CN1531843A - Novel cabbage type rape cytoplasmic male sterile line material of Chinese cabbage vegetable transformation and its obtaining method - Google Patents
Novel cabbage type rape cytoplasmic male sterile line material of Chinese cabbage vegetable transformation and its obtaining method Download PDFInfo
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Abstract
The method of obtaining cabbage type rape cytoplasmic male sterile line material of Chinese cabbage type vegetable transformation includes the following steps: artificial bud ripping in bud stage or blooming stage bagging hybridization of cabbage type rape cytoplasmic sterile line and Chinese cabbage type vegetable to obtain transformed first generation seed; generation-adding treatment of transformed first generation seed combined with field observation to select excellent sterile line plant, and multiple-generation backcross transformation with excellent Chinese cabbage plant; and obtaining stable genetic cytoplasmic sterile line material of Chinese cabbage after backcross for five generations. The sterile line material thus obtained has stable sterility, easy transformation, hearty growth, strong combining ability, high resistor and high yield. The said method is simple and low in cost.
Description
Technical field
The present invention relates to Chinese cabbage group transformation novel cabbage type rape cytoplasmic male sterile line material and preparation method thereof.
Background technology
Chinese cabbage group has tangible hybrid vigour.The domestic first generation of hybrid of extensively having carried out Chinese cabbage is utilized work, but generally adopt the method for self incompatible line or self compatible line to carry out first generation of hybrid production, this method exists and produces original seed cost height, technical sophistication, easily parents' vitality decline and occur pseudostationary easily takes place, and influences problems such as half-blood quality.Utilize male sterile line of cabbage to produce half-blood, can overcome the problems referred to above.Particularly the seed selection of cabbage cytoplasm male sterile line material and utilization are one of effective ways of Chinese cabbage breeding and heterosis utilization.Utilize the cytoplasm male sterility line production of hybrid seeds both can obtain high-quality hybrid generation, can protect breeder's interests again.For this reason, seek good cytoplasmic male sterile line, overcome the defective of utilizing the self incompatible line breeding and the production of hybrid seeds, be the important goal that the breeder pursues always.
Early seventies, Chinese scholar is found and has been initiated and utilizes male sterile dual-purpose system (male sterile of vouching recessive gene) to breed kind, and on producing, apply (as Chinese cabbage 156, the fast dish in Shenyang etc.), but owing to need repeatedly pull out 50% educated strain during the production of hybrid seeds, time-consuming taking a lot of work, and often hybrid seed yield is lower, thereby has limited the application and the development of this male sterile line.The Zhang Shufang of Agricultural Sciences Inst., Shenyang City (1989) cultivates nuclear gene first and makes male sterile line mutually and bred crossbreed large scale applications such as " No. one, fast dishes ".This male sterile line sterile degree and sterile plant rate all can reach 100%, can make the high-quality crossbreed, and the seed collecting amount is also higher, and its shortcoming is to be equivalent to triple hybrid, and hybrid purity is under some influence; The transformation of male sterile line is difficult in addition.Nineteen sixty-eight radish cytoplasmic sterility (Ogu CMS) is found, and wide-scale distribution worldwide, but low temperature yellow and nectary to degenerate be its deadly defect.This male sterile line of Li Guangchi etc. (1995) backcross transformation has only obtained the 134 Chinese cabbage cytoplasm male sterile lines that aetiolation weakens relatively after 16 generations, but because of a little less than the plant strain growth gesture, coordinate force is not strong etc. former thereby fail practicality.Over nearly 20 years, research both at home and abroad is Ogu cytoplasmic sterility (male sterile with radish cytoplasm) and Pol cytoplasmic sterility (pol cytoplasmic male sterility (pol cms)) with utilizing at most the most widely.
Early eighties, a large amount of research work to the Chinese cabbage transformation has been done to the Ogu male sterile with radish cytoplasm source of being introduced by the U.S. by China.But there are defectives such as blade yellow, growth retardation, nectary degeneration and coordinate force difference in this sterile source, so fail so far to obtain utilizing on producing.Cornell Univ USA carries out cytoplasm with Ogu male sterile with radish cytoplasm cauliflower and turnip type rape and merges, obtained synthetic male sterile brassica type rape burdo, show the advantage of not yellow of low temperature, obtained the Ogu cytoplasmic male sterility material of not yellow of Chinese cabbage low temperature after backcrossing with Chinese cabbage hybridization, many generations.1992, this radish kytoplasm male sterile line of cabbage 95A12,95A13 through improvement introduced at Vegetable Research center, Beijing.With Chinese cabbage continuous multi-generation backcross transformation after, though find that this sterile material is neat gradually with the increase of transformation algebraically, material degeneration is very fast, part seedling performance deformity, hybrid vigour is weak simultaneously, coordinate force is poor, the application prospect dimness.The Chinese Academy of Agricultural Sciences's vegetable or flower the radish cytoplasmic male sterilty material Nycms that is introduced has been carried out transformation and research, obtained same conclusion.
Academy of agricultural sciences, Shaanxi Province vegetable or flower utilizes cabbage type rape Polima sterile source to become allo-plasm Chinese cabbage male sterile line CMS for the material transformation
3411-7(patent No. is 92104134.9), its sterile plant rate can reach 100%, and sterile degree is more than 95%, but fatal weakness is to occur trace-pollen and slight flower bud abortion phenomenon under the hostile environment condition.Its fertility instability is found with Polima sterile source assorted 13, assorted 33 and the many researchs for backcross transformation of Chinese cabbage in Vegetable Research center, Beijing, has the powder strain easily, influences whole purity, is difficult to utilize.Responsive to temperature is that of Pol CMS is difficult to the thoroughly major issue of solution, the problem that the trace-pollen phenomenon of its existence (being the sterility instability) has become the breeder to be paid close attention to always.Because in hybrid seeding, bloom (flowering asynchronism) too early if run into male sterile line, and when having micro mist to take place, thereby self-crossing seedling reduction production of hybrid seeds hybrid rate just may appear (during flower synchronization, because the joining property late of Chinese cabbage selfing does not influence hybrid rate even micro mist occurs yet).Ogu-CMS exist many for plant growing way after the transformation degenerate, heterosis, hybrid vigor is poor, Pol CMS exists fertility to be subjected to the shortcomings such as defective of environmental influence.Therefore seeking the novel Chinese cabbage sterile line material that can overcome Ogu Chinese cabbage cytoplasmic sterile line and the above-mentioned shortcoming of Pol Chinese cabbage cytoplasmic sterile line is very important.
Summary of the invention
The purpose of this invention is to provide that a kind of sterility is stable, combining ability is strong, transformation is easy, growth potential is prosperous, disease resistance is strong, increase production the novel cabbage type rape cytoplasmic male sterile line of tangible Chinese cabbage group transformation material.
Another object of the present invention provides the preparation method of the novel cabbage type rape cytoplasmic male sterile line of this Chinese cabbage group transformation material.
For achieving the above object, the present invention takes following design:
The preparation method of the novel cabbage type rape cytoplasmic male sterile line of Chinese cabbage group transformation material, this method may further comprise the steps:
(1) cabbage type rape cytoplasmic sterile line and Chinese cabbage group are carried out artificial flower bud phase stripping flower bud or florescence bagging hybridization, expose column cap after, the pollen of Chinese cabbage is coated with thereon, column cap normally expands, and obtains transformation generation seed;
(2) transformation generation seed is adopted the method that adds the generation processing, in conjunction with field observation, selecting excellent sterile is individual plant, and carries out many for backcross transformation with the Chinese cabbage elite plant strain;
(3) backcross and obtain the Chinese cabbage cytoplasmic male sterile line material of genetic stability after five generations.
Described add for processing method be meant add every year the generation 1～2 generation.Specific practice is: the seed that will just accept soaked 20～40 minutes with clear water, was put in 2～4% the hydrogen peroxide to soak 2～4 hours again, and 23～27 ℃ of following vernalization, the seed that will sprout next day was put into 1～5 ℃ of refrigerator vernalization 20～30 days.Broadcast after the taking-up in 20～21 ℃ the greenhouse, it is long to adopt night intensity of illumination to be that the light source of 3000lux replenishes illumination to 16 hour day.Can bloom by bolting in 25～30 days, and carried out artificial back pollinating then.
The preparation method of the novel cabbage type rape cytoplasmic male sterile line of Chinese cabbage group transformation material, this method is: a variety of Chinese cabbage and stable Chinese cabbage sterile line material are carried out flower bud phase stripping flower bud and florescence bagging hybridization, obtain a variety of Chinese cabbage cytoplasmic sterility based material of genetic stability.The a variety of Chinese cabbage sterile line material also can use preceding method to obtain, but uses this method and more save time than using preceding method, efficiently.
Beneficial effect of the present invention: Chinese cabbage group sterile line material provided by the invention has following outstanding advantage: the male sterile line that (1) is obtained belongs to cytoplasmic sterility, and its sterility is stable, and many generations backcross and do not degenerate, and transformation is easy, and growth potential is prosperous, and combining ability is strong.
(2) in transformation generation early,, male sterile line is stronger than maintenance line plant strain growth gesture, has the interspecific cross advantage.
(3) in transformation generation early,, the late bolting of male sterile line is stronger than maintenance line.
(4) prepare F1 with male sterile line, the seed production height, breeding cost is low, the saving of labor.
(5) the male sterile line disease resistance is strong, and volume increase obviously.
The preparation method of this material provided by the invention is easy, and is with low cost, is convenient to promote, and market prospects are wide.
The invention will be further described below in conjunction with accompanying drawing.
Description of drawings
Fig. 1 is the flower and the flower pesticide figure of male sterile line of the present invention
Fig. 2 is that transformation is early for the bolting comparison diagram (left side: be male sterile line, be in squaring period of male sterile line with corresponding maintenance line; Right: as to be maintenance line, to be in peduncle-growing period for rapeseed)
Fig. 3-1 is transformation the male sterile line blade (left side) and maintenance line blade (right side) comparison diagram in generation morning
Early scheme by the balling situation (half armful of ball) and the siphonal lobe proterties (half decomposite leaf) of the male sterile line in generation for transformation for Fig. 3-2
Embodiment
The acquisition of embodiment 1, the novel cabbage type rape cytoplasmic male sterile line of Chinese cabbage transformation material
One, material and method:
1, material
Cabbage type rape cytoplasmic male sterile line material C MS96 derives from France, and its nectary is normal, not yellow of blade, and petal is big, and flower pesticide white is degenerated, and column cap is normal, and plant strain growth is vigorous.
56 parts of Chinese cabbage materials that are used for transformation, wherein 26 parts of Chinese cabbage materials are the needs improvement of some general types and the material that improves disease resistance.30 parts of remaining materials are Chinese cabbage materials that isozygoty, various types of.All material all derives from Vegetable Research center, Beijing.
2, method
(1) since spring in 1996, adopt artificial flower bud phase the stripping flower bud or florescence cover bag technique carry out cabbage type rape cytoplasmic male sterile line and Chinese cabbage crossing and its male sterility of Duo Dai backcross transformation.Method is: the bud of strip off cabbage type rape cytoplasmic male sterile line CMS96, expose column cap after, the pollen of 26 parts of Chinese cabbage strains system is coated with thereon, column cap normally expands, and obtains transformation 1 generation seed;
(2) transformation 1 generation seed is adopted the method that adds the generation processing, add every year, carry out many in conjunction with the little strain pollination of select and remain autumn old strain and spring simultaneously for backcross transformation for 1～2 time.Each of transformation is all investigated from generation to generation the botany proterties of plant, and superseded petiole has the intermediate materials such as decomposite leaf type of incising, being partial to cabbage type rape, the leaf individual plant similar of selecting and remain to Chinese cabbage, and more than carrying out with the Chinese cabbage elite plant strain for backcross transformation.The code name of spring sowing over the years or autumn sowing male sterile line is: 98-517,98-518,98-519,98-520; 99-327,99-329,99-331,99-332; 00.3-195,00.3-226,00.3-351; 01.3-178,01.3-204,01.3-307; 02.3-178,02.3-196,02.3-311.
The specific practice that adds generation is: the seed that will just accept soaked 30 minutes with clear water, was put in 3% the hydrogen peroxide to soak 3 hours again, and 25 ℃ of following vernalization, the seed that will sprout next day was put into 3 ℃ of refrigerator vernalization 25 days.Broadcast after the taking-up in 20 ℃～21 ℃ the greenhouse, adopting intensity of illumination night is that the light source of 3000lx carries out light filling.Can bloom by bolting in 25 days～30 days, and carried out artificial back pollinating with Chinese cabbage then.
(3) the cabbage cytoplasm sterile line material of acquisition genetic stability: backcrossing through 5 generations as stated above can the stable Chinese cabbage CMS96 cytoplasmic sterility based material of acquired character.Obtained at present that 5 parts of sterility are stable, economic characters are good, growth Chinese cabbage CMS96 cytoplasmic sterility based material neat and consistent, that isozygoty, (02.3-168,02.3-178,02.3-196,02.3-307,02.3-311).Continue that, the Chinese cabbage of isozygotying various types of with 30 parts and 2 parts isozygoty, stable Chinese cabbage male sterile line carries out backcross transformation, further enlarge the scope of utilizing of this male sterile line.Utilize stable cabbage cytoplasm male sterile line examination to join the first generation of hybrid simultaneously.
Two, result and analysis
1, carry out cross experiment with 26 parts of Chinese cabbage materials and Wild cabbage type cytoplasmic male sterile line CMS96, the result shows that the cross-fertile rate is high, reaches 100%, and wherein maximum pod knot number is 21 seed/pods, and minimum pod is 5 a/pod, and filial generation 100% is sterile.At microscopically, do not observe any pollen of this male sterile line.As shown in Figure 1, the flower of male sterile line, petal is bright-coloured, open, and can fully attract insects such as honeybee.The column cap of male sterile line is normal, is easy to accept foreign pollen, and flower pesticide white, empty flat degeneration simultaneously because the nectary of this male sterile line is normal, can be observed honeybee and visit flower and solid good branch under field conditions (factors).
Obtained at present the cabbage cytoplasm male sterile material of 5 parts of genetic stabilities, these male sterile lines and maintenance line or paternal hybrid solid normally and also seed production high.Carry out backcross transformation with 30 parts of common Chinese cabbage and 2 parts of stable Chinese cabbage male sterile lines, obtained the seed in 2 generations of transformation.
2, the feature of CMS96 cytoplasmic male sterile line
(1) CMS96 cytoplasmic male sterile line sterility is stable, and many generations backcross and do not degenerate, and transformation is easy, and growth potential is prosperous, and combining ability is strong.
Compared and utilized CMS96 male sterile line and corresponding maintenance line (this maintenance line and male sterile line are just had any different on fertility, other proterties is in full accord) botany proterties and nutrient composition content, the results are shown in Table 1, table 2, show: many CMS96 male sterile line growth potentials of isozygotying for the acquisition of backcrossing are not degenerated, its plant strain growth is vigorous, and advantage is strong.As shown in table 1, on the main plant proterties, male sterile line and maintenance line are basic identical.Early in generation, show the characteristics of interspecific cross advantage than the raising of maintenance line plant strain growth gesture in the male sterile line transformation.In addition, transformation is early strong than maintenance line for the late bolting of male sterile line.Early for male sterile line and corresponding maintenance line (seed of " broken tripe " is handled two weeks in 4 ℃ of refrigerators) after the identical low temperature treatment of process, male sterile line is than the late bolting of maintenance line in transformation.As shown in Figure 2, male sterile line is in squaring period, and maintenance line has been extracted very long colored a kind of sedge out.Transformation is early stage, and the male sterile line blade shape is the osculant (Fig. 3-1, Fig. 3-2) of cabbage type rape and Chinese cabbage.By the visible transformation of Fig. 3-1 early the male sterile line blade (left side) in generation be the osculant of cabbage type rape and Chinese cabbage, complete different with maintenance line blade (right side), the maintenance line blade edge does not split quarter.Fig. 3-2 shows as half armful of ball for the transformation male sterile line plant in generation morning, and siphonal lobe has and dark and big moderate is split quarter.For backcross transformation and selection, be easy to eliminate male sterile line intermediate form material through too much, final acquisition is with the Chinese cabbage blade and embrace the on all four sterile line material of ball proterties.
The comparative studies of table 1 CMS96 male sterile line and maintenance line and male sterile line F1, corresponding maintenance line F1
The heavy clean vegetables of the wide waste dish of item designation plant height degree of development number of sheets newel ball high ball is heavy
Title 2001-(cm) (cm) (sheet) length (cm) (cm) (kg) (kg)
(cm)
CMS96-1 3-178 36.6 62.6 9.0 3.4 32.8 18.0 2.7 2.1
Maintenance line 3-176 36.6 62.4 8.4 3.2 32.0 16.4 2.2 1.8
CMS96-2 3-304 53.6 81.2 12.0 5.9 37.6 21.0 5.5 3.6
Maintenance line 3-303 43.4 73.2 10.0 3.7 37.0 18.4 3.5 2.3
CMS96-3 3-307 50.0 72.6 10.8 4.7 35.2 18.6 3.8 2.5
Maintenance line 3-306 47.0 68.6 9.1 4.5 32.8 17.8 3.8 2.6
CMS96-4 205 42.8 66.2 9.4 3.3 28.3 18.3 3.4 2.6
Maintenance line 194 39.8 59.8 10.4 3.0 25.0 13.3 2.7 2.1
CMS96(F1) 3-378 38.0 63.8 9.4 5.6 28.6 23.6 5.1 4.1
Maintenance line (F1) 2-198 45.4 76.2 8.4 5.3 32.4 23.0 4.8 3.7
CMS96(F1) 3-383 41.6 69.4 7.8 5.1 29.6 20.2 4.2 3.2
Maintenance line (F1) 2-366 37.4 65.8 8.2 5.5 27.4 21.2 4.1 3.2
CMS96(F1) 3-395 50.2 82.0 10.2 5.4 36.2 26.8 6.3 4.1
Maintenance line (F1) 2-319 43.8 70.0 7.8 6.9 30.0 23.0 4.7 3.8
CMS96(F1) 3-398 53.2 83.6 11.2 6.1 39.8 18.2 5.5 4.1
Maintenance line (F1) 2-361 54.0 74.8 7.0 5.3 41.6 19.0 5.1 3.7
The comparative studies of table 2 CMS96 and maintenance line nutrient component mean value
Item designation Vc β carrot raw fiber Ca Fe thiocyanate radical
(mg/kg) (mg/kg) (mg/kg) for title 2001-(mg/100g) plain (%)
(mg/kg)
CMS96-1 3-178 20.0 -- 0.42 412 2.29 13.52
Maintenance line 3-176 25.4--0.50 452 2.51 11.86
CMS96-3 3-307 25.4 2.20 0.49 576 5.71 12.15
Maintenance line 3-306 22.5 1.90 0.48 434 3.15 13.42
CMS96-5 C27 19.6 0.70 0.50 571 5.92 17.44
Maintenance line P1 16.8 0.30 0.46 626 4.88 17.40
(2) CMS96 cytoplasmic sterility based material has that sterility is stable, sterile degree and sterile plant rate 100%, nectary are normal, the white degeneration of flower pesticide, transformation speed are fast, the advantage of plant neat and consistent.
Chinese cabbage male sterile line CMS96 is the cabbage type rape cytoplasmic sterility, and sterile plant rate and sterile degree are 100%, and sterility is stable.Acquired 5 parts of Chinese cabbage cytoplasm male sterile line CMS96 that isozygoty are also stable for sterility, sterile plant rate and sterile degree are 100%, the cytoplasmic male sterile line of genetic stability, and its nectary and column cap are normal, the white degeneration of flower pesticide and have outstanding advantages such as transformation is easy.Fast and the stable advantage of sterility of its transformation speed, be any in the past Chinese cabbage male sterile line can not compare.Can obtain the Chinese cabbage cytoplasm male sterile line of the required type of genetic stability with common Chinese cabbage and 5 generations of stable Chinese cabbage cytoplasm male sterile line CMS96 backcross transformation.
(3) this male sterile line is consistent with maintenance line on disease resistance, but volume increase obviously.
Male sterile line and the F1 that joins with male sterile line examination try the F1 that joins quite or increase to the disease resistance of the main diseases of Chinese cabbage such as virus disease, downy mildew, blackspot and black rot and corresponding maintenance line with maintenance line.Aspect output, by table 1 as seen, no matter male sterile line and its F1 be that waste dish and clean vegetables output are apparently higher than maintenance line and its F1.Male sterile line on average improves 33% than maintenance line clean vegetables output, is up to 57%.The F1 clean vegetables output that the F1 that joins with the male sterile line examination joins than the maintenance line examination on average improves 10%.
(4) the seed production height of the CMS96 cytoplasmic male sterile line first generation of hybrid is set seeds.
To 3 parts of male sterile lines and the vernalization at room temperature of corresponding maintenance line, treat that seed " broken tripe " is placed on that low temperature vernalization is after 25 days in 4 ℃ of refrigerators, sowing is in the greenhouse.The flower that other Chinese cabbage in full bloom same period is also arranged in the greenhouse simultaneously.Florescence allows insect nature supplementary pollination, results and species test behind the seed maturity, weighs, and the result is as shown in table 3, shows: male sterile line F1 is higher than the F1 seed production of maintenance line for seed production.
Three kinds of male sterile lines of table 3 and corresponding maintenance bind kind of production ratio
Code name characteristics seed production (gram)/strain difference
01-411 maintenance line 0.22
01-646 male sterile line 0.48 0.26
01-604 maintenance line 0.14
01-644 male sterile line 0.32 0.18
01-462 maintenance line 0.89
01-709 male sterile line 1.14 0.25
(5) the CMS96 cytoplasmic sterile line is a kind of novel and good cabbage cytoplasm male sterile material, has shown wide application prospect.
The CMS96 cabbage cytoplasm male sterile line has overcome Pol-CMS and has existed fertility to be subjected to the defective of environmental influence and Ogu-CMS many for shortcomings such as the degeneration of plant growing way, heterosis, hybrid vigor difference after the transformation.CMS96 male sterile line and maintenance line, male sterile line F1 and corresponding maintenance line F1 are being carried out analysis-by-synthesis in botany proterties and nutrient component, thinking that this male sterile line has obtained the proterties and the nutrient component of maintenance line after relatively, and vitality is not degenerated.Join hybrid combination with this male sterile line examination, showing yield potential preferably aspect the Chinese cabbage output, have the high characteristics of coordinate force; Male sterile line F1 is for the seed production height simultaneously, and the production of hybrid seeds is easy, cost is low.In sum, the CMS96 cabbage cytoplasm male sterile line be a have utilize very much be worth, good novel Chinese cabbage cytoplasm male sterile line, good application prospects is arranged.Further carry out the research of this male sterile line and utilize the improvement that will help the Chinese cabbage breeding method, will have breakthrough and improve aspect the Chinese cabbage breeding approach.
The acquisition of embodiment 2, the novel cabbage type rape cytoplasmic male sterile line of a variety of Chinese cabbage transformation material
Carry out the flower bud phase since 2000 with 5 portions of a variety of Chinese cabbages and 1 part of stable Chinese cabbage CMS96 male sterile line and shell flower bud and florescence bagging hybridization, choosing petiole from filial generation does not have the strain system of the leaf wing, leaf deflection a variety of Chinese cabbage, and continue to carry out backcross transformation with male sterile lines with these 5 parts good a variety of Chinese cabbages, (code name was respectively 03.N-3 in 2003 to have obtained the seed in 2 generations of transformation, 03.N-81,03.N-118,03.N-147,03.N-168).The result shows the two hybridization easily, can obtain sterile degree and sterile plant rate and be 100% a variety of Chinese cabbage male sterile line.This a variety of Chinese cabbage male sterile line growth potential is strong, sets seeds.Though also do not obtain at present a variety of Chinese cabbage male sterile line of genetic stability, this male sterile line shows after 2 generations of transformation stable sterility and vigorous advantages such as growth potential have shown its application prospect in a variety of Chinese cabbage breeding.
Claims (9)
1, the preparation method of the novel cabbage type rape cytoplasmic male sterile line of Chinese cabbage group transformation material is characterized in that this method may further comprise the steps:
(1) cabbage type rape cytoplasmic male sterile line and Chinese cabbage group are carried out artificial flower bud phase stripping flower bud or florescence bagging hybridization, expose column cap after, the pollen of Chinese cabbage is coated with thereon, column cap normally expands, and obtains transformation generation seed;
(2) transformation generation seed is adopted the method that adds the generation processing, by field observation, obtaining excellent sterile in conjunction with the little strain of select and remain autumn old strain and spring is individual plant, and carries out many for backcross transformation with the pollination of Chinese cabbage elite plant strain;
(3) backcross and obtain the Chinese cabbage cytoplasmic sterility based material of genetic stability after five generations.
2, method according to claim 1, it is characterized in that: describedly add generation and handle and to be meant and to add generation 1 time～2 times every year, specific practice is: the seed that will just accept soaked 20～40 minutes with clear water, be put into again in 2%～4% the hydrogen peroxide and soaked 2 hours～4 hours, 23 ℃～27 ℃ following vernalization, the seed that will sprout next day was put into 3 ℃ of refrigerator vernalization 20 days～30 days; Broadcast after the taking-up in 20 ℃～21 ℃ the greenhouse, adopting intensity of illumination night is that the light source of 3000lux replenishes illumination; Can bloom by bolting in 25 days～30 days, and carried out flower bud phase or artificial back pollinating of florescence with maintenance line then.
3, method according to claim 1 and 2 is characterized in that: described Chinese cabbage group is a Chinese cabbage.
4, method according to claim 1 and 2 is characterized in that: described Chinese cabbage group is an a variety of Chinese cabbage.
5, the preparation method of the novel cabbage type rape cytoplasmic male sterile line of Chinese cabbage group transformation material, it is characterized in that this method is: a variety of Chinese cabbage and stable Chinese cabbage sterile line material are carried out flower bud phase stripping flower bud and florescence bagging hybridization, obtain a variety of Chinese cabbage cytoplasmic sterility based material of genetic stability.
6, the novel cabbage type rape cytoplasmic male sterile line of the Chinese cabbage group transformation material of method acquisition according to claim 1 and 2.
7, male sterile line according to claim 6 is characterized in that: described Chinese cabbage group is a Chinese cabbage.
8, male sterile line according to claim 6 is characterized in that: described Chinese cabbage group is an a variety of Chinese cabbage.
9, the novel cabbage type rape cytoplasmic male sterile line of the Chinese cabbage group transformation material of method acquisition according to claim 5.
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| CN104620880A (en) * | 2015-02-02 | 2015-05-20 | 河南师范大学 | Chemical control method for facilitating BNS maintenance line fertility restoration |
| CN106472300A (en) * | 2016-10-26 | 2017-03-08 | 北京市农林科学院 | A kind of method that initiative can stablize the Chinese cabbage germplasm carrying Radix Raphani chromosome of heredity |
| CN106508669A (en) * | 2016-11-16 | 2017-03-22 | 青岛市农业科学研究院 | Breeding method of Chinese cabbages resistant to clubroot |
| CN110235774A (en) * | 2019-03-18 | 2019-09-17 | 长沙丰景大地农业科技有限公司 | A method of cultivating wax powder-free cabbage type rape tender flower stalk |
| CN110235774B (en) * | 2019-03-18 | 2022-08-19 | 长沙丰景大地农业科技有限公司 | Method for cultivating wax powder-free cabbage type rape bolts |
| CN111149682A (en) * | 2020-01-02 | 2020-05-15 | 贵州省油料研究所(贵州省香料研究所) | Breeding method of colorful leaf dwarfing ornamental rape line |
| CN111149682B (en) * | 2020-01-02 | 2022-10-04 | 贵州省油料研究所(贵州省香料研究所) | Breeding method of colorful leaf dwarfing ornamental rape line |
| CN112189563A (en) * | 2020-10-19 | 2021-01-08 | 西南大学 | Method for accelerating transformation of cytoplasmic male sterile line of brassica napus |
| CN115968771A (en) * | 2021-12-24 | 2023-04-18 | 苏州市农业科学院 | Breeding method of male sterile line of purple cabbage |
| CN115968771B (en) * | 2021-12-24 | 2024-05-14 | 苏州市农业科学院 | Breeding method of purple cabbage male sterile line |
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