Sesbania hybridizing method for improving pollination rate

Abstract

The invention discloses a sesbania hybridizing method for improving the pollination rate, and belongs to the technical field of cross breeding. The method comprises the following steps: (1) selecting parents; (2) collecting pollen; (3) emasculating the female parent; and (4) pollinating and listing. Standards are set for female parent flowers suitable for pollination, complete corolla structures are reserved after female parent emasculation, styles can be protected, and the hybridization efficiency and accuracy are effectively improved by placing the whole pollen sacs in the female parent flowers.

Description

Technical Field

[0001] The invention belongs to the technical field of hybrid breeding, and particularly relates to a sesbania hybridization method for improving the pollination rate. Background Art

[0002] Sesbania cannabina is an annual subshrubby herb. It belongs to the genus Sesbania of the Fabaceae family, and its Latin name is Sesbania cannabina. Sesbania cannabina is a leguminous plant that grows naturally in tropical areas and coastal areas. It has the characteristics of salt-alkali resistance and outstanding waterlogging resistance. It has a good nitrogen fixation effect as a green manure and helps to improve the soil. At present, the breeding method of new varieties of Sesbania cannabina is in its infancy, that is, the best of the existing varieties and germplasm are selected, and it is difficult for the selected varieties to achieve major breakthroughs in key shapes. Through hybrid breeding technology, more genetic mutation materials can be obtained, and at the same time, it is possible to enrich excellent traits. There are currently no reports on Sesbania cannabina hybridization technology. Sesbania cannabina hybridization technology does not have a standardized process and standard, resulting in low hybrid pollination efficiency and less fruiting. Therefore, it is necessary to provide a method for Sesbania cannabina hybridization that improves the pollination rate. Summary of the invention

[0003] In view of the above problems existing in the prior art, the technical problem to be solved by the present invention is to provide a method for hybridization of sesbania to improve the pollination rate. The method retains all petals and maintains the activity of the flower of the female parent when emasculating the female parent, and at the same time simulates the process of pollen sac drying and releasing pollen under natural conditions, effectively improving the hybridization efficiency and accuracy, and does not affect the hybridization pod setting rate.

[0004] In order to solve the above problems, the technical solution adopted by the present invention is as follows: A method for hybridizing sesbania to improve pollination rate, comprising the following specific steps: (1) Selecting parents: Planting and determining parent materials with consistent flowering periods, so that the parent materials enter the reproductive growth state at the same time to achieve consistency in flowering period; (2) Collecting pollen: removing pollen sacs from the male plant, storing the collected pollen in the form of pollen sacs, and collecting them in a centrifuge tube for later use; (3) Emasculation of female plants: Select unopened female plant flowers, wherein the calyx length: petal length of the female plant flowers is 1:1.2-1.5, gently pinch the petals and remove the stamens with tweezers, keeping the female plant flowers intact. The intact female plant flowers mean retaining the complete corolla structure, including the standard petal, wing petals, and keel petals; (4) Pollination and labeling: Place the pollen sacs collected in step (2) into the flower of the female parent to dry and release the pollen, and gently twist the petals to promote the release of the male parent pollen in the female parent flower.

[0005] Furthermore, in step (2), the pollen sacs collected from the male plant should be used up on the same day as far as possible, and the unused pollen sacs can be placed in a 4°C refrigerator for no more than 24 hours.

[0006] Furthermore, in step (3), when removing the stamens, ensure that the pollen sacs of the stamens of the mother plant flower do not come into contact with the air and thus scatter pollen, and at the same time, the pollen sacs should not be damaged during the operation to cause premature release of pollen, and a label with marking information should be hung on the pedicel.

[0007] Furthermore, in step (3), the operation method for keeping the mother flower intact is to cut the keel petals along the central axis to expose the stamens.

[0008] Furthermore, in step (3), the female parent flower is pollinated immediately after being emasculated.

[0009] Furthermore, in step (4), the pollen sac is placed as a whole in the pollinated female flower.

[0010] Beneficial effects of the present invention: (1) The present invention specifically describes the flower state of the optimal emasculation period of the Sesbania sesbania female plant, thereby avoiding the effects of premature emasculation on style development and late emasculation on plant self-pollination that affects hybridization results, thereby effectively improving hybridization efficiency and accuracy.

[0011] (2) The present invention retains the complete corolla structure, including the standard petal, wing petals, and keel petals, after the female plant is emasculated, which helps to protect the style and improve the hybridization efficiency.

[0012] (2) The present invention places the entire pollen sac into the mother plant flower, simulating the state of the pollen sac drying and spreading pollen in the natural state, which effectively improves the pollination success rate. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 The figure is a schematic diagram of selecting the emasculation period of the flowers of the female parent plant in the present invention; wherein A is the state of the flowers of Sesbania sesbania that have completed closed pollination, and such flowers cannot be selected as flowers for hybridization, and B is the state of flowers suitable for being a hybrid female parent, with a calyx to petal ratio of 1:1.2 to 1:1.5.

[0014] Figure 2 The figure is a schematic diagram of the steps of the sesbania pollination technology of the present invention; wherein A is a schematic diagram of collecting pollen sacs from male parent materials, B is a suitable female parent flower selected, C is the state of the female parent material after emasculation and the stripped stamens, D is the state of the flower after pollination, and E is the normal flowering of the flower after pollination.

[0015] Figure 3 The flower states mentioned in the comparative examples of the present invention; wherein A is the state of the female flower after emasculation in the comparative example, and B is the state of the female flower after pollination in the comparative example. DETAILED DESCRIPTION

[0016] The specific implementation modes of the present invention are described below so that those skilled in the art can understand the present invention. However, it should be clear that the present invention is not limited to the scope of the specific implementation modes. For those of ordinary skill in the art, as long as various changes are within the spirit and scope of the present invention as defined and determined by the attached claims, these changes are obvious, and all inventions and creations utilizing the concept of the present invention are protected.

[0017] The male variety used in the following examples is Zhongkejing 89, and the female variety is Zhongkejing 15. The variety information is shown in the following table.

[0018] Table 1 Variety information of all parent materials

[0019] Example 1: Hybridization and pollination of Sesbania sesbania The specific steps of the Sesbania hybridization method for improving the pollination rate are as follows: (1) Parent selection: The parent materials for hybridization need to be planted in advance and the corresponding flowering period must be determined so that the parents enter reproductive growth at the same time and have the same flowering period.

[0020] (2) Collecting pollen: Take the flowers of the male plant whose petals are about to open but whose keel petals are not yet exposed to the air, remove the pollen sacs with tweezers, and collect them in a 1.5 mL centrifuge tube for later use. The pollen sacs collected from the male plant should be used up on the same day as much as possible. The unused pollen sacs can be placed in a 4℃ refrigerator for no more than 24 hours.

[0021] (3) Female emasculation: Select flowers on the female plant whose petals have not yet opened and whose calyx length: petal length = 1:(1.2~1.5). Gently pinch the flower with your fingers to open the standard petal and wing petal. Use sharp tweezers to cut the keel petal along the central axis. While exposing all stamens, keep the integrity and activity of the standard petal, wing petal and keel petal in the butterfly corolla, and remove the stamens with sharp tweezers. Pollinate the female flower immediately after emasculation. In addition, when removing the stamens, ensure that the pollen sacs of the stamens of the female flower do not come into contact with the air and thus release pollen. At the same time, do not damage the pollen sacs during the operation to release pollen prematurely.

[0022] (4) Pollination and labeling: Place the pollen sac collected in step (2) into the mother plant's flower as a whole to simulate the natural drying process of pollen, allowing it to dry in the mother plant's petals and release pollen. At this time, the petals can be gently twisted to help release the pollen and complete pollination. Label the hybrid breeding information on the pedicel. The hybrid breeding information includes the hybrid parent information and the pollination date.

[0023] (5) Investigation on the effect of hybrid pollination Pollination was investigated 3 days after pollination. In this embodiment, 30 flowers were pollinated, 19 of which were successfully hybridized, with a success rate of 63%.

[0024] Example 2: Hybridization and pollination of Sesbania sesbania The specific steps of the Sesbania hybridization method for improving the pollination rate are as follows: (1) Parent selection: The parent materials for hybridization need to be planted in advance and the corresponding flowering period must be determined so that the parents enter reproductive growth at the same time and have the same flowering period.

[0025] (2) Collecting pollen: Take the flowers of the male plant whose petals are about to open but whose keel petals are not yet exposed to the air, remove the pollen sacs with tweezers, and collect them in a 1.5 mL centrifuge tube for later use. The pollen sacs collected from the male plant should be used up on the same day as much as possible. The unused pollen sacs can be placed in a 4℃ refrigerator for no more than 24 hours.

[0026] (3) Female emasculation: Select flowers on the female plant whose petals have not yet opened and whose calyx length: petal length = 1:(1.2~1.5). Gently pinch the flower with your fingers to open the standard petal and wing petal. Use sharp tweezers to cut the keel petal along the central axis. While exposing all stamens, keep the integrity and activity of the standard petal, wing petal and keel petal in the butterfly corolla, and remove the stamens with sharp tweezers. Pollinate the female flower immediately after emasculation. In addition, when removing the stamens, ensure that the pollen sacs of the stamens of the female flower do not come into contact with the air and thus release pollen. At the same time, do not damage the pollen sacs during the operation to release pollen prematurely.

[0027] (4) Pollination and labeling: Place the pollen sac collected in step (2) into the mother plant's flower as a whole to simulate the natural drying process of pollen, allowing it to dry in the mother plant's petals and release pollen. At this time, the petals can be gently twisted to help release the pollen and complete pollination. Label the hybrid breeding information on the pedicel. The hybrid breeding information includes the hybrid parent information and the pollination date.

[0028] (5) Investigation on the effect of hybrid pollination Pollination was investigated 3 days after pollination. In this embodiment, 30 flowers were pollinated, 17 flowers were successfully hybridized, and the success rate was 56%.

[0029] Example 3: Cross-pollination of Sesbania sesbania The specific steps of the Sesbania hybridization method for improving the pollination rate are as follows: (1) Parent selection: The parent materials for hybridization need to be planted in advance and the corresponding flowering period must be determined so that the parents enter reproductive growth at the same time and have the same flowering period.

[0030] (2) Collecting pollen: Take the flowers of the male plant whose petals are about to open but whose keel petals are not yet exposed to the air, remove the pollen sacs with tweezers, and collect them in a 1.5 mL centrifuge tube for later use. The pollen sacs collected from the male plant should be used up on the same day as much as possible. The unused pollen sacs can be placed in a 4℃ refrigerator for no more than 24 hours.

[0031] (3) Female emasculation: Select flowers on the female plant whose petals have not yet opened and whose calyx length: petal length = 1:(1.2~1.5). Gently pinch the flower with your fingers to open the standard petal and wing petal. Use sharp tweezers to cut the keel petal along the central axis. While exposing all stamens, keep the integrity and activity of the standard petal, wing petal and keel petal in the butterfly corolla, and remove the stamens with sharp tweezers. Pollinate the female flower immediately after emasculation. In addition, when removing the stamens, ensure that the pollen sacs of the stamens of the female flower do not come into contact with the air and thus release pollen. At the same time, do not damage the pollen sacs during the operation to release pollen prematurely.

[0032] (4) Pollination and labeling: Place the pollen sac collected in step (2) into the mother plant's flower as a whole to simulate the natural drying process of pollen, allowing it to dry in the mother plant's petals and release pollen. At this time, the petals can be gently twisted to help release the pollen and complete pollination. Label the hybrid breeding information on the pedicel. The hybrid breeding information includes the hybrid parent information and the pollination date.

[0033] (5) Investigation on the effect of hybrid pollination Pollination was investigated 3 days after pollination. In this embodiment, 20 flowers were pollinated, 14 of which were successfully hybridized, with a success rate of 70%.

[0034] Comparative Example 1 Conventional hybrid pollination of Sesbania sesbania (1) Parent selection: The parent materials for hybridization need to be planted in advance and the corresponding flowering period must be determined so that the parents enter reproductive growth at the same time and have the same flowering period.

[0035] (2) Collecting pollen: Take the flowers of the male plant whose petals are about to open but whose keel petals are not yet exposed to the air. Use tweezers to remove the pollen sacs and collect them in a 1.5 mL centrifuge tube for later use.

[0036] (3) Emasculation of the female plant: Select a female plant flower that has not yet bloomed, peel off the petals, and use tweezers to remove the stamens, avoiding damage to the style during the process.

[0037] (4) Pollination and labeling: Use a brush to gently apply the pollen collected in step (2) to the stigma of the female parent flower, or use tweezers to hold the dried pollen sac that has begun to shed pollen and gently touch the stigma to complete pollination. At this time, a label can be hung on the pedicel to mark the hybrid breeding information. The hybrid breeding information includes the hybrid parent information and the pollination date.

[0038] In this embodiment, a total of 50 flowers were pollinated, 8 flowers were successfully hybridized, and the success rate was 16%.

[0039] Comparative Example 2 Conventional hybridization pollination of Sesbania sesbania (1) Parent selection: The parent materials for hybridization need to be planted in advance and the corresponding flowering period must be determined so that the parents enter reproductive growth at the same time and have the same flowering period.

[0040] (2) Collecting pollen: Take the flowers of the male plant whose petals are about to open but whose keel petals are not yet exposed to the air. Use tweezers to remove the pollen sacs and collect them in a 1.5 mL centrifuge tube for later use.

[0041] (3) Emasculation of the female plant: Select a female plant flower that has not yet bloomed, peel off the petals, and use tweezers to remove the stamens, avoiding damage to the style during the process.

[0042] (4) Pollination and labeling: Use a brush to gently apply the pollen collected in step (2) to the stigma of the female parent flower, or use tweezers to hold the dried pollen sac that has begun to shed pollen and gently touch the stigma to complete pollination. At this time, a label can be hung on the pedicel to mark the hybrid breeding information. The hybrid breeding information includes the hybrid parent information and the pollination date.

[0043] In this embodiment, a total of 50 flowers were pollinated, 7 flowers were successfully hybridized, and the success rate was 14%.

[0044] Comparative Example 3 Conventional hybrid pollination of Sesbania sesbania (1) Parent selection: The parent materials for hybridization need to be planted in advance and the corresponding flowering period must be determined so that the parents enter reproductive growth at the same time and have the same flowering period.

[0045] (2) Collecting pollen: Take the flowers of the male plant whose petals are about to open but whose keel petals are not yet exposed to the air. Use tweezers to remove the pollen sacs and collect them in a 1.5 mL centrifuge tube for later use.

[0046] (3) Emasculation of the female plant: Select a female plant flower that has not yet bloomed, peel off the petals, and use tweezers to remove the stamens, avoiding damage to the style during the process.

[0047] (4) Pollination and labeling: Use a brush to gently apply the pollen collected in step (2) to the stigma of the female parent flower, or use tweezers to hold the dried pollen sac that has begun to shed pollen and gently touch the stigma to complete pollination. At this time, a label can be hung on the pedicel to mark the hybrid breeding information. The hybrid breeding information includes the hybrid parent information and the pollination date.

[0048] In this embodiment, a total of 40 flowers were pollinated, and 1 flower was successfully hybridized, with a success rate of 2.5%.

[0049] Compared with comparative examples 1 to 3, the hybridization success rate of embodiments 1 to 3 is significantly higher than that of comparative examples 1 to 3, thereby improving work efficiency.

[0050] Finally, it should be noted that the above is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for hybridizing sesbania to improve pollination rate, characterized in that: The specific steps are as follows: (1) Selecting parents: Planting and determining parent materials with consistent flowering periods, so that the parent materials enter the reproductive growth state at the same time to achieve consistency in flowering period; (2) Collecting pollen: removing pollen sacs from the male plant, storing the collected pollen in the form of pollen sacs, and collecting them in a centrifuge tube for later use; (3) Emasculation of female plants: Select unopened female plant flowers, wherein the calyx length: petal length of the female plant flowers is 1:1.2-1.5, gently pinch the petals and remove the stamens with tweezers, keeping the female plant flowers intact. The intact female plant flowers mean retaining the complete corolla structure, including the standard petal, wing petals, and keel petals; (4) Pollination and labeling: Place the pollen sacs collected in step (2) into the flower of the female parent to dry and release the pollen. Gently twist the petals to promote the release of the male parent pollen in the female parent flower, and label the information on the pedicel.

2. The method for hybridizing sesbania for improving pollination rate according to claim 1, characterized in that: In step (2), the pollen sacs collected from the male parent plant are used up on the same day, and the unused pollen sacs are placed in a 4°C refrigerator for no more than 24 hours.

3. The method for hybridizing sesbania for improving pollination rate according to claim 1, characterized in that: In step (3), when removing the stamens, ensure that the pollen sacs of the stamens of the mother flower do not come into contact with the air and thus scatter pollen, and at the same time, the pollen sacs should not be damaged during the operation to cause premature release of pollen.

4. The method for hybridizing sesbania for improving pollination rate according to claim 1, characterized in that: In step (3), the operation method for keeping the mother flower intact is to cut the keel petals along the central axis to expose the stamens.

5. The method for hybridizing sesbania for improving pollination rate according to claim 1, characterized in that: In step (3), the female flower is pollinated immediately after being emasculated.

6. The method for hybridizing sesbania for improving pollination rate according to claim 1, characterized in that: In step (4), the pollen sac is placed as a whole in the pollinated female flower.

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