Reagent for overcoming distant hybridization incompatibility of plants in the family Lamiaceae by using D-Ser

Abstract

The application discloses a reagent for overcoming distant hybridization incompatibility of plants in the Rutaceae family and a use method thereof. The reagent is used for spraying D-Ser solution onto the stigma of the plants in the Rutaceae family, and distant hybridization incompatibility of Citrus morrisonicola and Melia toosendron is overcome. The reagent has little harm to the plants, has remarkable effect in overcoming distant hybridization incompatibility, has wide application range, and can be further used for overcoming distant hybridization incompatibility of other plants in the Rutaceae family, and has important value for breeding and seed production of the plants in the Rutaceae family.

Description

TECHNICAL FIELD

[0001] The present application relates to the Rutaceae plant breeding technical field, specifically relates to a reagent for overcoming distant hybridization incompatibility of Rutaceae plants by D-Ser. BACKGROUND

[0002] Distant hybridization refers to the hybridization between different species, genera or more distant species in plant taxonomy. Through distant hybridization, the characteristics and properties of different species and genera can be combined to expand genetic variation and create new variation types or even new species. However, in order to hybridize between distant plants, pre-fertilization and post-fertilization reproductive barriers must be overcome. Pre-zygotic barriers refer to the fact that after pollination of two fertile species, the fertilization process cannot be carried out at all, i.e. there is incompatibility, including pollen that cannot germinate or germinate abnormally on the stigma, and pollen that can germinate normally but pollen tubes cannot grow normally in the style.

[0003] Rutaceae Citrus has undergone multiple intraspecific hybridization in the evolutionary process, and the Citrus species cultivated by humans are mostly hybrids of two or more original species. However, there have been no reports of hybridization between Rutaceae species such as Clausena and Melia toosendan and other species, including Citrus species.

[0004] Citrus is an important fruit tree in the world and plays an important role in the national economy, with great economic value. Citrus Huanglongbing bacteria mainly infect Rutaceae plants, and the cultivated Citrus, Ponicirus and Fortunella plants are all hosts. Clausena and Melia toosendan outside the Citrus genus have strong resistance to Huanglongbing, and if the barriers to distant hybridization can be overcome, the introduction of Huanglongbing resistance into Citrus species will have great research and economic value for citrus cultivation to overcome the Huanglongbing problem.

[0005] Distant pollen and stigma incompatibility is the first barrier to distant hybridization, which greatly limits the progress of distant hybridization breeding. Therefore, exploring an efficient and widely applicable method to overcome distant hybridization incompatibility is one of the important components of Rutaceae distant hybridization breeding. SUMMARY

[0006] In order to solve the defects and deficiencies in the prior art, the present application provides a reagent and method for overcoming distant hybridization incompatibility of Rutaceae plants, which can overcome distant hybridization incompatibility, is simple and convenient to operate, and is easy to popularize.

[0007] In order to achieve the purpose of the present application, the present application uses D-Ser (D-serine) solution as the main component of the reagent, and the specific technical scheme is as follows:

[0008] A reagent for overcoming the incompatibility of distant hybridization of plants in the Rutaceae family, the reagent is a D-Ser solution, and the concentration of the solution is 1-20 mmol / L.

[0009] Further, the distant hybridization includes interspecific distant hybridization and intergeneric distant hybridization.

[0010] Another object of the present application is to provide a method for overcoming the incompatibility of distant hybridization of plants in the Rutaceae family by using the reagent, the method comprising:

[0011] In the initial flowering stage of the plants in the Rutaceae family, the reagent is sprayed on the stigmas of unopened flower buds, and then pollination is performed.

[0012] Further, the reagent is sprayed on the stigmas of unopened flower buds twice.

[0013] Further, the interval between the two times of dropping the reagent is 30 min.

[0014] In addition, the present application also provides the application of the reagent in overcoming the incompatibility of distant hybridization of plants in the Rutaceae family.

[0015] Compared with the prior art, the present application has the following beneficial effects:

[0016] The present application is suitable for the incompatibility breeding of distant hybridization of plants in the Rutaceae family. The D-Ser solution effectively overcomes the incompatibility barrier of interspecific hybridization between the Citrus plants and the Melia plants, promotes the germination of the Melia pollen on the stigmas of the Citrus plants in the flowering stage, and makes the Melia pollen grow into the stigmas of the Citrus plants and complete the fertilization process. The method of the present application is simple and convenient to operate, easy to popularize, has little harm to plants, has remarkable effect in overcoming the incompatibility of distant hybridization, and has wide application range, and can be applied to the Citrus plants and the Melia plants. Moreover, the method can be further used to overcome the incompatibility of distant hybridization of other plants in the Rutaceae family, and has important value for the breeding and seed production of the plants in the Rutaceae family. BRIEF DESCRIPTION OF DRAWINGS

[0017] The disclosure of the present application will become more apparent with reference to the drawings. It is easy for those skilled in the art to understand that the drawings are only used to illustrate the technical solutions of the present application, and are not intended to limit the protection scope of the present application.

[0018] Figure 1 In order to treat the stigmas of the Citrus plants with the D-Ser solution, the stigmas of the Citrus plants are dyed with aniline blue after 4 days of pollination with the Melia pollen.

[0019] Figure 2 In order to treat the stigmas of the Citrus plants with the D-Ser solution, the stigmas of the Citrus plants are dyed with aniline blue after 4 days of pollination with the Melia pollen.

[0020] Figure 3 The stigmas of Citrus grandis were treated with water, and the stigmas of Citrus grandis were treated with pollen of C. poonensis after 4 days, and then the stigmas of Citrus grandis were dyed with aniline blue.

[0021] Figures 1-3 In the figure, pt is pollen tube, and vb is vascular bundle. DETAILED DESCRIPTION

[0022] In order to make the purpose, technical scheme and advantages of the present application more clear, the present application is further described in detail below in combination with the drawings and examples. It should be understood that the specific examples described herein are only used to explain the present application, and are not used to limit the present application. Based on the examples in the present application, all other examples obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0023] The so-called selective fertilization in the present application is a phenomenon that sperm nuclei with specific genetic basis preferentially fertilize oocytes, and selective fertilization mainly shows physiological and biochemical and genetic characteristics, including distant hybrid incompatibility, competition among multiple pollen sperm nuclei and the like. In the examples of the present application, distant hybridization between Citrus grandis and C. poonensis is taken as an example for illustration.

[0024] A reagent for overcoming distant hybridization incompatibility of plants in the Rutaceae family, the reagent is a D-Ser solution, and the concentration of the solution is 1-20 mmol / L.

[0025] In the plants in the Rutaceae family described in the present application, the distant hybridization incompatibility of plants in the Rutaceae family is overcome to promote the germination, growth, entry into the stigma and completion of the fertilization process on the stigma. In the present application, the concentration of the D-Ser solution used for different species of plants in the Rutaceae family is slightly different, and can be adjusted according to the actual situation.

[0026] For example, the concentration of the D-Ser solution used for Citrus grandis is 1-20 mmol / L. The use method of the reagent is to spray the reagent twice on the stigma of the bud stage of the plant in the Rutaceae family in the morning, and to add the reagent for the second time before pollination.

[0027] When spraying, the reagent is preferably sprayed in the time period of 6:00-8:00 in the morning on a sunny day, and the interval of adding the reagent is 30 min.

[0028] The reagent for overcoming distant hybridization incompatibility of plants in the Rutaceae family and the use method provided by the present application are described in detail below in combination with examples, but they should not be understood as limiting the protection scope of the present application.

[0029] Example 1

[0030] D-Ser solution was used to treat Citrus Moroc to overcome the cross-incompatibility between Citrus Moroc and Citrus Flavoviridis. Specifically, the day before spraying D-Ser solution, the opened flowers of Citrus Moroc were removed and then bagged for isolation. The next morning, before 8:00, the unopened flower buds were sprayed twice with D-Ser solution at a concentration of 1 mmol / L, and the interval between the two sprays was 30 minutes.

[0031] After the second treatment, the flowers were pollinated with Citrus Flavoviridis pollen and pollination bags were used to prevent contamination by other pollen. Four days later, the pollination bags were removed, the styles were removed and fixed with Carnoy's fixative, and aniline blue staining was used to observe pollen germination.

[0032] Example 2

[0033] The procedure was basically the same as in Example 1, except that the concentration of the D-Ser solution was 5 mmol / L.

[0034] Example 3

[0035] The procedure was basically the same as in Example 1, except that the concentration of the D-Ser solution was 10 mmol / L.

[0036] Example 4

[0037] The procedure was basically the same as in Example 1, except that the concentration of the D-Ser solution was 20 mmol / L.

[0038] Figure 1 The results of aniline blue staining of the stigmas of Citrus Moroc treated with different concentrations of D-Ser solution after 4 days of pollination with Citrus Flavoviridis pollen.

[0039] Comparative Example 1

[0040] The procedure was basically the same as in Example 1, except that the unopened flower buds of Citrus Moroc were not treated with the reagent.

[0041] Figure 2 The results of aniline blue staining of the stigmas of Citrus Moroc that were not treated with the reagent after 4 days of pollination with Citrus Flavoviridis pollen.

[0042] Comparative Example 2

[0043] The procedure was basically the same as in Example 1, except that water was used as the reagent to treat the unopened flower buds of Citrus Moroc.

[0044] Figure 3 The results of aniline blue staining of the stigmas of Citrus Moroc treated with water after 4 days of pollination with Citrus Flavoviridis pollen.

[0045] By Figures 1-3Compared with the contrast, after the D-Ser solution of the application is used to treat the Citrus morrisonicola, the pollen of the Citrus garrawensis can germinate, grow and enter the ovary to complete the fertilization process, and the distant hybridization incompatibility barrier between the Citrus morrisonicola and the Citrus garrawensis can be overcome.

[0046] The above is a further detailed description of the application in combination with specific / preferred embodiments, and cannot be deemed as limiting the specific implementation of the application to these descriptions. For those skilled in the art to which the application belongs, without departing from the concept of the application, some substitutions or modifications can be made to the described embodiments, and these substitutions or modifications shall be deemed as falling within the protection scope of the application.

Claims

1. Use of D-Ser in overcoming cross-species incompatibility in crosses of plants of the family Lamiaceae, characterized in that, The D-Ser refers to a D-Ser solution; at the beginning of flowering of the Rutaceae plant, the D-Ser solution is sprayed on the stigma of a flower at the unopened bud stage.

2. Use of D-Ser according to claim 1 to overcome distant hybridization incompatibility in the family Lamiaceae, characterized by the fact that, The concentration of the D-Ser solution is 1-20 mmol / L.

3. Use of D-Ser according to claim 1 or 2 to overcome distant hybridization incompatibility in the family Lamiaceae, characterized in that, The distant hybridization includes interspecific distant hybridization and intergeneric distant hybridization.

4. A method of using D-Ser as an agent to overcome Lathyrus sp. wide cross incompatibility, characterized by, The method comprises the following steps: at the beginning of flowering of the Rutaceae plant, a flower at the unopened bud stage is selected, a D-Ser solution is sprayed on the stigma of the flower, and then pollination is performed.

5. The method of using an agent to overcome distant hybridization incompatibility in the family Lamiaceae using D-Ser according to claim 4, characterized in that, The concentration of the D-Ser solution is 1-20 mmol / L.

6. The method for overcoming the distant hybridization incompatibility of the plants of the Lamiaceae family using D-Ser according to claim 4 or 5, characterized by the fact that, The D-Ser solution is sprayed on the stigma of the flower at the unopened bud stage twice.

7. The method of using an agent to overcome distant hybridization incompatibility in the family Lamiaceae using D-Ser according to claim 6, characterized in that, The interval between the two times of spraying of the reagent is 30 min. The interval between the two times of spraying of the reagent is 30 min.

Images