Method for shortening breeding period of clematis orientalis seeds and improving germination rate

Abstract

The invention discloses a method for seed production and seedling breeding of clematis orientalis. The method comprises five key steps of seed shell pretreatment, seed treatment, soil transferring and planting, plant transplanting and seedling culture management. According to the method, seed shells are creatively polished by abrasive paper until the seed shells are thinned but seed kernels are not damaged, and the seed germination time is shortened to 14.08 days and the germination rate is increased to 78.33% in combination with a synergistic treatment technology of soaking in a gibberellin solution. By designing a stepped seedling culture container system and matching with a peat-perlite-vermiculite matrix ratio, accurate matching between root development and container space is realized. By adopting a temperature and humidity intelligent regulation and control and staged water and fertilizer management scheme and combining with precise application of a water-soluble fertilizer and an aoGreen A5 slow-release fertilizer, the survival rate and growth vigor of seedlings are remarkably improved. The method breaks through the traditional breeding bottleneck, shortens the breeding period, improves the seed germination rate, realizes factory-like efficient production of clematis seedlings, and has remarkable economic benefits and ecological values.

Description

Technical Field

[0001] The present invention relates to the technical field of plant sowing and propagation, and in particular to oriental clematis ( Clematis intricata ) seed production and seedling factory breeding methods, especially the technical solutions for improving seed germination rate and shortening breeding cycle through physical breaking of seed shell, hormone synergistic treatment, stepped container system and precise water and fertilizer management. Background Art

[0002] Oriental Clematis ( Clematis intricata ) is a perennial herbaceous vine of the Ranunculaceae and Clematis genus. The fibrous roots are reddish brown and dense, the stems are cylindrical, and the surface is brown-black or dark red. It is an oriental resource plant widely distributed in Northeast my country, North China, Northwest China and other places. It generally grows in mountains, low wetlands, sandy land and field edges. The stems are climbing and branched; the leaves are opposite and twice pinnate, gray-green; the cymes are axillary, usually with 2-3 flowers, 4 yellow calyxes, and many stamens; the top of the achenes has a persistent feather-like style, up to 5cm long, and most achenes are gathered in a cotton ball shape; the flowering period is July-August, and the fruiting period is August-September. Oriental Clematis is used as an important medicinal plant in traditional Chinese medicine, Tibetan medicine, and Mongolian medicine. The whole plant is used as medicine to dispel rheumatism. It is mainly used for external treatment of rheumatic arthritis. People often mash the whole plant and add alum to the affected area to treat psoriasis. In addition, as an ornamental plant, Oriental Clematis is used for garden beautification and hedges, and has a high appreciation value. In short, Oriental Clematis is an important resource plant, and its development and utilization prospects are relatively broad.

[0003] At present, there are three main methods of reproduction: seed propagation, cutting propagation and layering propagation. Cutting propagation and layering propagation are limited by the size and maturity of Clematis plants, and the breeding amount is small. Clematis seeds are easy to pick and have a large yield. Therefore, seed propagation is the key to large-scale reproduction of Clematis.

[0004] However, the seed shell of oriental clematis is relatively thick, and it is difficult to germinate naturally. It needs to go through a low-temperature vernalization stage to germinate, and the germination time is long. It is impossible to stably propagate on a large scale, and it takes a long time. In the prior art, CN109287427B discloses a cutting propagation method for clematis, but the gibberellin concentration used is relatively high. At the same time, there is no comparative data on the seed germination time and germination rate under the coordinated treatment of gibberellin concentration and seed shell machinery, and there is no exploration of seedling containers, transplanting methods, sowing substrates and types of slow-release fertilizers.

[0005] Therefore, how to provide a method that can effectively improve the germination rate of oriental clematis seeds and shorten the breeding time of clematis seedlings is a problem that technical personnel in this field need to solve. There is an urgent need for a comprehensive breeding method that integrates physical shell breaking, hormone regulation, container adaptation and precise management. Summary of the invention

[0006] In view of the deficiencies of the above-mentioned prior art, the present invention aims to overcome the deficiency that Oriental Clematis cannot be stably propagated on a large scale in a short period of time, and to provide a method for improving the seed germination rate of Oriental Clematis and shortening the seedling breeding cycle of Oriental Clematis.

[0007] In order to achieve the above object, the present invention adopts the following technical solution: In one embodiment, the present invention provides a method for producing seeds and seedlings of Clematis orientalis, comprising the following steps: Step 1: Seed shell pretreatment: Select mature and plump Clematis seeds, and use sandpaper to grind the seed shell until it becomes thinner but does not damage the seed kernel; Step 2, seed treatment: soak the treated seeds in a solution of 10-80 mg / L gibberellin GA3 for 1 hour; Step 3: Planting in soil: sow the seeds in a transparent small seedling pot containing a sowing matrix, wherein the sowing matrix is ​​peat, perlite and vermiculite, with a sowing depth of 1-1.5 cm, and place in a greenhouse at 25±1°C and a humidity of 80-100%; Step 4, plant transplantation: When the root system is coiled around the substrate, move it into a medium-sized seedling pot. Water it when the moisture content of the substrate is lower than 20%. During this period, apply water-soluble fertilizer containing nitrogen, phosphorus and potassium. Specifically, apply 2500-3500 times of water-soluble fertilizer with a nitrogen, phosphorus and potassium ratio of 15-25:15-25:15-25, preferably apply 3000 times of water-soluble fertilizer with a nitrogen, phosphorus and potassium ratio of 20:20:20, and spray it once every 2 weeks; Step 5, seedling management: After the roots penetrate the medium-sized pot, transplant to the large seedling pot, set up a climbing support frame, and apply 5-55 grains / pot of Aolv A5 slow-release fertilizer, preferably 50 grains / pot.

[0008] In another embodiment, the present invention also includes that in step three, the sowing matrix is ​​pre-drenched and disinfected with 800 times diluted 50% carbendazim wettable powder, and the sowing matrix is ​​peat: perlite: vermiculite = 8:1:1.

[0009] In another embodiment, the present invention also includes that the concentration of the gibberellin GA3 solution is 20 mg / L.

[0010] In another embodiment, the present invention also includes that the seedling pot system includes: small seedling pot: top 7×7 cm, bottom 5.5×5.5 cm, height 10 cm; medium seedling pot: top 17×17 cm, bottom 13×13 cm, height 15.5 cm; large seedling pot: top 35×35 cm, bottom 27×27 cm, height 32 cm.

[0011] In another embodiment, the present invention also includes the step of spraying the water-soluble fertilizer in an amount sufficient to evenly wet each leaf until no droplets fall.

[0012] In another embodiment, the present invention further comprises that the pH value of the matrix is ​​5.5-5.6.

[0013] In another embodiment, the present invention further comprises applying the Aolv A5 slow-release fertilizer in a circular shape around the roots of the plants in step 5, 3-5 cm away from the root base.

[0014] In one embodiment, the present invention provides a method for shortening the germination period of oriental clematis seeds, comprising grinding the seed shell with sandpaper until it becomes thin but without damaging the seed kernel; soaking the polished seeds in a 20 mg / L gibberellin GA3 solution for 1 hour; and culturing in an environment of 25±1°C and humidity ≥80% after sowing.

[0015] In one embodiment, the present invention provides a composite regulation method for improving the survival rate of oriental clematis seedlings, comprising grinding the seed shell with sandpaper until it becomes thin but without damaging the seed kernel and co-treating with 10-80 mg / L gibberellin; step-by-step container transplantation based on the dynamics of root development; and phased application of water-soluble fertilizer with a nitrogen, phosphorus and potassium ratio of 20-20-20 and Aolv A5 slow-release fertilizer.

[0016] In one embodiment, the present invention provides a stepped seedling container system for Oriental Clematis suitable for the method described in the present invention, including: a container for the primary seedling stage, a container for the strong seedling stage and a container for the mature seedling stage, each container being filled with peat, perlite and vermiculite as a matrix.

[0017] Beneficial effects achieved by the present invention: It can be known from the technical solution of the present invention that compared with the prior art, the present invention discloses a method for shortening the breeding cycle of Oriental Clematis and improving the germination rate, combining the five key steps of seed shell pretreatment, seed treatment, soil planting, plant transplanting, and seedling management, innovatively using sandpaper to grind the seed shell until it becomes thinner but does not damage the seed kernel, combined with a collaborative treatment technology of soaking in a lower concentration of gibberellin solution, the seed germination time is shortened to 14.08 days, and the germination rate is increased to 78.33%. The root vitality is also significantly improved by designing a stepped seedling container system with a matrix ratio of peat-perlite-vermiculite, achieving a precise match between root development and container space, and the transplant survival rate is over 95%. The intelligent control of temperature and humidity and the staged water and fertilizer management scheme are adopted, combined with the precise application of water-soluble fertilizer and Aolv A5 slow-release fertilizer, which significantly improves the survival rate and growth potential of seedlings. It breaks through the bottleneck of traditional breeding, shortens the breeding cycle, improves the seed germination rate, and realizes the efficient factory production of Clematis seedlings, which has significant economic benefits and ecological value. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 : The stepped seedling container system of Oriental Clematis includes small, medium and large seedling pots.

[0019] In the figure: 1. Hydrophobic air holes at the bottom. DETAILED DESCRIPTION

[0020] The present invention is further described below in conjunction with specific embodiments, but the present invention is not limited by the embodiments.

[0021] Unless otherwise specified, the experimental materials, reagents, instruments and methods used in the following examples are conventional experimental materials, reagents, instruments and methods in the art and can be purchased through commercial channels.

[0022] Example 1 Clematis orientalis seed production and seedling breeding The embodiment of the invention discloses a method for improving the germination rate of oriental clematis seeds and shortening the breeding time of clematis seedlings. The breeding steps are simplified and quick, and the requirements of factory-based and batch breeding of clematis seedlings are met.

[0023] The experiment was carried out on November 27, 2023. With clean water as the control, the seeds were treated with different concentrations of gibberellin GA3 solution for 1 hour. Each treatment condition was set up with 3 replicates, and each group was tested independently with 40 seeds. The seed germination status was monitored at 10:00 every day according to the standard of obvious visible radicle. The experiment was carried out again on January 5, 2024. Healthy and dry Clematis seeds with mature, plump and uniform size were selected, and their seed coats were treated. Each treatment group had 3 biological replicates, and each replicate contained 40 seeds. Specifically, the seed coat was sanded with sandpaper until it became thin but did not damage the seeds. Then the seeds treated with seed coat were soaked in 20mg / L gibberellin GA3 solution for 1 hour. The seed germination status was also monitored at 10:00 every day according to the standard of obvious visible radicle, and the time node when the radicle first broke through the seed coat was recorded.

[0024] Specifically, the method for producing seeds and seedlings of oriental clematis comprises the following steps: Step 1: Seed shell pretreatment: Select healthy and dry clematis seeds that are mature, plump, uniform in size, and use sandpaper to slightly grind the clematis seed shell to make it thinner, but do not damage the seed coat; Step 2, seed treatment: soak the seeds treated with seed husk in step 1 in a solution of 10-80 mg / L gibberellin GA3 for 1 hour; Step 3, transfer to soil and plant: plant the seeds treated in step 2 into a small transparent seedling pot, which is square, with a top opening of 7cm, a bottom of 5.5cm, and a height of 10cm. The bottom is provided with a hydrophobic air hole, and the pot is filled with a sowing matrix, and the sowing depth is 1-1.5cm. The seedling matrix in the seedling pot is a uniform mixture of peat, perlite and vermiculite, and the mixing ratio is 8:1:1. The sowing matrix needs to be irrigated with water during sowing. After sowing, the seedling pot is placed in a greenhouse. The temperature of the greenhouse is maintained between 25±1℃ and the humidity is maintained between 80-100%. The amount of watering is determined every day according to the dryness and wetness of the substrate surface.

[0025] Step 4, plant transplantation: When the roots of the plants planted in step 3 can wrap around most of the substrate, move the plants into a medium-sized seedling pot with a square top opening side length of 17 cm, a square bottom side length of 13 cm, and a height of 15.5 cm, and cultivate them in a greenhouse. Water when the moisture content of the substrate is less than 20%. Fertilize the plants during the growth period. During the cultivation period in the medium-sized seedling pot, apply 3000 times of water-soluble compound fertilizer. The frequency of fertilization is once every two weeks. Use a spray bottle to spray on the surface of the leaves. The nitrogen, phosphorus and potassium ratio of the water-soluble compound fertilizer is 20-20-20.

[0026] Step 5, seedling management: After the roots of the Oriental Clematis plants cultivated in step 4 grow out of the water-repellent and air-permeable holes in the medium-sized seedling pot, transplant the Clematis plants in the medium-sized seedling pot into a large-sized seedling pot with a square top opening side of 35 cm, a square bottom side of 27 cm, and a height of 32 cm for cultivation. Set up a climbing support frame, and wrap the sprouting vines of the Clematis plants around the climbing support frame. During the cultivation period, apply an appropriate amount of Aolv A5 special slow-release fertilizer for promoting seedlings and roots. When fertilizing the clematis in the large seedling pot, apply 50 slow-release fertilizers per pot and place them around the roots to avoid placing them directly on the root base.

[0027] The size of the seedling pot is designed to suit the root development of the seedlings. Figure 1As shown, the first is a small transparent seedling pot (7cm×7cm on the top, 5.5cm×5.5cm on the bottom, 10cm high). In the early stage of rooting of oriental clematis seedlings (0-30 days), the main root length is usually 8-10cm, and the lateral root expansion radius is about 3-5cm. The height of the small transparent pot (10cm) matches the vertical growth space of the root system, and the bottom side length (5.5cm) provides a moderate lateral expansion space for the lateral roots to avoid root crowding and hypoxia. The small pot body (volume of about 350mL) can accurately control the water content of the substrate, and with the bottom hydrophobic holes, the substrate is kept moist but not waterlogged, reducing the risk of root rot. The second is a medium-sized seedling pot (17cm×17cm on the top, 13cm×13cm on the bottom, 15.5cm high), which supports explosive root growth: when transplanting (the root system fills the small transparent pot), the root volume increases 5-8 times compared to the initial volume. The volume of the medium-sized pot (about 3,500mL) provides 10 times the space for the roots than the small transparent pot. The height is increased by 50% (15.5cm) to accommodate roots 20-25cm long, and the bottom side length (13cm) meets the need for the lateral roots to expand horizontally to 10cm. And with the bottom drainage holes, it significantly promotes aerobic respiration. The third is the large seedling pot (top 35cm×35cm, bottom 27cm×27cm, height 32cm), which can meet the needs of adult roots. The taproot depth of the mature oriental clematis can reach 30cm, and the lateral root expansion radius is 15-20cm. The volume of the large pot (about 28,000mL) provides sufficient space, the height (32cm) is 100% higher than the medium pot, and the bottom side length (27cm) ensures that the lateral roots extend unimpeded, avoiding root distortion caused by space limitations.

[0028] Example 2 Effect of mechanical treatment of seed shells and GA on the germination process and germination rate of Clematis orientalis seeds The operation steps of oriental Clematis seed production and seedling propagation are basically the same as those in Example 1, except that: the seeds in step 1 were soaked in a solution of 10-80 mg / L gibberellin GA3 for 1 h, and the seed germination time and germination rate were compared (Table 1); then the effects of seed shell mechanical treatment and gibberellin GA3 on the germination process and germination rate of oriental Clematis seeds were investigated (Table 2).

[0029] Table 1 Comparison of GA3 concentration on germination time and germination rate of Clematis orientalis seeds (mean ± SD)

[0030] Note: Data in the same column without the same lowercase letter indicate that the two are statistically significant ( P < 0.05).

[0031] Table 2 Effects of seed husk mechanical treatment and GA3 on the germination process and germination rate of Clematis orientalis seeds (mean ± SD)

[0032] Note: Unpolished seeds + clean water (a); polished seeds + clean water (b); unpolished seeds + gibberellin GA3 (c); polished seeds + gibberellin GA3 (d); data in the same column without the same lowercase letter indicates that the two are statistically significant ( P <0.05).

[0033] The above two different implementation methods were used to produce seeds, sow, propagate and cultivate Clematis, and the results were statistically compared. The results in Table 1 show that 20 mg / L GA3 is the best concentration to break the dormancy of Clematis orientalis seeds. This treatment significantly shortened the seed germination time to 17.90 d and increased the germination rate to 72.50%.

[0034] The results in Table 2 show that the treated Clematis seeds were obtained by slightly grinding the seed shell of Oriental Clematis with sandpaper. The germination time of the treated Clematis seeds was significantly shortened and the germination rate was significantly improved. After the Oriental Clematis seeds with thinned seed shells were soaked in a solution of 20 mg / L gibberellin GA3 for 1 hour, the germination time of the Oriental Clematis seeds was shortened to 14.08 days, and the germination rate could reach 78.33%. These treatment methods shorten the breeding cycle and improve the seed germination rate, providing scientific basis and technical support for the large-scale propagation of Oriental Clematis, and contributing to the factory production and promotion of Clematis plants.

[0035] Although the present invention has been disclosed as above in the form of preferred embodiments, it is not intended to limit the present invention. Anyone familiar with this technology can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be based on the definition of the claims.

Claims

1. A method for producing seeds and seedlings of Clematis orientalis, characterized in that: The steps include: Step 1: Seed shell pretreatment: Select mature and plump Clematis seeds, and use sandpaper to grind the seed shell until it becomes thinner but does not damage the seed kernel; Step 2, seed treatment: soak the treated seeds in a solution of 10-80 mg / L gibberellin GA3 for 0.5h-2h, preferably 1h; Step 3: Planting in soil: sow the seeds in a transparent small seedling pot containing a sowing matrix, wherein the sowing matrix is ​​peat, perlite and vermiculite, with a sowing depth of 1-1.5 cm, and place in a greenhouse at 25±1°C and a humidity of 80-100%; Step 4, plant transplanting: When the root system is coiled around the substrate, move it into a medium-sized seedling pot. Water it when the moisture content of the substrate is lower than 20%. During this period, apply water-soluble fertilizer containing nitrogen, phosphorus and potassium. Specifically, apply 2500-3500 times of water-soluble fertilizer with a nitrogen, phosphorus and potassium ratio of 15-25:15-25:15-25, preferably apply 3000 times of water-soluble fertilizer with a nitrogen, phosphorus and potassium ratio of 20:20:20, and spray it once every 2 weeks; Step 5: Seedling management: After the roots penetrate the medium-sized pot, transplant them to the large-sized seedling pot, set up a climbing support frame, and apply 45-55 grains / pot of Aolv A5 slow-release fertilizer, preferably 50 grains / pot.

2. The method for producing seeds and seedlings of Clematis orientalis according to claim 1, characterized in that: In the step 3, the sowing matrix is ​​pre-drenched and disinfected with 800 times diluted 50% carbendazim wettable powder, and the sowing matrix is ​​peat: perlite: vermiculite = 8:1:

1.

3. The method for producing seeds and seedlings of Clematis orientalis according to claim 1 or 2, characterized in that: The concentration of the gibberellin GA3 solution is 15 mg / L-30 mg / L, preferably 20 mg / L.

4. The method for producing seeds and seedlings of Clematis orientalis according to any one of claims 1 to 3, characterized in that: The seedling pot system includes: a small seedling pot: 7×7 cm on the top, 5.5×5.5 cm on the bottom, and 10 cm high; a medium seedling pot: 17×17 cm on the top, 13×13 cm on the bottom, and 15.5 cm high; and a large seedling pot: 35×35 cm on the top, 27×27 cm on the bottom, and 32 cm high.

5. The method for producing seeds and seedlings of Clematis orientalis according to any one of claims 1 to 4, characterized in that: In step 4, the amount of water-soluble fertilizer sprayed is such that each leaf is evenly moistened until no droplets fall.

6. The method for producing seeds and seedlings of Clematis orientalis according to any one of claims 1 to 5, characterized in that: The pH value of the substrate is 5.5-5.

6.

7. The method for producing seeds and seedlings of Clematis orientalis according to any one of claims 1 to 6, characterized in that: In step 5, Aolv A5 slow-release fertilizer is applied in a circular shape around the roots of the plant, 3-5 cm away from the root base.

8. A method for shortening the germination period of Clematis orientalis seeds, characterized in that: include: Use sandpaper to grind the seed shell until it becomes thin but without damaging the seed kernel; soak the polished seeds in 20 mg / L gibberellin GA3 solution for 1 hour; after sowing, culture in an environment of 25±1℃ and humidity ≥80%.

9. A composite control method for improving the survival rate of Clematis orientalis seedlings, characterized in that: include: The seed shell was sanded with sandpaper until it became thin but without damaging the seed kernel, and then treated with 10-80 mg / L gibberellin; Stepped container transplantation based on the dynamics of root development; phased application of water-soluble fertilizer with a nitrogen, phosphorus and potassium ratio of 20:20:20 and Aolv A5 slow-release fertilizer.

10. A stepped seedling container system for Clematis orientalis suitable for the method of claims 1-9, characterized in that: include: The containers for the primary seedling stage, the strong seedling stage and the mature seedling stage are each filled with peat, perlite and vermiculite as a matrix.

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