Method for promoting rapid germination of grape seeds

Abstract

The invention relates to the technical field of grape seed germination methods, and discloses a method for promoting rapid germination of grape seeds, which comprises the following steps: collecting ripe grape fruits, stripping the seeds, washing with clear water for 3-5 times, drying the seeds in the shade, sub-packaging the seeds in centrifugal tubes, preserving at 4 DEG C for later use, placing the seeds in 75% alcohol, oscillating for 5 minutes, washing with sterile water for 3-5 times, and taking out the seeds. Transferring into a 1% sodium hypochlorite solution, oscillating for 30 minutes at the speed of 150r / min, cleaning with sterile water for 3-5 times, cutting off beaks and cotyledon ends of the seeds by using disinfection scissors, simultaneously cutting off the beaks and the cotyledon ends, not performing treatment, immersing the treated seeds into a 150mg / L gibberellin solution, oscillating for 8 hours at the room temperature at the speed of 150r / min, cleaning with sterile water for 3-5 times, absorbing moisture with filter paper, laying the filter paper and sterile gauze in a culture dish, and transplanting the seeds into the culture dish. Adding sterile water until the seeds are wet and free of accumulated water, setting three repetitions for each treatment group, uniformly placing 50 seeds for each repetition, culturing at 25 + / -2 DEG C, alternating illumination / darkness for 12 hours, supplementing water every 3 days, and recording the germination number and the root length.

Description

Technical Field

[0001] The invention relates to the technical field of grape seed germination, in particular to a method for promoting rapid germination of grape seeds. Background Art

[0002] Grapes (Vitis vinifera L.) are vine-growing fruit trees with rich nutritional value and high economic value. Cultivating new grape varieties and improving grape traits are key areas of grape breeding, with hybridization currently the primary approach. Therefore, breaking grape seed dormancy and promoting rapid germination of hybrid grape seeds can accelerate the grape breeding process.

[0003] Seeds are a crucial resource in fruit tree breeding and a crucial vehicle for selecting new varieties. Seed germination is an extremely complex process, influenced by multiple factors, including plant hormones, temperature, and the seed coat. Currently, the main methods for promoting grape seed germination include stratification and hormone treatment, such as cold stratification, normal temperature stratification, and variable temperature stratification. Plant hormone treatments, primarily gibberellins, are used. Stratification requires a long time, which can impact the breeding process. To overcome the inhibitory effect of the seed coat on seed germination, some studies have used concentrated sulfuric acid to treat the seed coat to promote seed germination. However, sulfuric acid treatment is risky and difficult to control for the duration of the treatment. Furthermore, the unique beak and seed coat structure of grape seeds often present obstacles to germination. Therefore, this study examined grape seed germination after various physical treatments to promote germination. The group treated with pruning the cotyledonary tip significantly shortened seed germination time and significantly increased the germination rate. The method of the present invention is a simple and effective method for promoting grape seed germination. Summary of the Invention

[0004] (1) Technical problems solved

[0005] In response to the shortcomings of the existing technology, the present invention provides a method for promoting rapid germination of grape seeds, which has the advantages of simple operation, high safety, and significantly improved germination efficiency. It solves the following technical problems: 1. Traditional stratification processes (such as cold stratification and variable temperature stratification) are too time-consuming (usually taking several months), seriously delaying the breeding process;

[0006] 2. Existing chemical methods (such as concentrated sulfuric acid treatment of seed coats) are highly dangerous, difficult to operate, and difficult to control;

[0007] 3. The problems of low germination rate (e.g. 10.3% in the control group) and long germination period (more than 14 days) under conventional physical or hormone treatments;

[0008] 4. The problem of grape seed dormancy being difficult to break due to the structure of the seed coat or endosperm.

[0009] (2) Technical solution

[0010] The present invention provides the following technical solution: a method for promoting rapid germination of grape seeds, comprising the following steps:

[0011] S1. Seed collection: Collect mature grape berries of 'Golden Field Blue Sapphire', peel the seeds, wash them 3-5 times with clean water, dry them in the shade, and then divide them into centrifuge tubes. Store them at 4°C until use (storage time ≤ 6 months).

[0012] S2. Seed disinfection: Place the seeds in 75% alcohol and shake for 5 minutes, rinse with sterile water 3-5 times, transfer to 1% sodium hypochlorite solution, shake at 150r / min for 30 minutes, and rinse with sterile water 3-5 times.

[0013] S3. Seed physical treatment:

[0014] Use sterilized scissors to cut off the beak of the seed (the protruding part of the radicle end);

[0015] Cut off the cotyledon ends;

[0016] Cut off the beak and cotyledon ends at the same time;

[0017] No treatment (control group).

[0018] S4. Gibberellic acid treatment:

[0019] Immerse the treated seeds in a 150mg / L gibberellin solution and shake at 150 rpm for 8 hours at room temperature.

[0020] Rinse with sterile water 3-5 times and dry with filter paper.

[0021] S5. Germination culture:

[0022] Place filter paper and sterile gauze in a culture dish and add sterile water until moistened without water accumulation.

[0023] Each treatment group was replicated three times, with 50 seeds per replicate, evenly spaced.

[0024] Incubate at 25±2℃ with 12-h light / dark cycles. Rehydrate every 3 days and record the number of sprouts, root length, and fresh weight.

[0025] Furthermore, the seed storage time in step S1 is 1-6 months.

[0026] Furthermore, the concentration of the sodium hypochlorite solution in step S2 is 1%, and the sodium hypochlorite treatment time in step S2 is 30 minutes.

[0027] Furthermore, the shaking speed in step S4 is 150 r / min, and the gibberellin treatment time in step S4 is 8 hours.

[0028] Furthermore, the culture temperature in step S5 is 25±2° C., the light cycle in step S5 is 12 hours of light / 12 hours of darkness, and sterile water is added once every 3 days in step S5.

[0029] Furthermore, the method further comprises the step of regularly measuring the root length of the seeds during the culture process, wherein the root length is measured 18 days after the seeds germinate.

[0030] Furthermore, the method can make grape seeds begin to germinate within 6-8 days, and the germination rate reaches more than 81.3% after 18 days. The method can make the average root length of the germinated seeds reach 2.2 cm.

[0031] Furthermore, the method can serve as a reference for seed germination of other grape plants.

[0032] (3) Beneficial effects

[0033] Compared with the prior art, the present invention provides a method for promoting rapid germination of grape seeds, which has the following beneficial effects:

[0034] 1. This method for promoting rapid germination of grape seeds, through the synergistic effect of cutting off the endosperm end (T2 treatment) and gibberellin, can enable seeds to germinate within 6 days, which is 57% shorter than the control group (14 days). The germination rate reaches 81.3% after 18 days, which is more than 8 times higher than the control group (10.3%). Physical treatment (cutting the beak or cotyledon end) directly breaks the mechanical barrier of the seed coat and accelerates the penetration of water and hormones, which is significantly better than single gibberellin or stratification treatment.

[0035] 2. This method for promoting rapid germination of grape seeds completely avoids chemical risks by replacing concentrated sulfuric acid treatment with physical shearing. It does not require special protection during operation and is suitable for large-scale application. The method can be extended to other grape plants such as Vitis amurensis and Vitis pubescens. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 This is a schematic diagram of different physical treatment methods for seeds of the present invention;

[0037] Figure 2 This is a statistical diagram of seed germination rates under different treatments of the present invention;

[0038] Figure 3 This is a comparison diagram of the root length of seeds treated differently according to the present invention. DETAILED DESCRIPTION

[0039] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0040] Example 1: Cutting off the cotyledonary tips of seeds (T2 treatment) to promote germination.

[0041] Seed collection and preservation

[0042] Mature 'Jintian Sapphire' grape berries were collected, the seeds were removed, washed 5 times with clean water, dried in the shade, and then divided into 10 mL centrifuge tubes and stored at 4°C for 5 months.

[0043] Seed disinfection

[0044] The seeds were placed in 75% alcohol and shaken for 5 minutes, and then washed 5 times with sterile water; transferred to 1% sodium hypochlorite solution, shaken at 150 r / min for 30 minutes, and then washed 5 times with sterile water.

[0045] Physical treatment

[0046] The cotyledonary tips of the seeds were cut off with sterilized scissors (T2 treatment), and the control group (CK) was not treated.

[0047] Gibberellic acid treatment

[0048] T2 and CK seeds were immersed in 150 mg / L gibberellin solution, shaken at 150 rpm for 8 h at room temperature, washed 5 times with sterile water, and dried with filter paper.

[0049] Germination culture

[0050] Line a Petri dish with filter paper and sterile gauze and add sterile water until moist. Set up three replicates per group, with 50 seeds per replicate. Incubate at 25 ± 2°C with a 12-hour light / dark cycle. Rehydrate every three days and record the number of germinations.

[0051] result

[0052] The seeds in the T2 treatment group began to germinate on the 6th day, and the germination rate reached 81.3% on the 18th day, which was significantly higher than that in the CK group (germination on the 14th day, germination rate 10.3% on the 18th day). The average root length of the germinated seeds was 2.4 cm (0.89 cm in the CK group).

[0053] Example 2: Comparison of cutting off the seed beak (T1 treatment) and the combined treatment (T3)

[0054] Seed treatment

[0055] The disinfection steps are the same as those in Example 1.

[0056] T1 group: beak was cut off; T3 group: beak + cotyledon end were cut off; CK group received no treatment.

[0057] Gibberellic acid and culture

[0058] Same conditions as Example 1.

[0059] result

[0060] The T1 group germinated on the 6th day, with a germination rate of 19.1% after 18 days; the T2 group germinated on the 8th day, with a germination rate of 40.6% after 18 days; and the T3 group germinated on the 6th day, with a germination rate of 35.2% after 18 days. These rates were significantly higher than the germination rate of the CK group (10.3%), but lower than the germination rate of the T2 group (81.3%). There were no significant differences in root length (T1: 2.2 cm; T2: 2.4 cm; T3: 2.6 cm).

[0061] The beneficial effects of the present invention are:

[0062] Efficient germination: Cutting the cotyledon tip (T2) makes the seeds germinate in 6 days, and the germination rate exceeds 81.3% in 18 days, which is 8 days shorter than the control group and increases the germination rate by more than 8 times.

[0063] High safety: physical treatment replaces concentrated sulfuric acid, which is easy to operate and non-hazardous.

[0064] Wide applicability: It can be extended to grape plants such as Vitis amurensis and Vitis pubescens to enhance the vitality of seedlings.

[0065] Parameter optimization: Clarify the optimal parameter ranges for disinfection, gibberellin, and culture conditions to ensure method stability and repeatability.

[0066] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A method for promoting rapid germination of grape seeds, characterized in that: The following steps are involved: S1. Seed collection: Collect mature grape berries of 'Golden Field Blue Sapphire', peel the seeds, wash them 3-5 times with clean water, dry them in the shade, and then divide them into centrifuge tubes. Store them at 4°C until use (storage time ≤ 6 months). S2. Seed disinfection: Place the seeds in 75% alcohol and shake for 5 minutes, rinse with sterile water 3-5 times, transfer to 1% sodium hypochlorite solution, shake at 150r / min for 30 minutes, and rinse with sterile water 3-5 times. S3. Seed physical treatment: Use sterilized scissors to cut off the beak of the seed (the protruding part of the radicle end); Cut off the cotyledon ends; Cut off the beak and cotyledon ends at the same time; No treatment (control group). S4. Gibberellic acid treatment: Immerse the treated seeds in a 150mg / L gibberellin solution and shake at 150 rpm for 8 hours at room temperature. Rinse with sterile water 3-5 times and dry with filter paper. S5. Germination culture: Place filter paper and sterile gauze in a culture dish and add sterile water until moistened without water accumulation. Each treatment group was replicated three times, with 50 seeds per replicate, evenly spaced. Incubate at 25±2℃ with 12-h light / dark cycle. Rehydrate every 3 days and record the number of sprouts and root length.

2. The method for promoting rapid germination of grape seeds according to claim 1, characterized in that: The seed storage time in step S1 is 1-6 months.

3. The method for promoting rapid germination of grape seeds according to claim 1, characterized in that: The concentration of the sodium hypochlorite solution in step S2 is 1%, and the sodium hypochlorite treatment time in step S2 is 30 minutes.

4. The method for promoting rapid germination of grape seeds according to claim 1, characterized in that: In step S3, the cotyledon ends are cut off.

5. The method for promoting rapid germination of grape seeds according to claim 1, characterized in that: The shaking speed in step S4 is 150 r / min, and the gibberellin treatment time in step S4 is 8 hours.

6. The method for promoting rapid germination of grape seeds according to claim 1, characterized in that: The culture temperature in step S5 is 25±2° C., the light cycle in step S5 is 12 hours of light / 12 hours of darkness, and sterile water is replenished every 2-4 days in step S5, that is, once every 3 days.

7. The method for promoting rapid germination of grape seeds according to claim 1, characterized in that: The method further comprises the step of regularly measuring the root length of the seedlings during the culture process, wherein the root length measurement is performed on the 18th day after the seeds germinate.

8. The method for promoting rapid germination of grape seeds according to claim 1, characterized in that: The method can make grape seeds begin to germinate within 6-8 days, and the germination rate reaches more than 81.3% after 18 days. The method can make the average root length of the germinated seeds reach 2.4 cm.

9. The method for promoting rapid germination of grape seeds according to claim 1, characterized in that: The method is applicable to seed germination of other grape plants.

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