Method for improving kiwifruit pollen germination rate and pollen storage
By subjecting kiwi male flower pollen to light treatment and optimizing the culture medium, the pollen germination rate and storage capacity were improved, solving the problem of low germination rate during pollen collection and storage and extending the storage period of pollen.
Patent Information
- Application Number
- CN202610582833.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-04-29
- Publication Date
- 2026-07-24
AI Technical Summary
Existing methods for collecting and storing kiwifruit pollen result in low pollen germination rates and weak pollen viability, affecting pollination effectiveness and fruit quality.
Kiwi male flower pollen was placed under light at 24-28℃ for 10-14 hours, and then cultured under light on an in vitro germination medium. The composition of the medium and the light conditions were optimized, the pollen germination status was observed, and the survival rate was calculated.
It significantly improves pollen germination rate, prolongs pollen storage period, reduces the decline in germination rate during storage, and enhances pollen storage capacity.
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Abstract
Description
Technical Field
[0001] This invention relates to the field of forest tree breeding technology, and in particular to a method for improving the pollen germination rate and pollen storage capacity of kiwifruit. Background Technology
[0002] Kiwifruit is a highly nutritious fruit, known as the "King of Fruits," and its industry is developing rapidly worldwide. Kiwifruit is a typical dioecious plant, and its fruit set rate, fruit size, quality, and yield largely depend on an effective pollination process. Therefore, high-quality pollen is one of the key factors ensuring successful pollination and high-quality, high-yield kiwifruit.
[0003] Artificial pollination has become an indispensable technique in the high-yield and high-efficiency cultivation of kiwifruit. During artificial pollination, the collection and processing techniques directly determine the quality of the pollen, thus affecting the pollination effect and fruit quality. Although research has been conducted on aspects such as the timing of kiwifruit pollen collection, anther separation time, drying methods, storage conditions, and testing methods, most collection and storage methods cause some degree of pollen viability loss, especially for kiwifruit varieties with weak pollen viability and low germination rates.
[0004] Therefore, improving the germination rate and storage capacity of kiwifruit pollen to obtain high-quality pollen is of great significance for improving the pollination effect and fruit quality of kiwifruit. Summary of the Invention
[0005] The purpose of this invention is to provide a method for improving the germination rate and storage capacity of kiwifruit pollen. The method of this invention makes the germination rate of kiwifruit pollen significantly higher than that of fresh pollen, and at the same time can improve the storage capacity of kiwifruit pollen, reducing the decline in the germination rate of kiwifruit pollen during storage.
[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides a method for improving the germination rate and storage capacity of kiwifruit pollen, comprising the following steps: S1. Place the pollen of male kiwifruit flowers in a light source at 24-28℃ for 10-14 hours; S2. Sprinkle the pollen treated in S1 onto the in vitro germination medium and culture it under light. S3. Observe the pollen germination status and calculate the pollen survival rate.
[0007] Preferably, the male flower is a flower whose petals have just opened or are about to open.
[0008] Preferably, in step S1, the intensity of the light is 2500~3500 LX.
[0009] Preferably, in step S2, the culture temperature is 24~26℃, the culture time is 6~7h, and the light intensity is 2500~3500LX.
[0010] Preferably, in step S2, the in vitro germination culture medium comprises the following components: agar 8~12g / L, white sugar 18~25g / L, boric acid 80~120mg / L; pH is 6.0~6.5.
[0011] Preferably, the method for preparing the in vitro germination medium includes first adding boric acid and agar, and after the agar is completely dissolved, adding white sugar, adjusting the pH to 6.0~6.5, and then setting it aside for use.
[0012] Preferably, in step S3, the pollen germination state is defined as the pollen tube length exceeding the pollen grain diameter.
[0013] Preferably, the formula for calculating the pollen survival rate is: pollen survival rate = number of germinating pollen ÷ total number of observed pollen × 100%.
[0014] Compared with the prior art, the present invention has the following beneficial effects: This invention improves the germination rate of collected kiwi male flower pollen by pre-treating it under 26°C light overnight for 12 hours. The germination rate is significantly higher than that of fresh pollen, effectively solving the problem of low pollen germination rate or weak pollen viability caused during pollen collection, which in turn affects pollination effect and fruit quality. This invention improves the germination rate of stored pollen by pre-treating it with light at 26°C overnight for 12 hours. Not only is the germination rate of the pollen higher than that of pollen not treated with light at 26°C overnight, but it also significantly reduces the decrease in germination rate when the pollen is stored at 4°C or -20°C, thereby improving the storage capacity of the pollen and extending its storage period. Detailed Implementation
[0015] This invention provides a method for improving the germination rate and storage capacity of kiwifruit pollen, comprising the following steps: S1. Place the pollen of male kiwifruit flowers in a light source at 24-28℃ for 10-14 hours; S2. Sprinkle the pollen treated in S1 onto the in vitro germination medium and culture it under light. S3. Observe the pollen germination status and calculate the pollen survival rate.
[0016] In this invention, the male kiwifruit flowers are preferably those whose petals have just opened or are about to open.
[0017] In this invention, the pollen light treatment temperature in step S1 is preferably 25~27℃, more preferably 26℃; the light treatment time is preferably 11~13h, more preferably 12h; and the light treatment intensity is preferably 2500~3500LX, more preferably 2800~3200LX, and more preferably 3000LX.
[0018] In this invention, the preferred temperature for pollen in vitro germination culture is 24~26℃, more preferably 26℃; the culture time is 6~7h, more preferably 7h; and the light intensity is 2500~3500LX, more preferably 3000~3200LX.
[0019] In this invention, the components of the in vitro germination culture medium are preferably 8-12 g / L agar, 18-25 g / L sucrose, and 80-120 mg / L boric acid, and more preferably 10 g / L agar, 20 g / L sucrose, and 100 mg / L boric acid.
[0020] In this invention, the pH of the in vitro germination medium is preferably 6.0 to 6.5, more preferably 6.2 to 6.4, and even more preferably 6.3.
[0021] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.
[0022] Example 1
[0023] S1. Pollen Collection and Pretreatment: Select male kiwifruit flowers and collect mature anthers. Collect male flowers (male flower variety: Moshan No. 4) whose petals are just opening or about to open, pack them in resealable bags and bring them indoors. First, place them on sulfuric acid paper to air dry. After the anthers naturally dehisce and release pollen, place them in petri dishes, sieve to remove impurities, and collect pure pollen. The collected pollen is placed in glass bottles and treated overnight in a 24℃ artificial climate chamber for 14 hours. The light intensity during the overnight treatment is 2800 LX. S2. Preparation of pollen in vitro germination medium: Weigh 4g agar, 9g sugar, and 40mg boric acid. First, add 450ml of tap water to a cup and heat it on an electric stove. Then add boric acid and agar. After the agar is completely dissolved, add sugar and adjust the volume to 500ml. Adjust the pH to 6.0 and set aside for later use. S3. Pollen inoculation and culture: Use a brush to take a small amount of pollen obtained in S1 and sprinkle it evenly on the culture medium. Culture it under light for 6 hours in an artificial climate chamber at 26℃, with a light intensity of 2500~3500 LX during culture. Then observe the germination of the pollen.
[0024] S4. Pollen germination observation and counting: Place the glass slide containing pollen under a microscope and observe it. Take two fields of view, with about 20 to 40 pollen grains in each field of view. Record the total number of pollen grains and the number of germinating pollen grains (the pollen tube length exceeding the pollen grain diameter is used as the germination standard). Then calculate the pollen germination rate.
[0025] S5. Formula for calculating survival rate: Pollen survival rate (%) = (Number of germinated pollen grains ÷ Total number of observed pollen grains) × 100%.
[0026] Example 2
[0027] S1. Pollen Collection and Pretreatment: Select male kiwifruit flowers and collect mature anthers. Collect male flowers (male flower variety: Moshan No. 4) with petals just opening or about to open, pack them in resealable bags and bring them indoors. First, place them on sulfuric acid paper to air dry. After the anthers naturally dehisce and release pollen, place them in petri dishes, then sieve to remove impurities and collect pure pollen. The collected pollen is placed in glass bottles and treated overnight in a 26℃ artificial climate chamber with light intensity of 3000 LX for 12 hours. S2. Preparation of pollen in vitro germination medium: Weigh 5g agar, 10g sugar, and 50mg boric acid. First, add 450ml of tap water to a cup and heat it on an electric stove. Then add boric acid and agar. After the agar is completely dissolved, add sugar and adjust the volume to 500ml. Adjust the pH to 6.3 and set aside for later use. S3. Pollen inoculation and culture: Use a brush to take a small amount of pollen obtained in S1 and sprinkle it evenly on the culture medium. Incubate in an artificial climate chamber at 26℃ for 7 hours under light. Then observe the germination of the pollen. The light intensity during culture is 2500~3500 LX.
[0028] S4. Pollen germination observation and counting: After pollen culture is completed, place the glass slide containing pollen under a microscope for observation. Take two fields of view, with about 20 to 40 pollen grains in each field of view for observation. Record the total number of pollen grains and the number of germinating pollen grains (the pollen tube length exceeding the pollen grain diameter is used as the germination standard). Then calculate the pollen germination rate.
[0029] S5. Formula for calculating survival rate: Pollen survival rate (%) = (Number of germinated pollen grains ÷ Total number of observed pollen grains) × 100%.
[0030] Example 3
[0031] S1. Pollen Collection and Pretreatment: Select male kiwifruit flowers and collect mature anthers. Collect male flowers whose petals are just opening or about to open (male flower variety is Moshan No. 4), pack them in resealable bags and bring them indoors. First, place them on sulfuric acid paper to air dry. After the anthers naturally dehisce and release pollen, place them in petri dishes, then sieve to remove impurities and collect pure pollen. The collected pollen is placed in glass bottles and treated overnight in a 28℃ artificial climate chamber with light intensity of 3200 LX for 10 hours. S2. Preparation of pollen in vitro germination medium: Weigh 6g agar, 12.5g sugar, and 60mg boric acid. First, add 450ml of tap water to a cup and heat it on an electric stove. Then add boric acid and agar. After the agar is completely dissolved, add sugar and adjust the volume to 500ml. Adjust the pH to 6.5 and set aside for later use. S3. Pollen inoculation and culture: Use a brush to take a small amount of pollen obtained in S1 and sprinkle it evenly on the culture medium. Culture it under light for 6 hours in an artificial climate chamber at 26℃, with a light intensity of 2500~3500 LX during culture.
[0032] S4. Pollen germination observation and counting: After pollen culture is completed, place the glass slide containing pollen under a microscope for observation. Take two fields of view, with about 20 to 40 pollen grains in each field of view for observation. Record the total number of pollen grains and the number of germinating pollen grains (the pollen tube length exceeding the pollen grain diameter is used as the germination standard). Then calculate the pollen germination rate.
[0033] S5. Formula for calculating survival rate: Pollen survival rate (%) = (Number of germinated pollen grains ÷ Total number of observed pollen grains) × 100%.
[0034] Experimental Example 1
[0035] The effect of culture time on pollen germination rate was observed: pollen treated with S1 in Example 2 and fresh pollen without any treatment were evenly sprinkled on the in vitro germination medium in Example 2 and cultured in an artificial climate chamber with light at 26℃. The germination of pollen was recorded every hour until there was no significant difference in the germination rate between the two times. This was the optimal culture time for kiwi pollen. The results are shown in Table 1.
[0036] Table 1. Culture time and pollen germination rate (%)
[0037] Table 1 shows that pollen germination was minimal or nonexistent during the first 3 hours of cultivation. Germination accelerated between 4 and 6 hours. After 6 hours, the germination rate stabilized, reaching a stable level after 7 hours. Therefore, the optimal time for kiwifruit pollen cultivation was determined to be 7 hours; beyond 7 hours, the germination rate showed no significant change. Furthermore, after 7 hours of cultivation, the germination rate of pollen pretreated overnight at 26℃ for 12 hours was significantly higher than that of fresh pollen, exceeding the fresh pollen germination rate by 13.48%. This indicates that overnight pretreatment at 26℃ for 12 hours can improve the pollen germination rate.
[0038] Experimental Example 2
[0039] The pollen storage capacity of Example 2 was tested. The pollen from step S1 of Example 2 was divided into two portions and subjected to two different treatments: A (treated with light overnight at 26°C for 12 hours and then stored at 4°C); B (treated with light overnight at 26°C for 12 hours and then stored at -20°C). The control group consisted of pollen that had not undergone the 12-hour light overnight treatment at 26°C and was subjected to the following two storage treatments: C (stored at 4°C); D (stored at -20°C). The pollen was taken out every other day and treated according to steps S2 to S5 of Example 2, and the germination rate was counted. The results are shown in Table 2.
[0040] Table 2 Pollen storage capacity of four different treatments
[0041] Table 2 shows that there was a significant difference between pollen treated with overnight light at 26℃ and pollen not treated with overnight light at 26℃. The germination rate and storage capacity of pollen treated with overnight light at 26℃ were both higher than those of pollen not treated with overnight light at 26℃. It can be seen that after storage, the germination rate of pollen treated with overnight light at 26℃ was higher than that of pollen not treated with overnight light at 26℃, with the highest germination rate being 21.26% higher. On the 4th day of storage, the germination rate of pollen in group A decreased by 32.4% compared to the previous day, the germination rate of pollen in group C decreased by 56.3% compared to the previous day, the germination rate of pollen in group B decreased by 40.3% compared to the previous day, and the germination rate of pollen in group D decreased by 59.7% compared to the previous day. It can be seen that compared with the control group, the decrease in germination rate of pollen treated with overnight light at 26℃ was significantly smaller. Furthermore, pollen that was not treated with overnight light exposure at 26°C almost completely lost its germination capacity on day 6, while pollen treated with overnight light exposure at 26°C showed germination rates of 15.87% (Group A) and 9.9% (Group B) on day 6, indicating that overnight light exposure at 26°C can improve pollen storage capacity. Therefore, the method provided by this invention can not only improve the pollen germination rate but also significantly reduce the decline in pollen germination rate during storage, thereby improving pollen storage capacity and extending the storage period.
[0042] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.
Claims
1. A method for improving the germination rate and storage capacity of kiwifruit pollen, characterized in that, Includes the following steps: S1. Place the pollen of male kiwifruit flowers in a light source at 24-28℃ for 10-14 hours; S2. Sprinkle the pollen treated in S1 onto the in vitro germination medium and culture it under light. S3. Observe the pollen germination status and calculate the pollen survival rate.
2. The method as described in claim 1, characterized in that, In step S1, the male flower is a flower whose petals have just opened or are about to open.
3. The method as described in claim 1, characterized in that, In step S1, the intensity of the illumination is 2500~3500 LX.
4. The method as described in claim 1, characterized in that, In step S2, the culture temperature is 24~26℃, the culture time is 6~7h, and the light intensity is 2500~3500LX.
5. The method as described in claim 1, characterized in that, In step S2, the in vitro germination culture medium comprises the following components: agar 8~12g / L, white sugar 18~25g / L, boric acid 80~120mg / L; pH is 6.0~6.
5.
6. The method as described in claim 5, characterized in that, The method for preparing the in vitro germination medium includes first adding boric acid and agar, and then adding white sugar after the agar has completely dissolved, adjusting the pH to 6.0~6.5, and then setting it aside for use.
7. The method as described in claim 1, characterized in that, In step S3, the pollen germination state is defined as the pollen tube length exceeding the pollen grain diameter.
8. The method as described in claim 1, characterized in that, In step S3, the formula for calculating the pollen survival rate is: Pollen survival rate = number of germinated pollen ÷ total number of observed pollen × 100%.