CN112450021A - Method for rapidly detecting purity of radish sterile line - Google Patents
Method for rapidly detecting purity of radish sterile line Download PDFInfo
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- CN112450021A CN112450021A CN202011420223.5A CN202011420223A CN112450021A CN 112450021 A CN112450021 A CN 112450021A CN 202011420223 A CN202011420223 A CN 202011420223A CN 112450021 A CN112450021 A CN 112450021A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/25—Root crops, e.g. potatoes, yams, beet or wasabi
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
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Abstract
The invention discloses a method for rapidly detecting the purity of a radish sterile line, which comprises the following steps: (1) selecting plump and robust radish sterile line seeds, then sterilizing the seeds, then spreading the seeds in moist gauze, placing in a germination box at 28 ℃ for accelerating germination for 12 hours, and keeping the seeds for later use after the seeds are exposed to white; (2) according to the character survey of the radish sterile line material in the early stage and the data arrangement statistics of the vernalization experiment, the bolting resistant radish sterile line seeds are placed in a refrigerator at 4 ℃ for 25 days, and the bolting easy radish sterile line seeds are placed in the refrigerator at 4 ℃ for 15 days. During the period, the temperature is controlled accurately and the humidity is controlled timely. (3) After the vernalization treatment is finished, the seeds are sown in the field. Drip irrigation under a film is adopted, water and fertilizer are integrated, and watering is carried out once every 5-10 days. The prevention and control of plant diseases and insect pests are required in the seedling stage, the phosphorus-potassium fertilizer is mainly used in the early flowering stage, and meanwhile, the boron fertilizer is required to be supplemented in the full flowering stage; (4) the material was observed immediately after sowing, and the number of flowers of each material was counted about 2 months after sowing, and the flowering rate (number of flowering plants/total number of seedlings × -100%) was calculated. In addition, the fertility of each plant was recorded at about 10 am on a fine day, and the sterility rate (number of sterile plants/total flowering plants) was calculated.
Description
Technical Field
The invention belongs to the technical field of vegetable genetic breeding, and particularly relates to a method for rapidly detecting the purity of a radish sterile line.
Background
Radish (Raphanus sativus L.) belongs to the genus Raphanus of the family Brassicaceae, and is a primary and secondary annual herbaceous plant. China is the biggest radish producing country in the world, and the planting area is about 1950 ten thousand mu each year. Although the radish cultivation area is large in China, excellent radish varieties are lacked, and the radish varieties are seriously depended on foreign imported varieties, so that the strength of radish breeding is increased by domestic breeding units, and the breeding of the excellent radish varieties is accelerated.
Radish has obvious heterosis, and the hybrid seeds are produced mainly with self-incompatible line and male sterile line. When the self-incompatible line is used for seed production, the self-incompatibility of the parents is easily influenced by high temperature and drought, so that a certain self-incompatibility rate is generated, and the purity of hybrid seeds is influenced. The use of male sterile lines for seed production can solve this problem, not only can improve the purity of hybrid seeds, but also can prevent parent loss and protect the interests of the breeder (Zhangli et al, 2020). The purity of sterile line materials can directly influence later-stage seed reproduction and production, while the conventional male sterile line breeding process has serious season limitation, the fertility identification of hybrid and backcross progeny needs to be carried out in the flowering period of radish in the spring of the second year, and the identification needs to be carried out in the time of one season. Therefore, a method for rapidly identifying the purity of the radish male sterile line material is necessary, the season limitation can be broken through by the method, the purity of the sterile line can be identified after the seeds of the current generation are harvested, and the identification time is greatly shortened.
Disclosure of Invention
The invention aims to overcome the original growth cycle of radish and promote the radish to bloom in advance by vernalizing the seeds of the radish sterile line material, thereby providing a method for rapidly detecting the purity of the radish sterile line, which has short time consumption and simple method.
A method for rapidly detecting the purity of a radish sterile line comprises the following steps:
(1) material preparation
(1.1) selecting radish sterile line material seeds.
And (1.2) accelerating germination of the prepared seeds and germinating the seeds.
(2) Vernalization treatment
The vernalization temperature and the vernalization time have great influence on late bolting and flowering of the radish, and the radish seeds can be subjected to the vernalization stage at 0-6 ℃ for 10-20 days generally, so that bolting and flowering are advanced. Too high or too low temperature and short vernalization time are not beneficial to earlier flowering of radishes, so that the control of the temperature and the time is a powerful measure for early bolting and flowering of radishes.
(2.1) accurate temperature control: the germinated seeds were placed in a refrigerator at a temperature set at 4 ℃.
(2.2) timely humidity control: after the seeds are placed in a refrigerator, the humidity of the surfaces of the seeds needs to be controlled, excessive humidity easily causes excessive growth of the seeds, and excessive humidity can also cause adverse effects on the seeds.
(2.3) according to the character survey of the radish sterile line material in the early stage and the data arrangement statistics of the vernalization experiment, placing the bolting-resistant radish sterile line seeds in a refrigerator at 4 ℃ for 25 days, and placing the radish sterile line seeds easy to bolt in the refrigerator at 4 ℃ for 15 days.
(3) Sowing in field
When sowing, seeds of different materials are respectively sown in different cells. Drip irrigation under a film is adopted, water and fertilizer are integrated, and watering is carried out once in 5-10 days.
(4) Index statistics
The materials were observed immediately after sowing, and the number of flowers of each material was counted about 2 months after sowing, and the flowering rate (number of flowering plants/total number of seedlings × 100%) was calculated. In addition, the fertility of each plant needs to be recorded in time, and the sterility rate (sterile plants/total flowering plants) needs to be calculated, wherein the higher the sterility rate is, the better the purity is.
Further, in the step (1), seeds with full grains and good quality are selected for later-stage operation;
further, observing the materials in the step (2) every day, paying attention to control the surface humidity of the seeds to keep the seeds in the optimal state, placing the bolting resistant radish seeds in a refrigerator for 10 days, then placing the bolting easy radish sterile line materials in the refrigerator for 15 days, and sowing the bolting easy radish sterile line materials at the same time for later statistics;
furthermore, in the step (3), the prevention and control of plant diseases and insect pests, particularly cabbage caterpillars, are required in the seedling stage, and the additional fertilizer is required in the later stage.
Further, the step (4) of observing and counting the sterility rate is performed at about ten am on a fine day.
Compared with the prior art, the invention has the following implementation effects:
(1) compared with the traditional mode, the original method has long time-consuming period and low efficiency; (2) compared with the rapid identification of the molecular marker, although the molecular marker technology can rapidly identify in the seedling stage, the method has the advantages of complex operation, large investment in the early stage and high cost; (3) compared with the common vernalization method, the common vernalization method is rough and has low accuracy.
In conclusion, the method is simple to operate, economical, efficient and short in time consumption, the purity of the sterile line of the radish can be rapidly detected within about 70-80 days, the breeding quality of radish breeding workers can be effectively improved, and the breeding progress is accelerated.
Drawings
FIG. 1 shows sterile radish plants photographed in the field;
FIG. 2 shows fertile plants found in sterile plant material of radish photographed in the field.
Detailed Description
The present invention will be further described with reference to the following detailed description:
(1) material preparation
(1.1) selecting seeds with good color and plump grains from the sterile line material of the current-generation propagated radishes.
(1.2) selecting plump and robust seeds, disinfecting the seeds by a 'warm soup seed soaking method', then spreading the seeds in wet gauze, placing the seeds in a germination box at 28 ℃ for accelerating germination for 12 hours, and placing the seeds in a culture dish paved with filter paper after the seeds are slightly exposed to white.
(2) Vernalization treatment
(2.1) accurate temperature control: a refrigerator with better temperature control condition is selected as the test equipment of this time, and the temperature is set to be 4 ℃. The petri dish containing the seeds was placed in a refrigerator, and an alcohol thermometer was also placed therein, so that the temperature in the refrigerator was monitored at any time at a later stage.
(2.2) timely humidity control: the filter paper should be wetted through before placing the petri dish in the refrigerator. The observation of the filter paper in the culture dish is ensured at least once a day during the whole test period, if the filter paper is dry, the filter paper can be sprayed with moisture so as to keep the filter paper moist, if the humidity is high, the water control is needed, and the observation is carried out at any time.
(2.3) according to the character survey of the radish sterile line material at the early stage and the arrangement statistics of experimental data, the radish sterile line material can be divided into bolting resistant type and bolting easy type, the bolting resistant radish sterile line seeds are firstly placed in a refrigerator at 4 ℃ for 10 days, and then the bolting easy radish sterile line seeds are placed in the refrigerator at 4 ℃ for continuous vernalization for 15 days.
(3) Sowing in field
After the vernalization treatment is finished, seeds of the two materials are respectively sown in different communities. Drip irrigation under a film is adopted, water and fertilizer are integrated, and watering is carried out once in 5-10 days. The prevention and control of plant diseases and insect pests are required in the seedling stage, the phosphorus-potassium fertilizer is mainly used in the early flowering stage, and the boron fertilizer is required to be supplemented in the full flowering stage.
(4) Index statistics
The number of flowering of each material was counted about 2 months after sowing, and the flowering rate (number of flowering plants/total number of seedlings generated ×. 100%) was calculated. In addition, fertility of all flowering plants of each material was counted at about 10 am on a sunny day, and the sterility rate (number of sterile plants/number of all flowering plants: 100%) was calculated (table 1).
TABLE 1 flowering and sterility rates of different materials
The above is only a specific application example of the present invention, and the protection scope of the present invention is not limited in any way. All the technical solutions formed by equivalent transformation or equivalent replacement fall within the protection scope of the present invention.
Claims (5)
1. A method for rapidly detecting the purity of a radish sterile line is characterized by comprising the following steps:
(1) material preparation
(1.1) selecting radish sterile line material seeds.
And (1.2) accelerating germination of the prepared seeds and germinating the seeds.
(2) Vernalization treatment
(2.1) accurate temperature control: the germinated seeds were placed in a refrigerator at a temperature set at 4 ℃.
(2.2) timely humidity control: after the seeds are placed in a refrigerator, the humidity of the surfaces of the seeds needs to be controlled, excessive humidity easily causes excessive growth of the seeds, and excessive humidity can also cause adverse effects on the seeds.
(2.3) according to the character survey of the radish sterile line material in the early stage and the data arrangement statistics of the vernalization experiment, placing the bolting-resistant radish sterile line seeds in a refrigerator at 4 ℃ for 25 days, and placing the radish sterile line seeds easy to bolt in the refrigerator at 4 ℃ for 15 days.
(3) Sowing in field
When sowing, seeds of different materials are respectively sown in different cells. Drip irrigation under a film is adopted, water and fertilizer are integrated, and watering is carried out once in 5-10 days.
(4) Index statistics
The materials were observed immediately after sowing, and the number of flowers of each material was counted about 2 months after sowing, and the flowering rate (number of flowering plants/total number of seedlings × 100%) was calculated. In addition, the fertility of each plant should be recorded in time and the sterility rate (number of sterile plants/number of full-flowering plants + 100%) calculated.
2. The method for rapidly detecting the purity of the radish sterile line according to claim 1, which is characterized in that: in the step (1), seeds with full grains and good quality are selected for later operation.
3. The method for rapidly detecting the purity of the radish sterile line according to claim 1, which is characterized in that: observing the materials in the step (2) every day, paying attention to control the surface humidity of the seeds to keep the seeds in the optimal state, placing the bolting resistant radish seeds in a refrigerator for 10 days, then placing the bolting easy radish sterile line materials in the refrigerator for 15 days, and sowing the bolting easy radish sterile line materials at the same time for later statistics.
4. The method for rapidly detecting the purity of the radish sterile line according to claim 1, which is characterized in that: in the step (3), attention is paid to prevention and control of plant diseases and insect pests, particularly cabbage caterpillars, in the seedling stage, and regular additional fertilization is carried out in the later stage.
5. The method for rapidly detecting the purity of the radish sterile line according to claim 1, which is characterized in that: in the step (4), the sterility rate is observed and counted about ten am in a fine day.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1054700A (en) * | 1990-03-07 | 1991-09-25 | 山西省农业科学院蔬菜研究所 | Practical hybrid seed production technology for high-yield and high-quality radishes |
| WO2003006622A2 (en) * | 2001-07-12 | 2003-01-23 | Mcgill University | Nuclear fertility restorer genes and methods of use in plants |
| CN1781357A (en) * | 2004-11-29 | 2006-06-07 | 中国农业大学 | Method for producing carrot hybrid seed and appraising purity |
| CN103609433A (en) * | 2013-11-01 | 2014-03-05 | 山东省农业科学院蔬菜花卉研究所 | Breeding method for bolting resistant radish selfing line |
| CN104170723A (en) * | 2014-08-12 | 2014-12-03 | 江苏沿海地区农业科学研究所 | Breeding method of heat-resistant red-skin radish |
| CN105830907A (en) * | 2016-02-29 | 2016-08-10 | 山东省农作物种质资源中心 | Genic male sterile line of radish and breeding method thereof |
| CN109691370A (en) * | 2018-12-18 | 2019-04-30 | 武汉市农业科学院 | A kind of radish quickly adds the method in generation |
-
2020
- 2020-12-07 CN CN202011420223.5A patent/CN112450021A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1054700A (en) * | 1990-03-07 | 1991-09-25 | 山西省农业科学院蔬菜研究所 | Practical hybrid seed production technology for high-yield and high-quality radishes |
| WO2003006622A2 (en) * | 2001-07-12 | 2003-01-23 | Mcgill University | Nuclear fertility restorer genes and methods of use in plants |
| CN1781357A (en) * | 2004-11-29 | 2006-06-07 | 中国农业大学 | Method for producing carrot hybrid seed and appraising purity |
| CN103609433A (en) * | 2013-11-01 | 2014-03-05 | 山东省农业科学院蔬菜花卉研究所 | Breeding method for bolting resistant radish selfing line |
| CN104170723A (en) * | 2014-08-12 | 2014-12-03 | 江苏沿海地区农业科学研究所 | Breeding method of heat-resistant red-skin radish |
| CN105830907A (en) * | 2016-02-29 | 2016-08-10 | 山东省农作物种质资源中心 | Genic male sterile line of radish and breeding method thereof |
| CN109691370A (en) * | 2018-12-18 | 2019-04-30 | 武汉市农业科学院 | A kind of radish quickly adds the method in generation |
Non-Patent Citations (2)
| Title |
|---|
| 李盛: "春化处理对萝卜抽薹及开花的影响", 《中国优秀博硕士学位论文全文数据库(硕士)农业科技辑》 * |
| 鲍文秀等: "《作物杂交制种新技术》", 31 January 2000, 北京出版社 * |
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