A method for propagating a strong winter self-incompatible brassica material

Abstract

The application discloses a kind of strong winter self-incompatible Brassica napus material's breeding method, the breeding method includes the following steps: S1: seed germination;S2: seed germination, cotyledon just transplants when unfolding;S3: seedling stage low temperature treatment, low temperature treatment time is 60-80 days;S4: field transplanting;S5: pollination;S6: harvest seed.The application is by seedling stage low temperature vernalization treatment, advantage is: (1) simple operation.It is only needed to be seedling to 10 leaf period low temperature vernalization treatment, then transplanting field can;(2) greatly shorten breeding cycle.The application is from August to May of next year, significantly shorten the breeding time, improve the breeding efficiency.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biotechnology, in particular to a propagation method of a strong winter self-incompatible Brassica oleracea material. BACKGROUND

[0002] Brassica oleracea L. is an important economic crop of Cruciferae, with many varieties, wide distribution and various functions. Curded cabbage is an important vegetable crop, which can provide dietary fiber needed by human body, and kale as a horticultural plant is also popular, in addition, some varieties of Brassica oleracea are also important research materials in the field of plant pathogen research, but the seed saving and propagation of Brassica oleracea has always been a problem faced by scientific researchers. There are several reasons: first, Brassica oleracea is a chloroplast vernalization plant, and the vegetative body needs to grow to a certain size before it can normally bloom through vernalization; second, Brassica oleracea has strong winter nature and needs a long time of continuous low temperature to normally pass through vernalization, and there are differences between different materials, and the operation of strong winter materials in the low temperature preservation stage is complex and has many details, and improper operation can easily cause material damage. The Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences has developed a set of Brassica oleracea propagation technology process, which selects seed plants at the mature stage (northern spring cabbage in May every year), stores at 4-6℃ for three months (with 2-3 times of airing), then cuts the top "ten" and then transplants to the open field after about 10 days of false planting and training, and blooms and pollinates in late March of the next year. Although this scheme process can ensure normal flowering and pollination of spring cabbage plants, it still has many limitations. First, the whole process lasts for a long time, from sowing seedlings in late January to harvesting seeds in June of the next year, which spans nearly a year and a half; second, the operation steps are complex, and need to go through seed plant selection, cold storage, false planting and training, and strict control of indoor environmental conditions, disinfectant spraying and other disease prevention treatments during cold storage. Therefore, there is an urgent need for a simple and short cycle propagation method of strong winter self-incompatible Brassica oleracea. SUMMARY

[0003] In view of the defects in the prior art, the present application provides a propagation method of a strong winter self-incompatible Brassica oleracea material, which greatly simplifies the propagation steps of Brassica oleracea material by exploring different characteristics of strong winter, self-incompatible Brassica oleracea material, and establishes a complete, efficient, simple and easy-to-operate Brassica oleracea propagation system, which provides a new reference method for similar Brassica oleracea material propagation.

[0004] The present application provides a propagation method of a strong winter self-incompatible Brassica oleracea material, comprising the following steps:

[0005] S1: seed germination;

[0006] S2: seedling training;

[0007] S3: low temperature treatment in seedling stage, and the low temperature treatment time is 60-80 days.

[0008] Further, the specific operation of the step S1 is as follows: placing filter paper at the bottom of a culture vessel, adding tap water, placing cabbage seeds in the culture vessel, and placing the culture vessel in an incubator and waiting for germination.

[0009] Further, the number of cabbage seeds is determined according to the size of the culture vessel, and generally 8-10 seeds per square centimeter are appropriate, and too high density will cause the roots to be intertwined after germination, which is not conducive to seedling separation and seedling training.

[0010] Further, the temperature of the incubator is 23-27℃.

[0011] Further, the specific operation of the step S2 is as follows: transplanting when the seeds germinate and the cotyledons just unfold, and placing in an artificial growth room after transplanting.

[0012] Further, the artificial growth room has a light illumination time of 16h, a light intensity of 5000 lux, and a temperature of 23-25℃ during the day, and a dark time of 8h and a temperature of 16-18℃ during the night.

[0013] Further, the specific operation of the step S3 is as follows: transplanting the seedlings to a low-temperature vernalization room for vernalization treatment when the cabbage seedlings grow to the 10-leaf stage.

[0014] Further, the temperature of the vernalization room is 4-6℃, the humidity is 60%-80%, the light illumination time is 16h during the day, and the soil is normally watered and fertilized during the period.

[0015] Further, the propagation method further includes the steps of field transplanting, pollination and harvesting seeds.

[0016] Further, the strong winter self-incompatible cabbage material includes any one of ECD11, ECD12, ECD13, ECD14 and ECD15. Among them, ECD11-ECD14 are four cabbage, and ECD15 is a kale. The five cabbages have different winter strengths, and are all self-incompatible materials, which are difficult to propagate and easy to lose materials if not operated properly.

[0017] In summary, compared with the prior art, the present application achieves the following technical effects:

[0018] (1) Simple operation. The prior art requires seedling screening, cold storage treatment, and false planting and training, while the present application only needs to vernalize the seedlings at the 10-leaf stage, and then transplant them to the field.

[0019] (2) Greatly shortens the breeding cycle. Existing technology harvests seeds from late January to June of the following year, which is a long time span. This invention, from August to May of the following year, significantly shortens the breeding time, improves efficiency, and facilitates further work. Attached Figure Description

[0020] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0021] Figure 1 This is a schematic flowchart of the reproduction method proposed in an embodiment of the present invention.

[0022] Figure 2 This is the result of cabbage transplanted to the field before overwintering in Experiment 3 of this invention.

[0023] Figure 3 This is the result of the full bloom period of cabbage in Experiment 3 of this embodiment of the invention. Detailed Implementation

[0024] To enable those skilled in the art to better understand the present invention, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of the present invention.

[0025] The European clubroot differential set is a set of materials specifically used to identify physiological races of clubroot fungi. The differential hosts were provided by the Warwick Genetic Resources Laboratory in the UK. This set consists of 15 different cruciferous varieties, including four heading cabbage varieties (ECD11, ECD12, ECD13, and ECD14) and one kale variety (ECD15). These five cabbage varieties exhibit varying degrees of winter hardiness and are all self-incompatible, making propagation difficult and prone to material loss if not handled properly. This invention uses these five cabbage varieties as examples to provide an efficient and simple method for propagating strong winter-hardy, self-incompatible cabbage varieties.

[0026] Example

[0027] The specific steps of the reproduction method of the present invention are as follows:

[0028] (1) Seed germination

[0029] Under the seasonal and climatic conditions of the Yangtze River Basin, in early August, a round filter paper is placed in a clean culture dish (10 cm in diameter), and the size is just suitable for sticking to the bottom of the culture dish. About 10 ml of tap water is added to the culture dish with a dropper, and 5 cabbage material seeds are respectively spotted in 5 different culture dishes. The number of seeds in each culture dish is about 200, and too dense will cause the roots to be intertwined and adhered after germination, which is not conducive to seedling separation and hardening-off. The culture dish with the spotted seeds is placed in a culture box at about 25°C, and germination is waited.

[0030] (2) Hardening-off

[0031] When the seeds germinate and the cotyledons just unfold, transplant them into the plastic flower pots filled with nutrient soil in advance, one plant per pot. Place the pots in a tray that can hold water and arrange them in an artificial growth room. The day and night temperatures of the artificial growth room are 25°C and 18°C respectively, the daily light time is 16h, the light intensity is 5000 lux, and normal watering management is maintained.

[0032] (3) Low temperature treatment

[0033] When the cabbage seedlings grow to the 10-leaf stage (late September), the seedlings are moved to a low-temperature vernalization room along with the tray, and the temperature in the vernalization room is controlled at 4-6°C and the humidity is 60%. The light time is 16h per day, and normal watering management is followed during this period, with appropriate supplementation of a small amount of compound fertilizer. Clean up the old leaves that naturally fall off during growth.

[0034] (4) Field transplanting

[0035] According to the difference in winter resistance, ECD15 is transplanted to the field with soil after 60 days of low-temperature treatment in the vernalization room (late November), and ECD11-ECD14 is transplanted to the field with soil after 80 days of low-temperature treatment (late December). The plant spacing and row spacing are both 30 cm. After transplanting, enough water is poured to fix the roots, and urea is applied once in the middle or late January to slow down the seedlings.

[0036] (5) Pollination

[0037] Under the seasonal and climatic conditions of the Yangtze River Basin, the five cabbage materials are in the early flowering stage in the middle of March. Before flowering, insect isolation is performed by covering each material with a mosquito net, and a beehive is placed in each mosquito net to assist pollination.

[0038] (6) Harvesting seeds

[0039] In late May, when all the plants have turned into "pi pa yellow" (a type of yellow), they are cut down and transferred to an open field for drying. When the pods are dry and ready to burst, they are threshed, and the seeds are dried and stored in a refrigerator for future use.

[0040] After 4 years of experimental exploration, the best operation scheme in the present application is summarized, and the specific experimental results are as follows:

[0041] Table 1 Reproduction results of cabbage with 30 days of vernalization time

[0042]

[0043] Table 2 Reproduction results of cabbage with 120 days of vernalization time

[0044]

[0045]

[0046] Table 3 Reproduction results of cabbage with 60 days of vernalization time

[0047]

[0048] Table 4 Reproduction results of cabbage with 80 days of vernalization time

[0049]

[0050] The vernalization time of the above experiment one is 30 days, and the vernalization time of the experiment two is 120 days, and except that ECD15 can produce a small amount of seeds (20 kg / acre) under the condition of the experiment two, the two vernalization times cannot normally produce cabbage seeds, which means that the reproduction fails. The vernalization time of the experiment three and the experiment four is 60 days and 80 days respectively, and the results show that the five strong winter self-incompatible cabbage materials ECD11, ECD12, ECD13, ECD14 and ECD15 can be normally reproduced, and the yield can reach more than 50 kg / acre, and the highest can reach 126.5 kg / acre, which shows that the reproduction method adopted by the present application is effective for strong winter self-incompatible cabbage materials.

[0051] The above only describes the preferred embodiments of the present application, and is not used to limit the present application, and any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A method of propagating a strong winter self-incompatible Brassica material, characterized by, Includes the following steps: S1: Seed germination; S2: Seedling hardening; S3: Low temperature treatment during the seedling stage, with a treatment time of 60-80 days; The specific operation of step S3 is as follows: when the cabbage seedlings have grown to 10 leaves, the seedlings are moved to a low-temperature vernalization room for vernalization treatment; The vernalization period should be maintained at a temperature of 4-6℃, a humidity of 60%-80%, and 16 hours of daylight. During this period, watering and fertilization should be carried out as usual. The strong winter-hardy self-incompatible cabbage material includes any one of ECD11, ECD12, ECD13, ECD14, and ECD15.

2. The propagation method according to claim 1, characterized in that, The specific operation of step S1 is as follows: Place filter paper at the bottom of the culture dish, add tap water, place the cabbage seeds in the culture dish, place the culture dish in an incubator, and wait for germination.

3. The propagation method according to claim 2, characterized in that, The number of cabbage seeds is 8 to 10 seeds per square centimeter.

4. The propagation method according to claim 2, characterized in that, The temperature of the incubator is 23-27℃.

5. The breeding method according to claim 1, characterized in that, The specific operation of step S2 is as follows: Transplant the seeds when they germinate and the cotyledons have just unfolded. After transplanting, place them in an artificial growing room to grow.

6. The breeding method according to claim 5, characterized in that, The artificial growth chamber has a daytime light duration of 16 hours, a light intensity of 5000 lux, and a temperature between 23 and 25°C; and a darkness duration of 8 hours, with a temperature between 16 and 18°C.

7. The breeding method according to claim 1, characterized in that, The propagation method also includes the steps of field transplanting, pollination, and seed harvesting.

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