Screening method of semi-lethal dose of Co-gamma ray mutagenesis beet seeds and method for promoting germination rate
The 60Co-γ ray mutagenesis technology was used to radiation treatment of beet seeds and screen out semi-lethal doses, which solved the problem of insufficient genetic diversity of beet germplasm resources and improved the mutation frequency and quality of beet seeds.
Patent Information
- Application Number
- CN202510255636.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-03-05
- Publication Date
- 2025-04-25
AI Technical Summary
The genetic background of existing beet varieties is relatively narrow, and the genetic diversity of germplasm resources is low, which limits the breeding of new varieties in terms of beet quality, disease resistance and stress resistance.
Through 60Co-γ ray mutagenesis technology, beet seeds were radiated and semi-lethal doses were screened to improve the germination rate and survival rate of beet seeds, thereby expanding the mutation spectrum of beet germplasm.
The mutation frequency of beet seeds has been improved, the genetic diversity of germplasm resources has been expanded, the breeding of beet varieties has been promoted, and the quality, disease resistance and stress resistance of beets have been improved.
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Figure CN119866928A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to a 60 The screening index and method of improving germination rate and half-lethal dose of Co-γ-ray induced mutagenesis of agricultural beet seeds were studied. 60 The median lethal dose of Co-γ-ray irradiation was determined by its germination rate and survival rate. Background Art
[0002] Beet (Beta vulgaris L.) is an important economic crop belonging to the genus Beta in the family Amaranthaceae. Beet is mainly divided into sugar beets, feed beets, vegetable beets and leaf beets. Beet is famous for its roots rich in sugar. The main use of sugar beets is to extract sucrose, and its output occupies an important position in the global sucrose production. Beet is not only widely grown as a raw material for sugar production, but also has multiple uses and economic value. The sugar extracted from beet roots can be used in the food industry, beverage production, and pharmaceutical manufacturing. Beet by-products, such as beet pulp and beet leaves, can also be used as feed. In addition, beets are rich in various vitamins and minerals, which have many benefits for human health.
[0003] Sugar beet cultivation in my country is mainly concentrated in the north and northwest regions, where the climate and soil conditions are suitable for sugar beet growth. The planting area of sugar beet in China is about 1 million hectares, with an annual output of about 15 million tons. China is one of the world's important sugar beet producers, and the sugar beet industry occupies an important position in the national sugar production. The progress of the sugar beet industry depends on the cultivation of high-quality varieties. However, the sugar beet strains currently planted in my country mainly rely on foreign imports, and the fundamental basis for cultivating new sugar beet varieties lies in the utilization of germplasm resources. The genetic background of existing sugar beet varieties is relatively narrow, and the genetic diversity of germplasm resources is low, which limits my country's selection and breeding of new varieties in terms of sugar beet quality, disease resistance and stress resistance. Therefore, it is particularly important to strengthen investment in artificial breeding of sugar beet germplasm resources.
[0004] Radiation breeding is a method of biological breeding. Under artificially controlled conditions, seeds are irradiated using physical radiation mutagenesis methods such as neutrons, protons or rays to induce changes in the number and structure of chromosomes, thereby affecting plant behavior variations, thereby obtaining mutants that can be used for production, and on this basis, further cultivating new germplasm breeding technology. γ-rays have become the most common radiation ray for constructing mutant libraries due to their strong penetrating properties. γ-ray mutagenesis has low cost and high mutation rate, which is 100 to 1,000 times higher than the natural mutation rate. It is easy to cause plant morphological structure and physiological and biochemical variations, and new varieties can be selected in a short time. The breeding prospects are broad. Radiation mutagenesis technology is a technology that uses various rays (such as X-rays, γ-rays, heavy ions, neutrons, etc.) to irradiate seeds, plants, organs and tissues of crops, prompting them to produce various mutations, and then screen out target mutants from them, and directly or indirectly cultivate new varieties.
[0005] The choice of radiation dose should be based on the specific conditions of the biological breeding goals and biological species. Too high a dose will kill a large number of cells and organisms, or produce more chromosomal aberrations, while too low a dose will make it difficult to obtain mutants. The dose rate is also an important factor affecting the mutation effect. A suitable dose rate may obtain more valuable mutants. In the selection of mutagenesis, the half-lethal dose is usually considered to be the optimal radiation dose. Summary of the invention
[0006] The purpose of the present invention is to determine the optimal radiation dose for radiating agricultural beet seeds to increase the germination rate of the beet seeds, and to provide a method for 60 A method for screening the semi-lethal dose of Co-γ ray induced mutagenesis in sugar beet seeds and a mutagenesis method for promoting the germination rate.
[0007] The present invention 60 The screening method of the half-lethal dose of Co-γ-ray induced mutagenesis beet seeds is implemented according to the following steps:
[0008] Step 1: Divide a number of diploid sugar beet (agricultural) seeds with full and uniform grains into N groups, of which N-1 group is the experimental group and one group is the control group (i.e., no experiment is performed). 60 Co-γ irradiation treatment), and the sugar beet seeds of each experimental group were 60 Co-γ irradiation treatment, each experimental group 60 The Co-γ radiation dose was different, and the radiation dose of each experimental group was controlled to be 100Gy to 400Gy, and the irradiated sugar beet seeds were obtained;
[0009] Step 2: using alcohol to disinfect the irradiated beet seeds and the control group beet seeds respectively, then repeatedly washing the beet seeds with distilled water, and immersing the washed beet seeds in distilled water to obtain washed beet seeds;
[0010] Step 3: Use the paper roll method to test the germination of the washed beet seeds. The ambient temperature is controlled at 25°C in a constant temperature incubator. The lighting conditions are 12 hours of light and 12 hours of darkness per day. The germination status is recorded and the seedling rates of the control group and each experimental group are counted.
[0011] Step 4: The control group and each experimental group 60 The Co-γ radiation dose is the horizontal axis, the seedling rate is the vertical axis, and the fitting curve is based on the corresponding value when the seedling rate is 50%. 60 The Co-γ radiation dose was taken as the median lethal dose.
[0012] The present invention 60 The method for promoting germination rate of beet seeds induced by Co-γ ray mutagenesis is achieved by the following steps:
[0013] Select a number of diploid beet seeds with full and uniform grains, sterilize them with alcohol, wash them with distilled water, and use 60 Co-γ ray irradiation treatment of sugar beet seeds to control 60 The Co-γ radiation dose is 90Gy to 110Gy, which promotes the germination rate of sugar beet seeds.
[0014] The invention uses radiation mutagenesis to treat beet seeds, so that the breeding efficiency is significantly increased. 60 Co-γ ray treatment can increase the mutation frequency of plants and expand their mutation spectrum. As the dose increases, the damage to the seeds gradually increases. Too large a radiation dose will kill the beet plants, and too small a radiation dose will not have a mutagenic effect on the plants. Therefore, the present invention determines the semi-lethal dose of beet seeds.
[0015] Radiation breeding is one of the breeding methods, which induces changes in the number and structure of chromosomes and then affects plant behavior variation, thereby obtaining mutants that can be used for production, because it has the advantages of convenience, easy operation and environmental friendliness. Compared with traditional breeding methods, physical mutagenesis has the characteristics of relatively simple methods, breaking trait linkage, completing gene recombination, multiple mutation types, high mutation frequency, stable traits, and a large number of mutants can be obtained in a short period of time. After the agricultural beet seeds are irradiated by the present invention, laboratory observations are carried out. The results of observations under the doses of 0Gy, 100Gy, 200Gy, 300Gy and 400Gy show that the germination indicators related to diploid beets all show an obvious trend of first increasing and then decreasing. When the radiation dose is 100Gy, the germination of seeds is promoted. When the dose reaches 300Gy, the growth of beet seeds is strongly inhibited; the mortality rate is positively correlated with the radiation dose. The results of linear regression equation analysis show that the half-lethal dose of diploid beet seeds is 348Gy. BRIEF DESCRIPTION OF THE DRAWINGS
[0016] Figure 1 The photos are photos of the growth of seedlings after being treated with different radiation doses in the embodiment;
[0017] Figure 2 The photos are of the germination of seedlings after being treated with different radiation doses in the embodiment;
[0018] Figure 3 It is the regression equation diagram of radiation dose and seedling rate in the embodiment. DETAILED DESCRIPTION
[0019] Specific implementation method 1: This implementation method 60 The screening method of the half-lethal dose of Co-γ-ray induced mutagenesis beet seeds is implemented according to the following steps:
[0020] Step 1: Divide a number of diploid sugar beet (agricultural) seeds with full and uniform grains into N groups, of which N-1 group is the experimental group and one group is the control group (i.e., no experiment is performed). 60 Co-γ irradiation treatment), and the sugar beet seeds of each experimental group were 60 Co-γ irradiation treatment, each experimental group 60 The Co-γ radiation dose was different, and the radiation dose of each experimental group was controlled to be 100Gy to 400Gy, and the irradiated sugar beet seeds were obtained;
[0021] Step 2: using alcohol to disinfect the irradiated beet seeds and the control group beet seeds respectively, then repeatedly washing the beet seeds with distilled water, and immersing the washed beet seeds in distilled water to obtain washed beet seeds;
[0022] Step 3: Use the paper roll method to test the germination of the washed beet seeds. The ambient temperature is controlled at 25°C in a constant temperature incubator. The lighting conditions are 12 hours of light and 12 hours of darkness per day. The germination status is recorded and the seedling rates of the control group and each experimental group are counted.
[0023] Step 4: The control group and each experimental group 60 The Co-γ radiation dose is the horizontal axis, the seedling rate is the vertical axis, and the fitting curve is based on the corresponding value when the seedling rate is 50%. 60 The Co-γ radiation dose was taken as the median lethal dose.
[0024] Specific implementation method 2: This implementation method is different from specific implementation method 1 in that the number of diploid sugar beet seeds in the experimental group in step 1 is 200 to 400.
[0025] Specific implementation method three: This implementation method is different from specific implementation methods one or two in that the control in step one 60 Co-γ radiation dose rate is 5Gy·min-1 .
[0026] Specific implementation method 4: This implementation method is different from specific implementation methods 1 or 2 in that in step 1, a number of diploid sugar beet (agricultural) seeds with full and uniform grains are divided into 5 to 8 groups.
[0027] Specific implementation method 5: This implementation method is different from specific implementation methods 1 to 4 in that the disinfection treatment time in step 2 is 5 minutes.
[0028] Specific implementation method 6: This implementation method is different from specific implementation methods 1 to 5 in that the soaking treatment time in step 2 is 40 to 50 hours.
[0029] Specific implementation method seven: This implementation method is different from any one of specific implementation methods one to six in that in step three, moistened paper is used to wrap the beet seeds to conduct a germination test on the washed beet seeds.
[0030] Specific implementation example eight: This implementation example is different from specific implementation examples one to seven in that the germination condition is recorded in step three, and the germination standard is that the radicle length exceeding 1 mm.
[0031] Specific implementation method 9: This implementation method 60 The method for promoting germination rate of beet seeds induced by Co-γ ray mutagenesis is implemented according to the following steps:
[0032] Select a number of diploid beet seeds with full and uniform grains, sterilize them with alcohol, wash them with distilled water, and use 60 Co-γ ray irradiation treatment of sugar beet seeds to control 60 The Co-γ radiation dose is 90Gy to 110Gy, which promotes the germination rate of sugar beet seeds.
[0033] Specific implementation method ten: This implementation method is different from the specific implementation method nine in that the 60 Co-γ ray irradiation treatment of sugar beet seeds to control 60 The Co-γ radiation dose was 100 Gy.
[0034] Embodiment 1: This embodiment 60 The screening method of the half-lethal dose of Co-γ-ray induced mutagenesis beet seeds is implemented according to the following steps:
[0035] Step 1: Select a number of diploid agricultural beet seeds with full and uniform grains and divide them into five groups (N=5), of which four groups are experimental groups and one group is a control group, with 300 seeds in each group. 60Co-γ irradiation treatment, the radiation doses of the four experimental groups were 100Gy, 200Gy, 300Gy and 400Gy respectively, and the control 60 Co-γ radiation dose rate is 5Gy·min -1 , without 60 Sugar beet seeds treated with Co-γ irradiation served as the control group (WT), and irradiated sugar beet seeds were obtained;
[0036] Step 2, using alcohol with a volume concentration of 75% to disinfect the irradiated beet seeds and the control group beet seeds for 5 minutes, then repeatedly washing the beet seeds with distilled water for 5 times, and immersing the washed beet seeds in distilled water for 48 hours to obtain washed beet seeds;
[0037] Step 3: Use the roll paper method to perform a germination test on the washed beet seeds, test 100 seeds in each group, repeat 3 times, control the ambient temperature at 25°C in a constant temperature incubator, and alternate the lighting conditions of 12 hours of light and 12 hours of darkness per day. Record the germination of the seeds every 24 hours, and use the radicle length exceeding 1 mm as the germination standard until all seeds have germinated. Statistic the germination rate (15th day), germination potential (germination rate on the 6th day), germination index and seedling rate (25th day) of the experimental group and the control group;
[0038] Step 4: According to the growth of plants, the control group and each experimental group 60 The Co-γ radiation dose is the horizontal axis, the seedling rate is the vertical axis, and the fitting curve is based on the corresponding value when the seedling rate is 50%. 60 The Co-γ radiation dose was taken as the median lethal dose.
[0039] The effects of different radiation doses on the seedling rate and germination index of sugar beet seeds are statistically shown in Tables 1 and 2 below.
[0040] Table 1 Effects of different radiation doses on the seedling rate of sugar beet seeds
[0041]
[0042] Table 2 Effects of different radiation doses on germination indexes of diploid sugar beet seeds
[0043]
[0044] Note: Different lowercase letters in the same column indicate significant differences at the P < 0.05 level
[0045] Embodiment 2: This embodiment 60 The method for promoting germination rate of beet seeds induced by Co-γ ray mutagenesis is implemented according to the following steps:
[0046] Select a number of beet seeds with full and uniform grains, sterilize them with alcohol, wash them with distilled water, and use 60 The sugar beet seeds were irradiated with Co-γ rays and the radiation dose was controlled to be 100 Gy, thereby increasing the germination rate of the sugar beet seeds.
[0047] The results of the observations under the treatments of 0Gy, 100Gy, 200Gy, 300Gy and 400Gy showed that the germination indexes of diploid beet showed an obvious trend of first increasing and then decreasing. When the radiation dose was 100Gy, the germination of seeds was promoted. When the dose reached 300Gy, the growth of beet seeds was strongly inhibited. The mortality rate of beet seedlings was positively correlated with the radiation dose. The results of linear regression equation analysis showed that the median lethal dose of diploid beet was 348Gy. Optimization of promoting germination rate 60 The Co-γ-ray radiation dose was 100 Gy.
Claims
1. A method for screening semi-lethal doses of Co-γ-ray induced beet seeds, characterized in that The screening method of the semi-lethal dose of Co-γ ray induced beet seeds is achieved by the following steps: Step 1: Divide a number of diploid sugar beet seeds with full and uniform grains into N groups, of which N-1 group is the experimental group and one group is the control group. 60 Co-γ irradiation treatment, each experimental group 60 The Co-γ radiation dose was different, and the radiation dose of each experimental group was controlled to be 100Gy to 400Gy, and the irradiated sugar beet seeds were obtained; Step 2: using alcohol to disinfect the irradiated beet seeds and the control group beet seeds respectively, then repeatedly washing the beet seeds with distilled water, and immersing the washed beet seeds in distilled water to obtain washed beet seeds; Step 3: Use the paper roll method to test the germination of the washed beet seeds. The ambient temperature is controlled at 25°C in a constant temperature incubator. The lighting conditions are 12 hours of light and 12 hours of darkness per day. The germination status is recorded and the seedling rates of the control group and each experimental group are counted. Step 4: The control group and each experimental group 60 The Co-γ radiation dose is the horizontal axis, the seedling rate is the vertical axis, and the fitting curve is based on the corresponding value when the seedling rate is 50%. 60 The Co-γ radiation dose was taken as the median lethal dose.
2. The method for screening the semi-lethal dose of Co-γ ray induced beet seeds according to claim 1, characterized in that The number of diploid sugar beet seeds in the experimental group in step 1 is 200 to 400.
3. The method for screening the semi-lethal dose of Co-γ ray induced beet seeds according to claim 1, characterized in that Step 1 Control 60 Co-γ radiation dose rate is 5Gy·min -1 .
4. The method for screening the semi-lethal dose of Co-γ ray induced beet seeds according to claim 1, characterized in that In step 1, a number of diploid sugar beet seeds with full and uniform grains are divided into 5 to 8 groups.
5. The method for screening the semi-lethal dose of Co-γ ray induced beet seeds according to claim 1, characterized in that The disinfection treatment time in step 2 is 5 minutes.
6. The method for screening the semi-lethal dose of Co-γ ray induced beet seeds according to claim 1, characterized in that The soaking treatment time in step 2 is 40 to 50 hours.
7. The method for screening the semi-lethal dose of Co-γ ray induced beet seeds according to claim 1, characterized in that In step three, the washed beet seeds are tested for germination by wrapping them in moistened paper.
8. The method for screening the semi-lethal dose of Co-γ ray induced beet seeds according to claim 1, characterized in that In step three, the germination status is recorded, and the germination standard is that the radicle length exceeds 1 mm.
9. 60 The method for promoting germination rate of beet seeds by Co-γ ray mutagenesis is characterized in that 60 The method for promoting germination rate of beet seeds induced by Co-γ ray mutagenesis is achieved by the following steps: Select a number of diploid beet seeds with full and uniform grains, sterilize them with alcohol, wash them with distilled water, and use 60 Co-γ ray irradiation treatment of sugar beet seeds to control 60 The Co-γ radiation dose is 90Gy to 110Gy, which promotes the germination rate of sugar beet seeds.
10. The method according to claim 9 60 The method for promoting germination rate of beet seeds by Co-γ ray mutagenesis is characterized in that use 60 Co-γ ray irradiation treatment of sugar beet seeds to control 60 The Co-γ radiation dose was 100 Gy.