CN109566401B - Method for quickly breeding bolting-resistant homozygous inbred line and inbred-incompatible line of black cabbage pole - Google Patents
Method for quickly breeding bolting-resistant homozygous inbred line and inbred-incompatible line of black cabbage pole Download PDFInfo
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- 238000009395 breeding Methods 0.000 title claims abstract description 16
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- 239000001963 growth medium Substances 0.000 claims abstract description 27
- 235000011303 Brassica alboglabra Nutrition 0.000 claims abstract description 25
- 235000011302 Brassica oleracea Nutrition 0.000 claims abstract description 25
- 230000035784 germination Effects 0.000 claims abstract description 18
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 claims abstract description 16
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- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims description 4
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- A—HUMAN NECESSITIES
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- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
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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
- A01G31/00—Soilless cultivation, e.g. hydroponics
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
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Abstract
The invention discloses a method for quickly breeding a bolting-resistant homozygous inbred line and a bolting-resistant incompatible line of a brassica oleracea, which relates to the technical field of genetic breeding and comprises the following steps: taking bolting-resistant brassica oleracea heterozygous plants as parents, carrying out artificial pollination in a flowering phase for 2 weeks, then, taking young embryos to carry out culture on a germination culture medium, carrying out vernalization treatment for 20 days after 2 weeks, carrying out normal culture in an artificial climate chamber, selecting the latest bolting single plant to carry out selfing, repeating the steps after artificial pollination in the flowering phase, and obtaining the bolting-resistant brassica oleracea homozygous selfing line after 3-4 generations; taking a late bolting cabbage heterozygous plant as a male parent, taking a cabbage self-incompatible line as a recurrent parent, selecting a young embryo with an affinity index meeting the requirement, and obtaining the bolting-resistant cabbage homozygous self-incompatible line after 4-5 generations through the same steps. By utilizing the method provided by the invention, the generation cultivation period is shortened, the bolting resistant homozygous inbred line and the self-incompatible line can be obtained within 18 months, the breeding period is greatly reduced, and the breeding cost is saved.
Description
Technical Field
The invention relates to the technical field of crop genetic breeding, in particular to a breeding method of a brassica oleracea bolting resistant homozygous inbred line and a brassica oleracea inbred incompatible line.
Background
Wucai is a variety of brassica seed Chinese cabbage subspecies, and the inbred line breeding of the Wuta-tsai is provided by Wang Chenggang, Cheng Yuan, Zhushidong and the like, for example, the method uses a Wuta-tsai heterozygous variety as an inbred line, carries out bud-stage inbred pollination, and obtains a homozygous inbred line through inbreeding of a plurality of generations; a breeding method of Wuta-tsai self-incompatible line, such as Wanchenggang, Zhouyanqian, Zhushidong, and the like, comprises the following steps: taking the Wuta-tsai self-compatible line as a recurrent parent, taking the small green vegetable self-incompatible line as a transformational source, and backcrossing the filial generation with the recurrent parent for a plurality of generations to obtain the stable Wuta-tsai self-incompatible line. However, these methods require 5-6 years to obtain a homozygous inbred line or a self-incompatible line, and the same breeding material cannot simultaneously obtain a self-compatible line and a self-incompatible line, which is not favorable for the rapid application of excellent parents. In addition, the prior brassica oleracea varieties are subjected to serious early bolting, so that the yield and the varieties are seriously influenced, and no report about an effective screening method for the bolting resistance of the brassica oleracea is available so far.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a method for quickly breeding a brassica oleracea bolting-resistant homozygous inbred line and an inbred-incompatible line so as to solve the problems that the prior art cannot quickly screen and the like.
The invention is realized by the following technical scheme:
the invention provides a method for quickly breeding a bolting-resistant homozygous inbred line and a bolting-resistant incompatible line of a brassica oleracea, which comprises the following steps:
step 1, taking the late bolting cabbage heterozygous plants as male parents, taking the cabbage self-incompatible line as recurrent parents, taking bolting resistant characters and parent characters of the self-incompatible line as screening target characters, manually hybridizing and pollinating the two parents in a bud stage, self-pollinating the late bolting cabbage heterozygous plants, and obtaining young and tender pods after 2 weeks;
step 2, taking young pods of 2 weeks in size, sterilizing the surfaces of the young pods with 70% alcohol for 30s under an aseptic condition, after the young pods are rinsed once with sterile water, sterilizing the young pods with 2% NaClO solution added with a drop of Tween 80 for 10min, then rinsing the young pods with the sterile water for 4 to 5 times, 5min each time, and after the young pods are sterilized, taking out the young embryos in the pods with a sterile scalpel and forceps;
step 3, placing the taken-out immature embryos on an immature embryo culture medium, and carrying out dark culture at the temperature of 25 +/-1 ℃;
step 4, culturing the immature embryos in an immature embryo culture medium until mature seeds are obtained, transferring the mature seeds into a germination culture medium after culturing for about 2 weeks, and performing dark vernalization treatment at the temperature of 4 +/-1 ℃;
step 5, after vernalization for 20 days, planting the germinated seeds into a plastic flowerpot filled with a solid matrix, gradually hardening the seedlings after the plastic preservative film is moisturized for 1-2 days, removing the preservative film after 5 days, and culturing in an artificial climate chamber until the seedlings bloom;
step 6, selecting the bolting plants in the last batch from the cultured plants, performing flowering phase selfing and bud phase selfing on the bolting plants, simultaneously hybridizing with recurrent parents, obtaining secondary young pods after 2 weeks, and calculating affinity indexes;
step 7, selecting secondary tender pods with the flower period and bud period affinity indexes both larger than 10 from selfed plants, selecting secondary tender pods with the bud period affinity index larger than 10 and the flower period affinity index smaller than 1 from plants hybridized with recurrent parents, and repeating the operation of the step 2 to the operation of the step 7;
and 8, repeating the step 7 for 4-5 times, wherein the plants prepared from secondary young pods with the florescence and bud period affinity indexes both greater than 10 in the finally obtained affinity indexes on the inbred plants are super bolting resistant brassica oleracea homozygous inbred lines, and the plants prepared from secondary young pods with the bud period affinity indexes greater than 10 and the florescence affinity indexes less than 1 in the plants hybridized with the recurrent parent are self-incompatible lines.
Further, the immature embryo culture medium in the step 3 consists of MS, 6-BA, NAA, sucrose and agar, wherein 2mg of 6-BA, 0.2mg of NAA, 30g of sucrose and 1.2mg of agar are added into each liter of MS, and the pH value is 5.8.
Further, the germination medium in step 4 consists of MS, 6-BA, sucrose and agar, wherein each liter of MS is added with 0.5mg of 6-BA, 30g of sucrose and 1.2mg of agar, and the pH is 5.8.
Further, the compatibility index in the step 7 meets the requirement that the selfing index of the selfing line in the flowering phase is greater than 10, and the compatibility index of the selfing incompatible line in the bud phase is greater than 10 and the flowering phase index is less than 1.
Further, the culture environment of the artificial climate chamber in the step 5 is specifically as follows: the temperature is 25 +/-1 ℃, the illumination is 1500-.
Further, the repetition times of the step 2-6 in the step 7 are 4-5 times.
Further, the bolting resistant brassica oleracea heterozygous plant and the self-incompatible line variety in the step 1 are selected from one of Huainan chrysanthemum heart, Huainan black heart black, combined fertilizer yellow heart black, combined fertilizer black heart black, Shanghai small eight leaves, collapsed anemone 1-6 and Hui Wu 1-13.
The invention also provides application of the method in breeding of the non-heading Chinese cabbages.
Compared with the prior art, the invention has the following advantages:
(1) the black-boned cabbage is generally planted in 9-10 months, blossoms and pods are generated in 3-4 months of the next year, seeds are harvested in 5 months, and the generation period is about 8 months. The invention utilizes embryo culture technology to carry out artificial pollination in the bud period or the flower period of the lindera aggregate, and the young embryo is taken for culture after 2 weeks, so that the young embryo is germinated and transferred to the next generation, thereby shortening the reproductive growth period.
(2) The black-boned cabbage can be bolting and flowering after low-temperature vernalization in winter. The invention uses low-temperature vernalization technology to treat the germination seeds of the black-flowered cabbage for 20 days at the temperature of 4 +/-1 ℃ so as to complete the vernalization; after the vernalized germinating seeds are planted for 2 weeks, bolting and flowering can be realized, and the period of vegetative growth is shortened.
(3) The invention combines embryo culture and vernalization technologies, completes generation cultivation in an artificial climate chamber, does not need field planting, shortens the cultivation period, reduces the labor and production cost, and has the advantages of easy operation and the like.
Drawings
FIG. 1 is the breeding process of bolting resistant homozygous inbred line and inbred incompatible line of cabbage.
Detailed Description
The present invention will be described below by way of examples, but the present invention is not limited to the following examples.
All the raw and auxiliary materials, reagents, instruments and equipment selected for use in the present invention are well known in the art and are not intended to limit the practice of the present invention, and other reagents and equipment known in the art may be used in the practice of the following embodiments of the present invention.
Example 1
1. Material
And (3) bolting heterozygous plants of the black-bone cabbage late: w13-2, which is a late bolting plant screened from the black-cabbage germplasm resources.
Self-incompatible line plant of black-bone vegetable: w02-8, is a self-incompatible line plant obtained by screening from black-bone cabbage germplasm resources and carrying out multi-generation self-copulation separation.
2. Seed selection step
a. Breeding of bolting-resistant homozygous inbred line of lindera aggregate
(1) Taking the late bolting cabbage heterozygous plant W13-2 as a parent, selfing and pollinating at the bud stage, and obtaining young and tender pods after 2 weeks;
(2) sterilizing pod of 2 weeks in size with 70% ethanol for 30s under aseptic condition, rinsing with sterile water, sterilizing with 2% NaClO solution containing Tween 80 for 10min, rinsing with sterile water for 5min for 4-5 times, and taking out embryo with sterile scalpel and tweezers.
(3) Placing the taken-out immature embryo on an immature embryo culture medium, and carrying out dark culture at the temperature of 25 +/-1 ℃;
(4) culturing the young embryo in a young embryo culture medium to mature seeds, transferring the young embryo into a germination culture medium after culturing for about 2 weeks, and carrying out dark vernalization treatment at the temperature of 4 +/-1 ℃;
(5) after 20 days of vernalization, planting the germinated seeds into a plastic flowerpot filled with a solid matrix (vermiculite: peat ═ 1:1), gradually hardening the seedlings after the plastic preservative film is moisturized for 1 to 2 days, removing the preservative film after 5 days, and culturing in an artificial climate chamber (the temperature is 25 +/-1 ℃, the illumination is 1500-;
(6) selecting the bolting plants of the last batch from the cultured plants, carrying out flowering phase selfing and bud phase selfing on the bolting plants, obtaining young and tender pods after 2 weeks, and calculating affinity indexes;
(7) obtaining plant immature embryos with 2-week-sized flowering phase and bud phase affinity indexes both larger than 10 by the same method as (2), and performing immature embryo culture and germination culture by the same methods as (3) and (4);
(8) repeating the steps (6) - (8) for 4 times to obtain a bolting cabbage homozygous inbred line W13-2-4;
the young pods in the steps (1) and (7) are pods with the size of 2 weeks;
the immature embryo culture medium in the step (3) consists of MS, 6-BA, NAA, sucrose and agar, wherein 2mg of 6-BA, 0.2mg of NAA, 30g of sucrose and 1.2mg of agar are added into each liter of MS, and the pH value is 5.8;
the germination culture medium in the step (4) consists of MS, 6-BA, sucrose and agar, wherein 0.5mg of 6-BA, 30g of sucrose and 1.2mg of agar are added into each liter of MS, and the pH value is 5.8.
b. Breeding of bolting resistant homozygous self-incompatible line of black cabbage pole
(1) Taking bolting-resistant brassica oleracea heterozygous plant W13-2 as a male parent, taking brassica oleracea self-incompatible line W02-8 as a recurrent parent, taking bolting-resistant and self-incompatible line parent traits as screening target traits, carrying out artificial hybrid pollination on an amphiphilic person in a bud period, and obtaining young and tender pods after 2 weeks;
(2) sterilizing pod of 2 weeks in size with 70% ethanol for 30s under aseptic condition, rinsing with sterile water, sterilizing with 2% NaClO solution containing Tween 80 for 10min, rinsing with sterile water for 5min for 4-5 times, and taking out embryo with sterile scalpel and tweezers after sterilization;
(3) placing the taken-out immature embryo on an immature embryo culture medium, and performing dark culture at the temperature of 25 +/-1 ℃;
(4) culturing the young embryo in a young embryo culture medium to mature seeds, transferring the young embryo into a germination culture medium after culturing for about 2 weeks, and carrying out dark vernalization treatment at the temperature of 4 +/-1 ℃;
(5) after 20 days of vernalization, planting the germinated seeds into a plastic flowerpot filled with a solid matrix (vermiculite: peat ═ 1:1), gradually hardening the seedlings after the plastic preservative film is moisturized for 1 to 2 days, removing the preservative film after 5 days, and culturing in an artificial climate chamber (the temperature is 25 +/-1 ℃, the illumination is 1500-;
(6) selecting the final bolting plant from the cultured plants, carrying out flowering phase selfing and bud phase selfing on the final bolting plant, simultaneously hybridizing the same plant with the recurrent parent, obtaining young and tender pods after 2 weeks, and calculating the affinity index.
(7) Obtaining 2-week plant immature embryos with the bud period affinity index of more than 10 and the flower period affinity index of less than 1 by the same method as (2), and performing immature embryo culture and germination culture by the same methods as (3) and (4);
(8) repeating the steps (6) - (8) for 5 times to obtain a bolting cabbage homozygous self-incompatible line W02-8-5;
the young pods in the steps (1) and (7) are pods with the size of 2 weeks;
the immature embryo culture medium in the step (3) consists of MS, 6-BA, NAA, sucrose and agar, wherein 2mg of 6-BA, 0.2mg of NAA, 30g of sucrose and 1.2mg of agar are added into each liter of MS, and the pH value is 5.8;
the germination culture medium in the step (4) consists of MS, 6-BA, sucrose and agar, wherein each liter of MS is added with 0.5mg of 6-BA, 30g of sucrose and 1.2mg of agar, and the pH value is 5.8;
experiment 1:
young pods 3, 7, 14, 21 and 28 days after pollination of the bolting resistant brassica oleracea hybrid plant W13-2 in the flowering phase and young pods 3, 7, 14, 21 and 28 days after pollination of the brassica oleracea self-incompatible line W02-8 in the bud phase are respectively taken, corresponding young embryos are taken out under the aseptic condition by the sterilization method in the embodiment 1a (2) as in the embodiment 1a (3), and dark culture is carried out on an embryo culture medium (the temperature is 25 +/-1 ℃). 30-35 immature embryos are treated for each treatment, the treatment is repeated three times, and the developmental maturity of the embryos is counted after 2 weeks.
TABLE 1 development of embryos on different days after embryo culture
The embryo culture medium is MS +30g/L sucrose +2 mg/L6-BA +0.2mg/L NAA +1.2mg/L agar, and the pH value is 5.8.
Embryo maturation rate (%) -. embryo developmental maturation number/inoculated embryo total number × 100%
As can be seen from the table, the mature rate of the embryos of the young embryos of 2 weeks after pollination of the lindera aggregate can reach more than 97 percent after embryo culture. Therefore, the young embryos of this size are the subject of embryo culture in the subsequent invention.
Experiment 2
Young pods 14 days after pollination of bolting resistant brassica oleracea hybrid plants W13-2 in flowering phase and young pods 14 days after pollination of brassica oleracea self-incompatible line W02-8 in bud phase are respectively taken, corresponding young embryos are taken out under the aseptic condition by the sterilization method in the embodiment 1a (2) and the sterilization method in the embodiment 1a (3), and dark culture (the temperature is 25 +/-1 ℃) is carried out on embryo culture media containing different 6-BA/NAA ratios (table 2). 30-35 immature embryos are treated for each treatment, the treatment is repeated three times, and the developmental maturity of the embryos is counted after 2 weeks.
TABLE 2 influence of different concentrations of 6-BA and NAA on the development of young embryos of Brassica campestris
The young embryo culture medium is MS +30g/L sucrose + (1-3) mg/L6-BA + (0.1-0.3) mg/L NAA +1.2mg/L agar, and the pH value is 5.8.
The embryo maturation rate (%) — the number of mature embryos developed/total number of inoculated embryos × 100%.
As can be seen from Table 2, both varieties developed best in the immature embryo medium containing 2 mg/L6-BA and 0.2mg/L NAA. Therefore, this medium was designated as a young embryo medium in the subsequent experiments.
Experiment 3
Young pods 14 days after pollination of the bolting resistant brassica oleracea hybrid plant W13-2 in the flowering phase and young pods 14 days after pollination of the brassica oleracea self-incompatible line W02-8 in the bud phase are respectively taken, under the aseptic condition, corresponding young embryos are taken out by the sterilization method in the embodiment 1a (2) and the method in the embodiment 1a (3), and dark culture is carried out in a young embryo culture medium containing 2 mg/L6-BA and 0.2mg/L NAA (the temperature is 25 +/-1 ℃). Embryos which have developed and matured after 2 weeks are taken and placed on embryo germination culture media containing different 6-BA/NAA ratios (shown in table 3) for culture (the illumination intensity is 3000Lux, the illumination time is 16h, the temperature is 25 +/-1 ℃/dark time is 8h, and the temperature is 22 +/-1 ℃). 30-35 mature embryos are treated for each time, the treatment is repeated three times, and the germination condition of the embryos is counted after 2 weeks.
TABLE 3 influence of different concentrations of 6-BA and NAA on germination of mature embryos of Brassica Oleracea
The embryo germination culture medium is MS +30g/L sucrose + (0.0-1.0) mg/L6-BA + (0.00-0.05) mg/L NAA +1.2mg/L agar, and the pH value is 5.8.
Embryo germination rate (%) -. embryo germination number/total number of inoculated mature embryos × 100%.
As can be seen from Table 3, the germination rates of both varieties were the highest in the embryo germination medium containing 0.5 mg/L6-BA. Therefore, this medium was designated as the embryo germination medium in the subsequent experiments.
Experiment 4
Mature embryos (seeds) which have germinated on W13-2 and W02-8 are placed at 4 +/-1 ℃ for different days of dark vernalization (Table 3); after the vernalization treatment, the germinated seeds are planted in a plastic flowerpot filled with a solid matrix (vermiculite: peat ═ 1:1), the plastic preservative film is gradually hardened after being moisturized for 1-2 days, the preservative film is removed after 5 days, the seeds are cultured in an artificial climate chamber (the temperature is 25 +/-1 ℃, the illumination is 1500-. 30-35 mature embryos (seeds) were treated each, in triplicate.
TABLE 3 influence of different vernalization days on bolting and flowering of black-boned vegetables
The bolting and flowering days are the vegetative growth days from the time when the germinated seeds are planted in the matrix to the time of budding.
As can be seen from Table 3, the vernalization time has a great influence on the vegetative growth time of the black-bone mustard, and the days for vegetative growth of the black-bone mustard gradually decrease with the prolongation of the vernalization treatment time; the number of vegetative growth days varied little over 20 days of vernalization treatment, and thus was defined as the number of vernalization days in the subsequent experiments.
Example 2
In the embodiment, the brassica oleracea bolting resistant heterozygous plant W9803-5 is taken as a male parent, the self-incompatible line S9801-2 is taken as a recurrent parent, other steps are the same as the embodiment 1, and the brassica oleracea bolting resistant homozygous inbred line W9803-5-4 and the homozygous self-incompatible line S9801-2-4 can be obtained after 4 generations.
Example 3
In the embodiment, the non-head Chinese cabbage bolting resistant heterozygous plant WC-02-08 is taken as a male parent, the self-incompatible line WC03-05 is taken as a recurrent parent, other steps are the same as the embodiment 1, and the non-head Chinese cabbage bolting resistant homozygous inbred line WC-02-08-04 and the homozygous self-incompatible line WC03-05-05 can be obtained after 4-5 generations.
Claims (3)
1. The fast selective breeding method of the bolting resistant homozygous inbred line and the inbred incompatible line of the lindera aggregate is characterized by comprising the following steps:
step 1, taking the late bolting cabbage heterozygous plants as male parents, taking the cabbage self-incompatible line as recurrent parents, taking bolting resistance and parent properties of the self-incompatible line as screening target properties, carrying out artificial hybrid pollination on the two parents in the bud period, carrying out self-pollination on the late bolting cabbage heterozygous plants, and obtaining young and tender pods after 2 weeks;
step 2, taking young pods of 2 weeks in size, sterilizing the surfaces of the young pods for 30s by 70% alcohol under an aseptic condition, rinsing the young pods once by using sterile water, sterilizing the young pods by using a 2% NaClO solution added with a drop of Tween 80 for 10min, rinsing the young pods for 4 to 5 times by using the sterile water for 5min each time, and taking out young embryos in the pods by using an aseptic scalpel and forceps after the young pods are sterilized;
step 3, placing the taken-out immature embryos on an immature embryo culture medium, and carrying out dark culture at the temperature of 25 +/-1 ℃;
step 4, culturing the immature embryos in an immature embryo culture medium until mature seeds are obtained, transferring the mature seeds into a germination culture medium after culturing for about 2 weeks, and performing dark vernalization treatment at the temperature of 4 +/-1 ℃;
step 5, after vernalization for 20 days, planting the germinated seeds into a plastic flowerpot filled with a solid matrix, gradually hardening the seedlings after the plastic preservative film is moisturized for 1-2 days, removing the preservative film after 5 days, and culturing in an artificial climate chamber until the seedlings bloom;
step 6, selecting the bolting plants in the last batch from the cultured plants, performing flowering phase selfing and bud phase selfing on the bolting plants, simultaneously hybridizing with recurrent parents, obtaining secondary young pods after 2 weeks, and calculating affinity indexes;
step 7, selecting secondary tender pods with the flower period and bud period affinity indexes both larger than 10 from selfed plants, selecting secondary tender pods with the bud period affinity index larger than 10 and the flower period affinity index smaller than 1 from plants hybridized with recurrent parents, and repeating the operation of the step 2 to the operation of the step 7;
8, repeating the step 7 for 4-5 times, wherein the plants prepared from secondary young pods with the florescence and bud period affinity indexes both greater than 10 in the finally obtained affinity indexes on the inbred plants are super bolting resistant brassica oleracea homozygous inbred lines, and the plants prepared from secondary young pods with the bud period affinity indexes greater than 10 and the florescence affinity indexes less than 1 in the plants hybridized with the recurrent parent are self-incompatible lines;
the immature embryo culture medium in the step 3 consists of MS, 6-BA, NAA, sucrose and agar, wherein 2mg of 6-BA, 0.2mg of NAA, 30g of sucrose and 1.2mg of agar are added into each liter of MS, and the pH value is 5.8;
the germination culture medium in the step 4 consists of MS, 6-BA, sucrose and agar, wherein 0.5mg of 6-BA, 30g of sucrose and 1.2mg of agar are added into each liter of MS, and the pH value is 5.8.
2. The method for rapidly breeding the bolting-resistant homozygous inbred line and the selfing-incompatible line of the brassica oleracea according to claim 1, wherein the repetition number in the step 8 is specifically 4.
3. The method for rapidly breeding the bolting-resistant homozygous inbred line and the self-incompatible line of the brassica oleracea according to claim 2, wherein the culture environment of the artificial climate chamber in the step 5 is specifically as follows: the temperature is 25 +/-1 ℃, the illumination is 1500-.
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