CN101810136A - Cauliflower cytoplasmic male sterile line breeding method and application of male sterile line - Google Patents
Cauliflower cytoplasmic male sterile line breeding method and application of male sterile line Download PDFInfo
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- CN101810136A CN101810136A CN 201010166573 CN201010166573A CN101810136A CN 101810136 A CN101810136 A CN 101810136A CN 201010166573 CN201010166573 CN 201010166573 CN 201010166573 A CN201010166573 A CN 201010166573A CN 101810136 A CN101810136 A CN 101810136A
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Abstract
The invention discloses a cauliflower cytoplasmic male sterile line breeding method and application of a male sterile line. A pure line cultured by isolated microspores of cauliflower is used as a recurrent parent to be carried out backcross of 4-5 generations with OgurDH cytoplasmic male sterile of European cauliflower to obtain a stable sterile line which is bred by using a tissue culture and rapid propagation technology, thereby avoiding character degeneration caused by multi-generation backcross. With the technical scheme, the invention overcomes the defects that the OgurDH cytoplasmic male sterile has yellowed plant seedlings at low temperature and undeveloped nectar, and ensures that the male sterile line is not influenced by temperature change, and the sterility rates and the male sterility reach 100 percent. Plants grow vigorously, have developed nectar and high combining ability, normally bloom and fruit, and reach the crossing rate of 100 percent when used for preparing crossbreeds in a strict isolation region. The first-generation crossbreeds have vigorous growth, disease resistance as well as high quality and yield.
Description
Technical field the present invention relates to a kind of vegetable breeding method, is specifically related to the application of a kind of cauliflower cytoplasmic male sterile line transfer method and male sterile line thereof.
The background technology cauliflower imports China in the middle of the 19th century, introduce some crossbreed from the U.S. seventies in 20th century, and its output obviously is better than conventional the kind, China breeding work person cauliflower cross breeding method that also begins one's study.It mainly adopts the self incompatible line production of hybrid seeds, but adopt this method hybrid rate to be difficult to reach 100%, and the self incompatible line production of hybrid seeds needs artificial bud pollination, cost is artificial greatly, breeding cost is high, and through after the continuous multi-generation selfing, economic characters can fail gradually, and hybrid seed yield reduces, and the advantage of crossbreed weakens.And utilize the male sterile line production of hybrid seeds can overcome the problems referred to above, the research of the person the pays attention to cauliflower male-sterile line breeding production of hybrid seeds that this impels the breeding work.
Cauliflower male sterile source mainly contains two kinds of nuclear gene male sterility and cytoplasmic male sterilities.Nuclear male sterility is through experimental results show that and can not using.Cytoplasmic male sterile line has Pol CMS (PolimDH CMS is sterile sources such as juncea, black mustard) and Ogu CMS (male sterile with radish cytoplasm), and what have value in cauliflower is the radish cytoplasmic male sterilty.Its sterility of Ogu CMS is to be controlled jointly by plasmon and 2 pairs of recessive karyon genes, exists problems such as low temperature yellow in following seedling stage, nectary be undeveloped, has influenced the application of the sterile type of Ogu CMS.U.S. Cornell University adopts the method for biotechnology that Ogu CMS sterile material gene is modified, overcome it to a certain extent in low temperature yellow in seedling stage, problems such as nectary is undeveloped, transformation success on some crops, but still have problems such as premunition, coordinate force, seed production and kytoplasm negative effect, therefore also do not enter the production phase at present.China's Tianjin Vegetable Research Institute, Foochow Vegetable Research Institute are introduced the OguCMS sterile material, breed " Tianjin product 65 (cauliflower variety that Tianjin Ke Run cultivates) " and " C50-2 " (cauliflower variety that Fuzhou City vegetables Science Institute cultivates) male sterile line through transformation respectively, but because production of hybrid seeds difficulty, also not large scale application.Therefore, the cauliflower cytoplasm male sterility line that seed selection sterility is stable, transformation is easy, coordinate force is strong, hybrid seed yield is high is breeding work person's the objective of the struggle.
Summary of the invention the objective of the invention is to overcome above-mentioned the deficiencies in the prior art, provides a kind of male sterility stable, plant with blossom and bear fruit normal, the cauliflower cytoplasm male sterility line transfer method that coordinate force is strong and the application of male sterile line thereof.
For realizing above purpose, the present invention adopts following transfer method: with the cauliflower pure lines is that 4-5 is carried out for backcrossing in recurrent parent and cauliflower cytoplasmic sterility source.Can obtain stable cauliflower cytoplasmic male sterile line.
Described cauliflower sterile source is that Europe utilizes cytoplasmic sterility source CMS2001DH, CMS2001B, the CMS2001C after biotechnology is modified;
Pure lines DH36-5, DH19-1, the DH32-2 of described cauliflower pure lines for breeding with free microspores culture method.
The method of described cauliflower pure line selection: free microspores culture was bred pure lines in 2 years.Being characterized in microspore 100% from nucleus, is the genotype of isozygotying, and breeding a plant from a microspore is exactly pure lines.
The resulting cauliflower cytoplasmic male sterile line of above-mentioned transfer method carries out vegetative propagation with group culturation rapid propagating technology.Be characterized in avoiding many and degenerate, keep the merit of male sterile line for the proterties of backcrossing.
The cauliflower cytoplasm male sterility line that resulting cauliflower cytoplasmic male sterile line of above-mentioned transfer method or group culturation rapid propagating technology are bred is for maternal, the cauliflower selfing is that male parent is hybridized the acquisition hybrid seed, and no brassicaceous vegetable district carries out hybrid seeding in 1500 meters scopes.
The present invention adopts above technical scheme, utilizing Europe to modify good cytoplasmic sterility source, back does maternal, the good pure lines of free microspores culture are made male parent, through 4 generation backcross transformation become plant strain growth normal, the proterties of blossoming and bearing fruit is normal, the cytoplasm male sterility line that coordinate force is strong, and utilize cytoplasmic male sterile line to make the cauliflower hybrid of strong advantage.The present invention has the following advantages: male sterility is controlled by cytoplasm, and transformation is easy, and any cauliflower inbred line can be as their maintenance line; Pure lines with free microspores culture are made backcross parent, backcross 4 generation proterties basicly stable, stable cauliflower cytoplasmic male sterile line is bred with group culturation rapid propagating technology, is characterized in avoiding many generations proterties of backcrossing to degenerate, and keeps the merit of male sterile line.The male sterile line stable fertility of breeding is not subjected to environmental influence, and sterile plant rate and sterile degree all can reach 100%; The male sterile line vine growth and development is normal, the nectary prosperity, and the proterties of blossoming and bearing fruit is normal, and coordinate force is strong; The male parent of crossbreed can be used inbred line, can spontaneous pollination in isolated area, and the output height, cost is low, the hybrid rate height, seed quality is guaranteed.The male sterile of breeding ties up in the production production of hybrid seeds and can not plagiarized.
Cauliflower pure lines DH36-5, DH19-1, the DH32-2 that embodiment the present invention breeds with the microspores culture method of dissociating is that recurrent parent utilizes cauliflower cytoplasmic sterility source CMS2001DH, CMS2001B, CMS2001C after biotechnology is modified to carry out 4-5 for backcrossing with Europe.Free microspores culture can be bred pure lines in 2 years.Can obtain stable cauliflower cytoplasmic male sterile line.
The resulting cauliflower cytoplasmic male sterile line of above-mentioned transfer method can adopt group culturation rapid propagating technology to carry out vegetative propagation.
The cauliflower cytoplasm male sterility line that resulting cauliflower cytoplasmic male sterile line of above-mentioned transfer method or group culturation rapid propagating technology are bred can be female parent, the cauliflower selfing is that male parent is hybridized the acquisition hybrid seed, and no brassicaceous vegetable district carries out hybrid seeding in 1500 meters scopes.
Embodiment: pure line selection:
Microspores culture to good crossbreed dissociates can obtain a large amount of double haploids, comprehensively identifies through the field, selects growth vigorous, bouquet is pure white, and is loose, the output height, coordinate force is strong, the strain system that index of affinity is high, i.e. DH36-5, DH32-2, DH19-1 (table 1).
The main economic characters of table 1 pure lines
The introduction of cytoplasmic male sterile line and transformation:
Calendar year 2001 is introduced cytoplasmic male sterility source CMS2001DH, CMS2001B, the CMS2001C of cauliflower after biotechnology is modified from Europe, identify through the field and select good individual plant.
With pure lines DH36-5 is that backcross parent and CMS2001DH carry out 4 generations backcross transformation acquisition cauliflower cytoplasmic male sterile line CMS36-5DH.
With pure lines DH32-2 is that backcross parent and CMS2001B carry out 4 generations backcross transformation acquisition cauliflower cytoplasmic male sterile line CMS32-2B.
With pure lines DH19-1 is that backcross parent and CMS2001C carry out 4 generations backcross transformation acquisition cauliflower cytoplasmic male sterile line CMS19-1C.
Can adopt simultaneously and highly carry out transformation for inbred line, but stable obviously slow than the pure lines transformation of proterties, as long as pure lines are 4 generations, and inbred line is wanted 5-6 generation, and the partial sterility that obtains a collection of cauliflower cytoplasmic male sterile line is proterties such as table 2.
The main economic characters of table 2 part male sterile line
With above cauliflower cytoplasm male sterility line CMS36-5DH, CMS32-2B, the CMS19-1C that breeds is maternal, can hybridize with cauliflower inbred line 1314,1422,1826, obtains hybrid seed 0606,0618,0625.
Below in conjunction with example the present invention is further described:
According to the advantage of European cauliflower OgurDH sterile source, introduce the cytoplasmic male sterilty source after modifying in a large number from Europe, carry out the field and comprehensively identify, filtering out does not have yellow seedling stage, the plant strain growth stalwartness, and bouquet is good, growth is bloomed normally, and nectary is flourishing, and solid good plant is as sterile source.Pure lines and the height cultivated with Isolated microspore are that backcross parent carries out extensive test cross for selfing, screening female parent and male parent, carrying out many generations simultaneously backcrosses, target is the yellow problem that solves male sterile line, optimize tree characteristics, improve resistance etc., make each proterties reach breeding objective, finally realize heterosis utilization near male parent.Carry out test cross simultaneously, breed dissimilar cauliflower cytoplasm male sterility lines, and carry out combining ability test, select the hybrid of strong advantage, be applied to produce.
The cauliflower cytoplasm male sterility line of breeding with said method is a female parent, is male parent with good cauliflower selfing, joins method with half wheel and hybridizes the preparation combination, and carry out the field and comprehensively identify, filter out the combination of strong advantage, and, breed the good first generation of hybrid by combination multiple spot comparative trial.
The hybridization achievement:
1, bred a collection of cauliflower cytoplasm male sterility line
Utilize cauliflower OgurDH sterile source, bred a collection of dissimilar male sterile line in order to last method transformation, the proterties of its non-yellow is a genetic stability, and the sterile plant rate of male sterile line, sterile degree all reach 100%, tree characteristics is near its corresponding maintenance line, and hybrid vigour is obvious.
2, preserved a collection of cauliflower germplasm resource
In the transformation of cauliflower cytoplasmic sterile line, we have compiled cauliflower germplasm resource both domestic and external, are tested and appraised, select, and have obtained a collection of dissimilar germ plasm resource.Carry out Isolated microspore simultaneously and cultivate, obtain a collection of good pure lines.
3, bred 3 morning, in ripe fine combination
Through combining ability test, comprehensive character test in field and variety comparative test filter out 0606,0618,0,625 three fine combination.It is characterized in that plant type compactness, disease-resistant, green stalk preserved egg, quality better, output height, be respectively breeding time 55 days, 60 days, 70 days.
4, produce the cauliflower crossbreed with cytoplasm male sterility line, its main points are as follows:
(1) determines sowing time, cultivate strong sprout
The male sterile line female parent: tissue cultivating seedling preceding 35 days at setting date, tissue cultivating seedling is transferred on the root media, after 25 days the bottle seedling is transplanted in the matrix and cultivates, shift out the greenhouse after 5 days, shading condition lower refining seedling 5 days.Male parent: arrange sowing time according to the maturing stage, early-maturing variety (in 60 days) is in sowing mid-September, and middle seasonal strain (60-70 days) was sowed in early September.Rough leaf is launched back thinning, heels in hardening when growing to 3 true leaves.
(2) field planting is rationally enhanced field management
The selection soil is fertile, and irrigation and drainage are convenient, and preceding plot of recommending the non-brassicaceous vegetable of plantation does not have the plantation brassicaceous vegetable in 1500 meters scopes.About every mu of plantation 3600 strains, maternal ratio with male parent is 2: 1.2 boron fertilizers of spray satisfy the demand of cauliflower to boron in vegetative period.
(3) the strict selection in time excised bouquet
In kind of strain vegetative period, note eliminating sick and weak strain.When bouquet begins to loose, the cut-out leaf-head, generally staying diameter is 6cm
2Bouquet.
(4) regulate the florescence, improve output
Male parent cans be compared to most maternally early bloomed in 2 days, and the parent of first bolting is pinched or sprays that N is fertile to be regulated, and the fertile promotion of the spray P of back bolting is bloomed.
(5) in time gather, guarantee quality
Plant the little Huang of pod, gather when seed changes brown, the seed on the male sterile line is exactly a hybrid seed.
Claims (7)
1. cauliflower cytoplasmic male sterile line transfer method is characterized in that: with the cauliflower pure lines is that 4-5 is carried out for backcrossing in recurrent parent and cauliflower cytoplasmic sterility source.
2. a kind of cauliflower cytoplasmic male sterile line transfer method as claimed in claim 1 is characterized in that: described cauliflower sterile source is that Europe utilizes cytoplasmic sterility source CMS2001DH, CMS2001B, the CMS2001C after biotechnology is modified.
3. a kind of cauliflower cytoplasmic male sterile line transfer method as claimed in claim 1 is characterized in that: pure lines DH36-5, DH19-1, the DH32-2 of described cauliflower pure lines for breeding with free microspores culture method.
4. a kind of cauliflower cytoplasmic male sterile line transfer method as claimed in claim 4 is characterized in that: the method for described cauliflower pure line selection: free microspores culture was bred pure lines in 2 years.
5. the application of the male sterile line of a kind of cauliflower cytoplasmic male sterile line as claimed in claim 1 transformation is characterized in that: the resulting cauliflower cytoplasmic male sterile line of above-mentioned transfer method carries out vegetative propagation with group culturation rapid propagating technology.
6. as the application of the male sterile line of claim 1 or 5 described a kind of cauliflower cytoplasmic male sterile line transformations, it is characterized in that: the cauliflower cytoplasm male sterility line that resulting cauliflower cytoplasmic male sterile line of above-mentioned transfer method or group culturation rapid propagating technology are bred is for maternal, and the cauliflower selfing is that male parent is hybridized the acquisition hybrid seed.
7. the application of the male sterile line of a kind of cauliflower cytoplasmic male sterile line as claimed in claim 6 transformation is characterized in that: hybridize in above-mentioned cross method no brassicaceous vegetable district in 1500 meters scopes.
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Cited By (5)
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| CN102726300A (en) * | 2012-07-06 | 2012-10-17 | 福建农林大学 | Anther culture method of cauliflower |
| CN103168681A (en) * | 2013-04-16 | 2013-06-26 | 镇江瑞繁农艺有限公司 | Breeding method of early-ripe and disease-resistant broccolini |
| CN110338053A (en) * | 2019-08-12 | 2019-10-18 | 浙江省农业科学院 | A kind of selection of orange pagoda cauliflower cenospecies |
| CN111789026A (en) * | 2020-08-24 | 2020-10-20 | 天津惠尔稼种业科技有限公司 | Ogura sterile source male sterile line generation-adding transformation technology |
| CN114938778A (en) * | 2022-07-25 | 2022-08-26 | 中国热带农业科学院三亚研究院 | Breeding method of passion flower with wing stem |
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| CN1565170A (en) * | 2003-07-02 | 2005-01-19 | 天津科润农业科技股份有限公司蔬菜研究所 | Hybrid cauliflower cultivation method utilizing male sterility line |
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| CN1565170A (en) * | 2003-07-02 | 2005-01-19 | 天津科润农业科技股份有限公司蔬菜研究所 | Hybrid cauliflower cultivation method utilizing male sterility line |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102726300A (en) * | 2012-07-06 | 2012-10-17 | 福建农林大学 | Anther culture method of cauliflower |
| CN103168681A (en) * | 2013-04-16 | 2013-06-26 | 镇江瑞繁农艺有限公司 | Breeding method of early-ripe and disease-resistant broccolini |
| CN110338053A (en) * | 2019-08-12 | 2019-10-18 | 浙江省农业科学院 | A kind of selection of orange pagoda cauliflower cenospecies |
| CN110338053B (en) * | 2019-08-12 | 2021-06-04 | 浙江省农业科学院 | Breeding method of orange pagoda cauliflower hybrid |
| CN111789026A (en) * | 2020-08-24 | 2020-10-20 | 天津惠尔稼种业科技有限公司 | Ogura sterile source male sterile line generation-adding transformation technology |
| CN114938778A (en) * | 2022-07-25 | 2022-08-26 | 中国热带农业科学院三亚研究院 | Breeding method of passion flower with wing stem |
| CN114938778B (en) * | 2022-07-25 | 2022-11-18 | 中国热带农业科学院三亚研究院 | Breeding method of passion flower with wing stem |
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Application publication date: 20100825 |