds and culture the spinach seeds in a dark place.
In one embodiment of the present invention, the carbon dots are spherical carbon nanoparticles having a size of 2 to 9 nm.
In one embodiment of the present invention, the molar ratio of the elements C and O in the carbon dots is: 1-6: 1, more preferably 2 to 4: 1; more preferably 2: 1.
in one embodiment of the present invention, the concentration of the aqueous solution of carbon dots is 1 to 50mg/L, more preferably 5 to 25mg/L, and still more preferably 10 to 20 mg/L.
In one embodiment of the present invention, the applying manner includes one of seed soaking, spraying and topdressing.
In one embodiment of the invention, the spraying is to apply 8-12mL of carbon dot aqueous solution in a 100 square centimeter container containing 80-100 spinach seeds for germination in the dark.
In one embodiment of the invention, the seed soaking is to soak spinach seeds in a carbon dot aqueous solution for 6 hours; then sprouting in dark is carried out.
In one embodiment of the invention, the top dressing is to arrange 80-100 spinach seeds on the surface of 100 square centimeters of soil and then spray 8-12mL of carbon dot aqueous solution; covering soil on the surface of the seeds, and culturing for germination.
In one embodiment of the invention, the spinach seed is sterilized prior to germination.
In one embodiment of the invention, the disinfection is carried out by soaking in a sodium chlorate solution for 20-40 minutes and then rinsing with water.
In one embodiment of the present invention, the sodium hypochlorite solution is a sodium hypochlorite aqueous solution with a mass concentration of 0.05%.
In one embodiment of the present invention, the temperature for the light-shielding culture is 20 to 30 ℃ (normal temperature).
A second object of the invention is the use of the method according to the invention in the field of agriculture.
The third purpose of the invention is to provide a method for improving crop seed germination based on aquaporin regulation, which comprises the steps of preparing carbon dots into a carbon dot aqueous solution, applying the carbon dot aqueous solution to spinach seeds, and culturing in a dark place; wherein the carbon dots are spherical carbon nano-particles, the size is 2-9nm, and the element molar ratio of C, O is as follows: 1-6: 1; the concentration of the carbon dot aqueous solution is 1-50 mg/L.
[ advantageous effects ]
(1) The method can obviously increase the water absorption of the seeds, improve the germination rate of the seeds, increase the biomass of the germinated seeds and be beneficial to ensuring the sound growth of crops in the future.
(2) According to the invention, the carbon dots are applied in the spinach seed germination stage, so that the opening of spinach seed aquaporins can be regulated and controlled, the moisture absorption is promoted, and the germination rate of crop seeds is improved.
(3) After spinach seeds are cultured for 10 days by the method, the germination rate reaches more than 63%, and compared with a method of solution culture without carbon spots, the method provided by the invention increases the fresh weight of the spinach seeds by more than 25% and increases the dry weight by more than 46.7%; the overground part is increased by more than 40.5 percent, and the root part is increased by more than 67.2 percent; the expression level of aquaporin gene is increased by 393.5% or more, and the water content of the seedling is increased by 25.9% or more.
Drawings
Fig. 1 is a TEM picture of the carbon dots used in example 1.
FIG. 2 shows the composition element ratio of carbon dots used in example 1.
FIG. 3 shows the germination percentage of spinach exposed to carbon spots in example 1.
FIG. 4 is the carbon dot exposed spinach biomass of example 1.
FIG. 5 is a scan of spinach exposed to carbon spots as in example 1.
FIG. 6 is the length of spinach exposed to carbon spots in example 1.
FIG. 7 is the surface area of the spinach exposed to the carbon spots in example 1.
FIG. 8 shows the expression levels of carbon-exposed spinach aquaporin PIPs in example 1.
FIG. 9 is the moisture content of spinach exposed to carbon spots in example 1.
Detailed Description
The following description of the preferred embodiments of the present invention is provided for the purpose of better illustrating the invention and is not intended to limit the invention thereto.
The test method comprises the following steps:
1. germination rate:
the germination rate (total number of germinated seeds at harvest/total number of test seeds) × 100%
The seed rootlet or cotyledon protrusion seed coat was considered to germinate during the experimental test, and only the rootlet exceeding 1mm was scored as root length.
2. Biomass
After the germination test was completed, the seedlings were collected, their fresh weight was measured using an electronic balance, and their dry weight was measured after drying in an oven.
3. Root parameter
After the germination test was completed, spinach seedlings were collected, and the length and surface area of the spinach seedlings were measured using a Scanner (Scanner, Epson 12000XL, Japan).
4. Determination of expression amount of aquaporins PIPs
And selecting specific primers of the PIPS genes to perform fluorescent quantitative PCR analysis on the PIPS genes.
Determination of the C/O ratio
The carbon points were measured by X-ray photoelectron spectroscopy (XPS, ESCALAB 250Xi, Saimer fly, USA), and the C/O ratio was calculated from the ratios of the different elements obtained.
EXAMPLE 1 spray application (CDs group)
A method for promoting spinach seed germination by using carbon dots comprises the following steps:
(1) spinach seeds from agricultural academy of sciences of Jiangsu province are disinfected in 0.05% sodium hypochlorite solution for 30 minutes, and then washed with deionized water for 3 times for disinfection; obtaining disinfected spinach seeds;
(2) uniformly distributing the disinfected spinach seeds obtained in the step (1) in a 100 square centimeter culture dish, wherein the density is 100 granules/100 square centimeters;
(3) preparing carbon dots into a carbon dot aqueous solution with the concentration of 10mg/L, and then spraying 10mL of the carbon dot aqueous solution into the culture dish in the step (2); wherein the size of the carbon dots is 2-9nm (figure 1), the molar ratio of the elements C/O is 2: 1 (fig. 2);
(4) and (4) sealing the culture dish in the step (3) by using a sealing film, and culturing for 4, 6, 8 and 10 days at 25 ℃ in a dark place. Comparative example 1 solution without carbon addition (CK group)
A spinach seed germination method comprises the following steps:
(1) spinach seeds from agricultural academy of sciences of Jiangsu province are disinfected in 0.05% sodium hypochlorite solution for 30 minutes, and then washed with deionized water for 3 times for disinfection; obtaining disinfected spinach seeds;
(2) uniformly distributing the disinfected spinach seeds obtained in the step (1) in a 100 square centimeter culture dish, wherein the density is 100 granules/100 square centimeters;
(3) and (3) sealing the culture dish in the step (2) by using a sealing film, and culturing for 4, 6, 8 and 10 days at 25 ℃ in a dark place.
The germinated spinach seeds obtained in example 1 and comparative example 1 were subjected to a performance test, and the test results were as follows:
the Carbon Spots (CDs) solution promoted well the germination of spinach seeds; on day 4, the spinach treated with the CDs solution germinated first, whereas the germination rates on days 6, 8 and 10 increased by 360.0%, 200.0% and 50.0%, respectively, compared to control 1 (fig. 3); CDs also significantly increased the dry fresh weight of spinach by 30.1% and 55.3% after 10 days of culture compared to control 1 (FIG. 4); meanwhile, CDs have promoting effect on the development of root and stem morphology of spinach, which is mainly reflected in the aspects of stem length and surface area (figure 5); CDs can increase the length of spinach seeds, and after 10 days of culture, the overground part is increased by 43.4%, and the root part is increased by 69.4% (FIG. 6); the CDs ENMs can increase the surface area of roots and stems of the spinach, and after 10 days of culture, the surface area of the roots is increased by 44.8 percent, and the surface area of the stems is increased by 38.7 percent (figure 7); the expression quantity of the aquaporin gene is increased by 409.0 percent (figure 8), and the water content of seedlings is increased by 29.2 percent (figure 9).
Example 2
The molar ratio of the elements C/O in step (3) of example 1 was adjusted to 1: 1. 3: 1. 6: 1, the mixture was incubated in the dark for 10 days, and the other conditions were the same as those in example 1, to obtain germinated spinach seeds.
The germinated spinach seeds were subjected to performance tests, the test results are given in table 1 below:
table 1 test results of example 2
Example 3
The concentration of the aqueous solution of carbon spots in step (3) of example 1 was adjusted to 1, 20, and 50mg/L, and the mixture was incubated in the dark for 10 days, and the rest was kept the same as in example 1, to obtain germinated spinach seeds.
The germinated spinach seeds were subjected to performance tests, the test results are given in table 1 below:
table 2 test results of example 3
Comparative example 2
Spinach of example 1 was changed to rape and tomato, and was cultured in the dark for 10 days, and the others were kept the same as those of example 1, to obtain a germinated spinach seed.
The germinated spinach seeds were subjected to performance tests, the test results are given in table 1 below:
table 3 test results of comparative example 2
Example 4 seed soaking
A method for promoting spinach seed germination by using carbon dots comprises the following steps:
(1) spinach seeds from agricultural academy of sciences of Jiangsu province are disinfected in 0.05% sodium hypochlorite solution for 30 minutes, and then washed with deionized water for 3 times for disinfection; obtaining disinfected spinach seeds;
(2) collecting 100 disinfected spinach seeds obtained in the step (1) into a test tube;
(3) preparing the carbon dots into a carbon dot aqueous solution with the concentration of 10mg/L, and then adding 100mL of carbon dot aqueous solution into the test tube in the step (2); wherein the size of the carbon dots is 2-9nm (FIG. 1) and the element molar ratio of C/O is 2 (FIG. 2);
(4) sealing the test tube in the step (3) by using a sealing film, and soaking seeds for 6 hours at 25 ℃ in a dark place;
(5) after seed soaking, the seeds are fished out, evenly distributed in a culture dish, sealed by a sealing film and cultured for 10 days at 25 ℃ in dark.
Comparative example 3
The carbon dot solution of example 4 was omitted and the rest was identical to example 4 to obtain germinated spinach seeds.
The germination rate of the control example 3 is found to be 53% through tests; the germination percentage of example 4 was 65%.
Example 5 soil culture
A method for promoting spinach seed germination by using carbon dots comprises the following steps:
(1) spinach seeds from agricultural academy of sciences of Jiangsu province are disinfected in 0.05% sodium hypochlorite solution for 30 minutes, and then washed with deionized water for 3 times for disinfection; obtaining disinfected spinach seeds;
(2) uniformly distributing 100 disinfected spinach seeds obtained in the step (1) on the surface of 100 square centimeters of soil;
(3) preparing carbon dots into a carbon dot aqueous solution with the concentration of 10mg/L, and then spraying 10mL of the carbon dot aqueous solution into the soil surface with the inner surface of 100 square parts in the step (2); wherein the size of the carbon dots is 2-9nm (FIG. 1) and the element molar ratio of C/O is 2 (FIG. 2);
(4) and (4) uniformly spreading a thin layer of fine soil on the seeds in the step (3), and culturing for 10 days.
Comparative example 4
The carbon dot solution of example 5 was omitted and the rest was identical to example 5 to obtain germinated spinach seeds.
The germination rate of the control example 4 is found to be 32% through tests; the germination percentage of example 5 was 63%.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (10)
1. A method for promoting spinach seed germination by using carbon dots is characterized in that the carbon dots are prepared into a carbon dot aqueous solution, and then the carbon dot aqueous solution is applied to spinach seeds for light-proof culture.
2. The method according to claim 1, wherein the molar ratio of the elements C and O in the carbon dots is: 1-6: 1.
3. the method according to claim 1 or 2, wherein the concentration of the aqueous carbon dot solution is 1 to 50 mg/L.
4. The method of claim 1, wherein the applying comprises one of seed soaking, spraying, and top dressing.
5. The method as claimed in claim 4, wherein the spraying is carried out by applying 8-12mL of carbon dot aqueous solution in a 100 cm square container containing 80-100 spinach seeds and germinating in the dark.
6. The method of claim 4, wherein the seed soaking is carried out by soaking spinach seeds in an aqueous carbon dot solution for 6 hours; then sprouting in dark is carried out.
7. The method of claim 4, wherein the top application comprises spreading 80-100 spinach seeds on a 100 cm square soil surface, and then spraying 8-12mL of an aqueous carbon dot solution; covering soil on the surface of the seeds, and culturing for germination.
8. The method according to any one of claims 1 to 7, wherein the temperature of the light-shielding culture is 20 to 30 ℃.
9. Use of the method according to any one of claims 1 to 8 in the agricultural field.
10. A method for improving crop seed germination based on regulation and control of aquaporin is characterized in that a carbon dot is prepared into a carbon dot aqueous solution, and then the carbon dot aqueous solution is applied to spinach seeds for light-tight culture; wherein the element mole ratio of C, O is: 1-6: 1; the concentration of the carbon dot aqueous solution is 1-50 mg/L.
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| CN114467653A (en) * | 2022-02-12 | 2022-05-13 | 厦门农运来农业技术服务有限公司 | Method for promoting female production and increasing yield of bitter gourds |
| CN115259138A (en) * | 2022-08-04 | 2022-11-01 | 江南大学 | Method for improving drought resistance and yield of crops based on carbon nano material |
| CN117204284A (en) * | 2023-09-21 | 2023-12-12 | 扬州大学 | Method for improving emergence rate of wheat, cultivating strong seedlings and increasing yield and application |
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Cited By (4)
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| CN114467653A (en) * | 2022-02-12 | 2022-05-13 | 厦门农运来农业技术服务有限公司 | Method for promoting female production and increasing yield of bitter gourds |
| CN115259138A (en) * | 2022-08-04 | 2022-11-01 | 江南大学 | Method for improving drought resistance and yield of crops based on carbon nano material |
| CN115259138B (en) * | 2022-08-04 | 2023-09-22 | 江南大学 | Method for improving drought tolerance and yield of crops based on carbon nanomaterial |
| CN117204284A (en) * | 2023-09-21 | 2023-12-12 | 扬州大学 | Method for improving emergence rate of wheat, cultivating strong seedlings and increasing yield and application |
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