A method for promoting bladderwort seed germination and seedling survival

Abstract

The invention discloses a method for promoting bladderwort seed germination and seedling survival, the steps of which are (1) collecting and preserving bladderwort seeds: collecting mature bladderwort seeds every year, placing the bladderwort seeds in a seed bottle with water, and refrigerating at low temperature; (2) determining the vitality of bladderwort seeds: taking out the bladderwort seeds that have been treated at low temperature, placing them at room temperature, treating the bladderwort seeds with a TTC reagent, dyeing and preserving them in the dark; (3) germinating bladderwort seeds under different conditions: taking out the remaining bladderwort seeds that have been placed at room temperature, using them for germination, and calculating the bladderwort seed germination rate based on the length of the embryo sprouting; (4) calculating the survival rate of bladderwort seedlings under different conditions: picking out the seeds that germinate in each group every day, placing them in new culture dishes respectively, the seedling culture conditions are consistent with the seed germination conditions, and calculating the survival rate of the seedlings after culture. The invention has the advantages of low cost, easy operation, convenient management, high survival rate, etc., and effectively improves the utilization efficiency of the ecology and scientific research of bladderwort seeds.

Description

Technical Field

[0001] The invention belongs to the technical field of plant breeding, and more specifically relates to a method for promoting bladderwort seed germination and seedling survival. Background Art

[0002] Reproduction is the key to changes in the distribution and abundance patterns of submerged plant populations in lakes, and successful reproduction is evidence of plant adaptation to complex environmental changes1. Under natural conditions, population reconstruction and ecosystem restoration of submerged vegetation mainly rely on their developed asexual reproduction system, but the role of sexual reproduction for aquatic plants is by no means secondary. Sexual reproduction is the main means of forming new genes and producing new genotypes. At the same time, sexual reproduction is also believed to play an important role in the long-term continuation of species, long-distance dispersal, and community recovery after extreme events2,3,4. In the process of sexual reproduction of freshwater submerged plants, seed germination is one of the key processes for successful reproduction. The seeds formed by sexual reproduction are very likely to be the basis for the recovery of submerged plants after extreme disturbance and destruction5. It has been found that the seeds produced by sexual reproduction of some submerged plants (such as Potamogetoncrispus and Potanogetonmaackianus) have extremely low germination rates under natural conditions, and their community maintenance and expansion are forced to rely on asexual reproductive bodies. There are some methods for promoting the germination of aquatic plants such as water horn and water shield. However, there is no technology for promoting the germination of seeds of bladderwort, which is a carnivorous plant among submerged plants. Summary of the invention

[0003] The invention aims to provide a method for promoting bladderwort seed germination and seedling survival, thereby improving the germination rate of bladderwort seeds produced by sexual reproduction under natural conditions, thereby improving the survival rate of plants.

[0004] In order to achieve the above technical purpose, the present invention adopts the following technical solution:

[0005] (1) Collection and preservation of bladderwort seeds

[0006] Mature bladderwort seeds were collected intermittently from May to September 2022. The geographical coordinates are the aquatic wetland plant resource garden of Poyang Lake Botanical Garden, Lianxi District, Jiujiang City, Jiangxi Province. The collected seeds were dried indoors in the shade, and then the bladderwort seeds were placed in a seed bottle with water and refrigerated for 30 days at a temperature range of 0-4°C.

[0007] (2) Determination of bladderwort seed vitality

[0008] Take out the treated bladderwort seeds from the refrigerator and place them at room temperature for 24 hours. Randomly select 50 bladderwort seeds. Prepare a 0.5% TTC dye solution, treat the bladderwort seeds with 0.5% TTC reagent, dye for 48 hours, and store in the dark. Then, count the staining under a stereo microscope and calculate the bladderwort seed vitality. Calculation formula (seed vitality = number of stained seeds / total number of seeds).

[0009] (3) Germination of bladderwort seeds under different conditions

[0010] Take out the remaining bladderwort seeds that have been treated and placed at room temperature, randomly select bladderwort seeds for germination, and randomly divide them into 7 groups. Each group of seeds is placed in a culture dish with a diameter of 0.9 cm, and 1 ml of sterile water is added. Set the germination condition range to: 10-45℃ (one germination condition for each 5℃ range), photoperiod 12:12h, temperature change varies with the photoperiod, daytime is 5℃ higher than night, light intensity 3000lux, and repeat three times. Effective germination is considered when the embryo sprouts 0.2cm in length. The number of germinations is counted every day, and the experimental period is 28 days. Finally, the germination rate of bladderwort seeds is counted, and the calculation formula (germination rate = number of germinated seeds / total number of seeds) is calculated.

[0011] (4) Survival rate of bladderwort seedlings under different conditions

[0012] The seeds germinated every day in each group were picked out and placed in new culture dishes respectively. The seedling culture conditions were consistent with the seed germination conditions. The survival rate of the seedlings was calculated after 20 days of culture. The calculation formula was (survival rate = number of surviving seedlings / total number of germinated seeds).

[0013] In the present invention, the result of the bladderwort seed vitality test is: it is found statistically that the bladderwort seed vitality obtained after TTC staining is 68%.

[0014] In the present invention, the germination rate test results of bladderwort seeds under different conditions show that the preferred germination conditions in step (3) are 15°C-35°C, a photoperiod of 12:12h, temperature change changes with the photoperiod, daytime is 5°C higher than nighttime, light intensity is 3000lux, germination start time: 3rd day to 18th day, and the preferred germination rate is: 20.00%-67.33%.

[0015] In the present invention, the germination rate test results of bladderwort seeds under different conditions show that the preferred germination conditions in step (3) are: 20-35°C, photoperiod 12:12h, temperature change changes with the photoperiod, daytime is 5°C higher than night, light intensity 3000lux; the survival rate reaches 100%.

[0016] In the present invention, the germination conditions in step (3) are: 30-35°C, photoperiod 12:12h, temperature change changes with the photoperiod, daytime is 5°C higher than nighttime, and light intensity is 3000lux. The seedling growth rate is the fastest, and the germination rate of live seeds at this time reaches 99.01%.

[0017] Compared with the prior art, the present invention has the following technical effects and advantages:

[0018] The after-ripening and dormancy of aquatic plant seeds are the key to the success of breeding and seedling cultivation. The present invention effectively solves the technical bottleneck of dormancy and after-ripening of aquatic plant seeds by first subjecting bladderwort seeds to low temperature and humidity treatment for 30 days, and then germinating and seedling cultivation under certain conditions, and mainly solves the technical problems of low germination rate of aquatic plant bladderwort in natural habitats. Through the technical measures, the germination rate of bladderwort seeds is as high as 67.33%, the germination rate of live seeds is as high as 99.01%, and the germination time is short and the germination time is consistent. Compared with the prior art, the present invention has the advantages of high efficiency, low cost, easy operation, convenient management, high survival rate, etc., and effectively improves the ecological and scientific research utilization efficiency of bladderwort seeds. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the technical solution of the present invention, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0020] Figure 1 This is a flow chart of the method for germinating bladderwort seeds;

[0021] Figure 2 This is the TTC staining image of the vitality of Bladderwort seeds;

[0022] Figure 3 is a curve chart of germination rate of Bladderwort seeds;

[0023] Figure 4 Germination of albino seedlings from bladderwort seeds;

[0024] Figure 5 This is the growth process of bladderwort seedlings;

[0025] Figure 6 The growth of bladderwort seedlings under T3, T4 and T5 culture conditions; DETAILED DESCRIPTION

[0026] The following will be combined with the attached embodiment of the present invention Figure 1-6, the technical solutions in the embodiments of the present invention are clearly and completely described. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0027] 1. Embodiment 1:

[0028] A method for promoting bladderwort seed germination and seedling survival, comprising the following steps:

[0029] (1) Collection and preservation of bladderwort seeds:

[0030] The author collected mature bladderwort seeds intermittently from May to September 2022. The geographical coordinates are the aquatic wetland plant resource garden of Poyang Lake Botanical Garden, Lianxi District, Jiujiang City, Jiangxi Province. The collected seeds were dried indoors in the shade, and then the bladderwort seeds were placed in a seed bottle with water and refrigerated for 30 days at a temperature range of 0-4°C.

[0031] (2) Determination of bladderwort seed vitality:

[0032] Take out the treated bladderwort seeds from the refrigerator and place them at room temperature for 24 hours. Randomly select 50 bladderwort seeds. Prepare a 0.5% TTC dye solution, treat the bladderwort seeds with 0.5% TTC reagent, dye for 48 hours, and store in the dark. Then, count the staining under a stereo microscope and calculate the bladderwort seed vitality. Calculation formula (seed vitality = number of stained seeds / total number of seeds).

[0033] (3) Germination of Bladderwort seeds:

[0034] The remaining bladderwort seeds placed at room temperature after the above treatment were taken out, and the total number of seeds was greater than 1100. The bladderwort seeds for germination were randomly selected, and 150 seeds were selected for each group, and each group was divided into three replicates, a total of 7 groups, and were placed in a culture dish with a diameter of 0.9 cm, and 1 ml of sterile water was added. Example 1 The germination conditions were set: T1 (10-15°C, photoperiod 12:12h, temperature change changed with the photoperiod, daytime was 5°C higher than night, light intensity 3000lux), and the treatment was repeated three times. The length of the embryo sprouting 0.2cm was regarded as effective germination. The number of germinations was counted every day, and the experimental period was 28 days. Finally, the germination rate of bladderwort seeds was counted, and the calculation formula (germination rate = number of germinated seeds / total number of seeds) was calculated.

[0035] (4) Survival rate of bladderwort seedlings under different conditions:

[0036] The seeds germinated every day in each group were picked out and placed in new culture dishes respectively. The seedling culture conditions were consistent with the seed germination conditions. The survival rate of the seedlings was calculated after 20 days of culture. The calculation formula was (survival rate = number of surviving seedlings / total number of germinated seeds).

[0037] Embodiment 2-7:

[0038] Examples 2-7 set the following germination conditions respectively:

[0039] Example 2: In step (3) of this example, the germination conditions are: T2 (15-20°C, the photoperiod is 12:12h, the temperature changes with the photoperiod, the daytime is 5°C higher than the nighttime, and the light intensity is 3000lux),

[0040] Example 3: In step (3) of this example, the germination conditions are: T3 (20-25°C, photoperiod 12:12h, temperature changes with the photoperiod, daytime is 5°C higher than nighttime, light intensity 3000lux),

[0041] Example 4: In step (3) of this example, the germination conditions are: T4 (25-30°C, photoperiod 12:12h, temperature changes with the photoperiod, daytime is 5°C higher than nighttime, light intensity 3000lux),

[0042] Example 5: In step (3) of this example, the germination conditions are: T5 (30-35°C, photoperiod 12:12h, temperature changes with the photoperiod, daytime is 5°C higher than nighttime, light intensity 3000lux),

[0043] Example 6: In step (3) of this example, the germination conditions are: T6 (35-40°C, photoperiod 12:12h, temperature changes with the photoperiod, daytime is 5°C higher than nighttime, light intensity 3000lux),

[0044] Example 7: In step (3) of this example, the germination conditions are: T7 (40-45°C, photoperiod 12:12h, temperature changes with the photoperiod, daytime is 5°C higher than nighttime, light intensity 3000lux);

[0045] The other implementation steps are the same as those in Example 1.

[0046] Example 8: Results and Analysis

[0047] 1. Vitality of Bladderwort Seeds

[0048] Statistics show that the vitality of Bladderwort seeds obtained after TTC staining is 68%. Figure 2 )

[0049] 2. Germination rate of bladderwort seeds under different conditions

[0050] The results showed that the bladderwort seeds did not germinate under T1 and T7 culture conditions, the bladderwort seeds germinated into albino seedlings under T6 culture conditions, and the bladderwort seeds began to germinate on the 18th, 8th, 5th and 3rd days under T2, T3, T4 and T5 culture conditions, respectively. The final germination rates of the bladderwort seeds under T2, T3, T4 and T5 culture conditions were 20.00%, 50.67%, 52.67% and 67.33%, respectively. (As Figure 3 (shown)

[0051] 3. Survival rate of bladderwort seedlings under different conditions

[0052] After 20 days of cultivation under the corresponding culture conditions, it was found that under T2 conditions, the seedlings stopped growing and finally died; under T6 culture conditions, the seedlings turned white and could not survive; under T3, T4, and T5 culture conditions, the survival rate of the seedlings reached 100%, and the seedling growth rate was the fastest under T5 culture conditions. Figure 4 — Figure 6 )

[0053] It is not difficult to find from the above results that the measured vitality of bladderwort seeds is 68%, and the germination rate of bladderwort seeds under T5 culture conditions is 67.33%, indicating that the germination rate of live seeds under T5 conditions is 99.01%. Therefore, in summary, the applicant believes that T5 (30-35°C, photoperiod 12:12h, temperature change changes with photoperiod, daytime is 5°C higher than night, light intensity 3000lux) conditions are the optimal conditions for germination of bladderwort seeds.

[0054] The above description is only a specific implementation of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be understood by anyone familiar with the technology within the technical scope disclosed by the present invention should be included in the scope of the present invention.

Claims

1. A method for promoting bladderwort seed germination and seedling survival, characterized in that: The following steps are involved: (1) Collection and preservation of bladderwort seeds: From May to September each year, mature bladderwort seeds were collected intermittently, dried indoors in the shade, and then placed in a seed bottle with water and refrigerated for 30 days at a temperature range of 0-4°C. (2) Determination of bladderwort seed vitality: Take out the low-temperature treated bladderwort seeds, place them at room temperature for 24 hours, randomly select several bladderwort seeds, prepare 0.5% TTC dye solution, treat the bladderwort seeds with 0.5% TTC reagent, dye for 48 hours, and store them away from light before calculating the vitality of the bladderwort seeds; (3) Germination of Bladderwort seeds: The remaining bladderwort seeds that had been placed at room temperature for 24 hours were taken out and placed in a culture dish with a diameter of 0.9 cm, and 1 mL of sterile water was added; the germination conditions were set to the range of 20-35°C, a photoperiod of 12:12h, a temperature change that varied with the photoperiod, a daytime temperature that was 5°C higher than a night temperature, a light intensity of 3000 lux, and effective germination when the embryo sprouted 0.2 cm in length, with a cycle of 28 days; (4) Survival rate of bladderwort seedlings: The seeds that germinated every day were picked out and placed in new culture dishes. The seedling culture conditions were consistent with the seed germination conditions. After 20 days of culture, the seedling survival rate was 100%.

2. A method for promoting bladderwort seed germination and seedling survival according to claim 1, characterized in that: The germination conditions described in step (3) are 30-35°C, a photoperiod of 12:12h, temperature changes with the photoperiod, daytime is 5°C higher than nighttime, and a light intensity of 3000 lux. When the seedling growth rate is the fastest, the live seed germination rate is 99.01%.

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