A seed propagation method for shortening the hydrangea seedling period

Abstract

The invention discloses a seed propagation method for shortening the hydrangea seedling period, and belongs to the technical field of hydrangea propagation. The method comprises the following steps: collecting hydrangea capsules, disinfecting and sterilizing them, soaking them in an activation solution, and then placing them for germination, then peeling out the seeds for aseptic sowing, disinfecting and sterilizing the capsules and allowing the seeds to germinate in the capsules, and then peeling out the seeds for aseptic sowing. The whole process can not only effectively reduce the risk of external microorganisms invading the seeds, but also reduce the contact time between the seeds and the disinfectant, thereby preventing damage to the seeds due to excessive disinfection and causing a decrease in the germination rate, thereby improving the seed germination rate, shortening the seedling period, and improving the seedling rate. In addition, the sowing and propagation management and operation of the hydrangea according to the invention are simple, and the hydrangea has important significance for promoting the development of the hydrangea industry.

Description

Technical Field

[0001] The invention relates to the technical field of hydrangea propagation, and in particular to a seed propagation method for shortening the hydrangea seedling period. Background Art

[0002] Hydrangea macrophylla, also known as hydrangea, is a plant of the genus Hydrangea in the family Hydrangeaceae. It has large and colorful flowers, a wide variety of varieties and a long flowering period. It is an important resource for landscaping, potted flowers, and dried and fresh cut flowers. China is the resource distribution center of the genus Hydrangea. Its ornamental traits and adaptability such as flower color, flower shape, leaf color and leaf shape have rich genetic diversity and great development potential. However, breeding work started late, and currently only a small number of varieties have obtained new plant variety rights. The breeding speed of new varieties is slow and the commercialization level is low.

[0003] The sowing propagation technology can quickly obtain a large number of offspring with rich genetic variation, and promote the cultivation of new hydrangea varieties. However, there are some problems with hydrangea seed propagation at present: First, hydrangea seeds need a certain amount of light to germinate. Burying the seeds directly in the soil will result in the seeds lacking light and failing to germinate or having an extremely low germination rate. If the seeds are scattered on the soil surface, a certain moisturizing device is required, otherwise the seeds may dry out and affect germination if not carefully covered. However, covering the moisturizing device will cause improper ventilation and cause pests and diseases. Second, hydrangea seeds are very small, and the seedlings after sowing and germination are also very small. The seedlings grow slowly and have high maintenance requirements. A slight lack of water may cause the seedlings to die, resulting in a very low seedling rate after hydrangea sowing.

[0004] Therefore, there is a need to find a seed propagation method that shortens the hydrangea seedling period, shortens the seedling period of sowing propagation, improves the growth of seedlings to increase the seedling rate, and thus better promotes the development and utilization of new hydrangea varieties. Summary of the invention

[0005] In view of this, the object of the present invention is to provide a seed propagation method for shortening the hydrangea seedling period, so as to solve the problem that the seedling period is long and difficult to maintain under the conventional sowing propagation method of hydrangea.

[0006] The present invention solves the above technical problems by the following technical means:

[0007] A seed propagation method for shortening the hydrangea seedling stage, the method being as follows:

[0008] (1) Collect full hydrangea capsules from August to November, disinfect them, soak them in activation solution for 2 to 5 days, store them at 4°C for 8 to 15 days, and then dry and peel out the seeds;

[0009] (2) sterilizing the seeds to obtain sterilized seeds; inoculating the sterilized seeds into a culture medium and culturing them in a tissue culture room;

[0010] (3) After the seedlings are cultured in the tissue culture room until they reach a height of 5 cm, they can be hardened and transplanted.

[0011] Under the conventional soil sowing propagation method, hydrangea seedlings grow slowly and have high requirements for maintenance operations. The present invention adopts the aseptic sowing method in the tissue culture room to cultivate hydrangeas, which can effectively shorten the growth time of hydrangea seedlings and improve the growth of seedlings. No other maintenance management is required during the entire seedling period, and the operation is relatively simple.

[0012] Furthermore, the capsule disinfection treatment in step (1) is performed as follows:

[0013] The hydrangea capsule is washed with water and then immersed in a 75% volume fraction ethanol solution for 5 to 10 minutes, then taken out and washed with sterile water for 5 times, then immersed in a 2wt% sodium hypochlorite solution for disinfection for 15 to 20 minutes, then washed with sterile water for 5 to 7 times, then drained, and subjected to ultraviolet sterilization for 10 minutes to obtain the disinfected capsule.

[0014] Furthermore, the activation solution in step (1) comprises the following raw materials in parts by weight:

[0015] 3 to 7 parts of n-decanol, 5 to 7 parts of ammonium succinate, 2 to 3 parts of methyl acetate, 0.1 to 0.3 parts of cinnamaldehyde, and 1000 to 2000 parts of a regulator.

[0016] Furthermore, the components of the regulator are: 0.2-0.5 mg / L naphthylacetic acid + 0.1-0.2 mg / L gibberellin GA3 + 100-200 mg / L salicylic acid, and the balance is water.

[0017] When using aseptic sowing to raise hydrangea seedlings, since hydrangea seeds are tiny and have thin seed coats, if the seeds are directly sterilized, if the sterilization time is too long, the seeds are easily damaged and inactivated, but if the sterilization time is too short, the culture medium will be contaminated due to incomplete sterilization. Therefore, the present invention collects hydrangea capsules, at which time the seeds are protected by the capsule skin, and the internal pathogenic bacteria are less contaminated. Directly performing a deeper degree of sterilization on the capsule will not damage the internal seeds, and can effectively kill most of the external pathogenic bacteria. Then, after soaking in an activation solution, cold storage treatment is carried out. The regulator in the activation solution is combined with the cold storage treatment to improve the enzyme activity inside the seeds, eliminate seed germination inhibitors, and promote the decomposition and transformation of storage substances inside the seeds. At the same time, the internal seeds can further absorb nutrients from the capsule skin, thereby better promoting seed germination, shortening the germination time, improving the growth of seedlings, and shortening the seedling period.

[0018] However, the presence of the capsule skin will hinder the components in the activation solution from entering the interior to exert their effects on the seeds. Therefore, ammonium succinate is added to the activation solution. Ammonium succinate acts on the capsule skin, interacts with the skin cell membrane and binds to the cell membrane. By destroying the ordered structure of the skin cell membrane, the permeability of the capsule skin is increased. At the same time, combined with the effect of methyl acetate, the affinity of the components in the activation solution to the capsule skin is improved, so that the components in the activation solution can enter the interior more smoothly to exert their effects. In addition, the treatment with the activation solution changes the internal physiological state of the seeds. At this time, the internal seeds consume oxygen, which reduces the dissolved oxygen content in the activation solution. In addition, the physical barrier effect of the capsule skin reduces the amount of oxygen available to the seeds, which can easily cause hypoxia stress to the seeds. Therefore, n-decanol is also added to the activation solution to increase the dissolved oxygen content of the activation solution, thereby better promoting seed germination.

[0019] Further, the activation solution preparation method is as follows:

[0020] A: Add methyl acetate and cinnamaldehyde into a 60% by volume ethanol solution to dissolve to obtain a methyl acetate-cinnamaldehyde solution for standby use; add ammonium succinate into water to dissolve, then add the methyl acetate-cinnamaldehyde solution and stir to obtain a mixed solution;

[0021] B: Add the mixed solution to the regulator, adjust the pH and stir well to mix, then add n-decanol and mix well to obtain the activation solution.

[0022] Furthermore, in step B, the pH is adjusted to 5.5-6.5.

[0023] Further, the seed disinfection operation in step (2) is as follows:

[0024] The seeds were wrapped in a cotton towel, disinfected with a 75% ethanol solution for 1 min in an ultra-clean working environment, then washed 5 times with sterile water, then sterilized with a 0.5wt% sodium hypochlorite solution for 2 min, and then washed 5 times with sterile water to obtain the disinfected seeds.

[0025] When disinfecting hydrangea capsules, most of the external invading pathogens have been killed. The disinfected capsules are then directly germinated. During this period, under the protection of the capsule skin, the risk of internal seeds contacting pathogens is low. Therefore, after the internal seeds are peeled out, only a short period of simple disinfection is needed to effectively reduce the contamination rate and improve the breeding efficiency.

[0026] Furthermore, the components of the culture medium are: 1 / 2MS+0.2mg / L 6-BA.

[0027] Furthermore, the culture temperature in the tissue culture room is 22-28° C., the lighting time is 12-16 h / d, the light intensity is 2000-3000 lx, and the relative humidity is 65-75%.

[0028] Beneficial effects:

[0029] 1. The present invention collects hydrangea capsules, fully disinfects them, soaks them in an activation solution, and then performs a germination treatment. The entire process is carried out while retaining the capsule skin, which reduces the contact time of the disinfectant on the seeds, prevents seed damage, and can effectively reduce the contamination rate.

[0030] 2. The present invention performs the germination operation while retaining the capsule skin, which can not only reduce the contamination rate of the culture medium, but also the seeds can further absorb nutrients in the skin inside to promote seed germination. The germinated seeds are peeled out and cultured in the culture medium, which can effectively improve the seed germination rate, shorten the cultivation time, and improve the growth of seedlings. It has good application prospects in the field of hydrangea breeding. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 : Pictures of the growth status of seedlings in experimental group 1 after 4 months of cultivation in the experiment of the present invention;

[0032] Figure 2 : Pictures of the growth status of seedlings in the conventional soil sowing method for 4 months in the experiment of the present invention;

[0033] Figure 3 : Pictures of the growth status of seedlings in experimental group 1 after 7 months of cultivation in the experiment of the present invention;

[0034] Figure 4 : Pictures of the growth status of seedlings for 7 months under the conventional soil sowing propagation method in the experiment of the present invention. DETAILED DESCRIPTION

[0035] The present invention will be described in detail below with reference to specific embodiments and accompanying drawings:

[0036] Example 1: Preparation of Activation Solution

[0037] Weigh 5 g of n-decanol, 6 g of ammonium succinate, 2.5 g of methyl acetate, 0.2 g of cinnamaldehyde, and 1500 g of a regulator, wherein the components of the regulator per liter are: 0.3 mg / L of naphthaleneacetic acid + 0.15 mg / L of gibberellin GA3 + 150 mg / L of salicylic acid, and the balance is water.

[0038] A: Add methyl acetate and cinnamaldehyde into 27g of 60% by volume ethanol solution to dissolve to obtain methyl acetate-cinnamaldehyde solution, and set aside; add ammonium succinate into 120g of water to dissolve, then add methyl acetate-cinnamaldehyde solution and stir to obtain a mixed solution;

[0039] B: Add the mixed solution to the regulator, adjust the pH to 6, stir at 1000 r / min for 6 min, and then add n-decanol and mix well to obtain the activation solution.

[0040] Example 2: Preparation of Activation Solution II

[0041] Weigh 3 g of n-decanol, 5 g of ammonium succinate, 2 g of methyl acetate, 0.1 g of cinnamaldehyde, and 1000 g of a regulator, wherein the components of the regulator per liter are: 0.2 mg / L of naphthaleneacetic acid + 0.1 mg / L of gibberellin GA3 + 100 mg / L of salicylic acid, and the balance is water.

[0042] A: Add methyl acetate and cinnamaldehyde into 21g of 60% by volume ethanol solution to dissolve to obtain methyl acetate-cinnamaldehyde solution, and set aside; add ammonium succinate into 100g of water to dissolve, then add methyl acetate-cinnamaldehyde solution and stir to obtain a mixed solution;

[0043] B: Add the mixed solution to the regulator, adjust the pH to 5.5, stir at 800 r / min for 5 minutes, and then add n-decanol and mix well to obtain the activation solution.

[0044] Example 3: Preparation of Activation Solution III

[0045] Weigh 7 g of n-decanol, 7 g of ammonium succinate, 3 g of methyl acetate, 0.3 g of cinnamaldehyde, and 2000 g of a regulator, wherein the components of the regulator per liter are: 0.4 mg / L of naphthaleneacetic acid + 0.2 mg / L of gibberellin GA3 + 200 mg / L of salicylic acid, and the balance is water.

[0046] A: Add methyl acetate and cinnamaldehyde into 33g of 60% by volume ethanol solution to dissolve to obtain methyl acetate-cinnamaldehyde solution, and set aside; add ammonium succinate into 140g of water to dissolve, then add methyl acetate-cinnamaldehyde solution and stir to obtain a mixed solution;

[0047] B: Add the mixed solution to the regulator, adjust the pH to 6.5, stir at 900 r / min for 7 minutes, and then add n-decanol and mix well to obtain the activation solution.

[0048] Comparative Example 1: Preparation of Activation Solution

[0049] In contrast to Example 1, the only difference is that methyl acetate is not added in step A when preparing the activation solution in Comparative Example 1, as follows:

[0050] A: Add 2 g of cinnamaldehyde to 60% ethanol solution to obtain a cinnamaldehyde solution for standby use; add ammonium succinate to 120 g of water to dissolve it, then add the cinnamaldehyde solution and stir to obtain a mixed solution;

[0051] B: Same as implementation 1.

[0052] Comparative Example 2: Preparation of Activation Solution

[0053] In contrast to Example 1, the only difference is that cinnamaldehyde is not added in step A when preparing the activation solution in Comparative Example 2, as follows:

[0054] A: Add methyl acetate to 25g of 60% by volume ethanol solution to dissolve to obtain a methyl acetate solution for standby use; add ammonium succinate to 120g of water to dissolve, then add the methyl acetate solution and stir to obtain a mixed solution;

[0055] B: Same as Example 1.

[0056] Comparative Example 3: Preparation of Activation Solution

[0057] In contrast to Example 1, the only difference is that ammonium succinate is not added in step A when preparing the activation solution in Comparative Example 3, as follows:

[0058] A: Add methyl acetate and cinnamaldehyde into 27g of 60% by volume ethanol solution to dissolve to obtain methyl acetate-cinnamaldehyde solution for standby use;

[0059] B: Add methyl acetate-cinnamaldehyde solution to the regulator, adjust the pH to 6, stir at 1000 r / min for 6 minutes, and then add n-decanol and mix well to obtain an activation solution.

[0060] Comparative Example 4: Preparation of Activation Solution

[0061] In contrast to Example 1, the only difference is that n-decanol is not added in step B when preparing the activation solution in Comparative Example 4, as follows:

[0062] A: Same as Example 1;

[0063] B: Add the mixed solution into the regulator, adjust the pH to 6, and stir at 1000 r / min for 6 minutes to obtain the activation solution.

[0064] Comparative Example 5: Preparation of Activation Solution

[0065] Preparation of activation solution

[0066] In contrast to Example 1, the only difference is that no regulator is added during the preparation of the activation solution in Comparative Example 6, as follows:

[0067] A: Add methyl acetate and cinnamaldehyde into 27g of 60% by volume ethanol solution to dissolve to obtain methyl acetate-cinnamaldehyde solution, and set aside; add ammonium succinate into 120g of water to dissolve, then add methyl acetate-cinnamaldehyde solution and stir to obtain a mixed solution;

[0068] B: After adjusting the pH of the mixed solution to 6, add n-decanol and mix well to obtain an activation solution.

[0069] Comparative Example 6: Preparation of Activation Solution

[0070] In contrast to Example 1, the only difference is that in Comparative Example 6, the pH is adjusted to 9 in step B during the preparation of the activation solution.

[0071] Embodiment 4: Hydrangea seed sowing and propagation method

[0072] The culture medium composition used in this embodiment is: 1 / 2MS+0.2mg / L 6-BA.

[0073] (1) In late September, full hydrangea capsules were collected, washed with water, and then soaked in a 75% by volume ethanol solution for 8 minutes, then taken out, washed with sterile water for 5 times, and then sterilized in a 2 wt % sodium hypochlorite solution for 20 minutes, then washed with sterile water for 7 times, drained, and sterilized by ultraviolet light for 10 minutes to obtain sterilized capsules, which were then soaked in the activation solution prepared in Example 1 for 3 days, then refrigerated at 4° C. for 10 days, and then placed in a silica gel dryer to dry completely and remove the seeds;

[0074] (2) Wrapping the seeds in a cotton towel, sterilizing them with a 75% ethanol solution for 1 min in an ultra-clean working environment, then washing them with sterile water for 5 times, sterilizing them with a 0.5wt% sodium hypochlorite solution for 2 min, and then washing them with sterile water for 5 times to obtain sterilized seeds; inoculating the sterilized seeds into a culture medium, and culturing them in a tissue culture room at a culture temperature of 24° C., a light exposure time of 15 h / d, a light intensity of 2500 lx, and a relative humidity of 65%;

[0075] (3) After the seedlings are grown indoors to a height of 5 cm, they can be hardened and transplanted to a greenhouse or outdoors.

[0076] Experiment: Hydrangea seed sowing and propagation experiment

[0077] 1. The activation solution prepared in Example 1 and Comparative Examples 1 to 6 was used to treat hydrangea seeds and then sowed. The experiment was divided into 9 groups, experimental group 1, control groups 1 to 6, and additional control groups 7 to 8. The activation solution and sowing method used in each group were as follows:

[0078] Experimental group 1: treated with the activation solution prepared by the method of Example 1 and aseptically sown and propagated according to the method of Example 4;

[0079] Control groups 1 to 6: treated with the activation solutions prepared by the methods of Comparative Examples 1 to 6 respectively and aseptically sowed and propagated according to the method of Example 4;

[0080] Control group 7: Hydrangea seeds were directly deeply disinfected and then subsequently treated as follows:

[0081] (1) Soaking hydrangea seeds in a 75% ethanol solution for 8 minutes, then taking them out, washing them with sterile water for 5 times, and then sterilizing them in a 2wt% sodium hypochlorite solution for 20 minutes, then washing them with sterile water for 7 times, draining the water, and sterilizing them with ultraviolet light for 10 minutes, then soaking them in the activation solution prepared according to the method of Example 1 for 3 days, refrigerating them at 4°C for 10 days, and then drying them completely in a silica gel dryer;

[0082] (2) Carry out according to the method of Example 4;

[0083] (3) Proceed according to the method of Example 4.

[0084] Control group 8: Soak the hydrangea seeds directly in cold storage, disinfect them slightly, and then sow them. The details are as follows:

[0085] (1) directly placing hydrangea seeds in the activation solution prepared according to Example 1 and soaking for 3 days, then refrigerating at 4° C. for 10 days and then placing in a silica gel dryer to dry completely;

[0086] (2) Carry out according to the method of Example 4;

[0087] (3) Proceed according to the method of Example 4.

[0088] 2. Result detection: The sowing amount of seeds in each group was 60 seeds. After 30 days of culture in the culture medium, the germination rate and contamination rate of each group were counted. The experiment was repeated three times, and the data obtained are shown in Table 1:

[0089] Table 1

[0090]

[0091]

[0092] According to the data analysis in Table 1:

[0093] (1) The hydrangea seeds of experimental group 1 of the present invention had a high germination rate and a low contamination rate, indicating that the sowing and propagation of hydrangea according to the method of the present invention can effectively improve the germination rate of hydrangea, and the hydrangea can be cultivated indoors in a culture medium without the need for additional management and maintenance of the seedlings, which has a good application prospect;

[0094] (2) Control groups 1 to 6 had a lack of raw materials for the preparation of the activation solution or a change in the conditions, which resulted in insufficient permeability of the activation solution, reduced utilization of the components of the activation solution by the seeds, or lack of oxygen, inappropriate pH values, and lack of regulators, which reduced the germination effect on the seeds and caused varying degrees of reduction in the germination rate.

[0095] (3) In control group 7, the seeds were directly deeply disinfected, which caused damage to the seeds. Although the contamination rate was low, the germination rate was significantly reduced. In control group 8, the seeds were directly lightly disinfected, the contact rate between the disinfectant and the seeds was reduced, the damage was small, the germination rate increased, but the contamination rate also increased significantly. This shows that the method of the present invention can better reduce the contamination rate without causing serious damage to the seeds, and better improve the germination rate.

[0096] Experiment 2: Seedling growth detection experiment

[0097] The aseptically sown seedlings of Experimental Group 1 in Experiment 1 were cultured for 4 months and 7 months, and the seedling height and leaf number were observed; at the same time, sterilized seeds were prepared according to the method of Example 4, and then sown in an ordinary peat soil matrix according to a conventional sowing method, and managed and maintained according to a conventional method. After culturing under natural conditions in a greenhouse for 4 months and 7 months, the seedling height and leaf number were observed, and the data obtained were shown in Table 2:

[0098] Table 2

[0099]

[0100]

[0101] According to the data analysis in Table 2:

[0102] The seedlings in the experimental group 1 were cultured for 4 months and 7 months, and the plant height and the number of leaves of the seedlings were observed. It was found that the number of leaves of the seedlings in the experimental group 1 could reach 5 to 7 pairs and the plant height could reach 2 to 3 cm after 4 months; the number of leaves could reach 7 to 8 pairs and the plant height could reach 4 to 5 cm after 7 months; while the number of leaves of the seedlings propagated by conventional soil sowing was only 2 pairs and the plant height was only 0.4 to 0.7 cm after 4 months; the number of leaves was only 3 to 4 pairs and the plant height was only 1 to 2 cm after 7 months. This indicates that the method of the present invention can effectively shorten the seedling period of hydrangea by aseptic sowing and propagation of hydrangea seeds, accelerate the growth of hydrangea seedlings, and more efficiently cultivate healthy hydrangea seedlings, which is of great significance for promoting the further development of the hydrangea industry.

[0103] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the present invention. Although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the purpose and scope of the technical solutions of the present invention, which should be included in the scope of the claims of the present invention. The techniques, shapes, and structural parts not described in detail in the present invention are all known technologies.

Claims

1. A seed propagation method for shortening the hydrangea seedling period, characterized in that: The breeding method is as follows: (1) Collect hydrangea capsules, disinfect them, soak them in activation solution, and then refrigerate them to dry and remove the seeds; (2) sterilizing seeds to obtain sterilized seeds; Inoculate the sterilized seeds into the culture medium and culture them in the tissue culture room; (3) After the seedlings are cultured in the tissue culture room until they are 5 cm tall, they can be hardened and transplanted; The activation solution in step (1) comprises the following raw materials in parts by weight: 3-7 parts of n-decanol, 5-7 parts of ammonium succinate, 2-3 parts of methyl acetate, 0.1-0.3 parts of cinnamaldehyde, 1000-2000 parts of a regulator; The components of the regulator are: 0.2-0.5 mg / L naphthylacetic acid + 0.1-0.2 mg / L gibberellin GA3 + 100-200 mg / L salicylic acid, and the balance is water; The activation solution preparation method is as follows: A: Add methyl acetate and cinnamaldehyde into a 60% by volume ethanol solution to dissolve to obtain a methyl acetate-cinnamaldehyde solution for standby use; add ammonium succinate into water to dissolve, then add the methyl acetate-cinnamaldehyde solution and stir to obtain a mixed solution; B: Add the mixed solution to the regulator, adjust the pH and stir well to mix, then add n-decanol and mix well to obtain the activation solution; In the step B, the pH is adjusted to 5.5-6.

5.

2. A seed propagation method for shortening the hydrangea seedling stage according to claim 1, characterized in that: The capsule disinfection treatment in step (1) is performed as follows: The hydrangea capsules were washed with water and immersed in a 75% by volume ethanol solution for 5 to 10 minutes, then taken out, washed with sterile water for 5 times, and disinfected in a 2wt% sodium hypochlorite solution for 15 to 20 minutes, then washed with sterile water for 5 to 7 times, drained, and sterilized with ultraviolet light for 10 minutes to obtain the disinfected capsules.

3. A seed propagation method for shortening the hydrangea seedling period according to claim 1, characterized in that: The seed disinfection operation in step (2) is as follows: The seeds were wrapped in a cotton towel, disinfected with a 75% volume fraction ethanol solution for 1 min, and then washed 5 times with sterile water. Then, they were sterilized with a 0.5wt% sodium hypochlorite solution for 2 min and then washed 5 times with sterile water to obtain the disinfected seeds.

4. A seed propagation method for shortening the hydrangea seedling period according to claim 3, characterized in that: The components of the culture medium in step (2) are: 1 / 2MS+0.2mg / L 6-BA.

5. A seed propagation method for shortening the hydrangea seedling period according to claim 4, characterized in that: The culture temperature in the tissue culture room is 22-28° C., the light duration is 12-16 h / d, the light intensity is 2000-3000 lx, and the relative humidity is 65-75%.

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