Method for promoting shape after-ripening and germination of fresh coptis chinensis seeds
By stratifying fresh Coptis chinensis seeds at 15-20℃ and treating them with gibberellin solution, the seed dormancy problem was solved, efficient seed germination was achieved, and the morphological after-ripening and germination of fresh Coptis chinensis seeds were promoted, providing technical support for artificial breeding.
Patent Information
- Application Number
- CN202511250857.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-09-03
- Publication Date
- 2025-11-04
AI Technical Summary
Fresh Coptis chinensis seeds suffer from seed dormancy due to internal factors, resulting in low germination rates, difficulty in germination, and uneven emergence. There is limited existing research on this issue, and there is an urgent need for methods to promote seed morphological after-ripening and germination.
Fresh Coptis chinensis seeds were stratified at 15-20℃ for 60-90 days, then soaked in a 200-400ppm gibberellin solution for 48 hours, and finally germinated at 10-20℃ using filter paper as a germination bed.
It significantly improved the germination rate of fresh Coptis chinensis seeds, reaching a maximum of 76.19%, providing technical support for the artificial propagation of fresh Coptis chinensis.
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Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of seed seedling, in particular to a method for promoting morphological after-ripening and germination of Plagiorhegma dubium Maxim. BACKGROUND
[0002] Plagiorhegma dubium Maxim. is a perennial herb of Plagiorhegma dubium Maxim. of Berberidaceae, which is dry root and rhizome, cold in nature, bitter in taste, and non-toxic. The whole grass of Plagiorhegma dubium Maxim. contains about 15% tannins, which can extract roasted glue and yellow dye; the tender stems and leaves can be eaten fresh and used as livestock feed. The root contains alkaloids, starch and other ingredients, and has the effects of clearing heat and detoxifying, cooling blood and stopping bleeding, and improving eyesight and stopping tears. Plagiorhegma dubium Maxim. is often used with licorice and atractylodes to treat stomach heat and acid reflux; the decoction of Plagiorhegma dubium Maxim. can be used to treat conjunctivitis.
[0003] Due to the high demand for medication, people have harvested a large amount of Plagiorhegma dubium Maxim., which has destroyed the resources of wild Plagiorhegma dubium Maxim. and cannot meet the market demand, so the artificial planting of Plagiorhegma dubium Maxim. is of great significance to resource protection, medication demand and adjustment of forestry economic structure. Plagiorhegma dubium Maxim. grows under coniferous forest, mixed wood forest, shrub or shady and humid place of hillside, and the natural propagation rate is low. The seed of Plagiorhegma dubium Maxim. is caused by internal factors, that is, the embryo development is incomplete and the physiological after-ripening is not completed. Even if the suitable conditions are provided, the seed cannot germinate, which leads to low germination rate, difficult germination and uneven emergence. At present, there are few researches on Plagiorhegma dubium Maxim. in China, mainly including extraction process, tissue culture, development morphology of inflorescence and flower and chemical component analysis, therefore, it is urgent to further study the morphological after-ripening, dormancy termination and germination of Plagiorhegma dubium Maxim. seed. SUMMARY
[0004] The purpose of the present application is to provide a method for promoting morphological after-ripening and germination of Plagiorhegma dubium Maxim. seed, which can effectively promote the embryo development of Plagiorhegma dubium Maxim. seed and complete morphological after-ripening, and further significantly promote seed germination, with the highest germination rate of 76.19%, which provides solid technical support for artificial breeding and industrial development of Plagiorhegma dubium Maxim..
[0005] To achieve the above purpose, the present application provides the following scheme:
[0006] The present application provides a method for promoting morphological after-ripening of Plagiorhegma dubium Maxim. seed, which comprises the step of stratifying the Plagiorhegma dubium Maxim. seed at 15-20℃.
[0007] Further, the stratification temperature is 15℃, and the time is 60-90 days.
[0008] The application further provides a method for promoting seed germination of fresh Coptis chinensis and / or increasing seed germination rate of fresh Coptis chinensis, comprising the following steps:
[0009] The fresh Coptis chinensis seeds are stratified at 10-20 DEG C to obtain stratified seeds;
[0010] The stratified seeds are soaked in a gibberellin solution, and after soaking, germination is carried out on filter paper as a germination bed.
[0011] Further, the stratification temperature is 15 DEG C, and the stratification time is 60-90 days.
[0012] Further, the content of gibberellin in the gibberellin solution is 200-400 ppm.
[0013] Further, the content of gibberellin in the gibberellin solution is 400 ppm.
[0014] Further, the soaking time is 48 h.
[0015] Further, the germination environment temperature is 10-20 DEG C.
[0016] Further, the germination environment temperature is 15 DEG C.
[0017] The application further provides application of the germinated seeds obtained by the above method in artificial breeding of fresh Coptis chinensis.
[0018] The application discloses the following technical effects:
[0019] The application provides a method for promoting morphological after-ripening of fresh Coptis chinensis seeds by constant temperature stratification at 15-20 DEG C for 60-90 days, and the experimental results show that the method can effectively promote embryo development of the fresh Coptis chinensis seeds and complete morphological after-ripening. Further, the seeds after morphological after-ripening are soaked in a 400 ppm gibberellin solution, and are placed on filter paper at a temperature of 10-20 DEG C, so that seed germination is promoted. The experimental results show that the germination rate of the fresh Coptis chinensis seeds is up to 76.19%. The application provides a rapid and efficient regulation method for terminating dormancy of fresh Coptis chinensis seeds and promoting seed germination, and provides solid technical support for artificial breeding and industrial development of fresh Coptis chinensis, and has outstanding practical application value and broad application prospect. BRIEF DESCRIPTION OF DRAWINGS
[0020] In order to more clearly illustrate the technical solutions in the embodiments of the application or the prior art, the following will briefly introduce the drawings needed in the embodiments. Obviously, the drawings in the following description only constitute some embodiments of the application, and for those skilled in the art, other drawings can also be obtained without creative labor.
[0021] Figure 1 Figure 1 is a diagram of the initial state of the fresh coptis seed; wherein A is the fresh coptis seed pod cracking; B is the fresh coptis seed with white appendages on the surface; C is the fresh coptis seed embryo state observation;
[0022] Figure 2 Figure 2 is a diagram of the fresh coptis seed embryo development state after 15 days of stratification; wherein A is the embryo development state after 15 days of stratification at 15°C, B is the embryo development state after 15 days of stratification at 20°C, C is the embryo development state after 15 days of stratification at 25°C, D is the embryo development state after 15 days of stratification at 30°C, and E is the embryo development state after 15 days of stratification at 35°C;
[0023] Figure 3 Figure 3 is a diagram of the fresh coptis seed embryo development state after 30 days of stratification; wherein A is the embryo development state after 30 days of stratification at 15°C, B is the embryo development state after 30 days of stratification at 20°C, C is the embryo development state after 30 days of stratification at 25°C, D is the embryo development state after 30 days of stratification at 30°C, and E is the embryo development state after 30 days of stratification at 35°C;
[0024] Figure 4 Figure 4 is a diagram of the fresh coptis seed embryo development state after 45 days of stratification; wherein A is the embryo development state after 45 days of stratification at 15°C, B is the embryo development state after 45 days of stratification at 20°C, C is the embryo development state after 45 days of stratification at 25°C, D is the embryo development state after 45 days of stratification at 30°C, and E is the embryo development state after 45 days of stratification at 35°C;
[0025] Figure 5 Figure 5 is a diagram of the fresh coptis seed embryo development state after 60 days of stratification; wherein A is the embryo development state after 60 days of stratification at 15°C, B is the embryo development state after 60 days of stratification at 20°C, C is the embryo development state after 60 days of stratification at 25°C, D is the embryo development state after 60 days of stratification at 30°C, and E is the embryo development state after 60 days of stratification at 35°C;
[0026] Figure 6 Figure 6 is a diagram of the fresh coptis seed embryo development state after 75 days of stratification; wherein A is the embryo development state after 75 days of stratification at 15°C, B is the embryo development state after 75 days of stratification at 20°C, C is the embryo development state after 75 days of stratification at 25°C, D is the embryo development state after 75 days of stratification at 30°C, and E is the embryo development state after 75 days of stratification at 35°C. DETAILED DESCRIPTION
[0027] The detailed description set forth below of various example implementations of the application describes and discloses the presently preferred embodiments of the application, and is intended to provide a description of certain aspects of the application, and is not intended to limit the scope of the application.
[0028] It is to be understood that the terminology used herein is for the purpose of describing particular embodiments of the application only and is not intended to limit the scope of the present application. In addition, where particular ranges of values are given, understand that each intervening value, to the upper or lower limit of the ranges is also specifically included. Each smaller range that falls within the broader ranges is also specifically included in the present application. The upper and lower limits of these smaller ranges can independently be included or excluded in the range.
[0029] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application, the preferred methods and materials are described. All patents, patent applications, publications, and descriptions mentioned herein are incorporated by reference to disclose and describe the methods and / or materials in connection with which the patents, patent applications, publications, and descriptions are cited.
[0030] Many modifications and variations of this application of the application can be made without departing from its spirit or scope, which will be apparent to those skilled in the art. Other embodiments of the application will be apparent to those skilled in the art from consideration of the specification and practice of the application disclosed herein. For example, the specification can be used to design other embodiments of the application. Consequently, it is intended that the specification and examples be considered as exemplary only with the true scope of the application being indicated by the appended claims.
[0031] As used herein, the terms "comprise", "comprising", "include", "including", "have", "having" and the like are open-ended and do not exclude additional elements or steps.
[0032] The method of preparing the substrate of the present application is as follows: desalt commercially available coconut husk, dry and sterilize, keep the moisture content at 60-80% (based on the weight of the dried coconut husk), and the substrate is obtained.
[0033] Example 1
[0034] 1. Materials and Methods
[0035] 1.1 Materials
[0036] The fresh seed of Plagiorhegma dubium Maxim. used in the experiment was collected from the microwave station of Dongchang District, Tonghua City, and identified as a perennial herb of the Berberidaceae family. The seeds were immediately removed from the seed clip after collection, mixed randomly, and stored in a 4°C refrigerator in the dark for later use. Unqualified seeds were removed by water selection, and the sinking seeds, which were full and round, were left for testing and named fresh seeds. The fresh seeds were naturally dried at room temperature and stored, and were named room temperature dried seeds.
[0037] Stemi 508 optical microscope (Zeiss), light incubator (Bo Xun, SPX-250B-G), electronic balance (Sartorius, BSA423S); pipette (Dalong, Dalong Xingchuang Experimental Instrument (Beijing) Co., Ltd.); electronic moisture meter (DHS-10A, Ningbo Yinzhou Huafeng Electronic Instrument Factory).
[0038] 1.2 Method
[0039] 1.2.1 Seed 100-grain mass
[0040] Randomly select 100 seeds and weigh them on an electronic balance to measure their 100-grain mass, repeating 5 times.
[0041] 1.2.2 Seed moisture content
[0042] Take an appropriate amount of fresh seed and accurately weigh it (G), place it in an electronic moisture meter, and dry it at 105°C until the weight is constant, i.e., the difference between two weights is not more than 1% of the seed weight, and record it as the dry weight (g), repeating 2 times. Within the allowable error range of 0.5% for the two test samples, the moisture content of the fresh seed is [(G-g) / G], the drying rate is (g / G), and the wet weight rate is (G / g).
[0043] 1.2.3 Seed water absorption rate
[0044] Take room temperature dried seeds (moisture content 6.02%), 100 seeds per group, weigh them on an electronic balance, and then place them in beakers containing 50 mL of distilled water at room temperature. After 6, 12, and 24 hours, respectively, remove them, absorb the surface water with filter paper, and quickly weigh them. Calculate the seed water absorption rate as (seed mass after soaking - seed mass before soaking) / seed mass before soaking x 100%.
[0045] 1.2.4 Fresh Plagiorhegma dubium seed stratification study at different temperatures
[0046] Mix the seeds with the substrate evenly, adjust the moisture to the appropriate level, put them into plastic boxes, and place them in 15℃, 20℃, 25℃, 30℃, and 35℃ light incubators, and provide oxygen by growing small Chinese cabbages in the incubators. Take the seeds at different temperatures every 15 days, cut them in half longitudinally, observe them under a stereomicroscope, take photos, and record the seed embryo development state at different temperatures.
[0047] 1.2.5 Fresh Coptis seed germination research
[0048] After the stratification, the seed coat naturally cracks, and further germination experiments are carried out. The fresh Coptis seeds are soaked in 100ppm, 200ppm, 300ppm, and 400ppm gibberellin for 48h, and the initial temperature is 40℃ sterile water as a control. After soaking, the surface adhering gibberellin is washed, and the germination experiment is carried out at 10℃, 15℃, 20℃, and 25℃ in filter paper as the germination bed.
[0049] After the seed embryo develops completely, the germination rate of the fresh Coptis seeds is counted, and the germination rate is calculated according to the following formula:
[0050] Germination rate = (number of germinated seeds / total number of test seeds) x 100%.
[0051] 2. Results and analysis
[0052] 2.1 Fresh Coptis seed initial state
[0053] The initial state of the fresh Coptis seed is shown in Figure 1 After the seed matures, the seed pod naturally cracks, exposing the seed, and the seed surface has white attachments. After being cut open, the seed embryo is observed under a stereomicroscope, and the initial state of the seed embryo is a globular embryo.
[0054] 2.2 Seed hundred-grain mass
[0055] The average hundred-grain mass is 1.376g.
[0056] 2.3 Seed moisture content
[0057] The moisture content of the fresh seed is 48.64%, the dry rate is 51.36%, and the wet weight rate is 194.69%.
[0058] 2.4 Seed water absorption rate
[0059] The fresh Coptis seed dries at room temperature and rapidly absorbs water within 6h, which is the rapid water absorption period. The water absorption rate rapidly rises from 0.00% to 108.75%, and the water absorption rate slowly rises from 108.75% to 114.77% in 6-12h, which is the slow water absorption period. After 12h, the absorption gradually tends to be balanced, and at this time, the water absorption basically reaches the saturation state, entering the water absorption saturation period.
[0060] 2.5 Different temperature seed layering of fresh Coptis chinensis
[0061] 2.5.1 Embryo development state of fresh Coptis chinensis seed layering for 15 days
[0062] The embryo development state of fresh Coptis chinensis seed layering for 15 days is shown in Table 2. The embryo developed into a heart-shaped embryo at 15°C, and into a globular embryo at the other temperatures. Figure 2
[0063] 2.5.2 Embryo development state of fresh Coptis chinensis seed layering for 30 days
[0064] The embryo development state of fresh Coptis chinensis seed layering for 30 days is shown in Table 3. The embryo developed into a late heart-shaped embryo at 15°C, and into a heart-shaped embryo at the other temperatures. Figure 3
[0065] 2.5.3 Embryo development state of fresh Coptis chinensis seed layering for 45 days
[0066] The embryo development state of fresh Coptis chinensis seed layering for 45 days is shown in Table 4. The embryo developed into a torpedo-shaped embryo at 15°C, and into a late heart-shaped embryo at the other temperatures. Figure 4
[0067] 2.5.4 Embryo development state of fresh Coptis chinensis seed layering for 60 days
[0068] The embryo development state of fresh Coptis chinensis seed layering for 60 days is shown in Table 5. The embryo developed into a cotyledon-shaped embryo at 15°C, and into a late heart-shaped embryo at the other temperatures. Figure 5
[0069] 2.5.5 Embryo development state of fresh Coptis chinensis seed layering for 75 days
[0070] The embryo development state of fresh Coptis chinensis seed layering for 75 days is shown in Table 6. The embryo developed into a cotyledon-shaped embryo at 15°C, and into a torpedo-shaped embryo at 20°C, and into a late heart-shaped embryo at the other temperatures. Figure 6
[0071] 2.6 Seed germination of fresh Coptis chinensis
[0072] The seed germination of fresh Coptis chinensis began on the third day of treatment and ended on the twelfth day of treatment. The germination results under different conditions are shown in Table 1. The results show that the germination rate of seeds treated with sterile water was 0% at each temperature, and seeds treated with gibberellin had different amounts of germination, indicating that the seeds may contain inhibitory factors causing physiological dormancy. After treatment with gibberellin, the seed dormancy was broken, and the germination trend increased with increasing gibberellin concentration. The optimal germination temperature was 15°C.
[0073] Table 1 Germination rate of fresh Coptis chinensis seeds under different conditions (%)
[0074]
[0075] The above embodiments are only to illustrate the preferred modes of the present application, and are not intended to limit the scope of the present application. Any modification and improvement made by those skilled in the art to the technical solutions of the present application without departing from the design spirit of the present application shall fall within the protection scope of the present application.
Claims
1. A method for promoting the morphological after-ripening of fresh Coptis chinensis seeds, characterized in that, This includes the step of stratifying the fresh Coptis chinensis seeds at 15-20°C.
2. The method as described in claim 1, characterized in that, The lamination temperature is 15°C, and the time is 60-90 days.
3. A method for promoting the germination of fresh Coptis chinensis seeds and / or increasing the germination rate of fresh Coptis chinensis seeds, characterized in that, Includes the following steps: The fresh Coptis chinensis seeds were laminated at 15-20℃ to obtain laminated seeds; The stratified seeds were soaked in gibberellin solution, and after soaking, they were germinated using filter paper as a germination bed.
4. The method as described in claim 3, characterized in that, The lamination temperature is 15°C, and the time is 60-90 days.
5. The method as described in claim 3, characterized in that, The gibberellin content in the gibberellin solution is 200-400 ppm.
6. The method as described in claim 5, characterized in that, The gibberellin solution contained 400 ppm of gibberellin.
7. The method as described in claim 3, characterized in that, The soaking time is 48 hours.
8. The method as described in claim 3, characterized in that, The germination environment temperature is 10-20℃.
9. The method as described in claim 8, characterized in that, The germination environment temperature is 15℃.
10. The application of germinating seeds obtained by the method according to any one of claims 3-9 in the artificial propagation of fresh Coptis chinensis.