WO2023005883A1 - 一种蓝标型两系法杂交小麦***的选育方法及应用 - Google Patents

一种蓝标型两系法杂交小麦***的选育方法及应用 Download PDF

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WO2023005883A1
WO2023005883A1 PCT/CN2022/107649 CN2022107649W WO2023005883A1 WO 2023005883 A1 WO2023005883 A1 WO 2023005883A1 CN 2022107649 W CN2022107649 W CN 2022107649W WO 2023005883 A1 WO2023005883 A1 WO 2023005883A1
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seeds
plants
plant
blue
dual
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French (fr)
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李中安
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西南大学
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Priority to GB2318620.8A priority Critical patent/GB2624547A/en
Priority to US17/998,341 priority patent/US20230329167A1/en
Priority to EP22848488.7A priority patent/EP4342289A1/en
Publication of WO2023005883A1 publication Critical patent/WO2023005883A1/zh

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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
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
    • A01H1/021—Methods of breeding using interspecific crosses, i.e. interspecies crosses
    • 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
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
    • 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
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
    • 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
    • A01H5/00—Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
    • A01H5/10—Seeds
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture

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  • the invention belongs to the field of crop genetics and breeding, and relates to a breeding method and application of a blue standard two-line hybrid wheat system.
  • Wheat is the most consumed food crop in the world. With the reduction of arable land and the increase of population, it is imperative to increase the yield of wheat per unit area.
  • the use of heterosis is an important way to increase the yield of crops. Rice, corn, sorghum and other crops have already been hybridized, and they have been widely used in production one after another, greatly increasing the yield.
  • the utilization of heterosis in wheat has always been a difficult problem in the world, and no breakthrough has been made in production until now.
  • the main reason is that wheat is polyploid, and it uses a large amount of seeds, and its heterosis is not as strong as that of rice, corn, sorghum and other crops. Therefore, in order to produce high-yield, low-cost hybrid seeds, a feasible hybrid wheat System is the key.
  • Driscoll successively established the XYZ system and the improved XYZ system. These two systems are both GMS systems, which utilize the dominant male fertility gene and the dominant marker gene for ear stem villi carried on the 5R chromosome of rye. Sexual markers can only be identified after wheat heading, which obviously lacks practicality for high-density planting wheat, but it has important guiding significance for the creation and application of GMS.
  • Huang Shousong et al. (1991) and Zhou Kuanji et al. (1998) successively established a blue standard type hybrid wheat system similar to the XYZ system with blue grains as the marker, which solved the problem of effective marking and distinguished the sterile line and the maintainer line through the color of the seeds.
  • the inventor previously established "a method for breeding two-line hybrid wheat with blue grains as a marker trait (ZL200610042629.8)", and has bred a batch of sterile lines and 3 provincial-level approved hybrids. A number of hybrid combinations are participating in the test, and more hybrids will be approved and promoted in succession. Seed sorting and seed production have met the requirements of hybrid wheat seeds, but the stability of the additional translocated chromosomes needs to be improved, and the break frequency of the additional chromosomes from the centromere is high (0.5-10%). The dual-purpose system with low fracture frequency has certain difficulties.
  • mutants at the ms1 gene locus including Pugsley's mutant (including allele ms1a), Probus mutant (including allele ms1b), Cornerstone mutant (including allele ms1c), FS2 mutant (including allele gene ms1d), FS3 mutants (including allele ms1e), FS24 mutants (including allele ms1f), LZ mutants (including allele ms1g) and Ningchun 4 series mutants (including allele series ms1h-p) etc.
  • Pugsley's mutant including allele ms1a
  • Probus mutant including allele ms1b
  • Cornerstone mutant including allele ms1c
  • FS2 mutant including allele gene ms1d
  • FS3 mutants including allele ms1e
  • FS24 mutants including allele ms1f
  • LZ mutants including allele ms1g
  • Ningchun 4 series mutants including allele series m
  • Morrison et al. (2004) described the origin and differences of the blue-grain gene (Ba).
  • blue-grain gene showing endosperm intuition and dosage effect can be used as a marker to judge whether there is blue-grain gene in wheat grain and how many copies of blue-grain gene are contained (endosperm and aleurone layer are triploid, 1 copy comes from the father, 2 copies from the mother side).
  • the color of the seeds formed is light blue, and the seeds contain 1 copy of Ba (seeds obtained by selfing are non-blue, light-blue, medium-blue and dark-blue); when The color of the seeds formed by only the female gametes containing the blue gene is medium blue, and the seeds contain 2 copies of Ba (seeds obtained by selfing are non-blue, light blue, medium blue and dark blue); when both male and female gametes have The color of the seeds formed by the blue-grain gene is dark blue, and the seeds contain 3 copies of Ba (seeds obtained by selfing are all blue-grained).
  • the present invention is carried out on the basis of "a two-line hybrid wheat breeding method with blue grains as a marker trait (ZL200610042629.8)" previously established by the inventor.
  • ZL200610042629.8 first creates blue grains as a marker trait To separate and maintain the original dual-purpose line of the karyotype sterile hybrid wheat system of the sterile line, and then establish the breeding method of the new dual-purpose line, which specifically includes the following steps:
  • the T4BS.4AgL translocation chromosome and the wheat 4B substitution derived from the 4th chromosome of T. thaoudar were identified by chromosome banding method and alcohol dehydrogenase electrophoresis analysis technology Line 4th′′, at the same time, the sterile gene ms1b and dwarf gene Rht3 located on the short arm of wheat 4B chromosome were linked together through genetic recombination; then the sterile gene ms1b and dwarf gene Rht3 were simultaneously recombined into the T4BS.4AgL translocation chromosome above, to obtain blue-grain dwarf sterile plants;
  • the blue-grained dwarf sterile strain was crossed with the wheat 4B substitution line 4th′′ of the 4th chromosome to obtain the 4th and T4BS.4AgL chromosome double-somotic plants carrying the ms1b and Rht3 genes, that is, the F 1 generation, and obtained sufficient For the F 2 generation white or red, light blue and dark blue seeds, only the F 2 generation dark blue seeds were sown on demand; all tall-stalk fertile plants in the F 2 population were numbered and used as male parents, respectively
  • the sterile plants in the line derived from the sterile gene ms1b of the "Probus" mutant were crossed, and only the hybrid ears with light blue seeds were harvested, and the F1 generation of hybrids with light blue seeds were planted, bagged after heading, and all single plants could be selected.
  • the plants grown from the selected white or red seeds are all sterile and similar All the fertile light blue-grained strains of the plants grown from the corresponding light blue-grained seeds are harvested from the light-blue-grained strains, wherein the light-blue-grained strains contain the required T4thS that integrates the exogenous restoration gene and the blue-grained gene .4AgL translocation chromosome;
  • T4thS.4AgL translocation chromosomal line Self-cross the above-mentioned T4thS.4AgL translocation chromosomal line or continue to cross with the sterile line without the Rht3 dwarf gene in the sterile ms1b derivative line containing the "Probus" mutant, and then self-cross and select from the offspring population
  • the ratio of blue grains to white grains or red grains is 1:2, and dual-purpose lines are initially screened out, and then confirmed by microscopic examination of chromosome pairing in the meiotic metaphase of pollen mother cells, so as to obtain the original dual-purpose lines of the hybrid wheat system.
  • Use 21II+I 4thS.4AgL Use 21II+I 4thS.4AgL ;
  • Method 1 Prepare a hybrid combination of the original dual-purpose line and an excellent line, select only blue-grained seeds for planting in the F1 generation, select an excellent combination according to the performance of each combination, and sort light blue-grained seeds from the F2 generation seeds of the selected combination Seed planting, select excellent individual plants; plant the white or red and light blue seeds in each line of the F 3 generation respectively, 50 plants each, and 25% or 100% of the white or red populations are selected and harvested
  • the light blue-grained line corresponding to the sterile plant and good agronomic properties, the latter will be a dual-purpose line after stabilization; and so on until the strain is stable and all the isolated white or red-grained populations are sterile; Test the yield of dual-purpose lines and conduct quality analysis and disease resistance identification of white or red seeds isolated from them, so as to obtain new dual-purpose lines with high yield, high quality, and multi-resistance;
  • Method 2 Select the superior sterile strain isolated from the F2 generation population of the hybridization of the sterile line and the conventional superior strain as the female parent to hybridize with the original dual-purpose strain, the same as the method 1 , select and retain the superior combination and Excellent single plant, F 3 generations or more to select excellent lines, but the difference is that only light blue seeds are planted in the remaining generations until the sterile line is needed; this method does not require emasculation when doing crosses, and is suitable for strong dominant combinations Utilization of superior sterile strains isolated from F2 populations.
  • the object of the present invention is to provide a breeding method and application of a blue standard two-line hybrid wheat system.
  • the invention provides a kind of breeding method of blue standard two-line method hybrid wheat, comprises the steps:
  • the white-grained sterile plant of the 06L6109-3 dual-purpose line was crossed with the wheat with the translocation chromosome T4AgL(Ba)-4BL ⁇ 4BS, and the dwarf and male sterile plants grown from blue-grained seeds were screened, namely It is a plant with translocation chromosome T4AgL(Ba)-4BL ⁇ 4BS(Rht3-ms1b), called T4AgL(Ba)-4BL ⁇ 4BS(Rht3-ms1b) translocation plant;
  • T4AgL(Ba)-4BL ⁇ 4BS(Rht3-ms1b) translocation line plants were crossed with wheat 4B substitution line 4th to obtain T4AgL(Ba)-4BL ⁇ 4BS(Rht3-ms1b) with 4th and translocation chromosome dual monomer seeds;
  • step (3) sowing and cultivating the double monomer seeds obtained in step (2) into plants, selfing, selecting dark blue seeds for planting and screening out tall and male fertile plants;
  • step (3) (4) hybridize the plant screened in step (3) with the white-grained sterile strain of the 14L6386 dual-purpose line, and screen the hybrid combination with blue-grained seeds;
  • the white-seed sterile plant of the 14L6386 dual-purpose line is a male sterile plant grown from the white-grained seeds obtained by selfing of the 14L6386 dual-purpose line;
  • the blue-grain dual-purpose line; 09L6034 dual-purpose line is a blue-grain dual-purpose line selected after backcrossing T0065-10B-2LB-4 dual-purpose line and the recurrent parent wheat line Zhou 88114 (named Zhoumai 11).
  • Use line; T0065-10B-2LB-4 is an addition line wheat containing translocation chromosome T4thS ⁇ 4AgL;
  • step (4) the blue grain seeds are sown and cultivated into plants. If the plants obtained from all the seeds are male fertile plants, the plants obtained by the development of the seeds of the corresponding hybrid combination form a potential easy Bit system;
  • step (5) The potential translocation line that step (5) obtains is carried out as follows: selfing and harvesting seeds, the seeds are divided into two groups of blue seeds and white seeds; Breeding as plants, if all the individual plants obtained from the blue seeds are male fertile plants and all the individual plants obtained from the white seeds are male sterile plants, then these male fertile individual plants obtained from the blue seeds FISH identification was carried out to screen the individual plants with translocation chromosome T4AgL segment (Ba)-4BL ⁇ 4thS(Rf), and these individuals constituted the T4AgL segment (Ba)-4BL ⁇ 4thS(Rf) translocation population;
  • step (6) From the selfing progeny of the translocation line that step (6) obtains, screen white seed: the plant that the quantity of blue seed is 2:1, namely contains translocation chromosome T4AgL fragment (Ba)-4BL.
  • the karyotype of the dual-purpose line is: 21II(msms)+I T4AgL fragment (Ba)-4BL ⁇ 4thS(Rf) .
  • the invention provides a kind of breeding method of blue standard two-line method hybrid wheat, comprises the steps:
  • 15L4167 dual-purpose line is selfed, and then the male sterile plant grown by the blue seed is selected, and the plant is crossed with the fertile plant of the dark blue seed, and then the male fertile plant grown by the dark blue seed is selected and carried out Selfing, and then selecting plants with male sterility from the plants developed from selfing seeds, and further screening by microscopy to obtain additional line plants with translocation end body T4AgL fragment (Ba)-4BL; dark blue grains are fertile Plant: It is a male fertile plant grown from dark blue seeds obtained by selfing of the dual-purpose line 15L4167, and all the seeds produced after selfing are blue seeds;
  • step (3) the seed obtained in step (3) is used as the male parent and the white seed as the female parent as a hybrid combination, cultivated as a plant and hybridized, and the blue seed is screened from the seeds obtained on the female plant; the white seed is obtained by The 15L4167 dual-purpose strain was obtained by selfing;
  • step (4) sow the blue seeds obtained in step (4) and cultivate them into plants, eliminate the male sterile plants, harvest the male fertile plants and test the seeds, and the selected seeds have both blue seeds and white seeds plant;
  • step (5) selected plant, sort out white seed and blue seed respectively by single plant;
  • the seeds obtained in sowing step (6) are cultivated into plants; if the plants obtained by the blue seeds of a certain individual plant are all male fertile plants, and the plants obtained by the white seeds of this individual plant are all male fertile plants. Breeding plants, microscopically inspecting the light blue seeds of the single plant, and obtaining a translocation line plant with a translocation chromosome T4AgL segment (Ba)-4BL ⁇ 4thS segment (Rf), also known as a dual-purpose line;
  • the 15L4167 dual-purpose line is the above-mentioned additional line containing the translocation chromosome T4AgL fragment (Ba)-4BL ⁇ 4thS(Rf).
  • the karyotype of the dual-purpose line is: 21II(msms)+I T4AgL fragment (Ba)-4BL ⁇ 4thS fragment (Rf) .
  • the invention provides a kind of breeding method of blue standard two-line method hybrid wheat, comprises the steps:
  • step (1) The male sterile plant screened in step (1) is hybridized with the male fertile plant obtained in step (1), the dark blue seeds are screened and cultivated into a plant, and the male sterile plant is screened out from its selfed offspring;
  • step (2) The male sterile plant screened in step (2) is crossed with the male fertile plant screened in step (3), the seeds are harvested and cultivated into plants, then selfed, and dark blue seeds are screened, and the terminal body 4AgL is screened by microscopy Double diplosome seeds of (Ba) and telosome 4thS(Rf);
  • step (4) The double doublet seeds obtained in step (4) are subjected to mutagenesis treatment, and then cultivated into plants;
  • step (4) The plant grown from the double diploid seeds obtained in step (4) is used as the male parent, hybridized with the white-grained sterile plant as the female parent, harvests the seeds on the female parent plant, and selects the blue-grained seeds to carry out mutagenesis treatment , and then cultivated into plants;
  • white-seed sterile plant a male sterile plant grown from white seeds obtained from selfing of the 16L6386 dual-purpose line;
  • step (7) The plant obtained in step (5) or step (6) is used as the male parent, hybridized with the white-grained sterile plant as the female parent, harvests the seeds on the female parent plant and screens out the blue-grained seeds therefrom; the white-grained sterile plant Plants: male sterile plants grown from white seeds obtained from selfing of the dual-purpose line 16L6386;
  • step (7) sowing the blue seeds obtained in step (7) and cultivating them into plants, eliminating male sterile plants, selfing the selected plants and harvesting seeds, and selecting the seeds that have both blue seeds and white seeds. plant;
  • step (8) For the plants screened in step (8), the white seeds and blue seeds obtained by selfing are sorted out by individual plants respectively;
  • the white seeds and blue seeds obtained in the sowing step (9) are cultivated into plants; if the plants obtained by the blue seeds of a certain individual plant are all male fertile plants, and all the white seeds of the individual plant It is a male sterile plant, and the blue-grained seeds of the single plant are examined under a microscope to obtain a plant with a translocated terminal body T4AgL fragment (Ba)-4thS fragment (Rf).
  • the light/medium blue-grained seeds are dual-purpose lines ;
  • 16L6386 dual-purpose line is an addition line of wheat containing translocation chromosome T4thS ⁇ 4AgL.
  • the karyotype of the dual-purpose line is: 21II(msms)+I T4AgL fragment (Ba)-4thS fragment (Rf) .
  • the invention provides a kind of breeding method of blue standard type two-line method hybrid wheat, comprises the steps:
  • step (1) The male sterile plant screened in step (1) is hybridized with the male fertile plant obtained in step (1), the dark blue seeds are screened and cultivated into a plant, and the male sterile plant is screened out from its selfed offspring;
  • step (2) The male sterile plant screened in step (2) is crossed with the male fertile plant screened in step (3), the seeds are harvested and cultivated into plants, then selfed, and dark blue seeds are screened, and the terminal body 4AgL is screened by microscopy Double diplosome seeds of (Ba) and telosome 4thS(Rf);
  • step (4) The double doublet seeds obtained in step (4) are subjected to mutagenesis treatment, and then cultivated into plants;
  • step (4) The plant grown from the double diploid seeds obtained in step (4) is used as the male parent, hybridized with the white-grained sterile plant as the female parent, harvests the seeds on the female parent plant, and selects the blue-grained seeds to carry out mutagenesis treatment , and then cultivated into plants;
  • white-seed sterile plant a male sterile plant grown from white seeds obtained from selfing of the 16L6386 dual-purpose line;
  • step (7) The plant obtained in step (5) or step (6) is used as the male parent, hybridized with the white-grained sterile plant as the female parent, harvests the seeds on the female parent plant and screens out the blue-grained seeds therefrom; the white-grained sterile plant Plants: male sterile plants grown from white seeds obtained from selfing of the dual-purpose line 16L6386;
  • step (7) sowing the blue seeds obtained in step (7) and cultivating them into plants, eliminating male sterile plants, selfing the selected plants and harvesting seeds, and selecting the seeds that have both blue seeds and white seeds. plant;
  • step (8) For the plants screened in step (8), the white seeds and blue seeds obtained by selfing are sorted out by individual plants respectively;
  • the seeds obtained in the sowing step (9) are cultivated into plants; if the plants obtained by the blue seeds of a certain individual plant are all male fertile plants, and the white seeds of the individual plant are all male sterile plants, Microscopically examine the blue-grained seeds of the single plant to obtain a plant with a translocation endosome T4thS fragment (Rf)-4AgL fragment (Ba), and the light/medium blue-grained seeds are dual-purpose lines;
  • 16L6386 dual-purpose line is an addition line of wheat containing translocation chromosome T4thS ⁇ 4AgL.
  • the karyotype of the dual-purpose line is: 21II (msms)+I T4thS fragment (Rf)-4AgL fragment (Ba) .
  • the invention provides a kind of breeding method of blue standard type two-line method hybrid wheat, comprises the steps:
  • step (1) The double-diende seeds obtained in step (1) are subjected to mutagenesis treatment, and then cultivated into plants;
  • step (3) The plant grown from the double-diende seeds obtained in step (1) is used as the male parent, hybridized with the white-grained sterile plant as the female parent, harvests the seeds on the female parent plant, and selects the blue-grained seeds to carry out mutagenesis treatment , and then cultivated into plants;
  • white-seed sterile plant a male sterile plant grown from white seeds obtained from selfing of the 15L4167 dual-purpose line; An additional line of Ba)-4BL ⁇ 4thS(Rf);
  • step (2) or step (3) The plant obtained in step (2) or step (3) is used as the male parent, and is hybridized with the white-grained sterile plant as the female parent, and the seeds on the female parent plant are harvested and the blue-grained seeds are screened out;
  • the white-grained sterile plant Plant a male sterile plant grown from white seeds obtained from selfing of the 15L4167 dual-purpose line;
  • the 15L4167 dual-purpose line is the above-mentioned one containing the translocation chromosome T4AgL fragment (Ba)-4BL ⁇ 4thS(Rf) additional department;
  • step (4) sowing the blue seeds obtained in step (4) and cultivating them into plants, eliminating male sterile plants, selfing the selected plants and harvesting seeds, and selecting the ones with both blue seeds and white seeds in the selected seeds plant;
  • step (5) For the plants screened in step (5), the white seeds and blue seeds obtained by selfing are sorted out by individual plants respectively;
  • the seeds obtained in sowing step (6) are cultivated into plants; if the plants obtained by the blue seeds of a certain individual plant are all male fertile plants, and the white seeds of this individual plant are all male sterile plants, The blue-grained seeds in these individual plants were examined under the microscope, and the plants with the translocation terminal body T4thS fragment (Rf)-4AgL fragment (Ba)-4BL ⁇ were obtained, and the light/medium blue-grained seeds were dual-purpose lines.
  • the karyotype of the dual-purpose line is: 21II(msms)+I T4thS fragment (Rf)-4AgL fragment (Ba)-4BL ⁇ .
  • Described method specifically comprises the steps:
  • the white-grained sterile plant of the 06L6109-3 dual-purpose line was crossed with the wheat with the translocation chromosome T4AgL(Ba)-4BL ⁇ 4BS, and the dwarf and male sterile plants grown from blue-grained seeds were screened, namely It is a plant with translocation chromosome T4AgL(Ba)-4BL ⁇ 4BS(Rht3-ms1b), called T4AgL(Ba)-4BL ⁇ 4BS(Rht3-ms1b) translocation plant;
  • translocation chromosome T4AgL(Ba)-4BL ⁇ 4BS is a tall, male-fertile wheat grown from blue-seeded seeds; translocation chromosome T4AgL(Ba)-4BL ⁇ 4BS is recombined 4AgL(Ba) into Obtained on 4B; white-grain sterile plant of 06L6109-3 dual-purpose line: it is a dwarf and male sterile plant grown from white-grained seeds obtained by selfing of 06L6109-3 dual-purpose line; 06L6109-3 dual-purpose line Blue grain with Rht3-ms1b produced by crossing ms1b-Rht3 containing ms1b-Rht3 wheat grown from white seeds with semi-dwarf and male sterility traits and additional line wheat containing translocation chromosome T4thS ⁇ 4AgL Dual-purpose system;
  • T4AgL(Ba)-4BL ⁇ 4BS(Rht3-ms1b) translocation line plants were crossed with wheat 4B substitution line 4th to obtain T4AgL(Ba)-4BL ⁇ 4BS(Rht3-ms1b) with 4th and translocation chromosomes - dimonomer seeds for ms1b);
  • step (3) sowing and cultivating the double monomer seeds obtained in step (2) into plants, selfing, selecting dark blue seeds for planting and screening out tall and male fertile plants;
  • step (3) (4) hybridize the plant screened in step (3) with the white-grained sterile strain of the 14L6386 dual-purpose line, and screen the hybrid combination with blue-grained seeds;
  • step (4) sow the blue-grained seeds of the hybrid combination and cultivate them into plants. If the plants obtained from all the seeds are male fertile plants, the plants obtained by the development of the seeds of the corresponding hybrid combination form a potential translocation lines;
  • step (5) The potential translocation line that step (5) obtains is carried out as follows: selfing and harvesting seeds, the seeds are divided into two groups of blue seeds and white seeds; Breeding as plants, if all the individual plants obtained from the blue seeds are male fertile plants and all the individual plants obtained from the white seeds are male sterile plants, then these male fertile individual plants obtained from the blue seeds FISH identification was carried out to screen the individual plants with translocation chromosome T4AgL segment (Ba)-4BL ⁇ 4thS(Rf), and these individuals constituted the T4AgL segment (Ba)-4BL ⁇ 4thS(Rf) translocation population;
  • step (6) From the selfing progeny of the translocation line that step (6) obtains, screen white seed: the plant that the quantity of blue seed is 2:1, namely contain translocation chromosome T4AgL fragment (Ba)-4BL.
  • the karyotype of the dual-purpose line is: 21II(msms)+I T4AgL fragment (Ba)-4BL ⁇ 4thS(Rf) .
  • Described method specifically comprises the steps:
  • the 15L4167 dual-purpose line is selfed, and then the male sterile plants grown from the blue-grained seeds are screened, that is, the plants with the translocation end body T4AgL fragment (Ba)-4BL ⁇ ; (Self-crossing of 15L4167 dual-purpose line, selection of male fertile plants grown from dark blue seeds, all seeds produced after selfing are blue seeds) hybridization, and then screening of plants grown from dark blue seeds.
  • the resulting male fertile plants were selfed, and then the plants with male sterility were screened from the plants developed from the selfed seeds, and further screened by microscopy to obtain the translocation end body T4AgL fragment (Ba)-4BL. add-on plants;
  • step (3) Use the seeds obtained in step (3) as the male parent and the white seed as the female parent (the white seed is obtained by selfing of the 15L4167 dual-purpose line) as a hybrid combination, and select from the seeds obtained on the female plant blue seed;
  • step (4) sowing and cultivating the blue-grained seeds obtained in step (4) into plants, eliminating male sterile plants, and harvesting male-fertile plants; testing the harvested male-fertile plants, and selecting the obtained seeds had blue-grained seeds A plant with white seeds in its seeds;
  • Step (5) selected individual plant, sort out white seed and blue seed respectively by individual plant;
  • the seeds obtained in sowing step (6) are cultivated into plants; if the plants obtained by the blue seeds of a certain individual plant are all male fertile plants, and the plants obtained by the white seeds of this individual plant are all male fertile plants. Breeding plants, microscopically inspecting the light blue seeds of the single plant, and obtaining translocation plants with translocation chromosome T4AgL fragment (Ba)-4BL ⁇ 4thS fragment (Rf), light/medium blue seeds are dual-purpose lines .
  • the karyotype of the dual-purpose line is: 21II(msms)+I T4AgL fragment (Ba)-4BL ⁇ 4thS fragment (RF) .
  • Described method specifically comprises the steps:
  • step (1) The male sterile plant screened in step (1) is hybridized with the male fertile plant obtained in step (1), the dark blue seeds are screened and cultivated into a plant, and the male sterile plant is screened out from its selfed offspring;
  • step (2) The male sterile plant screened in step (2) is crossed with the male fertile plant screened in step (3), and the harvest seeds (both are blue grains) are cultivated into plants, then selfed, and the dark blue grain seeds are screened, and passed through the mirror Detection and screening of double diplosome seeds with terminal body 4AgL(Ba) and terminal body 4thS(Rf);
  • step (4) The double doublet seeds obtained in step (4) are subjected to mutagenesis treatment, and then cultivated into plants;
  • step (4) The plant grown from the double diploid seeds obtained in step (4) is used as the male parent, and the male sterile plant (the white seed grown by selfing of the 16L6386 dual-purpose line) as the female parent Sterile plant) hybridization, harvest the seeds on the female plant, select the blue seed to carry out mutagenesis treatment, and then cultivate it into a plant;
  • the male sterile plant the white seed grown by selfing of the 16L6386 dual-purpose line
  • step (7) The plant obtained in step (5) or step (6) is used as the male parent, and the white-seeded sterile plant (the male sterile plant grown from the white-seeded seed obtained by selfing of the 16L6386 dual-purpose line) as the female parent ) hybridization, harvesting the seeds on the female plant and screening out the blue seeds therefrom;
  • step (7) sowing the blue seeds obtained in step (7) and cultivating them into plants, eliminating male sterile plants, selfing the selected plants and harvesting seeds, and selecting the seeds that have both blue seeds and white seeds. plant;
  • step (8) For the plants screened in step (8), the white seeds and blue seeds obtained by selfing are sorted out by individual plants respectively;
  • the seeds obtained in the sowing step (9) are cultivated into plants; if the plants obtained by the blue seeds of a certain individual plant are all male fertile plants, and the white seeds of the individual plant are all male sterile plants, These individual plants are potential plants with telomeres, and the blue-grained seeds of these individual plants are subjected to microscopic examination, and the plants with translocation telomeres T4AgL fragment (Ba)-4thS fragment (Rf) are selected, that is, they have translocation Translocation line of telosome T4AgL fragment (Ba)-4thS fragment (Rf) ⁇ , light/medium blue seed is dual-purpose line.
  • the karyotype of the dual-purpose line is: 21II(msms)+I T4AgL fragment (Ba)-4thS fragment (Rf) .
  • Described method specifically comprises the steps:
  • step (1) The male sterile plant screened in step (1) is hybridized with the male fertile plant obtained in step (1), the dark blue seeds are screened and cultivated into plants, and the male sterile plants are screened out from the plants developed from the selfed seeds;
  • step (2) The male sterile plant screened in step (2) is crossed with the male fertile plant screened in step (3), and the harvest seeds (both are blue grains) are cultivated into plants, then selfed, and the dark blue grain seeds are screened, and passed through the mirror Detection and screening of double diplosome seeds with terminal body 4AgL(Ba) and terminal body 4thS(Rf);
  • step (4) The double doublet seeds obtained in step (4) are subjected to mutagenesis treatment, and then cultivated into plants;
  • step (4) The plant grown from the double diploid seeds obtained in step (4) is used as the male parent, and the male sterile plant (the white seed grown by selfing of the 16L6386 dual-purpose line) as the female parent Sterile plant) hybridization, harvest the seeds on the female plant, select the blue seed to carry out mutagenesis treatment, and then cultivate it into a plant;
  • the male sterile plant the white seed grown by selfing of the 16L6386 dual-purpose line
  • step (7) The plant obtained in step (5) or step (6) is used as the male parent, and the white-seeded sterile plant (the male sterile plant grown from the white-seeded seed obtained by selfing of the 16L6386 dual-purpose line) as the female parent ) hybridization, harvesting the seeds on the female plant and screening out the blue seeds therefrom;
  • step (7) sowing the blue seeds obtained in step (7) and cultivating them into plants, eliminating male sterile plants, selfing the selected plants and harvesting seeds, and selecting the seeds that have both blue seeds and white seeds. plant;
  • step (8) For the plants screened in step (8), the white seeds and blue seeds obtained by selfing are sorted out by individual plants respectively;
  • the seeds obtained in the sowing step (9) are cultivated into plants; if the plants obtained by the blue seeds of a certain individual plant are all male fertile plants, and the white seeds of the individual plant are all male sterile plants, These individual plants are potential plants with telomeres, and the blue-grained seeds of these individual plants are examined under a microscope, and the plants with translocation telomer T4thS fragment (Rf)-4AgL fragment (Ba) are selected, that is, they have translocation Translocation line of telosome T4thS fragment (Rf)-4AgL fragment (Ba) ⁇ , light/medium blue seed is a dual-purpose line.
  • the karyotype of the dual-purpose line is: 21II (msms)+I T4thS fragment (Rf)-4AgL fragment (Ba) .
  • Described method specifically comprises the steps:
  • step (2) The double doublet seeds obtained in step (1) are subjected to mutagenesis treatment, and then cultivated into plants.
  • step (3) The plant grown from the double-diende seeds obtained in step (1) is used as the male parent, and the male sterile plant grown from the white-seed sterile plant (15L4167 dual-purpose line self-crossed) as the female parent Breeding plant) hybridization, harvest the seeds on the female parent plant, select the blue seed to carry out mutagenesis treatment, and then cultivate it into a plant;
  • step (2) or step (3) The plant obtained in step (2) or step (3) is used as the male parent, and the white-seeded sterile plant (the male sterile plant grown from the white seed obtained by selfing of the 15L4167 dual-purpose line) as the female parent hybridization, harvesting the seeds on the female parent plant and selecting blue seeds from them;
  • the white-seeded sterile plant the male sterile plant grown from the white seed obtained by selfing of the 15L4167 dual-purpose line
  • step (4) sowing the blue seeds obtained in step (4) and cultivating them into plants, eliminating male sterile plants, selfing the selected plants and harvesting seeds, and selecting the ones with both blue seeds and white seeds in the selected seeds plant;
  • step (5) For the plants screened in step (5), the white seeds and blue seeds obtained by selfing are sorted out by individual plants respectively;
  • the seeds obtained in sowing step (6) are cultivated into plants; if the plants obtained by the blue seeds of a certain individual plant are all male fertile plants, and the white seeds of this individual plant are all male sterile plants, Microscopically check the blue seeds in these individual plants, and select the plants with the translocation end body T4thS fragment (Rf)-4AgL fragment (Ba)-4BL ⁇ , that is, the translocation end body T4thS fragment (Rf)-4AgL fragment
  • the translocation line of (Ba)-4BL ⁇ , the light/medium blue seed is the dual-purpose line.
  • the karyotype of the dual-purpose line is: 21II(msms)+I T4thS fragment (Rf)-4AgL fragment (Ba)-4BL ⁇ .
  • the additional line wheat containing translocation chromosome T4thS ⁇ 4AgL is any wheat with this chromosome form, specifically, it can be the dual-purpose line wheat obtained by the method described in ZL200610042629.8 (that is, the authorized announcement number is CN 100420368C, page 5 For the dual-purpose system in line 7, both the original dual-purpose system and the new dual-purpose system are acceptable).
  • the karyotype of the addition line wheat (dual-purpose line) containing the translocation chromosome T4thS ⁇ 4AgL was 21II(msms)+I T4thS ⁇ 4AgL .
  • T4AgL(Ba)-4BL ⁇ 4BS(Rht3-ms1b) translocation plants homozygous: dwarf and male sterile plants grown from blue seeds.
  • the karyotype of diasomic seeds with 4th and translocation chromosome T4AgL(Ba)-4BL ⁇ 4BS(Rht3-ms1b) was: 20II+I T4AgL(Ba)-4BL ⁇ 4BS(Rht3-ms1b) +I 4th .
  • N4B wheat 4B substitution line 4th
  • Phenotype blue-grained (the blue-grained gene it contains does not express endosperm intuition), tall stem and male fertile.
  • White-seed sterile plant of 14L6386 dual-purpose line it is a male sterile plant grown from white-grained seeds obtained from selfing of 14L6386 dual-purpose line.
  • the 14L6386 dual-purpose line is a blue-grain dual-purpose line obtained by crossing the 09L6034 dual-purpose line (male parent) and the waxy wheat line (female parent) wheat).
  • the 09L6034 dual-purpose line is a blue-grain dual-purpose line selected from the wheat line Zhou 88114 (named Zhoumai 11) as the recurrent parent (female parent) and the T0065-10B-2LB-4 dual-purpose line after 3 backcrosses .
  • the T0065-10B-2LB-4 dual-purpose system is recorded in ZL200610042629.8 (that is, T0065-10B-2LB-4 in the 14th line on page 9 of the specification whose authorization announcement number is CN100420368C).
  • the karyotype of the dual-purpose line T0065-10B-2LB-4 is 21II(msms)+I T4thS ⁇ 4AgL , which is an additional line of wheat containing translocation chromosome T4thS ⁇ 4AgL.
  • the 15L4167 dual-purpose line is an addition line containing the translocation chromosome T4AgL fragment (Ba)-4BL ⁇ 4thS (Rf).
  • the karyotype of the 15L4167 dual-purpose line is: 21II(msms)+I T4AgL fragment (Ba)-4BL ⁇ 4thS(Rf) .
  • the karyotype of disomic addition line plants with translocation endosome T4AgL fragment (Ba)-4BL ⁇ was: 21II(msms)+II T4AgL fragment (Ba)-4BL ⁇ .
  • the phenotype of the disomic addition line with the translocation telosome T4AgL fragment (Ba)-4BL ⁇ was: the seeds were dark blue grains, and the plants showed male sterility.
  • the karyotype of disomic addition line plants with telosome 4thS(Rf) ⁇ is: 21II(msms)+II 4thS(Rf) ⁇ .
  • the phenotype of the disomic addition line with the terminal body 4thS(Rf) ⁇ is: the seeds are white, and the plants are male fertile.
  • the karyotype of double single-tetrasome seeds with translocated endosome T4AgL fragment (Ba)-4BL ⁇ and endosome 4thS(Rf) ⁇ is: 21II(msms)+I T4AgL fragment(Ba)-4BL ⁇ +I 4thS( Rf) ⁇ .
  • 16L6386 dual-purpose line is an addition line of wheat containing translocation chromosome T4thS ⁇ 4AgL.
  • the 16L6386 dual-purpose line is an additional line of wheat containing the translocation chromosome T4thS ⁇ 4AgL, and is a dual-purpose line selected through generation selection.
  • the 16L6386 dual-purpose line is a dual-purpose line cultivated in the invention patent (ZL200610042629.8), and a dual-purpose line obtained through generation selection.
  • the dual-purpose line cultivated in the invention patent (ZL200610042629.8) is an additional line of wheat containing the translocation chromosome T4thS ⁇ 4AgL (that is, the T0065-10B-2LB-4 material on the 14th line of page 9 in CN 100420368C with the authorized announcement number transformed wheat).
  • the karyotype of the add-on wheat dual-purpose line containing the translocation chromosome T4thS ⁇ 4AgL is: 21II(msms)+I T4thS ⁇ 4AgL .
  • the karyotype of double diplosome seeds with terminal body 4AgL(Ba) ⁇ and terminal body 4thS(Rf) ⁇ is: 21II(msms) +II 4AgL ⁇ +II 4thS ⁇ .
  • the karyotype of double diplosome seeds with translocated endosome T4AgL fragment(Ba)-4BL ⁇ and endosome 4thS(Rf) ⁇ is: 21II(msms)+II T4AgL fragment(Ba)-4BL ⁇ +II 4thS( Rf) ⁇ .
  • mutagenic treatments can specifically be treated with dimethyl sulfoxide (DMSO) chemical mutagen or ethyl methanesulfonate (EMS) chemical mutagen or radiation treatment.
  • DMSO dimethyl sulfoxide
  • EMS ethyl methanesulfonate
  • Light/medium blue seeds are planted, and the F 2 generation excellent individual plants with blue-white or blue-red seeds are selected.
  • 82 seeds were planted separately from blue seeds, white seeds or red seeds, each planted in 2 rows, 2m row length, row spacing 25cm, 5cm artificial on-demand sowing.
  • the proportion of white or red seeds in the individual plants or lines selected in each generation must be greater than or equal to 60%, and at the same time select the corresponding blue-grained individual plants or plants with high outcrossing seed setting rate of white/red sterile plants Tie.
  • the invention discloses the creation and application of a two-line hybrid wheat system with blue grains as a marker trait (abbreviated as a blue standard two-line hybrid wheat system).
  • T4thS Fram containing Rf
  • 4AgL Fram containing Ba
  • 4BL Frament close to centromere
  • the terminal body is used as an additional monomer (dual-purpose line) of the ms1 homozygous wheat line (line), thereby completing the creation of the improved blue standard two-line method hybrid wheat system.
  • the different additional exogenous translocation chromosomes or exogenous translocation end bodies are characterized in that they cannot be paired with any chromosome of common wheat, and can completely restore the ms1 sterile gene series (ms1a, ms1b, ms1c, ms1d ,...);
  • the different additional exogenous translocation chromosomes or exogenous translocation end bodies are characterized in that the Rf recovery gene is derived from the 4th chromosome of Triticum thaoudar and the Ba blue grain gene is derived from Agropyron elongatum, T4AgL (containing the fragment of Ba)- 4BL (fragment close to the centromere).
  • the break frequency of the 4thS translocation chromosome from the centromere is 0.5-5%, and the frequency of fertile plants in the sterile line is about half of that in the dual-purpose line; T4AgL ( Fragment containing Ba)-4BL (fragment close to centromere).
  • 4thS (fragment containing Rf) translocation chromosome breakage frequency from centromere is 0-1%, and the frequency of fertile plants in sterile lines is two About half of the sterile plants appear in the dual-purpose line of the use line; T4AgL (the fragment containing Ba)-4thS (the fragment containing Rf). The separation frequency of the two target genes of the terminal body is 0-1 ⁇ , and only the sterile line will appear 0-0.5 ⁇ of fertile plants, blue grain populations will not appear sterile plants; T4thS (fragment containing Rf)-4AgL (fragment containing Ba).
  • T4thS fragment containing Rf
  • 4AgL fragment containing Ba
  • 4BL fragment close to the centromere.
  • the separation frequency of the two target genes in the terminal body is 0-1 ⁇ , there will be no fertile plants in the sterile line, and there will be a 0-1 ⁇ infertility in the dual-purpose line. Breeding plants.
  • the proportion of seeds of different colors produced by selfing of different dual-purpose lines is different.
  • the range of white or red seeds (sterile lines) is 55-85%, and the range of light/medium blue seeds (monosome addition, dual-purpose lines) is 15%.
  • dark blue seed (additional double end body) is 2-6%, wherein, sterile line seed is used for producing hybrid, dual-purpose line seed is used for breeding sterile line and dual-purpose line, dark blue grain seed ( Double-end body addition) eliminated or used as a parent or used for purification and rejuvenation of dual-purpose lines.
  • This process can take the target sterile line as the recurrent parent and the improved dual-purpose line or dark blue grain homozygous line for more than 6 consecutive backcrosses in 2-3 years under artificial control, and screen the same dual-purpose line as the original one. line, thereby achieving replacement of additional translocated chromosomes or telomeres.
  • the above CMS lines have been backcrossed 4 times. In the same way, the exchange of new translocated chromosomes or terminal bodies can be realized.
  • the present invention also provides a translocation chromosome (heterotopic chromosome), which sequentially includes the following segments: 4AgL segment (Ba), 4BL, centromere, and 4thS (Rf).
  • translocation chromosome heterotopic chromosome
  • the present invention also provides a translocation chromosome (heterotopic chromosome), which sequentially includes the following segments: 4AgL segment (Ba), 4BL, centromere, and 4thS segment (Rf).
  • translocation chromosome heterotopic chromosome
  • the present invention also provides a translocation terminal body (ectopic telomere), which sequentially includes the following segments: 4AgL segment (Ba), 4thS segment (Rf), and centromere.
  • the present invention also provides a translocation terminal body (heterotopic telomere), which sequentially includes the following segments: 4thS fragment (Rf), 4AgL fragment (Ba), and centromere.
  • the present invention also provides a translocation terminal body (ectopic telomere), which sequentially includes the following segments: 4thS fragment (Rf), 4AgL fragment (Ba)-4BL, and centromere.
  • ectopic telomere which sequentially includes the following segments: 4thS fragment (Rf), 4AgL fragment (Ba)-4BL, and centromere.
  • 4AgL represents the long arm of chromosome 4 4Ag derived from Agropyron elongatum and has the Ba gene. 4AgL can also be represented by 4AgL(Ba). The 4AgL fragment retaining the Ba gene is represented by 4AgL fragment (Ba).
  • 4th represents the fourth chromosome derived from T. thaoudar.
  • 4thS represents the short arm of 4th and contains the gene (Rf gene) that restores male fertility.
  • 4thS can also be represented by 4thS(Rf).
  • the 4thS fragment retaining the Rf gene is represented by 4thS fragment (Rf).
  • 4B represents chromosome 4 of group B from hexaploid common wheat (T. aestivum).
  • 4BS represents the short arm of 4B.
  • 4BL represents the long arm of 4B.
  • 4BS represents 4BS with linkage of Rht3 gene and ms1b gene.
  • the Rht3 gene is a dwarf gene (from Nainari Rht3).
  • the ms1b gene is a male sterility gene (from the "Probus" mutant).
  • ⁇ stands for centromere.
  • T stands for translocated chromosomes or translocated telosomes (ie, chromosomes with the centromere at the end).
  • II represents a bivalent body. I stands for monovalent.
  • N4B stands for 4B chromosome deletion, that is, there is no 4B chromosome.
  • the ms1b-Rht3-containing wheat with semi-dwarf and male sterility traits is any wheat with this trait and chromosomal form, for example, the semi-dwarf and male sterility wheat obtained by the method described in ZL200610042629.8 Wheat containing ms1b-Rht3 with fertility traits (that is, ms-Rht3/Ms-rht3 (semi-dwarf) wheat in the 13th line on page 7 of the specification whose authorized announcement number is CN100420368C).
  • the present invention is based on the germplasm materials accumulated in the early stage, and makes full use of traditional biological technology, chromosome engineering technology and crop genetic breeding methods to create a more stable gene with exogenous restorer genes and blue grains. Tightly linked exogenous translocation chromosomes or translocation end bodies, so as to achieve the purpose of improving the blue standard type two-line hybrid wheat system.
  • the present invention improves the stability of the additional translocation chromosome in the production process of its sterile line, and creates a gene that has full recovery ability to the recessive sterility gene ms1 and contains expression dosage Effect and endosperm texture of the blue-grain gene Ba and the exogenous restorer gene Rf exogenous translocation chromosome [T4AgL (Ba-containing fragment)-4BL (closed to the silk point fragment). 4thS and T4AgL (Ba-containing fragment)- 4BL (fragment close to centromere).
  • White/red seed (sterile line) and blue seed mixed seeds are obtained through dual-purpose line selfing, and white/red seed (sterile line) and light/medium blue seed (dual-purpose line) are separated by color sorting machine And dark blue seeds, sterile lines are used for hybrid seed production, light/medium blue seeds (dual-purpose lines) are used for breeding sterile lines and dual-purpose lines, dark blue seeds are eliminated or used as parents to transfer new sterile It can be used as a dual-purpose system for purification and rejuvenation. Different translocation chromosomes or translocation end bodies can be replaced with each other as needed to meet the purity and stability requirements of sterile lines under different genetic backgrounds.
  • the invention has the characteristics of fast breeding of sterile lines, low production cost of hybrids, strong advantages, etc., and is better than the previous invention of "a method for breeding hybrid wheat with two-line method using blue grain as a marker trait (ZL200610042629.8 )” is more stable and reliable.
  • Figure 1 is the schematic diagram and FISH image of the translocated chromosome T4AgL fragment (Ba)-4BL ⁇ 4thS (Rf) (left: schematic diagram; right: FISH image).
  • Fig. 2 is the karyotype map of the additional line plants: 21II(msms)+I T4AgL fragment (Ba)-4BL ⁇ 4thS(Rf) .
  • Fig. 3 is a schematic diagram of translocated chromosome T4AgL fragment (Ba)-4BL ⁇ 4thS fragment (Rf) (left: schematic diagram; right: chromosome).
  • Fig. 4 is a schematic diagram of the translocation endosome T4AgL fragment (Ba)-4thS fragment (Rf) ⁇ (left: schematic diagram; right: chromosome endosome).
  • Figure 5 is a schematic diagram of the translocation endosome T4thS fragment (Rf)-4AgL fragment (Ba) ⁇ (left: schematic diagram; right: chromosome endosome).
  • Fig. 6 is a schematic diagram of the translocation endosome T4thS fragment (Rf)-4AgL fragment (Ba)-4BL ⁇ (left: schematic diagram; right: chromosome endosome).
  • Figure 7 is a schematic diagram of the breeding process of dual-purpose lines.
  • Fig. 8 is a schematic diagram of the breeding process of a strong dominant combination (hybrid).
  • 4AgL represents the long arm of the fourth chromosome 4Ag derived from Agropyron elongatum, which has the Ba gene, which is a blue-grain gene that exhibits dosage effects and endosperm intuition, and is a dominant gene, endosperm and aleurone layer
  • the grains with 3 copies of the Ba gene are dark blue, the grains with 2 copies of the Ba gene in the endosperm and aleurone layer are medium blue, and the grains with 1 copy of the Ba gene in the endosperm and aleurone layer are light blue , the endosperm and aleurone layer do not have the Ba gene kernels are white or red.
  • 4AgL can also be represented by 4AgL(Ba).
  • the 4AgL fragment retaining the Ba gene is represented by 4AgL fragment (Ba).
  • 4th represents the fourth chromosome derived from T. thaoudar.
  • 4thS represents the short arm of 4th and contains the gene (Rf gene) that restores male fertility.
  • 4thS can also be represented by 4thS(Rf).
  • the 4thS fragment retaining the Rf gene is represented by 4thS fragment (Rf).
  • 4B represents chromosome 4 of group B from hexaploid common wheat (T. aestivum).
  • 4BS represents the short arm of 4B.
  • 4BL represents the long arm of 4B.
  • 4BS represents 4BS with linkage of Rht3 gene and ms1b gene.
  • the Rht3 gene is a dwarf gene (from Nainari Rht3), which is a dominant gene. Plants with 2 copies of the Rht3 gene appear as short stems, and plants with 1 copy of the Rht3 gene appear as semi-dwarf stems without the Rht3 gene The plants appear as tall stems.
  • the ms1b gene is a male sterility gene (from the "Probus" mutant) and is a recessive gene.
  • ⁇ stands for centromere.
  • T stands for translocated chromosomes or translocated telosomes (ie, chromosomes with the centromere at the end).
  • II represents a bivalent body. I stands for monovalent.
  • N4B stands for 4B chromosome deletion, that is, there is no 4B chromosome.
  • non-blue grains including white grains or red grains
  • blue grains including light blue grains, medium blue grains or dark blue grains
  • short stem, semi-dwarf stem, and tall stem refer to plant height traits.
  • fertility and sterility refer to the male fertility traits of plants.
  • a plant having the traits of blue-grained, short stem and sterile means that the plant has the traits of dwarf stem and male sterility and the plant is grown from blue-grained seeds.
  • the male sterile plant is used as the female parent, and the other parental plant is used as the male parent.
  • White-grained sterile plant of 06L6109-3 dual-purpose line it is a dwarf and male sterile plant grown from white-grained seeds obtained from selfing of 06L6109-3 dual-purpose line.
  • the 06L6109-3 dual-purpose line is a hybrid of ms1b-Rht3-containing wheat grown from white seeds with semi-dwarf and male sterility traits and the dual-purpose line obtained by the method in ZL200610042629.8. Rht3-ms1b blue-grain dual-purpose line.
  • the karyotype of the dual-purpose line obtained by the method in ZL200610042629.8 is: 21II (msms)+I T4thS ⁇ 4AgL ; that is, an additional line of wheat containing the translocation chromosome T4thS ⁇ 4AgL.
  • the plants screened in step (2) are the plants with translocation chromosome T4AgL(Ba)-4BL ⁇ 4BS(Rht3-ms1b), called T4AgL(Ba)-4BL ⁇ 4BS(Rht3-ms1b) translocation line plants.
  • T4AgL(Ba)-4BL ⁇ 4BS(Rht3-ms1b) translocation plants homozygous: dwarf and male sterile plants grown from blue seeds.
  • step (3) The plants screened in step (3) are crossed with wheat (high stem and male fertile wheat grown from blue-grained seeds) with translocation chromosome T4AgL(Ba)-4BL ⁇ 4BS, and the obtained plants can be self-selected. Cross and save the seeds, and the dwarf and male sterile plants similar to those in step (3) can be continuously obtained.
  • T4AgL(Ba)-4BL ⁇ 4BS(Rht3-ms1b) translocation line plants (homozygous) were crossed with wheat 4B substitution line 4th, and more than 500 grains with 4th and translocation chromosome T4AgL(Ba)-4BL ⁇ 4BS were obtained (Rht3-ms1b) double monomeric seed.
  • the karyotype of dimeric seeds was: 20II+I T4AgL(Ba)-4BL ⁇ 4BS(Rht3-ms1b) +I 4th .
  • N4B wheat 4B substitution line 4th
  • Phenotype blue-grained (the blue-grained gene it contains does not express endosperm intuition), tall stem and male fertile.
  • step 2 Sowing and cultivating the double monomer seeds obtained in step 2 into plants, selfing, selecting dark blue seeds for planting and screening out tall and male fertile plants.
  • step 4 Hybridize the plant screened in step 3 with the white-grained sterile strain of the 14L6386 dual-purpose line, and prepare 561 hybrid combinations in total.
  • the hybrid combinations are numbered one by one, and the hybrid combinations with blue seeds are screened (the 4th chromosome can be screened out here blue-grain gene on
  • White-seed sterile plant of 14L6386 dual-purpose line it is a male sterile plant grown from white-grained seeds obtained from selfing of 14L6386 dual-purpose line.
  • the 14L6386 dual-purpose line is a blue-grain dual-purpose line obtained by crossing the 09L6034 dual-purpose line (male parent) and the waxy wheat line (female parent) wheat).
  • the 09L6034 dual-purpose line is a blue-grain dual-purpose line selected from Zhou 88114 (named Zhoumai 11) as the recurrent parent (female parent) and the T0065-10B-2LB-4 dual-purpose line after 3 backcrosses.
  • T0065-10B-2LB-4 dual-purpose system is recorded in ZL200610042629.8.
  • the karyotype of the dual-purpose line T0065-10B-2LB-4 is: 21II(msms)+I T4thS ⁇ 4AgL ; that is, an additional line of wheat containing translocation chromosome T4thS ⁇ 4AgL.
  • step 4 sow the blue seeds of different hybrid combinations in order and cultivate them into plants. If the plants obtained from all the seeds are male fertile plants, the corresponding hybrid combination seeds are developed. A potential translocation line. A total of 35 potential translocation lines were obtained, namely 14T1, 14T2, ..., 14T34, and 14T35.
  • the 35 potential translocation lines that step 5 obtains carry out the following operations respectively: selfing and harvesting seeds, seeds are divided into two groups of blue seeds and white seeds; blue seeds (100/potential translocation line) and white seeds (100 seeds/potential translocation line) were sown and cultivated into plants, if all the individual plants obtained from the blue seeds were male fertile plants and the individual plants obtained from all the white seeds were Male sterile plants, then these male fertile individual plants cultivated from blue seeds are identified by FISH, and the individual plants with translocation chromosome T4AgL fragment (Ba)-4BL ⁇ 4thS(Rf) are screened, and these individual plants form T4AgL Fragment (Ba)-4BL ⁇ 4thS(Rf) Translocation Line Population.
  • a total of 5 translocation lines meeting the requirements were obtained, namely 14T9, 14T12, 14T21, 14T31 and 14T34. At the same time, a less stable translocation line, namely 14T35, was obtained and used in Example 2.
  • step 7 From the selfing progeny of 5 translocation lines obtained in step 6, screen white seeds: plants with a blue seed quantity of about 2:1, which contain the translocation chromosome T4AgL fragment (Ba)-4BL ⁇ 4thS (Rf) additional system (dual-purpose system).
  • the karyotype of the additional line is: 21II(msms)+I T4AgL fragment (Ba)-4BL ⁇ 4thS(Rf) .
  • Figure 1 for the schematic diagram and FISH image of the translocation chromosome T4AgL fragment (Ba)-4BL ⁇ 4thS(Rf).
  • Figure 2 for the karyotype of the additional line plants.
  • the obtained additional line plants were planted in expanded populations, and the frequency of male sterile plants in blue-grained populations and male-fertile plants in white-grained populations in different individual plants was investigated.
  • the frequency of male sterile plants in the blue-grained population was 0.5-5%, and the frequency of male-fertile plants in the white-grained population was 0.2-3%.
  • the test population is more than 1000 plants.
  • the 15L4167 dual-purpose line was bred through generation selection (except for the poor stability of the additional chromosome, the rest of the agronomic traits have been stabilized).
  • the 15L4167 dual-purpose line is an addition line (dual-purpose line) containing the translocation chromosome T4AgL fragment (Ba)-4BL ⁇ 4thS(Rf).
  • the karyotype of the 15L4167 dual-purpose line is: 21II(msms)+I T4AgL fragment (Ba)-4BL ⁇ 4thS(Rf) .
  • the 15L4167 dual-purpose line was selfed, and then the male sterile plants grown from blue-grained seeds were screened, that is, the plants with the translocated terminal body T4AgL fragment (Ba)-4BL ⁇ .
  • This plant is crossed with dark blue grain fertile plants (15L4167 dual-purpose line selfing, screening male fertile plants grown from dark blue grain seeds, all the seeds formed after selfing are blue grain seeds), and then screened for dark blue grain seeds
  • the grown male fertile plants are selfed, and then the plants with male sterility are screened from the plants developed from the selfed seeds, and further screened by microscopy to obtain the T4AgL fragment (Ba)-4BL with the translocation end body. additional line of plants.
  • the karyotype of the additional line plants was: 21II(msms)+I T4AgL fragment (Ba)-4BL ⁇ .
  • the phenotype of the additional line is: the seeds are dark blue grains, and the plants are male sterile.
  • the 15L4167 dual-purpose line was selfed, and then the male fertile plants grown from the white seeds were selected, that is, the plants with the terminal body 4thS(Rf) ⁇ were further screened by microscopy to obtain the additional Department of plants.
  • the karyotype of the additional line plants was: 21II(msms)+I 4thS(Rf) ⁇ .
  • the phenotypes of the additional lines are: the seeds are white, and the plants are male fertile.
  • the seed obtained in step 2 is used as the male parent and the white seed as the female parent (the white seed is obtained by selfing of the 15L4167 dual-purpose line) as a hybrid combination, and is planted alternately by 2 rows than 2-4 rows and cultivated as Plants, artificially assisted pollination at flowering, blue-grained seeds selected from seeds obtained from the female parent plant.
  • step 4 Sow the blue seeds obtained in step 3 and cultivate them into plants, eliminate male sterile plants, and harvest male fertile plants; the male fertile plants harvested are tested indoors, and the selected seeds have both blue seeds and Plants with white seeds.
  • step 5 A total of 1480 individual plants were selected in step 4, and white seeds and blue seeds were sorted out according to individual plants.
  • the seed that sowing step 5 obtains (80 white grain seeds and 80 blue grain seeds of each single plant sowing) is cultivated as a plant; if the plants obtained by the blue grain seeds of a certain single plant are all male fertile plants, And the plants obtained from the white seeds of the single plant are all male sterile plants, and the light blue seeds of the single plant are subjected to microscopic examination to obtain the translocation chromosome T4AgL fragment (Ba)-4BL ⁇ 4thS fragment (Rf). Translocation plants are also known as dual-purpose lines.
  • 15yi 3357 had a translocation chromosome T4AgL fragment (Ba)-4BL ⁇ 4thS fragment (Rf).
  • FIG. 3 The schematic diagram of the translocated chromosome T4AgL fragment (Ba)-4BL ⁇ 4thS fragment (Rf) is shown in Fig. 3 .
  • the obtained additional line plants were planted in expanded populations, and the frequency of male sterile plants in blue-grained populations and male-fertile plants in white-grained populations in different individual plants was investigated.
  • the break frequency of the translocation chromosome T4AgL segment (Ba)-4BL ⁇ 4thS segment (Rf) from the centromere is less than 2%, and the frequency of fertile plants in the sterile line is about half of that of the sterile plant in the dual-purpose line (less than 1 %).
  • the test population is more than 1000 plants.
  • the karyotype of the dual-purpose line cultivated in the invention patent is: 21II (msms)+I T4thS ⁇ 4AgL ; that is, the addition line wheat (16L6386 dual-purpose line) containing the translocation chromosome T4thS ⁇ 4AgL.
  • the 16L6386 dual-purpose line was selfed, and dark blue seeds were harvested, planted and cultivated into plants, and these plants were male fertile plants.
  • step (2) The plants screened in step (2) are crossed with the plants obtained in step (3), the dark blue seeds are screened and cultivated into plants, and male sterile plants are screened from their selfed offspring.
  • the above-screened dark blue seeds are self-propagating enough seeds (seeds are all dark blue).
  • the 16L6386 dual-purpose line was selfed, the white seeds were harvested, planted and cultivated into plants, the male fertile plants were selected, and the plants with the terminal body 4thS(Rf) ⁇ were screened by microscopy. Self-propagation enough seeds for future use.
  • the male sterile plant screened in step (4) is crossed with the male fertile plant screened in step (5), and the harvested seeds (both are blue grains) are cultivated into plants, then selfed, and the dark blue grain seeds are screened through a microscope.
  • the double-diende seeds with the terminal body 4AgL(Ba) ⁇ and the terminal body 4thS(Rf) ⁇ were detected and screened, which was the double-diende addition line.
  • the karyotype of the double doublet seeds was: 21II(msms)+II 4AgL(Ba) ⁇ +II 4thS(Rf) ⁇ . Seeds (all dark blue seeds) sufficient for self-propagation are used for future use.
  • step 2 The double-diende seeds obtained in step 1 are treated with dimethyl sulfoxide (DMSO) chemical mutagen or ethyl methanesulfonate (EMS) chemical mutagen or radiation treatment, and then cultivated into plants.
  • DMSO dimethyl sulfoxide
  • EMS ethyl methanesulfonate
  • the plant grown from the heavy double-end body seeds obtained in step 1 is used as the male parent, and the white-seeded sterile plant (the male sterile plant grown from the white-seeded seed obtained by selfing of the 16L6386 dual-purpose line) as the female parent ) hybridization, harvest the seeds on the female plant, select the blue seed to treat with dimethyl sulfoxide (DMSO) chemical mutagen or treat with ethyl methanesulfonate (EMS) chemical mutagen or radiation treatment, then cultivate for plants.
  • DMSO dimethyl sulfoxide
  • EMS ethyl methanesulfonate
  • step 2 or step 3 The plant obtained in step 2 or step 3 is used as the male parent, and is crossed with the white-grained sterile plant (the male sterile plant grown from the white-grained seed obtained by selfing of the 16L6386 dual-purpose line) as the female parent, and the female parent is harvested. Seeds on this plant and select blue seeds from them.
  • the white-grained sterile plant the male sterile plant grown from the white-grained seed obtained by selfing of the 16L6386 dual-purpose line
  • step 4 Sow the blue seeds obtained in step 4 and cultivate them into plants, eliminate the male sterile plants, and select 26567 individual plants; the selected plants will be selfed and harvested seeds, and the selected seeds have both blue seeds and Plants with white seeds.
  • step 5 For the plants screened in step 5, the white seeds and blue seeds obtained by selfing were collected respectively.
  • step 6 Sow the seeds obtained in step 6 (sort out 100 white seeds and 100 blue seeds for each individual plant) and cultivate them into plants; if the blue seeds of a certain individual plant obtain plants that are all male fertile Plants, and the white seeds of the individual plants are all male sterile plants, these individual plants are potential plants with telomeres, the blue seeds of these individual plants are subjected to microscopic examination, and the T4AgL fragments with translocation telomeres are selected (Ba)-4thS fragment (Rf) ⁇ or translocation endosome T4thS fragment (Rf)-4AgL fragment (Ba) ⁇ , light/medium blue seeds are dual-purpose lines.
  • the male sterile strains are 0 lines, that is, the translocation lines with the translocation end body T4AgL fragment (Ba)-4thS fragment (Rf), which are 20DT1, 20DT3, 20DT7, and 20DT9.
  • the male fertile line is 0, that is, the translocation line with the translocation terminal body T4thS fragment (Rf)-4AgL fragment (Ba), which is 20DT2, 20DT4, 20DT5, 20DT6, 20DT8 .
  • the schematic diagram of the translocation terminal body T4AgL fragment (Ba)-4thS fragment (Rf) is shown in FIG. 4 .
  • the schematic diagram of the translocation terminal body T4thS fragment (Rf)-4AgL fragment (Ba) is shown in FIG. 5 .
  • step 2 The double-diende seeds obtained in step 1 are treated with dimethyl sulfoxide (DMSO) chemical mutagen or ethyl methanesulfonate (EMS) chemical mutagen or radiation treatment, and then cultivated into plants.
  • DMSO dimethyl sulfoxide
  • EMS ethyl methanesulfonate
  • the plant grown from the double-diende seeds obtained in step 1 is used as the male parent, and the male sterile plant grown from the white seed grown by selfing of the 15L4167 dual-purpose line as the female parent Crossbreeding, harvest the seeds on the female plant, select blue grain seeds to process with dimethyl sulfoxide (DMSO) chemical mutagen or process or radiation treatment with ethyl methanesulfonate (EMS) chemical mutagen, then cultivate into plants.
  • DMSO dimethyl sulfoxide
  • EMS ethyl methanesulfonate
  • step 2 or step 3 The plant obtained in step 2 or step 3 is used as the male parent, and is crossed with the white-grained sterile plant (the male sterile plant grown from the white-grained seeds obtained by selfing of the 15L4167 dual-purpose line) as the female parent, and the female parent is harvested Seeds on the plant and sifting the blue seed from it.
  • the white-grained sterile plant the male sterile plant grown from the white-grained seeds obtained by selfing of the 15L4167 dual-purpose line
  • step 4 Sowing the blue seeds obtained in step 4 and cultivating them into plants, eliminating the male sterile plants, selfing the selected plants and harvesting seeds, and selecting the plants whose seeds have both blue seeds and white seeds.
  • step 5 For the plants screened in step 5, the white seeds and blue seeds obtained by selfing were collected respectively.
  • step 6 Sow the seeds obtained in step 6 (sort out 100 white seeds and 100 blue seeds for each individual plant) and cultivate them into plants; if the blue seeds of a certain individual plant obtain plants that are all male fertile Plants, and the white seeds of the individual plants are all male sterile plants, the blue seeds in these individual plants are examined under the microscope, and selected into the T4thS fragment (Rf)-4AgL fragment (Ba)-4BL with translocation terminal body.
  • the strain that is, the translocation line with the translocation end body T4thS fragment (Rf)-4AgL fragment (Ba)-4BL ⁇ .
  • One translocation line, namely 21DT1 was obtained.
  • the schematic diagram of the translocation terminal body T4thS fragment (Rf)-4AgL fragment (Ba)-4BL ⁇ is shown in FIG. 6 .
  • Translocation telosome T4AgL fragment (Ba)-4thS fragment (Rf) The frequency of separation of the two target genes is 0-1 ⁇ , only the sterile line will have 0-0.5 ⁇ fertile plants, and the blue-grain population will not appear sterile plants.
  • the separation frequency of the two target genes is 0-1 ⁇ , sterile There will be no fertile plants in the line, and 0-1 ⁇ sterile plants will appear in the dual-purpose line.
  • Embodiment 4 the breeding method of new dual-purpose line (sterile line)
  • the dual-purpose lines cultivated in Examples 1 to 3 can be used to cultivate new dual-purpose lines.
  • the flow chart is shown in FIG. 7 .
  • the excellent line and the blue-grain dual-purpose line or dark blue-grain hybrid combination of the new system are selected.
  • the blue-grained seeds are selected for planting.
  • the blue-white or blue-red seeds are finally selected.
  • Light/medium blue seeds are planted, and the F 2 generation excellent individual plants with blue-white or blue-red seeds are selected.
  • 82 seeds were planted separately from blue seeds, white seeds or red seeds, each planted in 2 rows, 2m row length, row spacing 25cm, 5cm artificial on-demand sowing.
  • the existing wheat color sorter is designed for processing wheat to remove impurities and mildewed grains. After research and experimentation by the machine manufacturer, it was found that the color sorter produced by Zhongrui Microvision is more suitable for sorting dark blue, light/medium blue (dual-purpose), white or red seeds. After 2-3 times of continuous sorting, the sorting accuracy of blue, white or red seeds can reach more than 99.9%.
  • the color sorter model 6SXZ-68 produced by Zhongrui Microvision has been purchased and used. Use this color sorter to complete the sorting of white or red seeds, light/medium blue seeds (dual-purpose lines) and dark blue seeds to satisfy the breeding of dual-purpose lines (sterile lines). Light/medium blue seeds are used for propagation to produce CMS and dual-purpose seeds.
  • the white-grained sterile lines produced by using multiple dual-purpose lines that have been bred have been respectively compared with varieties (lines) Weilong 121, Weilong 166, 13W8-11, Gaoda 1, and Shannong 22 that are suitable as restorer lines.
  • varieties (lines) Weilong 121, Weilong 166, 13W8-11, Gaoda 1, and Shannong 22 that are suitable as restorer lines.
  • Chuanmai 93, 14th grade 16, 13th grade 6, Mianmai 827, Mianmai 903, MY6381, Chuanyu 36, Shumai 1671, Chuannong 39, Chuanfumai 14, Mianmai 906 and other excellent strains in different ecological regions Hybrid combinations are being formulated, and strong dominant combination screening and new hybrid wheat varieties are being selected in different ecological zones.
  • the invention discloses a breeding method and application of a blue standard two-line hybrid wheat system.
  • the blue standard two-line method hybrid wheat system of the present invention obtains white/red grain (sterile line) and blue grain mixed seeds through self-crossing of dual-purpose lines, and white/red grain seeds (sterile line) are separated by a color sorter , Light/Medium Blue Seeds (Dual Purpose) and Dark Blue Seeds, CMS for Hybrid Seed Production, Light/Medium Blue Seeds (Dual Purpose) for Propagation of CMS and Dual Purpose, Dark Blue
  • the seeds are eliminated or used as parents to breed new sterile lines or to purify and rejuvenate dual-purpose lines.
  • the invention has the characteristics of fast breeding of sterile lines, low production cost of hybrids, strong advantages and the like.

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Abstract

本发明公开了一种蓝标型两系法杂交小麦***的选育方法及应用。本发明创建了对隐性核不育基因ms1具有完全恢复能力且含表现剂量效应和胚乳质感的蓝粒基因Ba和外源恢复基因Rf的外源易位染色体【T4AgL(含Ba的片段)-4BL(靠着丝点的片段).4thS和T4AgL(含Ba的片段)-4BL(靠着丝点的片段).4thS(含Rf的片段)】以及外源易位端体【T4AgL(含Ba的片段)-4thS(含Rf的片段).、T4thS(含Rf的片段)-4AgL(含Ba的片段).和T4thS(含Rf的片段)-4AgL(含Ba的片段)-4BL(靠着丝点的片段).】。完成了蓝标型两系法杂交小麦***的改进。

Description

一种蓝标型两系法杂交小麦***的选育方法及应用 技术领域
本发明属于作物遗传育种领域，涉及一种蓝标型两系法杂交小麦***的选育方法及应用。
背景技术
小麦是世界食用量最大的粮食作物，随着耕地面积减少、人口增加，提高小麦单产势在必行。杂种优势的利用是提高作物单产的重要途径，水稻、玉米、高粱等作物早已杂交化，相继在生产上广泛应用，大幅度提高了产量。小麦杂种优势的利用一直是一个世界难题，直到现在尚未在生产上取得突破。主要原因是小麦是多倍体，其用种量大，本身的杂种优势不如水稻、玉米、高粱等作物的杂种优势强，因而要想生产出高产、低成本的杂交种子，切实可行的杂交小麦***是关键。
自1951年日本科学家Kihara首次发现小麦细胞质雄性不育系以来，科学家们和一些知名的种子公司或生物技术育种公司先后建立和尝试了多种杂交小麦***，包括细胞质雄性不育***(Cytoplasm Male Sterility，CMS)、化学杀雄(Chemical Hybridization Agent，CHA)、光温敏雄性不育***(Photo-thermo-sensitive Cytoplasm Male Sterility，PCMS)和核型不育***(Genetic Male Sterility，GMS)。但由于这些***存在一些缺陷或不足而难于规模化生产。
Driscoll先后建立了XYZ体系和改良XYZ体系，这两个***均是GMS体系，是利用黑麦的5R染色体上携带的显性雄性可育基因和穗茎绒毛显性标记基因，因穗茎绒毛显性标记只能在小麦抽穗后才能识别，对于高密度种植的小麦显然缺乏实用性，但对核不育***的创建和应用具有重要的指导意义。黄寿松等(1991)和周宽基等(1998)先后建立了类似XYZ体系以蓝粒为标记的蓝标型杂交小麦体系，解决了有效标记，通过种子颜色区分不育系和保持系，但因蓝粒两用系自交结实不理想而难以培育出强优势杂交种应用于生产，后者虽然蓝粒植株在自交结实率(30-80％)有所提高，但出现白化苗、黄化苗等不良影响(刘忠祥等，2004)。以蓝粒为标记的杂交小麦***还有澳大利亚的专利“Genetically modified wheat plants and progeny and method for production of hybrid wheat【PCT/AU93/00017(WO93/13649)】”和以色列的专利“Methods for production of hybrid wheat【PCT/IL98/00220(WO98/51142)】”，后面未见其研究和应用的报道。本发明人先前建立的“一种以蓝粒为标记性状的两系法杂交小麦的选育方法(ZL200610042629.8)”，已培育出一批不育系和3个省级审定的杂交种，一批杂交组合正在参加试验，陆续会有更多的杂交种审定推广。在种子分拣、制种都达到了杂交小麦种子要求，但附加的易位染色体在稳定性上还需要改进，该附加染色体从着丝点的断裂频率偏高(0.5-10％)，选育低断裂频率的两用系 有一定的难度。
小麦雄性不育基因多数是由自然突变或人工诱导产生的，现已发现和诱导的雄性核不育材料较多，所含的不育基因包括ms1(隐性)、Ms2(显性)、Ms3(显性)、Ms4(显性)和ms5(隐性)，分别定位在4BS、4DS、5AS、4DS和3AL染色体臂上。ms1基因位点有很多突变体，包括Pugsley's突变体(含等位基因ms1a)、Probus突变体(含等位基因ms1b)、Cornerstone突变体(含等位基因ms1c)、FS2突变体(含等位基因ms1d)、FS3突变体(含等位基因ms1e)、FS24突变体(含等位基因ms1f)、LZ突变体(含等位基因ms1g)和宁春4号系列突变体(含等位基因系列ms1h-p)等。
Wazuddin和Driscoll(1986)将二倍体小麦(一粒小麦)Triticum thaoudar(＝T.boeoticum)、T.urartu和T.monococcum的第4条染色体引入的六倍体普通小麦(T.aestivum)中，发现该染色体不能与普通小麦的任何染色体配对，证明普通小麦中并不存在一粒小麦的第4条染色体，该染色体所带有恢复基因(Rf)可以完全恢复ms1不育基因。Morrison等(2004)描述了蓝粒基因(Ba)的起源和不同，表现胚乳直感和剂量效应的蓝粒基因来源于Agropyron elongatum(Host)Beauv.和A.intermedium(Host)Beauv.(A.tricophorum K.Richter)(2n＝10x＝70)的4Ag染色体，而不表现胚乳直感的蓝粒基因来自T.boeoticum(＝T.thaoudar)第4条染色体。T.boeoticum第4条染色体长臂上存在一个不表现胚乳直感蓝粒基因，也间接证明该染色体不属于普通小麦的染色体。只有表现胚乳直感和剂量效应的蓝粒基因才可作为标记判断小麦籽粒中是否含有蓝粒基因及含有几个拷贝的蓝粒基因(胚乳和糊粉层为三倍体，1个拷贝来自父方，2个拷贝来自母方)。当只有雄配子(花粉)含有蓝粒基因所形成的种子颜色为浅蓝，种子中含1个Ba拷贝(自交所得种子为非蓝粒、浅蓝粒、中蓝粒和深蓝粒)；当只有雌配子含有蓝粒基因所形成的种子颜色为中蓝，种子中含2个Ba拷贝(自交所得种子为非蓝粒、浅蓝粒、中蓝粒和深蓝粒)；当雌雄配子均有蓝粒基因所形成的种子颜色为深蓝，种子中含3个Ba拷贝(自交所得种子全部为蓝粒)。我们的研究表明T.thaoudar(＝T.boeoticum)第4条染色体4th在对小麦农艺性状的影响上要优于T.urartu和T.monococcum中的第4条染色体。
诱导染色体易位的方法很多，主要有以下五种方法:①利用外源染色体单体的错***和不稳定性自发易位；②利用辐射处理；③利用DMSO、EMS等物质化学诱变；④诱导部分同源染色体配对产生易位；⑤组织培养。实践证明，一般染色体易位发生频率都在1％以下。胡英考描述了37个小麦-异源易位系，其中15个易位系是通过辐射的方法获得的，11个易位系是通过部分同源重组的方法获得的，6个自发易位，3个组织培养获得的。Friebe等阐述了57个自发或诱导的小麦-异源易位染色体，有10个是在着丝点发生易位，45个在染色体顶端发 生易位，2个在染色体中间发生易位。这为我们创建对不育基因ms1具有完全恢复能力，含表现胚乳直感和剂量效应的蓝粒基因Ba的外源易位染色体提供了依据。
发明公开
本发明在本发明人先前建立的“一种以蓝粒为标记性状的两系法杂交小麦的选育方法(ZL200610042629.8)”的基础上进行，ZL200610042629.8首先创建以蓝粒为标记性状来分离、保持不育系的核型不育杂交小麦***的原始两用系，然后建立新两用系的选育方法，具体包括下列步骤：
1)创建本***中的原始浅蓝粒两用系
①T4thS.4AgL(ZL200610042629.8中记为4thS.4AgL)外源附加易位染色体的创建
为确保基础材料的真实性和可靠性，通过染色体分带方法及酒精脱氢酶电泳分析技术鉴定T4BS.4AgL易位染色体和来源于一粒小麦(T.thaoudar)的4th染色体的小麦4B代换系4th″，同时将位于小麦4B染色体短臂上的不育基因ms1b和矮秆基因Rht3通过基因重组连锁在一起；再将不育基因ms1b与矮秆基因Rht3同时重组到T4BS.4AgL易位染色体上，获得蓝粒矮秆不育株；
蓝粒矮秆不育株与4th染色体的小麦4B代换系4th″杂交，获得小麦的4th和携带ms1b和Rht3基因的T4BS.4AgL染色体双单体植株即F 1代，经自交得到足够的F 2代白粒或红粒、浅蓝粒和深蓝粒三种种子，只点播F 2代深蓝粒种子；对该F 2代群体中所有高秆可育株编号并作为父本，分别与含“Probus”突变体的不育基因ms1b衍生系中的不育株杂交，只收获具有浅蓝粒种子的杂交穗，种植杂种F 1代浅蓝粒种子，抽穗后套袋，入选全部单株可育的杂交组合，选留较好的单株，分拣出浅蓝粒、白粒或红粒种子继续种植，观察育性；选留白粒或红粒种子长出的植株全部不育且相对应的浅蓝粒种子长出的植株全部可育的浅蓝粒株系，收获浅蓝粒株系，其中浅蓝粒株系含有所需的集外源恢复基因和蓝粒基因为一体的T4thS.4AgL易位染色体；
或者直接以4th和携带ms1b和Rht3基因的T4BS.4AgL染色体双单体植株为父本与含“Probus”突变体的不育基因ms1b衍生系中不含Rht3矮秆基因的不育株杂交，获得足够的F1代蓝粒种子种植，对高秆可育株编号，再分别与含“Probus”突变体的不育基因ms1b衍生系中不含Rht3矮秆基因的不育株杂交，同样选留蓝粒种子种植，观察育性，筛选出所有植株都可育的组合，即获得含有T4thS.4AgL易位染色体株系；
②原始两用系的筛选与鉴定
将上述含T4thS.4AgL易位染色体株系自交或继续与含“Probus”突变体的不育基因ms1b衍生系中的不含Rht3矮秆基因不育株杂交后自交，从后代群体中入选蓝粒比白粒或红粒为1∶2的单株，初步筛选出两用系，再通过对花粉母细 胞减数***中期染色体配对情况镜检进行确认，从而获得该杂交小麦***的原始两用系21Ⅱ+Ⅰ 4thS.4AgL；
2)新两用系选育方法
方法一：原始两用系与优良品系配制杂交组合，在F1代只选留蓝粒种子种植，根据各组合的表现情况，决选优良组合，从入选组合F2代籽粒中分拣出浅蓝粒种子种植，选择优良单株；将F 3代各株系中白粒或红粒和浅蓝粒种子分别种植，各50株，入选并收获白粒或红粒群体中有25％或100％的不育株且农艺性状好的相对应的浅蓝粒株系，后者稳定后即为两用系；以此类推，直到株系稳定、分离出的白粒或红粒群体全部不育为止；测试两用系的产量并对其分离出的白粒或红粒种子进行品质分析、抗病性鉴定，以便获得高产、优质、多抗的新两用系；
方法二：选取不育系与常规优良品系杂交的F2代群体中分离出的优良不育株为母本与原始两用系杂交，与方法一相同，F 1及F 2代选留优良组合和优良单株，F 3代以上筛选优良株系，但不同的是其余各代只种植浅蓝粒种子直到需要不育系之时为止；该方法做杂交时不用去雄，适用于对强优势组合F2代群体中分离出的优良不育株的利用。
本发明的目的是提供一种蓝标型两系法杂交小麦***的选育方法及应用。
1、本发明提供了一种蓝标型两系法杂交小麦的选育方法，包括如下步骤：
(1)06L6109-3两用系的白粒不育株与具有易位染色体T4AgL(Ba)-4BL·4BS的小麦杂交，筛选由蓝粒种子长成的矮杆且雄性不育的植株，即为具有易位染色体T4AgL(Ba)-4BL·4BS(Rht3-ms1b)的植株，称为T4AgL(Ba)-4BL·4BS(Rht3-ms1b)易位系植株；
具有易位染色体T4AgL(Ba)-4BL·4BS的小麦由蓝粒种子长成且具有高杆且雄性可育的性状；06L6109-3两用系的白粒不育株：是06L6109-3两用系自交得到的白粒种子长成的矮杆且雄性不育的植株；06L6109-3两用系为由白粒种子长成的具有半矮秆且雄性不育性状的含ms1b-Rht3的小麦与含有易位染色体T4thS·4AgL的附加系小麦杂交并选育而成的具有Rht3-ms1b的蓝粒两用系；
(2)T4AgL(Ba)-4BL·4BS(Rht3-ms1b)易位系植株与小麦4B代换系4th杂交，获得具有4th和易位染色体T4AgL(Ba)-4BL·4BS(Rht3-ms1b)的双单体种子；
(3)将步骤(2)获得的双单体种子播种并培育成植株，自交，选深蓝粒种子种植并筛选出高杆且雄性可育的植株；
(4)将步骤(3)筛选的植株与14L6386两用系的白粒不育株杂交，筛选结有蓝粒种子的杂交组合；
14L6386两用系的白粒不育株：是14L6386两用系自交得到的白粒种子长成的雄性不育的植株；14L6386两用系为09L6034两用系与糯小麦品系杂交并选育而成的蓝粒两用系；09L6034两用系是T0065-10B-2LB-4两用系与作为轮回亲本的小麦品系周88114(定名为周麦11)回交后选育而成的蓝粒两用系； T0065-10B-2LB-4为含有易位染色体T4thS·4AgL的附加系小麦；
(5)根据步骤(4)的筛选结果，将蓝粒种子播种并培育成植株，如果所有种子得到的植株均为雄性可育株，相应的杂交组合的种子发育得到的植株组成一个潜在的易位系；
(6)步骤(5)得到的潜在的易位系，进行如下操作：自交并收获种子，将种子分为蓝粒种子和白粒种子两组；分别将蓝粒种子和白粒种子播种并培育为植株，如果所有蓝粒种子得到的单株均为雄性可育株且所有白粒种子得到的单株均为雄性不育株，那么将这些由蓝粒种子培育得到的雄性可育单株进行FISH鉴定，筛选具有易位染色体T4AgL片段(Ba)-4BL·4thS(Rf)的单株，这些单株组成T4AgL片段(Ba)-4BL·4thS(Rf)易位系群体；
(7)从步骤(6)得到的易位系的自交后代中，筛选白粒种子:蓝粒种子的数量为2:1的植株，即为含有易位染色体T4AgL片段(Ba)-4BL·4thS(Rf)的附加系，又称为两用系。
两用系的核型为：21Ⅱ(msms)+Ⅰ T4AgL片段(Ba)-4BL·4thS(Rf)。
2、本发明提供了一种蓝标型两系法杂交小麦的选育方法，包括如下步骤：
(1)15L4167两用系自交，然后筛选由蓝粒种子长成的雄性不育植株，将该植株与深蓝粒可育株杂交，然后筛选由深蓝粒种子长成的雄性可育植株并进行自交，然后从自交种子发育的植株中筛选具有雄性不育性状的植株，进一步通过镜检筛选，获得具有易位端体T4AgL片段(Ba)-4BL·的附加系植株；深蓝粒可育株：是15L4167两用系自交得到的深蓝粒种子长成的雄性可育植株，自交后所结种子全为蓝粒种子；
(2)15L4167两用系自交，然后筛选由白粒种子长成的雄性可育植株，进一步通过镜检筛选，获得具有端体4thS(Rf)·的附加系植株；
(3)将具有端体4thS(Rf)·的附加系植株作为父本，具有易位端体T4AgL片段(Ba)-4BL·的附加系植株作为母本，进行杂交，获得具有易位端体T4AgL片段(Ba)-4BL·和端体4thS(Rf)·的双单端体种子；
(4)将步骤(3)得到的种子作父本与作为母本的白粒种子作为杂交组合，培育为植株并杂交，从母本植株上获得的种子中筛选蓝粒种子；白粒种子由15L4167两用系自交而得；
(5)将步骤(4)得到的蓝粒种子播种并培育为植株，淘汰雄性不育株，收获雄性可育株并进行考种，入选得到的种子既有蓝粒种子又有白粒种子的植株；
(6)步骤(5)入选的植株，按单株分别分拣出白粒种子和蓝粒种子；
(7)播种步骤(6)得到的种子，培育为植株；如果某个单株的蓝粒种子得到的植株全部为雄性可育植株，且该单株的白粒种子得到的植株全部为雄性不育植株，将该单株的浅蓝粒种子进行镜检，获得具有易位染色体T4AgL片段(Ba)-4BL·4thS片段(Rf)的易位系植株，又称为两用系；
所述15L4167两用系即以上所述的含有易位染色体T4AgL片段 (Ba)-4BL·4thS(Rf)的附加系。
两用系的核型为：21Ⅱ(msms)+Ⅰ T4AgL片段(Ba)-4BL·4thS片段(Rf)。
3、本发明提供了一种蓝标型两系法杂交小麦的选育方法，包括如下步骤：
(1)16L6386两用系自交，收取蓝粒种子，种植并培育为植株，选择其中的雄性不育株，通过镜检筛选具有端体4AgL(Ba)·的植株；16L6386两用系自交，收取深蓝粒种子，种植并培育为植株，这些植株为雄性可育株；
(2)步骤(1)筛选的雄性不育株与步骤(1)获得的雄性可育株杂交，筛选深蓝粒种子并培育为植株，从其自交后代中筛选出雄性不育株；
(3)16L6386两用系自交，收取白粒种子，种植并培育为植株，选择其中的雄性可育株，通过镜检筛选具有端体4thS(Rf)·的植株；
(4)步骤(2)筛选的雄性不育株与步骤(3)筛选的雄性可育株杂交，收获种子并培育为植株，然后自交，筛选深蓝粒种子，通过镜检筛选具有端体4AgL(Ba)·和端体4thS(Rf)·的重双端体种子；
(5)步骤(4)获得的重双端体种子进行诱变处理，然后培育为植株；
(6)步骤(4)获得的重双端体种子长成的植株作为父本，与作为母本的白粒不育植株杂交，收获母本植株上的种子，选取蓝粒种子进行诱变处理，然后培育为植株；白粒不育植株：由16L6386两用系自交而得的白粒种子长成的雄性不育植株；
(7)将步骤(5)或步骤(6)得到植株作为父本，与作为母本的白粒不育植株杂交，收获母本植株上的种子并从中筛选出蓝粒种子；白粒不育植株：由16L6386两用系自交而得的白粒种子长成的雄性不育植株；
(8)将步骤(7)得到的蓝粒种子播种并培育为植株，淘汰雄性不育株，将入选的植株自交并收获种子，入选所结种子既有蓝粒种子又有白粒种子的植株；
(9)步骤(8)筛选的植株，分别按单株分拣出其自交获得的白粒种子和蓝粒种子；
(10)播种步骤(9)得到的白粒种子和蓝粒种子，培育为植株；如果某个单株的蓝粒种子得到的植株全部为雄性可育植株，且该单株的白粒种子全部为雄性不育植株，将该单株的的蓝粒种子进行镜检，获得具有易位端体T4AgL片段(Ba)-4thS片段(Rf)·的植株，浅/中蓝粒种子为两用系；
16L6386两用系即含有易位染色体T4thS·4AgL的附加系小麦。
两用系的核型为：21Ⅱ(msms)+Ⅰ T4AgL片段(Ba)-4thS片段(Rf)·。
4、本发明提供了一种蓝标型两系法杂交小麦的选育方法，包括如下步骤：
(1)16L6386两用系自交，收取蓝粒种子，种植并培育为植株，选择其中的雄性不育株，通过镜检筛选具有端体4AgL(Ba)·的植株；16L6386两用系自交，收取深蓝粒种子，种植并培育为植株，这些植株为雄性可育株；
(2)步骤(1)筛选的雄性不育株与步骤(1)获得的雄性可育株杂交，筛选深蓝粒种子并培育为植株，从其自交后代中筛选出雄性不育株；
(3)16L6386两用系自交，收取白粒种子，种植并培育为植株，选择其中的雄性可育株，通过镜检筛选具有端体4thS(Rf)·的植株；
(4)步骤(2)筛选的雄性不育株与步骤(3)筛选的雄性可育株杂交，收获种子并培育为植株，然后自交，筛选深蓝粒种子，通过镜检筛选具有端体4AgL(Ba)·和端体4thS(Rf)·的重双端体种子；
(5)步骤(4)获得的重双端体种子进行诱变处理，然后培育为植株；
(6)步骤(4)获得的重双端体种子长成的植株作为父本，与作为母本的白粒不育植株杂交，收获母本植株上的种子，选取蓝粒种子进行诱变处理，然后培育为植株；白粒不育植株：由16L6386两用系自交而得的白粒种子长成的雄性不育植株；
(7)将步骤(5)或步骤(6)得到植株作为父本，与作为母本的白粒不育植株杂交，收获母本植株上的种子并从中筛选出蓝粒种子；白粒不育植株：由16L6386两用系自交而得的白粒种子长成的雄性不育植株；
(8)将步骤(7)得到的蓝粒种子播种并培育为植株，淘汰雄性不育株，将入选的植株自交并收获种子，入选所结种子既有蓝粒种子又有白粒种子的植株；
(9)步骤(8)筛选的植株，分别按单株分拣出其自交获得的白粒种子和蓝粒种子；
(10)播种步骤(9)得到的种子，培育为植株；如果某个单株的蓝粒种子得到的植株全部为雄性可育植株，且该单株的白粒种子全部为雄性不育植株，将该单株的的蓝粒种子进行镜检，获得具有易位端体T4thS片段(Rf)-4AgL片段(Ba)·的植株，浅/中蓝粒种子即为两用系；
16L6386两用系即含有易位染色体T4thS·4AgL的附加系小麦。
两用系的核型为：21Ⅱ(msms)+Ⅰ T4thS片段(Rf)-4AgL片段(Ba)·。
5、本发明提供了一种蓝标型两系法杂交小麦的选育方法，包括如下步骤：
(1)将2、(3)步骤中所获得的具有易位端体T4AgL片段(Ba)-4BL·和端体4thS片段(Rf)·的重双端体种子繁殖；
(2)步骤(1)获得的重双端体种子进行诱变处理，然后培育为植株；
(3)步骤(1)获得的重双端体种子长成的植株作为父本，与作为母本的白粒不育植株杂交，收获母本植株上的种子，选取蓝粒种子进行诱变处理，然后培育为植株；白粒不育植株：15L4167两用系自交而得的白粒种子长成的雄性不育植株；所述15L4167两用系即以上所述的含有易位染色体T4AgL片段(Ba)-4BL·4thS(Rf)的附加系；
(4)将步骤(2)或步骤(3)得到植株作为父本，与作为母本的白粒不育植株杂交，收获母本植株上的种子并从中筛选出蓝粒种子；白粒不育植株：15L4167两用系自交而得的白粒种子长成的雄性不育植株；所述15L4167两用系即以上所述的含有易位染色体T4AgL片段(Ba)-4BL·4thS(Rf)的附加系；
(5)将步骤(4)得到的蓝粒种子播种并培育为植株，淘汰雄性不育株，将 入选的植株自交并收获种子，入选所结种子既有蓝粒种子又有白粒种子的植株；
(6)步骤(5)筛选的植株，分别按单株分拣出其自交获得的白粒种子和蓝粒种子；
(7)播种步骤(6)得到的种子，培育为植株；如果某个单株的蓝粒种子得到的植株全部为雄性可育植株，且该单株的白粒种子全部为雄性不育植株，将这些单株中的蓝粒种子进行镜检，获得具有易位端体T4thS片段(Rf)-4AgL片段(Ba)-4BL·的植株，浅/中蓝粒种子即为两用系。
两用系的核型为：21Ⅱ(msms)+Ⅰ T4thS片段(Rf)-4AgL片段(Ba)-4BL·。
所述方法具体包括如下步骤：
(1)06L6109-3两用系的白粒不育株与具有易位染色体T4AgL(Ba)-4BL·4BS的小麦杂交，筛选由蓝粒种子长成的矮杆且雄性不育的植株，即为具有易位染色体T4AgL(Ba)-4BL·4BS(Rht3-ms1b)的植株，称为T4AgL(Ba)-4BL·4BS(Rht3-ms1b)易位系植株；
具有易位染色体T4AgL(Ba)-4BL·4BS的小麦为由蓝粒种子长成的高杆且雄性可育的小麦；易位染色体T4AgL(Ba)-4BL·4BS是将4AgL(Ba)重组到4B上得到的；06L6109-3两用系的白粒不育株：是06L6109-3两用系自交得到的白粒种子长成的矮杆且雄性不育的植株；06L6109-3两用系为由白粒种子长成的具有半矮秆且雄性不育性状的含ms1b-Rht3的小麦与含有易位染色体T4thS·4AgL的附加系小麦杂交并选育而成的具有Rht3-ms1b的蓝粒两用系；
(2)T4AgL(Ba)-4BL·4BS(Rht3-ms1b)易位系植株(纯合体)与小麦4B代换系4th杂交，获得具有4th和易位染色体T4AgL(Ba)-4BL·4BS(Rht3-ms1b)的双单体种子；
(3)将步骤(2)获得的双单体种子播种并培育成植株，自交，选深蓝粒种子种植并筛选出高杆且雄性可育的植株；
(4)将步骤(3)筛选的植株与14L6386两用系的白粒不育株杂交，筛选结有蓝粒种子的杂交组合；
(5)根据步骤(4)的筛选结果，将杂交组合的蓝粒种子播种并培育成植株，如果所有种子得到的植株均为雄性可育株，相应的杂交组合的种子发育得到的植株组成一个潜在的易位系；
(6)步骤(5)得到的潜在的易位系，进行如下操作：自交并收获种子，将种子分为蓝粒种子和白粒种子两组；分别将蓝粒种子和白粒种子播种并培育为植株，如果所有蓝粒种子得到的单株均为雄性可育株且所有白粒种子得到的单株均为雄性不育株，那么将这些由蓝粒种子培育得到的雄性可育单株进行FISH鉴定，筛选具有易位染色体T4AgL片段(Ba)-4BL·4thS(Rf)的单株，这些单株组成T4AgL片段(Ba)-4BL·4thS(Rf)易位系群体；
(7)从步骤(6)得到的易位系的自交后代中，筛选白粒种子:蓝粒种子的数量为2:1的植株，即为含易位染色体T4AgL片段(Ba)-4BL·4thS(Rf)的附加系 (两用系)。
两用系的核型为：21Ⅱ(msms)+Ⅰ T4AgL片段(Ba)-4BL·4thS(Rf)。
所述方法具体包括如下步骤：
(1)15L4167两用系自交，然后筛选由蓝粒种子长成的雄性不育植株，即具有易位端体T4AgL片段(Ba)-4BL·的植株；将该植株与深蓝粒可育株(15L4167两用系自交，筛选由深蓝粒种子长成的雄性可育植株，自交后所结种子全为蓝粒种子)杂交，然后从自交种子发育的植株中筛选由深蓝粒种子长成的雄性可育植株并进行自交，然后从自交种子发育的植株中筛选具有雄性不育性状的植株，进一步通过镜检筛选，获得具有易位端体T4AgL片段(Ba)-4BL·的附加系植株；
(2)15L4167两用系自交，然后筛选由白粒种子长成的雄性可育植株，即具有端体4thS(Rf)·的植株，进一步通过镜检筛选，获得具有端体4thS(Rf)·的附加系植株；
(3)将具有端体4thS(Rf)·的附加系植株作为父本，具有易位端体T4AgL片段(Ba)-4BL·的附加系植株作为母本，进行杂交，获具有易位端体T4AgL片段(Ba)-4BL·和端体4thS(Rf)·的双单端体种子；
(4)将步骤(3)得到的种子作父本与作为母本的白粒种子(白粒种子由15L4167两用系自交而得)作为杂交组合，从母本植株上获得的种子中筛选蓝粒种子；
(5)将步骤(4)得到的蓝粒种子播种并培育为植株，淘汰雄性不育株，收获雄性可育株；对收获的雄性可育株进行考种，入选得到的种子既有蓝粒种子又有白粒种子的植株；
(6)步骤(5)入选的单株，按单株分别分拣出白粒种子和蓝粒种子；
(7)播种步骤(6)得到的种子，培育为植株；如果某个单株的蓝粒种子得到的植株全部为雄性可育植株，且该单株的白粒种子得到的植株全部为雄性不育植株，将该单株的浅蓝粒种子进行镜检，获得具有易位染色体T4AgL片段(Ba)-4BL·4thS片段(Rf)的易位系植株，浅/中蓝粒种子为两用系。
两用系的核型为：21Ⅱ(msms)+Ⅰ T4AgL片段(Ba)-4BL·4thS片段(RF)。
所述方法具体包括如下步骤：
(1)16L6386两用系自交，收取蓝粒种子，种植并培育为植株，选择其中的雄性不育株，通过镜检筛选具有端体4AgL(Ba)·的植株；16L6386两用系自交，收取深蓝粒种子，种植并培育为植株，这些植株为雄性可育株；
(2)步骤(1)筛选的雄性不育株与步骤(1)获得的雄性可育株杂交，筛选深蓝粒种子并培育为植株，从其自交后代中筛选出雄性不育株；
(3)16L6386两用系自交，收取白粒种子，种植并培育为植株，选择其中的雄性可育株，通过镜检筛选具有端体4thS(Rf)·的植株；
(4)步骤(2)筛选的雄性不育株与步骤(3)筛选的雄性可育株杂交，收 获种子(均为蓝粒)并培育为植株，然后自交，筛选深蓝粒种子，通过镜检筛选具有端体4AgL(Ba)·和端体4thS(Rf)·的重双端体种子；
(5)步骤(4)获得的重双端体种子进行诱变处理，然后培育为植株；
(6)步骤(4)获得的重双端体种子长成的植株作为父本，与作为母本的白粒不育植株(由16L6386两用系自交而得的白粒种子长成的雄性不育植株)杂交，收获母本植株上的种子，选取蓝粒种子进行诱变处理，然后培育为植株；
(7)将步骤(5)或步骤(6)得到植株作为父本，与作为母本的白粒不育植株(由16L6386两用系自交而得的白粒种子长成的雄性不育植株)杂交，收获母本植株上的种子并从中筛选出蓝粒种子；
(8)将步骤(7)得到的蓝粒种子播种并培育为植株，淘汰雄性不育株，将入选的植株自交并收获种子，入选所结种子既有蓝粒种子又有白粒种子的植株；
(9)步骤(8)筛选的植株，分别按单株分拣出其自交获得的白粒种子和蓝粒种子；
(10)播种步骤(9)得到的种子，培育为植株；如果某个单株的蓝粒种子得到的植株全部为雄性可育植株，且该单株的白粒种子全部为雄性不育植株，这些单株为潜在的具有端体的植株，将这些单株的的蓝粒种子进行镜检，入选具有易位端体T4AgL片段(Ba)-4thS片段(Rf)·的植株，即具有易位端体T4AgL片段(Ba)-4thS片段(Rf)·的易位系，浅/中蓝粒种子为两用系。
两用系的核型为：21Ⅱ(msms)+Ⅰ T4AgL片段(Ba)-4thS片段(Rf)·。
所述方法具体包括如下步骤：
(1)16L6386两用系自交，收取蓝粒种子，种植并培育为植株，选择其中的雄性不育株，通过镜检筛选具有端体4AgL(Ba)·的植株；16L6386两用系自交，收取深蓝粒种子，种植并培育为植株，这些植株为雄性可育株；
(2)步骤(1)筛选的雄性不育株与步骤(1)获得的雄性可育株杂交，筛选深蓝粒种子并培育为植株，从自交种子发育的植株中筛选出雄性不育株；
(3)16L6386两用系自交，收取白粒种子，种植并培育为植株，选择其中的雄性可育株，通过镜检筛选具有端体4thS(Rf)·的植株；
(4)步骤(2)筛选的雄性不育株与步骤(3)筛选的雄性可育株杂交，收获种子(均为蓝粒)并培育为植株，然后自交，筛选深蓝粒种子，通过镜检筛选具有端体4AgL(Ba)·和端体4thS(Rf)·的重双端体种子；
(5)步骤(4)获得的重双端体种子进行诱变处理，然后培育为植株；
(6)步骤(4)获得的重双端体种子长成的植株作为父本，与作为母本的白粒不育植株(由16L6386两用系自交而得的白粒种子长成的雄性不育植株)杂交，收获母本植株上的种子，选取蓝粒种子进行诱变处理，然后培育为植株；
(7)将步骤(5)或步骤(6)得到植株作为父本，与作为母本的白粒不育植株(由16L6386两用系自交而得的白粒种子长成的雄性不育植株)杂交，收获母本植株上的种子并从中筛选出蓝粒种子；
(8)将步骤(7)得到的蓝粒种子播种并培育为植株，淘汰雄性不育株，将入选的植株自交并收获种子，入选所结种子既有蓝粒种子又有白粒种子的植株；
(9)步骤(8)筛选的植株，分别按单株分拣出其自交获得的白粒种子和蓝粒种子；
(10)播种步骤(9)得到的种子，培育为植株；如果某个单株的蓝粒种子得到的植株全部为雄性可育植株，且该单株的白粒种子全部为雄性不育植株，这些单株为潜在的具有端体的植株，将这些单株的的蓝粒种子进行镜检，入选具有易位端体T4thS片段(Rf)-4AgL片段(Ba)·的植株，即具有易位端体T4thS片段(Rf)-4AgL片段(Ba)·的易位系，浅/中蓝粒种子为两用系。
两用系的核型为：21Ⅱ(msms)+Ⅰ T4thS片段(Rf)-4AgL片段(Ba)·。
所述方法具体包括如下步骤：
(1)具有易位端体T4AgL片段(Ba)-4BL·和端体4thS片段(Rf)·的重双单端体种子繁殖；
(2)步骤(1)获得的重双端体种子进行诱变处理，然后培育为植株。
(3)步骤(1)获得的重双端体种子长成的植株作为父本，与作为母本的白粒不育植株(15L4167两用系自交而得的白粒种子长成的雄性不育植株)杂交，收获母本植株上的种子，选取蓝粒种子进行诱变处理，然后培育为植株；
(4)将步骤(2)或步骤(3)得到植株作为父本，与作为母本的白粒不育植株(15L4167两用系自交而得的白粒种子长成的雄性不育植株)杂交，收获母本植株上的种子并从中筛选出蓝粒种子；
(5)将步骤(4)得到的蓝粒种子播种并培育为植株，淘汰雄性不育株，将入选的植株自交并收获种子，入选所结种子既有蓝粒种子又有白粒种子的植株；
(6)步骤(5)筛选的植株，分别按单株分拣出其自交获得的白粒种子和蓝粒种子；
(7)播种步骤(6)得到的种子，培育为植株；如果某个单株的蓝粒种子得到的植株全部为雄性可育植株，且该单株的白粒种子全部为雄性不育植株，将这些单株中的蓝粒种子进行镜检，入选具有易位端体T4thS片段(Rf)-4AgL片段(Ba)-4BL·的植株，即具有易位端体T4thS片段(Rf)-4AgL片段(Ba)-4BL·的易位系，浅/中蓝粒种子即为两用系。
两用系的核型为：21Ⅱ(msms)+Ⅰ T4thS片段(Rf)-4AgL片段(Ba)-4BL·。
含有易位染色体T4thS·4AgL的附加系小麦为任意具有该染色体形式的小麦，具体可为ZL200610042629.8中记载的方法获得的两用系小麦(即为授权公告号为CN 100420368C中说明书第5页第7行的两用系，原始两用系、新两用系均可)。含有易位染色体T4thS·4AgL的附加系小麦(两用系)的核型为21Ⅱ(msms)+Ⅰ T4thS·4AgL。
T4AgL(Ba)-4BL·4BS(Rht3-ms1b)易位系植株(纯合体)的相关表型：由蓝粒种子长成的矮秆且雄性不育的植株。
具有4th和易位染色体T4AgL(Ba)-4BL·4BS(Rht3-ms1b)的双单体种子的核型为：20Ⅱ+Ⅰ T4AgL(Ba)-4BL·4BS(Rht3-ms1b)+I 4th。
小麦4B代换系4th即来源于一粒小麦T.thaoudar(二倍体，2n＝14)的第4条染色体4th的小麦4B代换系4th″(N4B)。小麦4B代换系4th的相关表型：蓝粒(其含有的蓝粒基因不表现胚乳直感)、高杆且雄性可育。
14L6386两用系的白粒不育株：是14L6386两用系自交得到的白粒种子长成的雄性不育的植株。
14L6386两用系为09L6034两用系(父本)与糯小麦品系(母本)杂交并选育而成的蓝粒两用系(除附加染色体稳定性较差外，其余农艺性状已稳定,糯小麦)。
09L6034两用系是以小麦品系周88114(定名为周麦11)作为轮回亲本(母本)与T0065-10B-2LB-4两用系回交3次后选育而成的蓝粒两用系。T0065-10B-2LB-4两用系记载于ZL200610042629.8(即为授权公告号为CN100420368C说明书中第9页第14行的T0065-10B-2LB-4)。T0065-10B-2LB-4两用系的核型为21Ⅱ(msms)+Ⅰ T4thS·4AgL，即含有易位染色体T4thS·4AgL的附加系小麦。
15L4167两用系为含易位染色体T4AgL片段(Ba)-4BL·4thS(Rf)的附加系。
15L4167两用系的核型为：21Ⅱ(msms)+Ⅰ T4AgL片段(Ba)-4BL·4thS(Rf)。
具有易位端体T4AgL片段(Ba)-4BL·的二体附加系植株的核型为：21Ⅱ(msms)+Ⅱ T4AgL片段(Ba)-4BL·。
具有易位端体T4AgL片段(Ba)-4BL·的二体附加系的表型为：种子为深蓝粒，植株表现为雄性不育。
具有端体4thS(Rf)·的二体附加系植株的核型为：21Ⅱ(msms)+Ⅱ 4thS(Rf)·。
具有端体4thS(Rf)·的二体附加系的表型为：种子为白粒，植株表现为雄性可育。
具有易位端体T4AgL片段(Ba)-4BL·和端体4thS(Rf)·的双单端体种子的核型为：21Ⅱ(msms)+Ⅰ T4AgL片段(Ba)-4BL·+Ⅰ 4thS(Rf)·。
16L6386两用系为含有易位染色体T4thS·4AgL的附加系小麦。
16L6386两用系为含有易位染色体T4thS·4AgL的附加系小麦，通过世代选择选育得到的两用系。
16L6386两用系为发明专利(ZL200610042629.8)中培育的两用系，通过世代选择选育得到的两用系。发明专利(ZL200610042629.8)中培育的两用系为含有易位染色体T4thS·4AgL的附加系小麦(即为授权公告号为CN 100420368C中第9页第14行的T0065-10B-2LB-4材料转育而成的小麦)。
含有易位染色体T4thS·4AgL的附加系小麦两用系的核型为：21Ⅱ(msms)+Ⅰ T4thS·4AgL。
具有端体4AgL(Ba)·和端体4thS(Rf)·的重双端体种子的核型为：21Ⅱ(msms) +Ⅱ 4AgL·+Ⅱ 4thS·。
具有易位端体T4AgL片段(Ba)-4BL·和端体4thS(Rf)·的重双端体种子的核型为：21Ⅱ(msms)+Ⅱ T4AgL片段(Ba)-4BL·+Ⅱ 4thS(Rf)·。以上任一所述筛选白粒种子:蓝粒种子的数量为2:1的植株，指的的分子遗传学中的筛选白粒种子:蓝粒种子的数量约为2:1的植株，并非严谨意义上的2:1，可上下浮动5％或10％。
以上任一所述诱变处理具体可为用二甲亚砜(DMSO)化学诱变剂处理或者用甲基磺酸乙酯(EMS)化学诱变剂处理或辐射处理。
以上任一所述方法还可包括如下步骤(方法一)：
优良品系与新***的蓝粒两用系或深蓝粒配制杂交组合，在F 1代只选留蓝粒种子种植，经田间表现和室内考种情况，决选出蓝白粒或蓝红粒种子颜色区分明显的优良组合。浅/中蓝粒种子种植，入选蓝白粒或蓝红粒种子颜色区分明显的F 2代优良单株。按单株将蓝粒和白粒或红粒种子分开各自种植82粒种子，每个种2行，2m行长，行距25cm，5cm人工点播。入选白粒或红粒群体中有约25％或100％的不育株且农艺性状好的对应的蓝粒优良单株，直至所选单株的白粒或红粒群体全部不育为止且农艺性状稳定一致后，获得新两用系(浅/中蓝粒种子)。也可利用本发明设计人所建立的分子检测方法(专利申请号：201810110738.1)对不育基因ms1b进行跟踪监测，有目的的完成在人工控制条件下一年多代的两用系定向快速转育。注意各代入选的单株或株系中的白粒或红粒种子所占比例必须大于或等于60％，同时筛选白/红粒不育株异交结实率高的相应蓝粒单株或株系。
以上任一所述方法还可包括如下步骤(方法二)：
选取不育系与新***的蓝粒两用系或深蓝粒配制杂交组合，除了各代中只种植蓝粒种子外，其余的选育方式与方法一相同。
本发明公开了以蓝粒为标记性状的两系法杂交小麦***(简称蓝标型两系法杂交小麦***)的创制及应用，其特征在于，该发明创建了具备对小麦4B染色体上的隐性核不育基因ms1完全恢复能力和具有胚乳质感的蓝粒基因的外源易位染色体【T4AgL(含Ba的片段)-4BL(靠着丝点的片段).4thS和T4AgL(含Ba的片段)-4BL(靠着丝点的片段).4thS(含Rf的片段)】以及外源易位端体【T4AgL(含Ba的片段)-4thS(含Rf的片段).、T4thS(含Rf的片段)-4AgL(含Ba的片段).和T4thS(含Rf的片段)-4AgL(含Ba的片段)-4BL(靠着丝点的片段).】，并将创建的任一易位染色体或端体作为ms1纯合体小麦品(系)的附加单体(两用系)，从而完成改进型蓝标型两系法杂交小麦***创制。
所述的不同附加外源易位染色体或外源易位端体，其特征在于，均不能与普通小麦的任何染色体配对，而且均可完全恢复ms1不育基因系列(ms1a，ms1b，ms1c，ms1d，……)；
所述的不同附加外源易位染色体或外源易位端体，其特征在于Rf恢复基因来源于Triticum thaoudar第4条染色体和Ba蓝粒基因来源于Agropyron elongatum， T4AgL(含Ba的片段)-4BL(靠着丝点的片段).4thS易位染色体从着丝点的断裂频率为0.5-5％，不育系出现可育株的频率为两用系出现不育株的一半左右；T4AgL(含Ba的片段)-4BL(靠着丝点的片段).4thS(含Rf的片段)易位染色体从着丝点的断裂频率为0-1％，不育系出现可育株的频率为两用系两用系出现不育株的一半左右；T4AgL(含Ba的片段)-4thS(含Rf的片段).端体两个目的基因分开的频率为0-1‰，仅不育系会出现0-0.5‰的可育株，蓝粒群体不会出现不育株；T4thS(含Rf的片段)-4AgL(含Ba的片段).和T4thS(含Rf的片段)-4AgL(含Ba的片段)-4BL(靠着丝点的片段).端体两个目的基因分开的频率为0-1‰，不育系中不会出现可育株，两用系中会出现0-1‰的不育株。
不同两用系自交产生不同颜色的种子比例有差异，白粒或红粒(不育系)种子变幅55-85％、浅/中蓝粒种子(单体附加，两用系)为15-40％、深蓝粒种子(双端体附加)为2-6％，其中，不育系种子用于生产杂交种，两用系种子用于繁殖不育系和两用系，深蓝粒种子(双端体附加)淘汰或作亲本或用于两用系的提纯复壮。
为了加速蓝标型两系法杂交小麦***产业化，使用“一种以蓝粒为标记性状的两系法杂交小麦的选育方法(ZL200610042629.8)”培育审定的杂交种和强优势组合中所用的两用系(不育系)的稳定性需要进一步提高，将新创建的外源易位染色体或端体替换掉原来的附加染色体，这些两用系包括08L5070、12L8012、12L8015、12L8016、18L7077等。这个过程可在人工控制环境下2-3年时间以目标不育系为轮回亲本与改进型两用系或深蓝粒纯合系连续回交6次以上，筛选不育系与原来一样的两用系，从而实现附加易位染色体或端体的替换。以上不育系已获得回交4次的群体。同理可实现新易位染色体或端体的互换。
本发明还提供了一种易位染色体(异位染色体)，依次包括如下区段：4AgL片段(Ba)、4BL、着丝粒、4thS(Rf)。
本发明还提供了一种易位染色体(异位染色体)，依次包括如下区段：4AgL片段(Ba)、4BL、着丝粒、4thS片段(Rf)。
本发明还提供了一种易位端体(异位端粒)，依次包括如下区段：4AgL片段(Ba)、4thS片段(Rf)、着丝粒。
本发明还提供了一种易位端体(异位端粒)，依次包括如下区段：4thS片段(Rf)、4AgL片段(Ba)、着丝粒。
本发明还提供了一种易位端体(异位端粒)，依次包括如下区段：4thS片段(Rf)、4AgL片段(Ba)-4BL、着丝粒。
4AgL代表来源于长穗偃麦草(Agropyron elongatum)的第4条染色体4Ag的长臂，具有Ba基因。4AgL也可以用4AgL(Ba)表示。保留有Ba基因的4AgL片段用4AgL片段(Ba)表示。
4th代表来源于一粒小麦T.thaoudar的第4条染色体。
4thS代表4th的短臂，含有恢复雄性育性的基因(Rf基因)。4thS也可以用4thS(Rf)表示。保留有Rf基因的4thS片段用4thS片段(Rf)表示。
4B代表来自于六倍体普通小麦(T.aestivum)的B组第4条染色体。
4BS代表4B的短臂。
4BL代表4B的长臂。
4BS(Rht3-ms1b)代表具有Rht3基因和ms1b基因连锁的4BS。Rht3基因为矮杆基因(来自Nainari Rht3)。ms1b基因为雄性不育基因(来自“Probus”突变体)。
“·”代表着丝点。T代表易位染色体或易位端体(即着丝点位于端部的染色体)。Ⅱ代表二价体。I代表单价体。N4B代表4B染色体缺失，即无4B染色体。
本文中，所述具有半矮秆且雄性不育性状的含ms1b-Rht3的小麦为任意具有该性状和染色体形式的小麦，例如以ZL200610042629.8中记载的方法得到的具有半矮秆且雄性不育性状的含ms1b-Rht3的小麦(即为授权公告号为CN100420368C说明书中第7页第13行的ms-Rht3/Ms-rht3(半矮秆)小麦)。
本发明在理论和实践的指导下，以前期积累的种质材料为基础，充分利用传统的生物学技术、染色体工程技术以及作物遗传育种方法，创建更加稳定的具有外源恢复基因和蓝粒基因紧密连锁的外源易位染色体或易位端体，从而达到改进蓝标型两系法杂交小麦***的目的。
本发明以专利(ZL200610042629.8)为基础，针对其不育系生产过程中的附加易位染色体的稳定性进行了改进，创建了对隐性核不育基因ms1具有完全恢复能力且含表现剂量效应和胚乳质感的蓝粒基因Ba和外源恢复基因Rf的外源易位染色体【T4AgL(含Ba的片段)-4BL(靠着丝点的片段).4thS和T4AgL(含Ba的片段)-4BL(靠着丝点的片段).4thS(含Rf的片段)】以及外源易位端体【T4AgL(含Ba的片段)-4thS(含Rf的片段).、T4thS(含Rf的片段)-4AgL(含Ba的片段).和T4thS(含Rf的片段)-4AgL(含Ba的片段)-4BL(靠着丝点的片段).】。并将创建的任一易位染色体或易位端体作为ms1纯合体品(系)的附加单体(即两用系，种子颜色为中蓝和浅蓝)，从而完成蓝标型两系法杂交小麦***的改进。
通过两用系自交获得白/红粒(不育系)和蓝粒混合种子，经色选机分离出白/红粒种子(不育系)、浅/中蓝粒种子(两用系)和深蓝粒种子，不育系用于杂交种制种，浅/中蓝粒种子(两用系)用于繁殖不育系和两用系，深蓝粒种子淘汰或用作亲本转育新不育系或用于两用系提纯复壮。不同易位染色体或易位端体可根据需要相互替换，以满足不同遗传背景下不育系纯度和稳定性需求。本发明具有不育系选育快、杂交种生产成本低、优势强等特点，而且比先前本人发明的“一种以蓝粒为标记性状的两系法杂交小麦的选育方法 (ZL200610042629.8)”更加稳定可靠。
附图说明
图1为易位染色体T4AgL片段(Ba)-4BL·4thS(Rf)的示意图和FISH图(左：示意图；右：FISH图)。
图2为附加系植株的核型图：21Ⅱ(msms)+Ⅰ T4AgL片段(Ba)-4BL·4thS(Rf)。
图3为易位染色体T4AgL片段(Ba)-4BL·4thS片段(Rf)的示意图(左：示意图；右：染色体)。
图4为易位端体T4AgL片段(Ba)-4thS片段(Rf)·的示意图(左：示意图；右：染色体端体)。
图5为易位端体T4thS片段(Rf)-4AgL片段(Ba)·的示意图(左：示意图；右：染色体端体)。
图6为易位端体T4thS片段(Rf)-4AgL片段(Ba)-4BL·的示意图(左：示意图；右：染色体端体)。
图7为两用系选育流程示意图。
图8为强优势组合(杂交种)的选育流程示意图。
实施发明的最佳方式
下面结合具体实施方式对本发明进行进一步的详细描述，给出的实施例仅为了阐明本发明，而不是为了限制本发明的范围。以下提供的实施例可作为本技术领域普通技术人员进行进一步改进的指南，并不以任何方式构成对本发明的限制。
下述实施例中的实验方法，如无特殊说明，均为常规方法，按照本领域内的文献所描述的技术或条件或者按照产品说明书进行。下述实施例中所用的材料、试剂等，如无特殊说明，均可从商业途径得到。如无特殊说明，以下实施例中的定量试验，均设置三次重复实验，结果取平均值。
4AgL代表来源于长穗偃麦草(Agropyron elongatum)的第4条染色体4Ag的长臂，具有Ba基因，该基因为表现剂量效应和胚乳直感的蓝粒基因，是显性基因，胚乳和糊粉层具有3个拷贝的Ba基因的籽粒为深蓝色，胚乳和糊粉层具有2个拷贝的Ba基因的籽粒为中蓝色，胚乳和糊粉层具有1个拷贝的Ba基因的籽粒为浅蓝色，胚乳和糊粉层不具有Ba基因的籽粒为白色或红色。4AgL也可以用4AgL(Ba)表示。保留有Ba基因的4AgL片段用4AgL片段(Ba)表示。
4th代表来源于一粒小麦T.thaoudar的第4条染色体。
4thS代表4th的短臂，含有恢复雄性育性的基因(Rf基因)。4thS也可以用4thS(Rf)表示。保留有Rf基因的4thS片段用4thS片段(Rf)表示。
4B代表来自于六倍体普通小麦(T.aestivum)的B组第4条染色体。
4BS代表4B的短臂。
4BL代表4B的长臂。
4BS(Rht3-ms1b)代表具有Rht3基因和ms1b基因连锁的4BS。Rht3基因为矮杆基因(来自Nainari Rht3)，是显性基因，具有2个拷贝的Rht3基因的植株表现为矮杆，具有1个拷贝的Rht3基因的植株表现为半矮杆，不具有Rht3基因的植株表现为高杆。ms1b基因为雄性不育基因(来自“Probus”突变体)，为隐性基因。
“·”代表着丝点。T代表易位染色体或易位端体(即着丝点位于端部的染色体)。Ⅱ代表二价体。I代表单价体。N4B代表4B染色体缺失，即无4B染色体。
实施例中，非蓝粒(包括白粒或红粒)、蓝粒(包括浅蓝粒、中蓝粒或深蓝粒)指的是种子颜色性状。实施例中，矮杆、半矮秆、高杆指的是植株株高性状。实施例中，可育、不育指的是植株雄性育性性状。例如，具有蓝粒、矮杆且不育性状的植株指的是该植株具有矮杆和雄性不育的性状且该植株由蓝粒种子长成。
如无特殊说明，实施例中的杂交，雄性不育植株作为母本，另一亲本植株作为父本。
实施例1、易位染色体T4AgL片段(Ba)-4BL·4thS(Rf)的创建
1、获得易位染色体T4AgL(Ba)-4BL·4BS(Rht3-ms1b)。
(1)将4AgL(Ba)重组到4B上，获得易位染色体T4AgL(Ba)-4BL·4BS。
(2)06L6109-3两用系的白粒不育株与具有易位染色体T4AgL(Ba)-4BL·4BS的小麦(由蓝粒种子长成的高杆且雄性可育的小麦)杂交，筛选由蓝粒种子长成的矮杆且雄性不育的植株。
06L6109-3两用系的白粒不育株：是06L6109-3两用系自交得到的白粒种子长成的矮杆且雄性不育的植株。
06L6109-3两用系为由白粒种子长成的具有半矮秆且雄性不育性状的含ms1b-Rht3的小麦与ZL200610042629.8中的方法获得的两用系杂交并选育而成的具有Rht3-ms1b的蓝粒两用系。ZL200610042629.8中的方法获得的两用系的核型为：21Ⅱ(msms)+Ⅰ T4thS·4AgL；即含有易位染色体T4thS·4AgL的附加系小麦。
(3)步骤(2)筛选的植株即为具有易位染色体T4AgL(Ba)-4BL·4BS(Rht3-ms1b)的植株，称为T4AgL(Ba)-4BL·4BS(Rht3-ms1b)易位系植株。T4AgL(Ba)-4BL·4BS(Rht3-ms1b)易位系植株(纯合体)的相关表型：由蓝粒种子长成的矮秆且雄性不育的植株。
(4)步骤(3)筛选的植株与具有易位染色体T4AgL(Ba)-4BL·4BS的小麦(由蓝粒种子长成的高杆且雄性可育的小麦)杂交，获得的植株可进行自交并保存种子，可源源不断地获得与步骤(3)中相似的矮秆且雄性不育的植株。
2、T4AgL(Ba)-4BL·4BS(Rht3-ms1b)易位系植株(纯合体)与小麦4B代换 系4th杂交，获得500粒以上具有4th和易位染色体T4AgL(Ba)-4BL·4BS(Rht3-ms1b)的双单体种子。双单体种子的核型为：20Ⅱ+Ⅰ T4AgL(Ba)-4BL·4BS(Rht3-ms1b)+I 4th。
小麦4B代换系4th即来源于一粒小麦T.thaoudar(二倍体，2n＝14)的第4条染色体4th的小麦4B代换系4th″(N4B)。小麦4B代换系4th的相关表型：蓝粒(其含有的蓝粒基因不表现胚乳直感)、高杆且雄性可育。
3、将步骤2获得的双单体种子播种并培育成植株，自交，选深蓝粒种子种植并筛选出高杆且雄性可育的植株。
4、将步骤3筛选的植株与14L6386两用系的白粒不育株杂交，共配制561个杂交组合，杂交组合逐一编号，筛选结有蓝粒种子的杂交组合(此处可以筛除4th染色体上的蓝粒基因)。
14L6386两用系的白粒不育株：是14L6386两用系自交得到的白粒种子长成的雄性不育的植株。
14L6386两用系为09L6034两用系(父本)与糯小麦品系(母本)杂交并选育而成的蓝粒两用系(除附加染色体稳定性较差外，其余农艺性状已稳定,糯小麦)。
09L6034两用系是以周88114(定名为周麦11)作为轮回亲本(母本)与T0065-10B-2LB-4两用系回交3次后选育而成的蓝粒两用系。T0065-10B-2LB-4两用系记载于ZL200610042629.8。T0065-10B-2LB-4两用系的核型为：21Ⅱ(msms)+Ⅰ T4thS·4AgL；即含有易位染色体T4thS·4AgL的附加系小麦。
5、根据步骤4的筛选结果，将不同杂交组合的蓝粒种子按顺序分别播种并培育成植株，如果所有种子得到的植株均为雄性可育株，相应的杂交组合的种子发育得到的植株组成一个潜在的易位系。共获得35个潜在的易位系，即14T1、14T2、…、14T34、和14T35。
6、步骤5得到的35个潜在的易位系，分别进行如下操作：自交并收获种子，将种子分为蓝粒种子和白粒种子两组；分别将蓝粒种子(100粒/个潜在易位系)和白粒种子(100粒/个潜在易位系)播种并培育为植株，如果所有蓝粒种子得到的单株均为雄性可育株且所有白粒种子得到的单株均为雄性不育株，那么将这些由蓝粒种子培育得到的雄性可育单株进行FISH鉴定，筛选具有易位染色体T4AgL片段(Ba)-4BL·4thS(Rf)的单株，这些单株组成T4AgL片段(Ba)-4BL·4thS(Rf)易位系群体。共获得符合要求的5个易位系，即14T9、14T12、14T21、14T31和14T34。同时获得了1个稳定较差的易位系，即14T35，用于实施例2。
7、分别从步骤6得到的5个易位系的自交后代中，筛选白粒种子:蓝粒种子的数量约为2:1的植株，即为含易位染色体T4AgL片段(Ba)-4BL·4thS(Rf)的附加系(两用系)。附加系的核型为：21Ⅱ(msms)+Ⅰ T4AgL片段(Ba)-4BL·4thS(Rf)。易位染色体T4AgL片段(Ba)-4BL·4thS(Rf)的示意图和FISH图见图1。附加系植株的 核型图见图2。
将得到的附加系植株进行扩大的群体种植并调查不同单株中的蓝粒群体中雄性不育株和白粒群体中的雄性可育株出现频率。蓝粒群体中出现雄性不育株的频率为0.5-5％，白粒群体中出现雄性可育株的频率为0.2-3％。测试群体在1000株以上。
实施例2、易位染色体T4AgL片段(Ba)-4BL·4thS片段(Rf)的创建
1、含Rf基因染色体端体附加系和含Ba基因的染色体端体附加系的创建。
实施例1中获得的易位系14T35，通过世代选择选育出15L4167两用系(除附加染色体稳定性较差外，其余农艺性状已稳定)。15L4167两用系为含易位染色体T4AgL片段(Ba)-4BL·4thS(Rf)的附加系(两用系)。15L4167两用系的核型为：21Ⅱ(msms)+Ⅰ T4AgL片段(Ba)-4BL·4thS(Rf)。
15L4167两用系自交，然后筛选由蓝粒种子长成的雄性不育植株，即具有易位端体T4AgL片段(Ba)-4BL·的植株。将该植株与深蓝粒可育株(15L4167两用系自交，筛选由深蓝粒种子长成的雄性可育植株，自交后所结种子全为蓝粒种子)杂交，然后筛选由深蓝粒种子长成的雄性可育植株并进行自交，然后从自交种子发育的植株中筛选具有雄性不育性状的植株，进一步通过镜检筛选，获得具有易位端体T4AgL片段(Ba)-4BL·的附加系植株。附加系植株的核型为：21Ⅱ(msms)+Ⅰ T4AgL片段(Ba)-4BL·。附加系的表型为：种子为深蓝粒，植株表现为雄性不育。
15L4167两用系自交，然后筛选由白粒种子长成的雄性可育植株，即具有端体4thS(Rf)·的植株，进一步通过镜检筛选，获得具有端体4thS(Rf)·的附加系植株。附加系植株的核型为：21Ⅱ(msms)+Ⅰ 4thS(Rf)·。附加系的表型为：种子为白粒，植株表现为雄性可育。
2、将具有端体4thS(Rf)·的附加系植株作为父本，具有易位端体T4AgL片段(Ba)-4BL·的附加系植株作为母本，进行杂交，获得500粒以上具有易位端体T4AgL片段(Ba)-4BL·和端体4thS(Rf)·的双单端体种子。双单端体种子的核型为：21Ⅱ(msms)+Ⅰ T4AgL片段(Ba)-4BL·+Ⅰ 4thS(Rf)·。
3、将步骤2得到的种子作父本与作为母本的白粒种子(白粒种子由15L4167两用系自交而得)作为杂交组合，按2行比2-4行相间播种并培育为植株，开花时人工辅助授粉，从母本植株上获得的种子中筛选蓝粒种子。
4、将步骤3得到的蓝粒种子播种并培育为植株，淘汰雄性不育株，收获雄性可育株；对收获的雄性可育株进行室内考种，入选得到的种子既有蓝粒种子又有白粒种子的植株。
5、步骤4共入选1480个单株，按单株分别分拣出白粒种子和蓝粒种子。
6、播种步骤5得到的种子(每个单株播种80粒白粒种子和80粒蓝粒种子)，培育为植株；如果某个单株的蓝粒种子得到的植株全部为雄性可育植株，且该 单株的白粒种子得到的植株全部为雄性不育植株，将该单株的浅蓝粒种子进行镜检，获得具有易位染色体T4AgL片段(Ba)-4BL·4thS片段(Rf)的易位系植株，又称为两用系。
田间选择共入选2个单株，编号分别为15易3357和15易3359，通过镜检，其中15易3357具有易位染色体T4AgL片段(Ba)-4BL·4thS片段(Rf)。
易位染色体T4AgL片段(Ba)-4BL·4thS片段(Rf)的示意图见图3。
将得到的附加系植株进行扩大的群体种植并调查不同单株中的蓝粒群体中雄性不育株和白粒群体中的雄性可育株出现频率。易位染色体T4AgL片段(Ba)-4BL·4thS片段(Rf)从着丝点的断裂频率小于2％，不育系出现可育株的频率为两用系出现不育植株的一半左右(小于1％)。测试群体在1000株以上。
实施例3、易位端体T4AgL片段(Ba)-4thS片段(Rf)·、易位端体T4thS片段(Rf)-4AgL片段(Ba)·或易位端体T4thS片段(Rf)-4AgL片段(Ba)-4BL·的创建
一、易位端体T4AgL片段(Ba)-4thS片段(Rf)·和易位端体T4thS片段(Rf)-4AgL片段(Ba)·的创建
1、获得重双端体附加系。
(1)发明专利(ZL200610042629.8)中培育的两用系的核型为：21Ⅱ(msms)+Ⅰ T4thS·4AgL；即含有易位染色体T4thS·4AgL的附加系小麦(16L6386两用系)。
(2)16L6386两用系自交，收取蓝粒种子，种植并培育为植株，选择其中的雄性不育株，通过镜检筛选具有端体4AgL(Ba)·的植株。
(3)16L6386两用系自交，收取深蓝粒种子，种植并培育为植株，这些植株为雄性可育株。
(4)步骤(2)筛选的植株与步骤(3)获得的植株杂交，筛选深蓝粒种子并培育为植株，从其自交后代中筛选出雄性不育株。上述筛选的深蓝粒种子自交繁殖足够的种子(种子均为深蓝粒)。
(5)16L6386两用系自交，收取白粒种子，种植并培育为植株，选择其中的雄性可育株，通过镜检筛选具有端体4thS(Rf)·的植株。自交繁殖足够的种子备用。
(6)步骤(4)筛选的雄性不育植株与步骤(5)筛选的雄性可育植株杂交，收获种子(均为蓝粒)并培育为植株，然后自交，筛选深蓝粒种子，通过镜检筛选具有端体4AgL(Ba)·和端体4thS(Rf)·的重双端体种子，即为重双端体附加系。重双端体种子的核型为：21Ⅱ(msms)+Ⅱ 4AgL(Ba)·+Ⅱ 4thS(Rf)·。自交繁殖足够的种子(均为深蓝粒)备用。
2、步骤1获得的重双端体种子用二甲亚砜(DMSO)化学诱变剂处理或者用甲基磺酸乙酯(EMS)化学诱变剂处理或辐射处理，然后培育为植株。
3、步骤1获得的重双端体种子长成的植株作为父本，与作为母本的白粒不育植株(由16L6386两用系自交而得的白粒种子长成的雄性不育植株)杂交， 收获母本植株上的种子，选取蓝粒种子用二甲亚砜(DMSO)化学诱变剂处理或者用甲基磺酸乙酯(EMS)化学诱变剂处理或辐射处理，然后培育为植株。
4、将步骤2或步骤3得到植株作为父本，与作为母本的白粒不育植株(由16L6386两用系自交而得的白粒种子长成的雄性不育植株)杂交，收获母本植株上的种子并从中筛选出蓝粒种子。
5、将步骤4得到的蓝粒种子播种并培育为植株，淘汰雄性不育株，共入选26567个单株；将入选的植株自交并收获种子，入选所结种子既有蓝粒种子又有白粒种子的植株。
6、步骤5筛选的植株，分别收集其自交获得的白粒种子和蓝粒种子。
7、播种步骤6得到的种子(每个单株分拣出100粒白粒种子和100粒蓝粒种子)，培育为植株；如果某个单株的蓝粒种子得到的植株全部为雄性可育植株，且该单株的白粒种子全部为雄性不育植株，这些单株为潜在的具有端体的植株，将这些单株的的蓝粒种子进行镜检，入选具有易位端体T4AgL片段(Ba)-4thS片段(Rf)·或易位端体T4thS片段(Rf)-4AgL片段(Ba)·的株系，浅/中蓝粒种子为两用系。共获得9个株系，即20DT1,20DT2,…,20DT9。蓝粒种子得到的植株中雄性不育株为0的株系，即具有易位端体T4AgL片段(Ba)-4thS片段(Rf)·的易位系，为20DT1、20DT3、20DT7、20DT9。白粒种子得到的植株中雄性可育株为0的株系，即具有易位端体T4thS片段(Rf)-4AgL片段(Ba)·的易位系，为20DT2、20DT4、20DT5、20DT6、20DT8。
易位端体T4AgL片段(Ba)-4thS片段(Rf)·的示意图见图4。
易位端体T4thS片段(Rf)-4AgL片段(Ba)·的示意图见图5。
二、易位端体T4thS片段(Rf)-4AgL片段(Ba)-4BL·的创建
1、实施例2中得到的具有易位端体T4AgL片段(Ba)-4BL·和端体4thS片段(Rf)·的重双端体种子繁殖。
2、步骤1获得的重双端体种子用二甲亚砜(DMSO)化学诱变剂处理或者用甲基磺酸乙酯(EMS)化学诱变剂处理或辐射处理，然后培育为植株。
3、步骤1获得的重双端体种子长成的植株作为父本，与作为母本的白粒不育植株(15L4167两用系自交而得的白粒种子长成的雄性不育植株)杂交，收获母本植株上的种子，选取蓝粒种子用二甲亚砜(DMSO)化学诱变剂处理或者用甲基磺酸乙酯(EMS)化学诱变剂处理或辐射处理，然后培育为植株。
4、将步骤2或步骤3得到植株作为父本，与作为母本的白粒不育植株(15L4167两用系自交而得的白粒种子长成的雄性不育植株)杂交，收获母本植株上的种子并从中筛选出蓝粒种子。
5、将步骤4得到的蓝粒种子播种并培育为植株，淘汰雄性不育株，将入选的植株自交并收获种子，入选所结种子既有蓝粒种子又有白粒种子的植株。
6、步骤5筛选的植株，分别收集其自交获得的白粒种子和蓝粒种子。
7、播种步骤6得到的种子(每个单株分拣出100粒白粒种子和100粒蓝粒 种子)，培育为植株；如果某个单株的蓝粒种子得到的植株全部为雄性可育植株，且该单株的白粒种子全部为雄性不育植株，将这些单株中的蓝粒种子进行镜检，入选具有易位端体T4thS片段(Rf)-4AgL片段(Ba)-4BL·的株系，即具有易位端体T4thS片段(Rf)-4AgL片段(Ba)-4BL·的易位系。获得1个易位系，即21DT1。
易位端体T4thS片段(Rf)-4AgL片段(Ba)-4BL·的示意图见图6。
三、易位端体T4AgL片段(Ba)-4thS片段(Rf)·、易位端体T4thS片段(Rf)-4AgL片段(Ba)·或易位端体T4thS片段(Rf)-4AgL片段(Ba)-4BL·的性能
易位端体T4AgL片段(Ba)-4thS片段(Rf)·两个目的基因分离的频率为0-1‰，仅不育系会出现0-0.5‰的可育株，蓝粒群体不会出现不育株。易位端体T4thS片段(Rf)-4AgL片段(Ba)·和易位端体T4thS片段(Rf)-4AgL片段(Ba)-4BL·两个目的基因分开的频率为0-1‰，不育系中不会出现可育株，两用系中会出现0-1‰的不育株。
上述不同两用系自交产生不同颜色的种子比例有差异，白粒或红粒种子(不育系)变幅55-85％、浅/中蓝粒种子(两用系)为15-40％、深蓝粒种子(双端体附加)为2-6％，其中，不育系用于生产杂交种，两用系用于繁殖不育系和两用系，深蓝粒种子淘汰或作亲本或用于两用系提纯复壮。
实施例4、新***两用系(不育系)的选育方法
可采用实施例1至实施例3培育的两用系培育新的两用系，流程示意图见图7。
一、方法一
优良品系与新***的蓝粒两用系或深蓝粒配制杂交组合，在F 1代只选留蓝粒种子种植，经田间表现和室内考种情况，决选出蓝白粒或蓝红粒种子颜色区分明显的优良组合。浅/中蓝粒种子种植，入选蓝白粒或蓝红粒种子颜色区分明显的F 2代优良单株。按单株将蓝粒和白粒或红粒种子分开各自种植82粒种子，每个种2行，2m行长，行距25cm，5cm人工点播。入选白粒或红粒群体中有约25％或100％的不育株且农艺性状好的对应的蓝粒优良单株，直至所选单株的白粒或红粒群体全部不育为止且稳定一致后，获得新两用系(除深蓝粒外)。也可利用本发明设计人所建立的分子检测方法(专利申请号：201810110738.1)对不育基因ms1b进行跟踪监测，有目的的完成在人工控制条件下一年多代的两用系定向快速转育。注意各代入选的单株或株系中的白粒或红粒种子所占比例必须大于或等于60％，同时筛选白/红粒不育株异交结实率高的相应蓝粒单株或株系。
二、方法二
选取不育系与新***的蓝粒两用系或深蓝粒配制杂交组合，除了各代中只种植蓝粒种子外，其余的选育方式与方法一相同。
三、两用系(不育系)分拣和繁育方法
现有的小麦色选机是为加工小麦去除杂质、霉变籽粒等而设计的，色选机生产厂家有中瑞微视、捷讯光电、嘉禾光电、美亚光电等，通过去多家色选机生产厂家调研和尝试，发现中瑞微视生产的色选机比较适合深蓝粒、浅/中蓝粒(两用系)、白粒或红粒种子的分拣。经2-3次连续分选可对蓝、白或红籽粒的分拣精度达到99.9％以上。已购买和使用中瑞微视生产的色选机型号为6SXZ-68。用该色选机完成白粒或红粒、浅/中蓝粒(两用系)和深蓝粒种子的分拣来满足两用系(不育系)的繁殖。浅/中蓝粒种子用于繁殖生产不育系和两用系种子。
四、强优势组合的筛选
流程示意图见图8。
现已利用选育出的多个两用系所产生的白粒不育系分别与适合做恢复系的品种(系)伟隆121、伟隆166、13W8-11、高大1号、山农22、川麦93、14品16、13品6、绵麦827、绵麦903、MY6381、川育36、蜀麦1671、川农39、川辐麦14、绵麦906等不同生态区域的优良品系配制杂交组合，正在不同生态区进行强优势组合筛选和杂种小麦新品种选育。
以上对本发明进行了详述。对于本领域技术人员来说，在不脱离本发明的宗旨和范围，以及无需进行不必要的实验情况下，可在等同参数、浓度和条件下，在较宽范围内实施本发明。虽然本发明给出了特殊的实施例，应该理解为，可以对本发明作进一步的改进。总之，按本发明的原理，本申请欲包括任何变更、用途或对本发明的改进，包括脱离了本申请中已公开范围，而用本领域已知的常规技术进行的改变。按以下附带的权利要求的范围，可以进行一些基本特征的应用。
工业应用
本发明公开了一种蓝标型两系法杂交小麦***的选育方法及应用。本发明的蓝标型两系法杂交小麦***通过两用系自交获得白/红粒(不育系)和蓝粒混合种子，经色选机分离出白/红粒种子(不育系)、浅/中蓝粒种子(两用系)和深蓝粒种子，不育系用于杂交种制种，浅/中蓝粒种子(两用系)用于繁殖不育系和两用系，深蓝粒种子淘汰或用作亲本转育新不育系或用于两用系提纯复壮，本发明具有不育系选育快、杂交种生产成本低、优势强等特点。

Claims (10)

  1. 一种蓝标型两系法杂交小麦的选育方法，包括如下步骤：
    (1)06L6109-3两用系的白粒不育株与具有易位染色体T4AgL(Ba)-4BL·4BS的小麦杂交，筛选由蓝粒种子长成的矮杆且雄性不育的植株，即为具有易位染色体T4AgL(Ba)-4BL·4BS(Rht3-ms1b)的植株，称为T4AgL(Ba)-4BL·4BS(Rht3-ms1b)易位系植株；
    具有易位染色体T4AgL(Ba)-4BL·4BS的小麦由蓝粒种子长成且具有高杆且雄性可育的性状；06L6109-3两用系的白粒不育株：是06L6109-3两用系自交得到的白粒种子长成的矮杆且雄性不育的植株；06L6109-3两用系为由白粒种子长成的具有半矮秆且雄性不育性状的含ms1b-Rht3的小麦与含有易位染色体T4thS·4AgL的附加系小麦杂交并选育而成的具有Rht3-ms1b的蓝粒两用系；
    (2)T4AgL(Ba)-4BL·4BS(Rht3-ms1b)易位系植株与小麦4B代换系4th杂交，获得具有4th和易位染色体T4AgL(Ba)-4BL·4BS(Rht3-ms1b)的双单体种子；
    (3)将步骤(2)获得的双单体种子播种并培育成植株，自交，选深蓝粒种子种植并筛选出高杆且雄性可育的植株；
    (4)将步骤(3)筛选的植株与14L6386两用系的白粒不育株杂交，筛选结有蓝粒种子的杂交组合；
    14L6386两用系的白粒不育株：是14L6386两用系自交得到的白粒种子长成的雄性不育的植株；14L6386两用系为09L6034两用系与糯小麦品系杂交并选育而成的蓝粒两用系；09L6034两用系是T0065-10B-2LB-4两用系与作为轮回亲本的小麦回交后选育而成的蓝粒两用系；T0065-10B-2LB-4为含有易位染色体T4thS·4AgL的附加系小麦；
    (5)根据步骤(4)的筛选结果，将蓝粒种子播种并培育成植株，如果所有种子得到的植株均为雄性可育株，相应的杂交组合的种子发育得到的植株组成一个潜在的易位系；
    (6)步骤(5)得到的潜在的易位系，进行如下操作：自交并收获种子，将种子分为蓝粒种子和白粒种子两组；分别将蓝粒种子和白粒种子播种并培育为植株，如果所有蓝粒种子得到的单株均为雄性可育株且所有白粒种子得到的单株均为雄性不育株，那么将这些由蓝粒种子培育得到的雄性可育单株进行FISH鉴定，筛选具有易位染色体T4AgL片段(Ba)-4BL·4thS(Rf)的单株，这些单株组成T4AgL片段(Ba)-4BL·4thS(Rf)易位系群体；
    (7)从步骤(6)得到的易位系的自交后代中，筛选白粒种子:蓝粒种子的数量为2:1的植株，即为含有易位染色体T4AgL片段(Ba)-4BL·4thS(Rf)的附加系，又称为两用系。
  2. 一种蓝标型两系法杂交小麦的选育方法，包括如下步骤：
    (1)15L4167两用系自交，然后筛选由蓝粒种子长成的雄性不育植株，将 该植株与深蓝粒可育株杂交，然后筛选由深蓝粒种子长成的雄性可育植株并进行自交，然后从自交种子发育的植株中筛选具有雄性不育性状的植株，进一步通过镜检筛选，获得具有易位端体T4AgL片段(Ba)-4BL·的附加系植株；深蓝粒可育株：是15L4167两用系自交得到的深蓝粒种子长成的雄性可育植株；
    (2)15L4167两用系自交，然后筛选由白粒种子长成的雄性可育植株，进一步通过镜检筛选，获得具有端体4thS(Rf)·的附加系植株；
    (3)将具有端体4thS(Rf)·的附加系植株作为父本，具有易位端体T4AgL片段(Ba)-4BL·的附加系植株作为母本，进行杂交，获得具有易位端体T4AgL片段(Ba)-4BL·和端体4thS(Rf)·的双单端体种子；
    (4)将步骤(3)得到的种子作父本与作为母本的白粒种子作为杂交组合，培育为植株并杂交，从母本植株上获得的种子中筛选蓝粒种子；白粒种子由15L4167两用系自交而得；
    (5)将步骤(4)得到的蓝粒种子播种并培育为植株，淘汰雄性不育株，收获雄性可育株并进行考种，入选得到的种子既有蓝粒种子又有白粒种子的植株；
    (6)步骤(5)入选的植株，按单株分别分拣出白粒种子和蓝粒种子；
    (7)播种步骤(6)得到的种子，培育为植株；如果某个单株的蓝粒种子得到的植株全部为雄性可育植株，且该单株的白粒种子得到的植株全部为雄性不育植株，将该单株的浅蓝粒种子进行镜检，获得具有易位染色体T4AgL片段(Ba)-4BL·4thS片段(Rf)的易位系植株，又称为两用系；
    所述15L4167两用系即权利要求1得到的含有易位染色体T4AgL片段(Ba)-4BL·4thS(Rf)的附加系。
  3. 一种蓝标型两系法杂交小麦的选育方法，包括如下步骤：
    (1)16L6386两用系自交，收取蓝粒种子，种植并培育为植株，选择其中的雄性不育株，通过镜检筛选具有端体4AgL(Ba)·的植株；16L6386两用系自交，收取深蓝粒种子，种植并培育为植株，这些植株为雄性可育株；
    (2)步骤(1)筛选的雄性不育株与步骤(1)获得的雄性可育株杂交，筛选深蓝粒种子并培育为植株，从其自交后代中筛选出雄性不育株；
    (3)16L6386两用系自交，收取白粒种子，种植并培育为植株，选择其中的雄性可育株，通过镜检筛选具有端体4thS(Rf)·的植株；
    (4)步骤(2)筛选的雄性不育株与步骤(3)筛选的雄性可育株杂交，收获种子并培育为植株，然后自交，筛选深蓝粒种子，通过镜检筛选具有端体4AgL(Ba)·和端体4thS(Rf)·的重双端体种子；
    (5)步骤(4)获得的重双端体种子进行诱变处理，然后培育为植株；
    (6)步骤(4)获得的重双端体种子长成的植株作为父本，与作为母本的白粒不育植株杂交，收获母本植株上的种子，选取蓝粒种子进行诱变处理，然后培育为植株；白粒不育植株：由16L6386两用系自交而得的白粒种子长成的雄性不育植株；
    (7)将步骤(5)或步骤(6)得到植株作为父本，与作为母本的白粒不育植株杂交，收获母本植株上的种子并从中筛选出蓝粒种子；白粒不育植株：由16L6386两用系自交而得的白粒种子长成的雄性不育植株；
    (8)将步骤(7)得到的蓝粒种子播种并培育为植株，淘汰雄性不育株，将入选的植株自交并收获种子，入选所结种子既有蓝粒种子又有白粒种子的植株；
    (9)步骤(8)筛选的植株，按单株分拣出其自交获得的白粒种子和蓝粒种子；
    (10)播种步骤(9)得到的白粒种子和蓝粒种子，培育为植株；如果某个单株的蓝粒种子得到的植株全部为雄性可育植株，且该单株的白粒种子全部为雄性不育植株，将该单株的的蓝粒种子进行镜检，获得具有易位端体T4AgL片段(Ba)-4thS片段(Rf)·的植株，浅/中蓝粒种子为两用系；
    16L6386两用系即含有易位染色体T4thS·4AgL的附加系小麦。
  4. 一种蓝标型两系法杂交小麦的选育方法，包括如下步骤：
    (1)16L6386两用系自交，收取蓝粒种子，种植并培育为植株，选择其中的雄性不育株，通过镜检筛选具有端体4AgL(Ba)·的植株；16L6386两用系自交，收取深蓝粒种子，种植并培育为植株，这些植株为雄性可育株；
    (2)步骤(1)筛选的雄性不育株与步骤(1)筛选的雄性可育株杂交，筛选深蓝粒种子并培育为植株，从其自交后代中筛选出雄性不育株；
    (3)16L6386两用系自交，收取白粒种子，种植并培育为植株，选择其中的雄性可育株，通过镜检筛选具有端体4thS(Rf)·的植株；
    (4)步骤(2)筛选的雄性不育株与步骤(3)筛选的雄性可育株杂交，收获种子并培育为植株，然后自交，筛选深蓝粒种子，通过镜检筛选具有端体4AgL(Ba)·和端体4thS(Rf)·的重双端体种子；
    (5)步骤(4)获得的重双端体种子进行诱变处理，然后培育为植株；
    (6)步骤(4)获得的重双端体种子长成的植株作为父本，与作为母本的白粒不育植株杂交，收获母本植株上的种子，选取蓝粒种子进行诱变处理，然后培育为植株；白粒不育植株：由16L6386两用系自交而得的白粒种子长成的雄性不育植株；
    (7)将步骤(5)或步骤(6)得到植株作为父本，与作为母本的白粒不育植株杂交，收获母本植株上的种子并从中筛选出蓝粒种子；白粒不育植株：由16L6386两用系自交而得的白粒种子长成的雄性不育植株；
    (8)将步骤(7)得到的蓝粒种子播种并培育为植株，淘汰雄性不育株，将入选的植株自交并收获种子，入选所结种子既有蓝粒种子又有白粒种子的植株；
    (9)步骤(8)筛选的植株，按单株分拣其自交获得的白粒种子和蓝粒种子；
    (10)播种步骤(9)得到的种子，培育为植株；如果某个单株的蓝粒种子得到的植株全部为雄性可育植株，且该单株的白粒种子全部为雄性不育植株，将该单株的的蓝粒种子进行镜检，获得具有易位端体T4thS片段(Rf)-4AgL片段 (Ba)·的植株，浅/中蓝粒种子为两用系；
    16L6386两用系即含有易位染色体T4thS·4AgL的附加系小麦。
  5. 一种蓝标型两系法杂交小麦的选育方法，包括如下步骤：
    (1)将具有易位端体T4AgL片段(Ba)-4BL·和端体4thS(Rf)·的重双端体种子繁殖；
    (2)步骤(1)获得的重双端体种子进行诱变处理，然后培育为植株；
    (3)步骤(1)获得的重双端体种子长成的植株作为父本，与作为母本的白粒不育植株杂交，收获母本植株上的种子，选取蓝粒种子进行诱变处理，然后培育为植株；白粒不育植株：15L4167两用系自交而得的白粒种子长成的雄性不育植株；所述15L4167两用系即权利要求1得到的含有易位染色体T4AgL片段(Ba)-4BL·4thS(Rf)的附加系；
    (4)将步骤(2)或步骤(3)得到植株作为父本，与作为母本的白粒不育植株杂交，收获母本植株上的种子并从中筛选出蓝粒种子；白粒不育植株：15L4167两用系自交而得的白粒种子长成的雄性不育植株；所述15L4167两用系即权利要求1得到的含有易位染色体T4AgL片段(Ba)-4BL·4thS(Rf)的附加系；
    (5)将步骤(4)得到的蓝粒种子播种并培育为植株，淘汰雄性不育株，将入选的植株自交并收获种子，入选所结种子既有蓝粒种子又有白粒种子的植株；
    (6)步骤(5)筛选的植株，按单株分拣出其自交获得的白粒种子和蓝粒种子；
    (7)播种步骤(6)得到的种子，培育为植株；如果某个单株的蓝粒种子得到的植株全部为雄性可育植株，且该单株的白粒种子全部为雄性不育植株，将这些单株中的蓝粒种子进行镜检，获得具有易位端体T4thS片段(Rf)-4AgL片段(Ba)-4BL·的植株，浅/中蓝粒种子即为两用系；
    所述双单端体种子为权利要求2的步骤3获得的具有易位端体T4AgL片段(Ba)-4BL·和端体4thS片段(Rf)·的重双单端体种子。
  6. 一种异位染色体，依次包括如下区段：4AgL片段(Ba)、4BL、着丝粒、4thS(Rf)。
  7. 一种异位染色体，依次包括如下区段：4AgL片段(Ba)、4BL、着丝粒、4thS片段(Rf)。
  8. 一种异位端体，依次包括如下区段：4AgL片段(Ba)、4thS片段(Rf)、着丝粒。
  9. 一种异位端体，依次包括如下区段：4thS片段(Rf)、4AgL片段(Ba)、着丝粒。
  10. 一种异位端体，依次包括如下区段：4thS片段(Rf)、4AgL片段(Ba)-4BL、着丝粒。
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116326480A (zh) * 2023-03-21 2023-06-27 黑龙江省农业科学院作物资源研究所 一种重离子与组织培养相结合进行小麦育种的方法

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113557957B (zh) * 2021-07-28 2023-03-14 西南大学 一种蓝标型两系法杂交小麦***的选育方法及应用

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993013649A1 (en) 1992-01-16 1993-07-22 Pacific Seeds Pty. Ltd. Genetically modified wheat plants and progeny and method for production of hybrid wheat
WO1998051142A1 (en) 1997-05-15 1998-11-19 Yeda Research And Development Co. Ltd. Methods for production of hybrid wheat
WO2002052924A2 (en) * 2001-01-04 2002-07-11 Yeda Research And Development Co. Ltd. Method to maintain a genic male-sterile female parental lines for the production of hybrid wheat
CN1826875A (zh) * 2006-04-06 2006-09-06 西北农林科技大学 一种以蓝粒为标记性状的两系法杂交小麦的选育方法
CN103270935A (zh) * 2013-02-07 2013-09-04 马士芳 一种培育蓝色矮败小麦的方法
CN104719129A (zh) * 2015-03-27 2015-06-24 西南大学 一种小麦新品种的选育方法
CN108496791A (zh) * 2018-03-20 2018-09-07 西南大学 Rht3显性矮秆基因在杂种优势上的利用方法
CN111511199A (zh) * 2017-08-29 2020-08-07 科沃施种子欧洲股份两合公司 改良蓝色糊粉及其他分离***
CN113557957A (zh) * 2021-07-28 2021-10-29 西南大学 一种蓝标型两系法杂交小麦***的选育方法及应用

Patent Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993013649A1 (en) 1992-01-16 1993-07-22 Pacific Seeds Pty. Ltd. Genetically modified wheat plants and progeny and method for production of hybrid wheat
WO1998051142A1 (en) 1997-05-15 1998-11-19 Yeda Research And Development Co. Ltd. Methods for production of hybrid wheat
WO2002052924A2 (en) * 2001-01-04 2002-07-11 Yeda Research And Development Co. Ltd. Method to maintain a genic male-sterile female parental lines for the production of hybrid wheat
CN1826875A (zh) * 2006-04-06 2006-09-06 西北农林科技大学 一种以蓝粒为标记性状的两系法杂交小麦的选育方法
CN100420368C (zh) 2006-04-06 2008-09-24 西北农林科技大学 一种以蓝粒为标记性状的两系法杂交小麦的选育方法
CN103270935A (zh) * 2013-02-07 2013-09-04 马士芳 一种培育蓝色矮败小麦的方法
CN104719129A (zh) * 2015-03-27 2015-06-24 西南大学 一种小麦新品种的选育方法
CN111511199A (zh) * 2017-08-29 2020-08-07 科沃施种子欧洲股份两合公司 改良蓝色糊粉及其他分离***
CN108496791A (zh) * 2018-03-20 2018-09-07 西南大学 Rht3显性矮秆基因在杂种优势上的利用方法
CN113557957A (zh) * 2021-07-28 2021-10-29 西南大学 一种蓝标型两系法杂交小麦***的选育方法及应用

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
BUREŠOVÁ VERONIKA; KOPECKÝ DAVID; BARTOŠ JAN; MARTINEK PETR; WATANABE NOBUYOSHI; VYHNÁNEK TOMÁŠ; DOLE: "Variation in genome composition of blue-aleurone wheat", THEORETICAL AND APPLIED GENETICS, vol. 128, no. 2, 16 November 2014 (2014-11-16), Berlin/Heidelberg, pages 273 - 282, XP035434255, ISSN: 0040-5752, DOI: 10.1007/s00122-014-2427-3 *
LI ZHONG AN , ZHANG JIE: "Selection and Identification of Wheat Alien Translocation Chromosome 4BS·4AgL", JOURNAL OF XI'AN UNITED UNIVERSITY, vol. 4, no. 2, 25 May 2001 (2001-05-25), pages 16 - 21, XP093028268, ISSN: 1008-777x *
XING CUI-PING,AN LIN-LI,HAN DONG-CUI,SHI MIN-FANG: "Study on Blue Seed Dual-purpose Wheat", SHAANXI JOURNAL OF AGRICULTURAL SCIENCES, vol. 37, no. 3, 20 March 2009 (2009-03-20), pages 33 - 35, XP093028261, ISSN: 1002-2481 *
ZHENG, Q. ; LI, B. ; LI, H. ; LI, Z.: "Utilization of blue-grained character in wheat breeding derived from Thinopyrum poticum", JOURNAL OF GENETICS AND GENOMICS, vol. 36, no. 9, 1 September 2009 (2009-09-01), AMSTERDAM, NL , pages 575 - 580, XP026644220, ISSN: 1673-8527, DOI: 10.1016/S1673-8527(08)60149-6 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116326480A (zh) * 2023-03-21 2023-06-27 黑龙江省农业科学院作物资源研究所 一种重离子与组织培养相结合进行小麦育种的方法

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