CN111903505B - Replacement method of cytoplasm of blue dwarf male sterile line - Google Patents

Replacement method of cytoplasm of blue dwarf male sterile line Download PDF

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CN111903505B
CN111903505B CN202010800904.8A CN202010800904A CN111903505B CN 111903505 B CN111903505 B CN 111903505B CN 202010800904 A CN202010800904 A CN 202010800904A CN 111903505 B CN111903505 B CN 111903505B
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germplasm
blue
sterile
male
cytoplasm
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CN111903505A (en
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耿爱民
于聪
樊庆琦
王京京
刘玉东
徐学亮
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Zibo Aimin Seed Co ltd
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/02Methods or apparatus for hybridisation; Artificial pollination ; Fertility
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/04Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection

Abstract

The invention discloses a replacement method of cytoplasm of a blue dwarf male sterile line, belonging to the field of wheat breeding. The invention obtains the blue dwarf through autonomous breedingFailure Ms2The germplasm material with the characteristics having the complete inhibition function can ensure that blue grains on the blue dwarf-male-sterile plants are all fertile, and completely breaks through the current situation that the blue grains formed by the existing blue dwarf-male-sterile plants are not fertile; and the germplasm is used as the middle bridge germplasm, so that the cytoplasm replacement of the blue dwarf male-sterile line is realized, and the difficult problem of the blue dwarf male-sterile cytoplasm replacement is solved.

Description

Replacement method of cytoplasm of blue dwarf male sterile line
Technical Field
The invention relates to the field of wheat breeding, in particular to a replacement method of cytoplasm of a blue dwarf male sterile line.
Background
The blue dwarf male sterile line is a novel wheat dominant genic male sterile line, and the male sterile line is used as a translocation addition line monomer and comprises blue grains (4D/4E) and short stems (Rht)10) Anther abortion (Ms)2) Three dominant characters are linked and locked at 4D#The genetic behavior of the dominant genic male sterile line is in accordance with the genetic rule of the monomer on the monomer addition line of the chromosome, namely the blue dwarf male sterile line is a dominant genic male sterile line with blue grains, short stems and two dominant character markers. The sterile line is successively researched by Tianning, Pu zong monarch, Gunn people and other teams, so that a plurality of advantages of the sterile line in the aspect of wheat heterosis utilization are fully displayed, and hope is brought to the wheat heterosis utilization.
Sterile gene Ms for blue dwarf male sterile2The gene germplasm has the advantages of thorough anther abortion, large glume opening angle and the like. The study time of blue dwarfing and breeding is relatively short, and people are always looking for Ms from blue dwarfing and breeding2The suppressor gene of (2) can realize cytoplasm replacement of dominant genic male sterile line, overcome many adverse effects caused by single cytoplasm, and create conditions for utilization of double heterosis (heterosis between different cytoplasm and nucleus, and between nucleus and nucleus), thereby providing possibility.
Disclosure of Invention
In order to make up for the defects of the prior art, the invention provides.
The technical scheme of the invention is as follows:
a method for replacing cytoplasm of blue dwarf male sterile line comprises the following steps:
a selective breeding pair blue dwarf-male-sterile Ms2Germplasm material with complete inhibition of character
A1) Selecting germplasm materials with high ratio of blue grain fertile plants formed by the test blue dwarf-male-sterile pollination in various wheat germplasm resources to obtain germplasm A, germplasm B, germplasm C and germplasm D;
A2) single crossing between germplasm A and germplasm B, single crossing between germplasm C and germplasm D, and then double crossing;
A3) continuously selfing the backcross progeny, firstly carrying out mixed selection and then carrying out systematic breeding to obtain a single plant which has a complete inhibition effect on the blue dwarf male sterile;
A4) retaining single plant progeny with complete inhibition effect on blue dwarf male sterile, and selecting for several successive generations to breed stable blue dwarf male sterile Ms2Germplasm with complete inhibition effect on trait expression;
b, performing cytoplasm replacement on the blue dwarf male sterile by using the germplasm bred in the step A
B1) B, pollinating and hybridizing the blue dwarf male sterile with the cytoplasm A by using the germplasm bred in the step A, and selecting blue seeds with all fertile generations;
B2) the pollen of blue-grain plant which becomes self-fertile is used, the wheat germ plasm containing B cytoplasm is used as female parent, the wheat germ plasm containing B cytoplasm is pollinated by using artificial emasculation method, the blue grains therein are continuously sorted for planting, and the blue dwarf-male-sterile germ plasm which is newly selected from the progeny segregation population is the blue dwarf-male-sterile germ plasm containing B cytoplasm.
Preferably, the germplasm materials obtained by screening in the step A1) are all parent materials containing exogenous blood sources.
Preferably, in the step A1), the germplasm A is Pricel 143, the germplasm B is Pricel 4094, the germplasm C is Tianchou 46 and the germplasm D is Elytrigia 503.
Further, in step A4), pairs Ms are retained2The single plant progeny with complete inhibition effect on character expression is bred for several generations through continuous system breeding to obtain stable Ms for blue dwarf2The character expression has the advantages ofGermplasm material Ms with full inhibition effect2-R101。
Preferably, in step B1), the blue dwarf male-sterile with formazan cytoplasm is blue dwarf male-sterile black horse No. 1. Of course, any other stable transformation blue dwarf or abortive can be used.
Preferably, in step B2), the female parent is aegilops tauschii D2Chenchenge number 4 for the type cytoplasm. Other germplasm not having the same cytoplasm as proto-blue dwarf could also be used.
Step A breeding blue dwarf-male-sterile Ms2Germplasm material (Ms) with complete inhibition of character2-R101) rendering blue dwarf-male-sterile monosdominant genic male sterile gene Ms2The expression is inhibited, and the breeding can inhibit the blue dwarf male sterile single dominant genic male sterile gene Ms2The expressed wheat germ plasm resource is obtained by widely screening various germ plasms which are suspected to contain pairs of Ms2The character expression has inhibition or dominant non-allele, and the four germplasm resources are the germplasm resources containing partial exogenous blood source.
Then use Ms2The R101 germplasm is blue dwarf male-sterile pollinated hybrid with first cytoplasm, about 72 percent of blue dwarf male-sterile seeds are white grains and about 28 percent of blue grains, the blue grains are planted and sorted, the blue grains are all fertile, but the blue grains and the white grains are still remained in the same plants of the blue fertile plants, then the blue grains are sorted and planted, the blue grain plant pollen is used for manually emasculating and pollinating common white grain wheat varieties (second cytoplasm) with cytoplasm different from the original blue dwarf male-sterile cytoplasm, the harvested hybrid seeds with the displaced cytoplasm are further planted, the blue grains and the white grains are still grown on the plants of self-bred offspring, only the blue grains are sorted and planted in each generation, the blue grain dwarf sterile plant is reselected in the separated offspring group, namely the blue dwarf male-sterile plant with the displaced cytoplasm is blue dwarf male-sterile, and the cytoplasm of the blue dwarf male-sterile plant is already displaced from the first cytoplasm to the second cytoplasm. Simultaneous discovery of Ms2-R101 has double effect and can inhibit Ms simultaneously2And Rht10Trait expression of two genes, for Ms2Has complete inhibitory effect on Rht10Has partial inhibiting effect.
The beneficial effects of the invention are as follows:
the method obtains the blue dwarf-male-sterile Ms by autonomous breeding2Germplasm material (Ms) with complete inhibition of character2R101), the germplasm can ensure that blue grains formed on the blue dwarf-male-sterile plants can be all fertile, and completely breaks through the current situation that the blue grains formed by the existing blue dwarf-male-sterile plants cannot be fertile; and the germplasm is used as the intermediate bridge germplasm, so that the cytoplasm replacement of the blue dwarf male-sterile line is realized, and the difficult problem of the blue dwarf male-sterile line cytoplasm replacement is solved.
Detailed Description
The present invention will be further explained with reference to the following embodiments.
The replacement method of the cytoplasm of the blue dwarf male-sterile line comprises the following steps:
one, the screening may contain the energy pairs Ms2Germplasm material with gene trait expression inhibiting effect (i.e. germplasm material capable of containing non-allelic inhibitor gene or dominant episome gene)
1. Widely collecting a large amount of various types of wheat germplasm resources, particularly focusing on collecting various types of wheat germplasm resources containing exogenous genetic materials (a heterotransposition line, a heterotopic line, an outer edge osmotic line and a heterotransition line);
2. constructing a hybridization identification nursery:
selecting any several (or one) stable blue dwarf-male-sterile backcross populations as test materials, respectively sowing collected wheat varieties beside the blue dwarf-male-sterile populations as male parents by using a proper blue dwarf-male-sterile population scale (usually two rows of seeding are reserved, a row of male parent seeding positions are reserved, and the walk positions between about 1 m and each isolation region are reserved), building the isolation regions by using bamboo supports, iron wires, plastic woven cloth and other materials before ear-picking, and building a blue dwarf-sterile isolation region for each material, thereby building the blue dwarf-sterile hybridization identification nursery. Naturally pollinating blue dwarf male sterile plants by a male parent in each isolation area, timely pulling out fertile plants (blue-cultivated plants) appearing in blue dwarf male sterile lines in the heading period, sorting blue grains from seeds (grains with blue and white colors of the same plant) on the blue dwarf male sterile plants according to grain colors, independently harvesting and threshing each isolation area, respectively sorting according to grain colors (counting the blue grain proportion), and respectively and independently bagging and numbering;
3. establishing a screening and identifying garden:
the method comprises the steps of uniformly and respectively numbering the sorted blue seeds after the blue dwarf-male-sterile pollination is fruited by each variety, respectively sowing the blue seeds in sequence in autumn, establishing respective isolation regions, generally sowing 2-3 rows of the sorted blue seeds, sowing 1 row of male parents (backcrossing with the same variety in the last year) beside the blue seeds, reserving walkways among the isolation regions, and arranging at least 3-5 backcrossing cells of the blue dwarf-male-sterile original variety in the whole screening and identifying nursery for comparison. Before heading, isolation areas are built by using materials such as bamboo supports, iron wires, plastic woven cloth and the like, pollination is carried out naturally, after the isolation areas are mature, the isolation areas are collected and placed separately, threshing is carried out separately, blue grains and white grains are sorted according to grain colors, and the ratio of each blue grain to the white grain is counted.
4. Analysis, evaluation and screening:
observing and recording the quantity and percentage of fertile and sterile plants (or single ears) of blue seed plants sowed in each isolation region in heading stage of the population, screening male parent variety materials with high ratio of fertile (selfing and fructification) plants in the isolation region by comparing with a control (blue dwarf male sterile original variety uniform backcross cell), analyzing and screening the male parent variety materials which may contain the Ms inhibitor2Germplasm material for character expression or fertility restorer material with dominant superior relation.
Second, the hybrid polymerization suppressor gene (or dominant episomal gene) is synchronously identified and tested for fertility restoration inhibition effect, and the breeding is carried out on blue dwarf-male-sterile Ms2Germplasm material with complete inhibition of character
Selecting different varieties with high ratio of the screened fertile plants, and combining the varieties into a hybrid combination. Will consider possible to contain pairs Ms2The germplasm materials with the character expression inhibiting effect (or dominant epistatic effect) are combined pairwise. Artificial emasculation, artificial pollination and hybridization are adopted, then repeated hybridization is carried out, each subsequent generation is selfed, mixed selection is firstly carried out to expand a basic population, and then systematic breeding is carried out. From the beginning of selecting single plant, all generations are used as pollination male parent material of a selected blue dwarf male-sterile (any blue dwarf male-sterile) to pollinate the blue dwarf male-sterile, all generations of materials used as pollination male parents are still independently harvested and utilized,and continuing to perform subsequent breeding according to a pedigree seed selection procedure, sorting blue grains from the pollinated and fructified blue dwarf male sterile seeds, continuing to plant, observe and record the quantity and proportion of fertile and sterile plants, and comparing the quantity and proportion with the selected blue dwarf male sterile backcross control population, and finally breeding into a germplasm material capable of enabling the blue dwarf male sterile seeds to be all fertile.
Thirdly, the cytoplasm replacement is carried out on the blue dwarf male sterile by using the germplasm material which is bred to ensure that the blue dwarf male sterile seeds can be all bred
Pollinating and hybridizing the blue dwarf male sterile with cytoplasm A by using the germplasm material which is bred to ensure that the blue white seed produced by the blue dwarf male sterile is all fertile, and sorting out the blue seed with all fertile generations;
the pollen of the blue-grain plant which becomes self-fertile is utilized, the wheat germplasm containing the B cytoplasm is used as a female parent, the wheat germplasm containing the B cytoplasm is pollinated by an artificial emasculation method, the blue grains in the wheat germplasm are continuously sorted for planting, and the newly selected blue dwarf male-sterile germplasm in the progeny segregation population is the blue dwarf male-sterile germplasm containing the B cytoplasm.
Example 1
The method for replacing the cytoplasm of the blue dwarf male sterile line comprises the following steps:
screening of blue dwarf and male-sterile Ms2Germplasm material with inhibiting property
Screening according to the above may contain pairs of Ms2The process of the germplasm materials with the inhibiting effect on the gene character expression takes the Cimin blue dwarf-male-sterile 66 (also can be any one or more other stable blue dwarf-male-sterile) as the test cross blue dwarf-male-sterile, four germplasm materials with the highest ratio of blue-grain fertile plants formed by pollination of blue dwarf-male-sterile are screened from the germplasm resources by planting and screening various massive germplasm resources, namely germplasm A, germplasm B, germplasm C and germplasm D; the four germplasm materials obtained by screening are all discovered to be parent materials containing partial exogenous blood sources.
Screening out:
the germplasm A is Pubing 143, examined and determined variety in Shaanxi province;
germplasm B is a clinical 4094, selected breeding line of wheat in Shanxi province;
germplasm C is selected from 46 days, and examined and determined by Gansu province;
the germplasm D is Xiao elytrigia 503, examined and identified variety in Shaanxi province.
Second, the hybrid polymerization suppressor gene (or dominant episomal gene) is synchronously identified and tested for fertility restoration inhibition effect, and the breeding is carried out on blue dwarf-male-sterile Ms2Germplasm material Ms with complete inhibition of characters2-R101
2.1, crossing germplasm A with germplasm B (A/B), crossing germplasm C with germplasm D (C/D), and then crossing again (A/B)// (C/D);
2.2, continuously selfing the backcross progeny, selecting only a few poor individuals in the first two generations according to a mixed seed selection program, mixedly harvesting and expanding the rest of individuals, expanding a seed selection basic group, selecting individual plants from the selfing separation group according to a system seed selection program from the third generation after backcross, selecting individual plants from the selected individual plants to be independently planted (and used as male parents), synchronously pollinating the Aimin blue dwarf and sterile 66, respectively establishing respective isolation regions, counting the proportion of blue fertile plants in the next generation, comparing the proportion with the proportion of blue fertile plants in the backcross (Liang star 66) control group of the Aimin blue dwarf and sterile 66, and selecting and reserving a plant line with high proportion of fertile plants, wherein the next generation tests show that the Ms of the selected individual plants are selected in the current generation2The subsequent breeding adopts a systematic breeding method, only selfing progeny of the single plant are tested and selected, single plant progeny with complete inhibition on blue dwarf fertility and excellent comprehensive characters are reserved, other all single plants are eliminated, and after continuous 4-generation selection, the germplasm material Ms which is stable and has inhibition on blue dwarf fertility is bred2-R101 and a plurality of sister series materials.
By Ms2R101 is blue dwarf male sterile pollination and can also completely inhibit the expression of the sterile character of blue dwarf male sterile.
Ms2-R101 germplasm, a hexaploid common wheat variety carrying partial exogenic chromosome fragments or genes, still having the AABBDD genome and having the following basic characteristics: the plant height of the awn glume is about 75 cm, and blue dwarf pollination can ensure that blue is whiteThe seeds (white grains and blue grains) grown on the dwarf-male-sterile plants can be fully cultivated in the next generation.
Wherein the Liangxing 66 is a certified variety of Shandong province,
the Aimin blue dwarf male-sterile 66 is winter blue dwarf male-sterile germplasm formed by self-transfer of Zibo Aimin breed limited company, and is formed by utilizing a good star 66 of a certified variety in Shandong province as a recurrent male parent, carrying out continuous backcross on blue dwarf male-sterile 97-866 (note: the fact that the applicant has a condition of obtaining the germplasm under a cooperative agreement with the national ministry of the national city of Yi nationality of Sichuan Liang mountain, the germplasm is called 'wheat dwarf male-sterile blue standard sterile line', published and published in various publications) and carrying out breeding transfer (obtaining compensation) in the process of genetic background winterization.
III, Ms2Cytoplasmic displacement of blue dwarf by-R101 (or its sister line) germplasm material
By Ms2-R101 material is blue dwarf male sterile pollinated hybrid: 4 rows of blue dwarf male-sterile black horses are sown in autumn, each two rows are one group, the row spacing of the 2 rows in the group is 20 centimeters, 25 grains/meter, the row length is 2 meters, the two groups are spaced by 40 centimeters (the position of a reserved partition wall), and the two groups are arranged side by side. Sowing Ms on one side of blue dwarf male-sterile black horse No. 12-R101 as male parent, 50 grains/m. The other side still uses backcross male parent black horse No. 1 with blue dwarf male as a male parent (contrast), 50 grains/meter and small walkways are arranged around the male parent black horse. Before heading, bamboo-wood piles, iron wires and plastic woven cloth are used for building a partition wall to form two isolation areas. Timely pulling out fertile plants separated from the blue dwarf male sterile, and carrying out natural pollination or shifting wheat plants by using a small bamboo rod for artificial supplementary pollination. After the mature, the blue dwarf male sterile is obtained in the two isolated districts, and the blue and white grains are sorted according to grain color.
Observing and comparing the fertility changes of the two male parent groups to breed blue-grain plants: and (3) sorting blue grains and white grains of seeds from blue dwarf aborts harvested from two isolated cells, and sowing the seeds respectively, wherein one section is sown in each step. Last year Ms2R101 as male parent material, which is sorted by sowing 2 rows of blue grains, with a row spacing of 20 cm, a row length of 2 m, 20 grains/m, and sowing Ms on one side thereof2-R101 is prepared as a backcross male parent, white grains which are sorted in 3 rows are sowed on the other side in parallel, the spacing between the white grains and blue grains is 40 centimeters (reserved as a building partition wall), and 1 meter of small walkway is reserved on the periphery. Spacing 50 cm in the forward direction, and pressingThe same field layout and method are adopted to sow contrast blue dwarf-male-sterile black horse No. 1 and backcross male parent black horse No. 1 and white grains after sorting. Before heading, 4 isolation areas are built by building isolation walls with bamboo-wood piles, iron wires and plastic woven cloth. The obvious difference of the two treatments can be obviously seen from the period from the node extraction to the flag picking period: the basic characteristics of normal blue dwarf male sterility are still maintained in the first-stage contrast of pollination black horse No. 1 in the last year, all the white grain sorting isolation regions can be selfed and fructified, the blue dwarf male sterility in the blue grain sorting isolation regions is still dwarf and sterile, and the backcrossed male parent black horse No. 1 is used for natural pollination; last year pollination Ms2All white grains sorted by R101 can be selfed and fruited, but the plant height is basically normal in the sorted blue grain isolation area, all the white grains have anthers, and the white grains can be selfed and fruited normally (the inhibition of MS is shown)2The existence of the gene expressed by the sterile character realizes fertility restoration and simultaneously shows that Rht is inhibited10Expression of characters), but the seeds still bear blue and white seeds from the same strain, and the blue seeds (code number DY 104) are sorted for standby.
Cytoplasm replacement: sowing artificial hybrid parent garden in autumn, planting D-containing aegilops tauschii210 rows of chenchenmai No. 4 as parent material, 25 cm row spacing, 2 m row length, 20 grains/m; 20 rows of the sorted blue granules DY104 are planted side by side, 10 rows are planted on each side of the blue granules DY104 as a spike pollinated variety, the row length is 2 m, the row spacing is 25 cm, and the number of the blue granules DY is 30 granules/m. 1 m of field walkway is respectively arranged at both ends. In the early stage of ear pulling, the ears of the Chinese goosebeery No. 4 are selected and manually castrated and sleeved with bags, 50 ears are manually castrated and sleeved with bags, the blue-grain male parent variety DY104 arranged in the flowering period is cut and selected to carry out ear twisting and pollination for the Chinese goosebeery No. 4 which is manually castrated, in order to improve the fructification and repeat pollination, harvesting is carried out in time in the mature period, threshing and preserving are carried out, and autumn sowing is waited.
In autumn, about 600 hybrid seeds obtained by single-seed dibbling and artificial emasculation hybridization are sown, 20 hybrid seeds are sown in a 6-row district with the row length of 5 m, the hybrid seeds are harvested in the whole area of the maturation period, blue and white grains exist on the harvested wheat ears, the blue grains and the white grains are sorted according to the grain colors, and the blue grains are selected and remained for continuous planting.
In autumn, the blue grains sorted and retained are all single-grain dibbled with 20 grains/m, the line spacing adopts 30 cm large line spacing (convenient for observation and discovery of separated blue dwarf and abortive), and in the communityTwo rows of conventional variety wheat 22 were sown on each side (reserved for the discovered blue dwarf pollination). After wheat nodes are pulled out and before ear emergence, the discovered short stalk plants are marked (small branches are inserted), the adjacent high stalk plants are pulled out, a growing space is reserved for growing nodes, the separated blue dwarf-male-sterile whole plants are bagged, ears are taken from the sowed Jimai 22 for pollination in the flowering period, and the wheat is marked by listing. Harvesting at maturity the separated blue dwarf-male-sterile, sorted blue grains (blue dwarf-male-sterile), which have been replaced by the sequence of the order of the Chinese Caterpillar fungus No. 42Blue stunt of type cytoplasm. And subsequently, keeping or transferring the blue dwarf male sterile by using common white-grain varieties such as the Jimai 22 and the like for blue dwarf male sterile pollination, wherein the blue dwarf male sterile is replaced by cytoplasm.
The germplasm of wheat used in the cytoplasm replacement step:
jimai 22 is a approved variety in Shandong province;
the serial No. 4 of the Chinese patent specification is examined and classified in Hebei province;
no. 1 of the black horse is a certified variety of Shandong province;
blue dwarf male-sterile black horse No. 1 (germplasm is disclosed in publication Shandong agricultural science); the germplasm used in the method is Zibo lovemin germplasm Limited company, which utilizes the Black horse No. 1 as a recurrent male parent and continuously backcrosses blue dwarf-rancidity 97-866 (note: the germplasm is obtained conditionally by a cooperative agreement between the applicant and the agricultural departments of autonomous State West Chang of Yi nationality in Liangshan, Sichuan, is called a wheat dwarf-rancid blue standard sterile line and published and disclosed in various publications) and winter blue dwarf-rancidity germplasm (which can be obtained for compensation) bred in the winterization process of a genetic background.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; such modifications and substitutions do not depart from the spirit and scope of the present invention, and they should be construed as being included in the following claims and description.

Claims (3)

1. A method for replacing cytoplasm of a blue dwarf male sterile line is characterized by comprising the following steps: the method comprises the following steps:
a selective breeding pair blue dwarf-male-sterile Ms2Germplasm material with complete inhibition of character
A1) Selecting germplasm materials with high ratio of blue grain fertile plants formed by the test blue dwarf-male-sterile pollination in various wheat germplasm resources to obtain germplasm A, germplasm B, germplasm C and germplasm D;
A2) single crossing between germplasm A and germplasm B, single crossing between germplasm C and germplasm D, and then double crossing;
A3) continuously selfing the backcross progeny, firstly carrying out mixed selection and then carrying out systematic breeding to obtain a single plant which has a complete inhibition effect on the blue dwarf male sterile;
A4) retaining single plant progeny with complete inhibition effect on blue dwarf male sterile, and selecting for several successive generations to breed stable blue dwarf male sterile Ms2Germplasm with complete inhibition effect on trait expression;
b, performing cytoplasm replacement on the blue dwarf male sterile by using the germplasm bred in the step A
B1) B, pollinating and hybridizing the blue dwarf male sterile with the cytoplasm A by using the germplasm bred in the step A, and selecting blue seeds with all fertile generations;
B2) the pollen of the blue-grain plant which becomes self-fertile is utilized, the wheat germplasm containing the B cytoplasm is used as a female parent, the wheat germplasm containing the B cytoplasm is pollinated by an artificial emasculation method, the blue grains in the wheat germplasm are continuously sorted for planting, and the newly selected blue dwarf male-sterile germplasm in the progeny segregation population is the blue dwarf male-sterile germplasm containing the B cytoplasm;
in the step A1), the germplasm A is Pricel 143, the germplasm B is Ningyuan 4094, the germplasm C is Tianchong 46 and the germplasm D is Xiaoyan 503;
step B1), the blue dwarf-male-sterile with the formazan cytoplasm is blue dwarf-male-sterile black horse No. 1;
in the step B2), the female parent is the female parent containing aegilops tauschii D2Chenchenge number 4 for the type cytoplasm.
2. The method for replacing cytoplasm of blue dwarf male sterile line according to claim 1, wherein the method comprises the following steps: the germplasm materials obtained by screening in the step A1) are all parent materials containing exogenous blood sources.
3. The method for replacing cytoplasm of blue dwarf male sterile line according to claim 1, wherein the method comprises the following steps: in step A4), the pairs Ms are retained2The single plant progeny with complete inhibition effect on character expression is bred for several generations through continuous system breeding to obtain stable Ms for blue dwarf2Germplasm material Ms with complete inhibition effect on character expression2-R101。
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