Method for rapidly breeding new variety of medicinal locust tree

Abstract

The invention discloses a method for rapidly breeding a new variety of medicinal locust tree, which comprises the following steps: (1) collecting locust tree branches in different regions, grafting in the same region, and keeping the stock age and the later-period management measure consistent; (2) performing continuous regular monitoring, adaptive observation and botanical shape continuous observation after grafting survival; (3) according to monitoring and observation results of 3-5 years, excellent plants with fast growth vigor, strong resistance and obvious biological characters are screened out; (4) determining the rutin content in the sophora flowers of the screened plants, and screening out the plants with high rutin content; and (5) detecting the expression levels of rutin synthesis related genes and degradation genes in the sophora flower buds of the screened plants, and finally determining the sophora flower buds with high synthetic gene expression quantity and low degradation gene expression quantity as excellent varieties. According to the method for rapidly breeding the new variety of the medicinal locust tree, evaluation is more comprehensive, excellent varieties can be screened out more easily, and the breeding time is greatly shortened.

Description

Technical Field

[0001] The invention relates to the technical field of new plant variety breeding, and in particular to a method for rapidly breeding new medicinal locust tree varieties. Background Art

[0002] Sophora japonica is a tree species that combines timber and ornamental purposes. It is widely planted in southern and northern my country for its strong adaptability, long life, and beautiful tree shape. It is especially used as the main greening and ornamental tree species in northern my country. Sophora japonica L. is a dried flower bud of the leguminous plant Sophora japonica L. It is a special Chinese medicinal material rich in chemical components such as rutin and flavonoids. Compared with other plants, Sophora japonica L. has a high rutin content and low extraction cost, and is the main raw material for industrial extraction of rutin. The most prominent problem of medicinal Sophora japonica is that although the Sophora japonica germplasm resources in the country are extremely rich, the content of active ingredients in many germplasms does not meet the national standard (the Chinese Pharmacopoeia requires that the rutin content in Sophora japonica L. is higher than 15%), and the development and utilization value is low. At present, the commonly used breeding method is to monitor the rutin in its Sophora japonica L. and then breed new varieties. Sophora japonica takes more than ten years to bloom, so the breeding time of traditional new varieties is extremely long. How to shorten the breeding time of new varieties has become an urgent problem to be solved.

[0003] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention and should not be regarded as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art. Summary of the invention

[0004] The purpose of the present invention is to provide a method for rapidly breeding new varieties of medicinal Sophora japonica, thereby overcoming the disadvantage that the existing breeding time of new Sophora japonica varieties is long.

[0005] To achieve the above object, the present invention provides a method for rapidly breeding new varieties of medicinal Sophora japonica, comprising the following steps:

[0006] (1) Collect locust tree branches from different regions and graft them in the same area, keeping the rootstock age and subsequent management measures consistent;

[0007] (2) Continuous regular monitoring, adaptation observation and botany morphological observation after grafting;

[0008] (3) Based on the monitoring and observation results of step (2) for 3-5 years, select excellent plants with fast growth, strong resistance and obvious biological traits;

[0009] (4) measuring the rutin content in the flowers of Sophora japonica of the plants screened in step (3) to screen out plants with high rutin content;

[0010] (5) detecting the expression levels of rutin synthesis-related genes and degradation genes in the Sophora japonica seeds of the plants selected in step (3), selecting those with high expression levels of synthesis genes and low expression levels of degradation genes, and finally determining them as excellent varieties.

[0011] Preferably, in the above technical solution, step (1) screens healthy branches, removes branches that are too thick or too thin, and keeps the thickness and age of branches in different areas basically consistent.

[0012] Preferably, in the above technical solution, the selected Sophora japonica branches have a stem thickness of 0.6-1 cm and are annual branches that bloom in the current year.

[0013] Preferably, in the above technical solution, the rootstock is selected from seedlings grown from local Sophora japonica seeds, and the seedling age is 1-2 years.

[0014] Preferably, in the above technical solution, the grafting is carried out by branch grafting in March during the Spring Festival.

[0015] Preferably, in the above technical solution, pest and disease control is carried out in time in the later stage management, compound fertilizer is applied once in April and June each year, the amount of fertilizer applied each time is 30-50g / plant, and the new shoots are toppled once in May each year when they are 15-25cm long.

[0016] Preferably, in the above technical solution, the regular detection in step (2) includes detecting the diameter and number of branches of the plant.

[0017] Preferably, in the above technical solution, the adaptability observation in step (2) includes adaptability to high temperature weather in summer, adaptability to low temperature weather in winter and the occurrence of diseases and insect pests. That is, observe the resistance traits of the locust tree such as cold resistance, drought resistance, disease and insect pest resistance, etc.

[0018] Preferably, in the above technical solution, the continuous observation of botanical traits in step (2) includes plant branch color, spots on branches, leaf traits, leaf color, upright growth, new shoot germination ability, early flowering, length of flowering period, flower spike size, sophora japonica seed size or other relatively stable and specific shapes. That is, observe whether the sophora japonica tree has special spots, upright growth, new shoot germination ability, early flowering, length of flowering period and other biological traits; economic traits such as the size of the flower spike and the size of the sophora japonica seed.

[0019] Preferably, in the above technical solution, the observation of step (2) also includes the content of rutin compounds and brass compounds in Sophora japonica. The content of flavonoid compounds such as rutin is a chemical property. The specific parameters are compared macroscopically based on the measurement results.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] (1) The method for rapidly breeding new varieties of medicinal Sophora japonica trees of the present invention not only takes into account growth potential, resistance and rutin content, but also takes into account regulatory genes in the rutin synthesis and accumulation pathways. The combination of external traits and internal regulatory mechanisms makes the evaluation more comprehensive and the subsequent performance more stable.

[0022] (2) The method of the present invention takes into account the regulatory genes of rutin synthesis during the breeding process. In addition to the high expression of the synthetic genes, the increase in rutin content also takes into account the role of the degradation genes, thereby evaluating the synthesis mechanism of rutin from the two aspects of synthesis and accumulation, making it easier to screen out excellent varieties. By revealing the differences in the synthesis mechanisms of different germplasms, the breeding time is greatly shortened. DETAILED DESCRIPTION

[0023] The specific implementation modes of the present invention are described in detail below in conjunction with specific embodiments, but it should be understood that the protection scope of the present invention is not limited by the specific implementation modes.

[0024] Example

[0025] A method for rapidly breeding new varieties of medicinal Sophora japonica trees comprises the following steps:

[0026] (1) Collect healthy branches of Sophora japonica trees from different regions, remove branches that are too thick or too thin, and keep the thickness and age of branches in different regions basically the same. Specifically, select one-year-old branches that are about 0.8 cm thick and bloom in the same year.

[0027] (2) When grafting is carried out in the same area, the rootstock age and subsequent management measures should remain consistent.

[0028] The rootstock is selected from the seedlings cultivated from local locust tree seeds, with an age of 1-2 years. The grafting method is uniformly used, and the grafting is uniformly carried out in March in spring. After grafting, the later management is carried out, which mainly includes timely prevention and control of diseases and insect pests (conventional methods); compound fertilizer is applied once in April and June every year, and the amount of fertilizer applied each time is 30-50g / plant. Top it once in May every year when the new shoots are about 20cm long.

[0029] (3) After the graft is established, regular monitoring should be carried out to monitor the diameter, number of branches, etc.

[0030] (4) Continuous observation after the grafting is successful, including the adaptability to high temperatures in summer, low temperatures in winter, and the occurrence of diseases and insect pests.

[0031] (5) After the grafting is successful, the botanical traits are continuously observed, including the color of the plant branches, spots on the branches, leaf traits, leaf color, upright growth, new shoot germination ability, early or late flowering, duration of flowering, flower spike size, Sophora japonica bud size, and other relatively stable and specific traits.

[0032] During the monitoring and observation process of steps (3) to (5), the following aspects are observed as key points: resistance traits such as cold resistance, drought resistance, and resistance to diseases and insect pests. Biological traits such as the presence or absence of special spots, upright growth, new shoot germination ability, early or late flowering, and length of flowering period; economic traits such as the size of the flower spike and the size of the Sophora japonica seed. Chemical traits such as the content of flavonoids such as rutin. Specific parameters are compared macroscopically based on the measurement results.

[0033] (6) Based on the above 3-5 years of monitoring and observation results, select excellent plants with fast growth, strong resistance and obvious biological traits.

[0034] (7) The rutin content in the flowers of the selected excellent plants is measured to select excellent plants with high rutin content.

[0035] (8) The expression levels of rutin synthesis-related genes and degradation genes in the sophora japonica seeds of the selected excellent plants with high rutin content were tested, and the ones with high expression of synthesis genes and low expression of degradation genes were selected, and finally determined as excellent varieties.

[0036] Control example: The existing conventional breeding of new Sophora japonica varieties includes: collecting Sophora japonica seeds from different regions, then using the seeds to cultivate seedlings, observing their biological characteristics during the growth of the seedlings, and only after the seedlings bloom can their flower spikes and effective ingredient content be determined. However, it takes more than 10 years for the seedlings to bloom, so the breeding cycle is extremely long.

[0037] The method for rapidly breeding new varieties of medicinal Sophora japonica of the present invention, in addition to considering growth potential, resistance and rutin content, also considers the regulatory genes in the synthesis and accumulation pathways of rutin, combines the external traits with the internal regulatory mechanism, and its evaluation is more comprehensive, and its later performance is also more stable. In the breeding process, the regulatory genes of rutin synthesis are considered, and the increase of rutin content, in addition to the high expression of synthetic genes, also considers the role of degradation genes, thereby evaluating the synthetic mechanism of rutin from two aspects of synthesis and accumulation, and it is easier to screen out excellent varieties. By revealing the differences in the synthetic mechanisms of different germplasms, the breeding time is greatly shortened.

[0038] The foregoing description of specific exemplary embodiments of the present invention is for the purpose of illustration and demonstration. These descriptions are not intended to limit the present invention to the precise form disclosed, and it is clear that many changes and variations can be made based on the above teachings. The purpose of selecting and describing the exemplary embodiments is to explain the specific principles of the present invention and its practical application, so that those skilled in the art can realize and utilize various different exemplary embodiments of the present invention and various different selections and changes. The scope of the present invention is intended to be limited by the claims and their equivalents.

Claims

1. A method for rapidly breeding new varieties of medicinal Sophora japonica, characterized in that: The following steps are involved: (1) Collect locust tree branches from different regions and graft them in the same area, keeping the rootstock age and subsequent management measures consistent; (2) Continuous regular monitoring, adaptation observation and botany morphological observation after grafting; (3) Based on the monitoring and observation results of step (2) for 3-5 years, select excellent plants with fast growth, strong resistance and obvious biological traits; (4) measuring the rutin content in the flowers of Sophora japonica of the plants screened in step (3) to screen out plants with high rutin content; (5) detecting the expression levels of rutin synthesis-related genes and degradation genes in the Sophora japonica seeds of the plants selected in step (3), selecting those with high expression levels of synthesis genes and low expression levels of degradation genes, and finally determining them as excellent varieties.

2. The method for rapidly breeding new varieties of medicinal Sophora japonica according to claim 1, characterized in that: Step (1) screen healthy branches, remove branches that are too thick or too thin, and keep the thickness and age of branches in different areas basically the same.

3. The method for rapidly breeding new varieties of medicinal Sophora japonica according to claim 2, characterized in that: The selected Sophora japonica branches have a stem thickness of 0.6-1 cm and are annual branches that bloom in the same year.

4. The method for rapidly breeding new varieties of medicinal Sophora japonica according to claim 1, characterized in that: Step (1) The rootstock is selected from seedlings grown from local Sophora japonica seeds, which are 1-2 years old and are grafted in March.

5. The method for rapidly breeding new varieties of medicinal Sophora japonica according to claim 1, characterized in that: The grafting in step (1) is carried out by branch grafting and is carried out in March during the Spring Festival.

6. The method for rapidly breeding new varieties of medicinal Sophora japonica according to claim 1, characterized in that: Later management includes timely disease and pest control, applying compound fertilizer once in April and June each year, with each application amount of 30-50g / plant, and topping once in May each year when the new shoots are 15-25cm long.

7. The method for rapidly breeding new varieties of medicinal Sophora japonica according to claim 1, characterized in that: The regular inspection in step (2) includes inspecting the diameter and number of branches of the plants.

8. The method for rapidly breeding new varieties of medicinal Sophora japonica according to claim 1, characterized in that: The adaptability observation in step (2) includes adaptability to high temperature weather in summer, adaptability to low temperature weather in winter and the outbreak of diseases and insect pests.

9. The method for rapidly breeding new varieties of medicinal Sophora japonica according to claim 1, characterized in that: The continuous observation of botanical traits in step (2) includes plant branch color, spots on branches, leaf traits, leaf color, upright growth, new shoot germination ability, early or late flowering, length of flowering period, flower spike size, Sophora japonica bud size or other relatively stable and specific shapes.

10. The method for rapidly breeding new varieties of medicinal Sophora japonica according to claim 1, characterized in that: The observation in step (2) also includes the content of rutin compounds and brass compounds in Sophora japonica flowers.