CN105660361A - Breeding method of rice common genic male sterile line - Google Patents

Breeding method of rice common genic male sterile line Download PDF

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Publication number
CN105660361A
CN105660361A CN201610012557.6A CN201610012557A CN105660361A CN 105660361 A CN105660361 A CN 105660361A CN 201610012557 A CN201610012557 A CN 201610012557A CN 105660361 A CN105660361 A CN 105660361A
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molecular marker
strain
selection
type
sterile
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李莉
李新奇
袁定阳
邝翡婷
宋书锋
王天抗
李懿星
史江伟
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Hunan Hybrid Rice Research Center
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Hunan Hybrid Rice Research Center
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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
    • 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

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  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
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  • Developmental Biology & Embryology (AREA)
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  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention relates to a breeding method of a rice common genic male sterile line. The method comprises the steps of: 1) conducting molecular marker development on common genic male sterile gene A in a common genic male sterile mutant to obtain a molecular marker S of A and a corresponding fertile gene molecular marker R; 2) taking a common genic male sterile mutant as a female parent, adopting rice with a molecular marker of S/R or R/R rice as a recurrent parent, i.e. a male parent, and conducting hybridization to obtain F1; 3) in F1, selecting S/R as the foreground, selecting a partial recurrent parent molecular marker type whole-genome marker, and performing multi-generation backcross to obtain BCnF1, wherein n is greater than or equal to 2; 4) in BCnF1, selecting S/R as the foreground, and performing inbreeding to obtain BCnF2; 5) in BCnF2, selecting S/S as the foreground, breeding a rice sterile line with A as the sterile gene source, and carrying out multi-generation inbreeding to obtain BCnFm, wherein m is greater than or equal to 3; and 6) carrying out sterile line identification on BCnFm to obtain a common genic male sterile line according with identification standards. The method provided by the invention has the advantages of wide sterile gene source, simple breeding procedure, short period and simple molecular marker development process.

Description

A kind of selection of the common line with genic sterile of Oryza sativa L.
Technical field
The present invention relates to the selection of the common line with genic sterile of a kind of Oryza sativa L., particularly relate to a kind of sterile gene wide material sources, selection-breeding program is simple, the cycle is shorter and the selection of the common line with genic sterile of the simple Oryza sativa L. of molecular markers development process, belong to heterosis utilization field, particularly in Rice intersubspecific hybrid use of advantage field.
Background technology
Hybrid rice selects two to have different in heredity exactly, the rice varieties that their merit again can be complementary simultaneously, hybridizes, produces and have heterotic first generation cenospecies, be used for producing. Oryza sativa L. has long-grained nonglutinous rice and two subspecies of japonica rice (including java rice), the Yang Shouren research report of China, there is between indica and japonica subspecies stronger hybrid vigor, it is mainly manifested in plant tall and big, big panicle many grains per panicle, germination percentage is high, tiller gesture is strong, stem stalk is sturdy anti-fall, well developed root system, the feature that regeneration premunition is strong. The basic reason that the strong advantage of inter subspecific hybrid produces: the hereditary difference of Hybrids of Indica and Japonica combination is bigger.
The research of hybrid rice has three important concepts:
Male sterility line: be that a kind of male organs grow imperfection, it is impossible to forming normal pollen, its female organ is grown normal. Thus can not self-reproduction, it is necessary to seed just can be born by means of external paddy pollen. Therefore, by this female Oryza sativa L. as genetic tool, by the way of artificial supplementary pollination, hybrid seed can just be produced in a large number.
Keep system: being a kind of normal rice varieties, its specific function is the sterility that can keep corresponding sterile line, award after sterile line with its pollen, produced offspring, remain male sterile. Therefore, by keeping system, sterile line just can be bred down from generation to generation.
Restorer: be a kind of normal rice varieties, its specific function is to award the male recovery of cenospecies produced by sterile line normally with its pollen, and energy self-fertility, if this cenospecies has superiority, so that it may be used for producing.
Wherein, sterile line provides probability for producing a large amount of hybrid seeds, simultaneously by keeping system to carry out breeding male sterile lines, and utilizes restorer to produce male recovery and the hybrid paddy rice having superiority to sterile line pollination.
And japonica Cross sterile line breeding method is usually in prior art: utilize wide compatibility gene, Xian round-grained rice is built bridge, and cultivates Xian round-grained rice middle type and keeps system, then by sterile gene transformation to maintenance system, is bred as and keeps system.
Such as, the selection-breeding of training C311 (round-grained rice type) and samsara 422 (Java type), its technology path is shown in shown in Figure of description 2.
Again such as, the selection-breeding of indica type Peiai 64 and Peiai 64S, its technology path is shown in shown in Figure of description 3.
The gene of objective trait also would generally be selected by paddy rice cross breeding sterile line breeding method by molecular marking technique. Molecular marker compared with traditional Phenotypic Selection, the advantage mainly having the following aspects: (1) on the selection of objective trait by the impact of gene expression and environmental condition. (2) can select in any stage of generation morning and plant strain growth, thus shortening breeding cycle. (3) segregating generation can identify the genotype of plant rapidly and accurately. (4) can chain efficiently against target gene and unfavorable gene. (5) pyramiding breeding is carried out rapidly and accurately. In crop breeding, mainly application molecular marker assisted selection carries out foreground selection and Foreground selection, and its reliability depends primarily on tightness degree chain between labelling and target gene. If only target gene being selected with a labelling, then chain between labelling and target gene closely, must can reach significantly high accuracy. If be tracked selecting to target gene with two labellings that both sides are adjacent simultaneously, it is greatly improved the accuracy of selection.
At present, mainly utilizing molecular marker labelling wide compatibility gene S 5 n and Restore gene Rf (6), Rf (4), Rf (3), its technology path is shown in shown in Figure of description 4.
More than comprehensive, the selection of existing rice sterile line there is problems in that
(1) program is complicated, excessive cycle. Owing to Asia rice-cultivating indica-japonica hybrid exists sterile phenomenon, therefore when sterile line handed over by selection-breeding Xian round-grained rice, phenotype be difficult to distinguish be because Xian round-grained rice hand over cause sterile, or it is sterile that sterile gene causes, therefore, the breeding route that Xian round-grained rice is handed over is all from educate after the maintenance system that can educate, then import sterile gene, transformation becomes sterile line. The method, program is complicated, and the cycle is extremely long, at least needs the more than ten years could be bred as a Xian round-grained rice and hands over sterile line.
(2) sterile gene source is single. The sterile genes such as the sterile gene of the sterile line of hybridized rice of current main-stream loses essentially from open country, red lotus A, Nongken58s, peace agriculture S, sterile gene source is relatively easy, limits the degree of hybrid rice heterosis utilization largely. And the molecule mechanism relative complex of these genes, molecular markers development process relative complex.
(3) common Genetic Sterility exists extensively at nature, and abundance is controlled its fertility by Recessive genes, and mechanism is relatively easy, and molecular markers development process is simple, but is never utilized aborning.
CN103477971B discloses the selection of a kind of rice sterile line, with high-yield disease resisting sterile line land 18S for female parent, with high-quality and there is the precocious power of heredity extensively to account for by force 63-4S for paternal hybrid, selects two-line sterile line.
CN103947541A discloses the selection of a kind of cytoplasmic male sterile rice. The method includes: with Dongxiang wild rice for female parent, hybridizes with the first rice varieties respectively, obtains the first hybrid generation; Using the first hybrid generation as female parent, and hybridize with the second rice varieties, obtain the second hybrid generation; Second hybrid generation is isolated male sterile plant, and as maternal, hybridize with the 3rd rice varieties respectively, obtain triple-crossing filial generation; Using triple-crossing filial generation as male parent, using male sterile plant as female parent, backcrossing, transformation becomes cytoplasmic male sterile rice.
Summary of the invention
The technical problem to be solved is to provide a kind of sterile gene wide material sources, selection-breeding program is simple, the cycle is shorter and the selection of the common line with genic sterile of the simple Oryza sativa L. of molecular markers development process.
For solving the technical problem of the present invention, the technical scheme adopted is as follows:
The selection of the common line with genic sterile of a kind of Oryza sativa L., specifically includes following steps:
(1) the Male sterile gene A in common Genetic Sterility mutant is carried out molecular markers development, obtain with the closely linked molecular marker S of common Genetic Sterility Gene A, and with the educated Gene A corresponding to Male sterile gene A ' closely linked molecular marker R;
(2) with the rice strain containing Male sterile gene A for female parent, its molecular marker is S/S type or S/R type; With molecular marker type be S/R type or R/R type Oryza sativa L. for recurrent parent, i.e. male parent, hybridize to obtain F1;
(3) carrying out Markers for Detection in F1, selection prospect is S/R genotype, and full-length genome labelling selects the strain that deflection wheel returns parent molecules type to be female parent, carries out many for backcrossing with recurrent parent, obtains BCnF1, wherein: n >=2;
(4) carrying out Markers for Detection in BCnF1, selection prospect is S/R genotype, and full-length genome labelling selects deflection wheel to return the strain of parent molecules type, and selfing obtains BCnF2;
(5) carrying out Markers for Detection in BCnF2, selection prospect is S/S genotype, and full-length genome labelling selects partially homozygous strain, and inbreeding of more generation obtains BCnFm, wherein: m >=3;
(6) BCnFm obtained in step (5) is carried out sterile line qualification, it is thus achieved that meet the common line with genic sterile of sterile line standard of perfection.
In such scheme, described step (3) can be:
Plantation F1For hybrid seed, it is thus achieved that F1Markers for Detection is carried out for hybrid plant and to it, selection prospect molecular marker is S/R, its full-length genome molecular marker type is identical with recurrent parent molecular marker type or more identical strain, in conjunction with traditional breeding way, selecting the strain that field character meets breeding objective in above-mentioned strain is female parent, with recurrent parent for male parent, repeatedly backcrosses, obtain to obtain BCnF1, wherein: n >=2;
Or it is:
To F1Markers for Detection is carried out for seed, selection prospect molecular marker is S/R, its full-length genome molecular marker type is identical with recurrent parent molecular marker type or more identical seed plantation, obtaining F1 generation hybridization strain, in conjunction with traditional breeding way, selecting the strain that field character meets breeding objective in above-mentioned strain is female parent, with recurrent parent for male parent, repeatedly backcross, it is thus achieved that obtain BCnF1, wherein: n >=2;
Or it is:
Plantation F1For hybrid seed, it is thus achieved that F1For hybrid plant, fertile plant system therein is selected to carry out Markers for Detection, selection prospect molecular marker is S/R, its full-length genome molecular marker type is identical with recurrent parent molecular marker type or more identical strain, and in conjunction with traditional breeding way, selecting the strain that field character meets breeding objective in above-mentioned strain is female parent, with recurrent parent for male parent, repeatedly backcross, it is thus achieved that obtain BCnF1, wherein: n >=2.
Described step (4) can be:
Plantation BCnF1For hybrid seed, it is thus achieved that BCnF1Markers for Detection is carried out for hybrid plant and to it, selection prospect molecular marker is S/R, its full-length genome molecular marker type is identical with recurrent parent molecular marker type or more identical strain, in conjunction with traditional breeding way, above-mentioned strain select field character meet the strain selfing of breeding objective, it is thus achieved that BCnF2Seed;
Or it is:
To BCnF1Carrying out Markers for Detection for seed, selecting prospect molecular marker is S/R, and its full-length genome molecular marker type is identical with recurrent parent molecular marker type or more identical seed is planted, it is thus achieved that BCnF1Generation hybridization strain, in conjunction with traditional breeding way, selects field character to meet the strain selfing of breeding objective, it is thus achieved that BC in above-mentioned strainnF2Seed;
Or it is:
Plantation BCnF1For hybrid seed, it is thus achieved that BCnF1For hybrid plant, fertile plant system therein is selected to carry out Markers for Detection, selection prospect molecular marker is S/R, the strain that its full-length genome molecular marker type is complete or more identical with recurrent parent molecular marker type, in conjunction with traditional breeding way, above-mentioned strain select field character meet the strain selfing of breeding objective, it is thus achieved that BCnF2。
Described step (5) can be:
Plantation BCnF2For hybrid seed, it is thus achieved that BCnF2Markers for Detection is carried out for hybrid plant and to it, selection prospect molecular marker is S/S, its full-length genome molecular marker type is entirely pure and mild type or homozygous more strain, in conjunction with traditional breeding way, above-mentioned strain select field character meet the strain selfing of breeding objective, it is thus achieved that BCnFm seed;
Or it is:
To BCnF1Carrying out Markers for Detection for seed, selecting prospect molecular marker is S/S, and its full-length genome molecular marker type is entirely pure and mild type or homozygous more seed plantation, it is thus achieved that BCnF1Generation hybridization strain, in conjunction with traditional breeding way, selects field character to meet the strain selfing of breeding objective, it is thus achieved that BCnFm seed in above-mentioned strain;
Or it is:
Plantation BCnF1For hybrid seed, it is thus achieved that BCnF1For hybrid plant, fertile plant system therein is selected to carry out Markers for Detection, selection prospect molecular marker is S/S, its full-length genome molecular marker type is entirely pure and mild type or homozygous more strain, in conjunction with traditional breeding way, above-mentioned strain select field character meet the strain selfing of breeding objective, it is thus achieved that BCnFm.
Further,
N=2～7 described in described step (3), it may be assumed that n is 2 or 3 or 4 or 5 or 6 or 7, and namely preferably backcrossed 2～7 generations.
Further,
N=2 in described step (3), namely backcrossed 2 generations.
Further,
M=2 in described step (5), i.e. selfing 2 generation.
Further,
Described step (1) Middle molecule is labeled as and can reflect between bion or population the DNA fragment specific of certain species diversity in genome.
Preferably,
Described step (1) Middle molecule labelling is specially restriction fragment length polymorphism molecular marker (RFLP), simple repeated sequence molecular marker (SSR), single nucleotide polymorphism molecular marker (SNP).
Preferably,
Described step (1) Middle molecule marker development preferably employs single nucleotide polymorphism molecular marker (SNP) exploitation.
Third generation SNP (nucleotide polymorphisms molecular marker) molecular marker refers to the not difference of individual nucleotide between iso-allele in same site, this species diversity includes disappearance or the insertion of single base, the more commonly replacement of single core thuja acid, and often occur between purine bases (A and G) and pyrimidine bases (C and T), SNP marker can help distinguish between the difference of two individual hereditary materials.
SNP compares the difference of the RFLP (restriction fragment length polymorphism molecular marker) of the first generation and SSR (simple repeated sequence molecular marker) labelling of the second filial generation 2 aspects: one, SNP no longer changes as detection means using the length of DNA fragmentation, and directly using sequence variations as labelling; Its two, SNP marker analysis abandoned completely classics gel electrophoresis, instead up-to-date DNA chip technology.
Adopt single nucleotide polymorphism molecular marker (SNP) exploitation, be more conducive to realize automation mechanized operation, reproducible.
In this programme:
Prospect molecular marker: refer to that the target gene to wanting transformation carries out molecular marker.
Full-length genome labelling selects: be exactly the marker assisted selection within the scope of full-length genome, it is utilize covering whole genomic labelling that chromosome is divided into several fragments, namely two often adjacent labellings are exactly a chromosome segment, then pass through marker genetype and estimate the effect of each chromosome segment in conjunction with phenotypic character and pedigree information respectively, finally utilize individual entrained label information that the phenotypic information of its unknown is predicted, the effect being about to each chromosome segment that individuality carries adds up, and then estimates genomic breeding value and select.
Full-length genome labelling includes multiple labelling, and each labelling is it is possible that three kinds of situations: identical with female parent, and hybrid identical with male parent. Full-length genome Marker-assisted selection deflection wheel returns the strain of parent molecules type, namely select the strain that full-length genome molecular marker type and recurrent parent (i.e. male parent) molecular marker type are complete or more identical, namely select in full-length genome molecular marker type as far as possible how identical with recurrent parent (i.e. male parent) molecular marker type as far as possible. And homozygous to be just to try to select in full-length genome molecular marker type as far as possible many with recurrent parent (i.e. male parent) or identical with female parent molecular marker type.
Recurrent parent: it is maternal that backcross (hybrid of two parents generations hybridizes with one of parent again) selects the kind with good characteristic to make when being typically in first time hybridization, and when backcrossing, making male parent afterwards for each time, this parent is recurrent parent when backcrossing. The purpose backcrossed is that the good characteristic making parent is slowly strengthened in hybrid generation, and a certain advantage of nonrecurrent parent is transferred to hybrid.
There is advantages that
1, the sterile line breeding method of the present invention utilizes the Male sterile gene of wide material sources to originate as sterile gene, can expand and heterotic utilize scope, and comprehensive raising is heterotic utilizes level.
2, the present invention chooses Male sterile gene and originates as sterile gene, owing to common Genetic Sterility exists extensively at nature, and abundance, and it controls its fertility by Recessive genes, molecule mechanism is relatively easy, therefore, can simplify the process of molecular markers development.
3, the sterile line breeding method of the present invention is directly to hand over selection-breeding sterile line either directly through Xian round-grained rice, rather than the maintenance by Xian round-grained rice friendship selection-breeding Cheng Keyu is that transformation becomes sterile line again, the procedure of breeding is simple, breeding cycle can be shortened greatly, needing the sterile line that the Xian round-grained rice that could realize in more than ten years (even more than 20 generation) is handed over before this, the present invention is the shortest can be able to be realized in 5 generations (first-filial generation+two generations that backcross+selfing two generation).
Accompanying drawing explanation
In order to be illustrated more clearly that the technical scheme of the embodiment of the present invention, the accompanying drawing used required in embodiment will be briefly described below, apparently, accompanying drawing in the following describes is only some embodiments of the present invention, for those of ordinary skill in the art, under the premise not paying creative work, it is also possible to obtain other accompanying drawing according to these accompanying drawings.
Fig. 1 is the technology path flow chart of the selection of the common line with genic sterile of Oryza sativa L. of the embodiment of the present invention 1;
Fig. 2 is the technology path flow chart of the selection training C311 (round-grained rice type) and samsara 422 (Java type) in prior art;
Fig. 3 is the technology path flow chart of the selection of indica type Peiai 64 and Peiai 64S in prior art;
Fig. 4 utilizes molecular marking technique labelling wide compatibility gene S 5 n and Restore gene Rf (6), Rf (4), the technology path flow chart of Rf (3) in prior art.
Detailed description of the invention
Below in conjunction with accompanying drawing and specific embodiment, the invention will be further described:
The Male sterile gene related in following example can be inquired about on " Oryza sativa L. data center of country " website: http://www.ricedata.cn/gene/
Embodiment 1:
As it is shown in figure 1,
The selection of the common line with genic sterile of a kind of Oryza sativa L. of the present embodiment, specifically includes following steps:
(1) first, the Male sterile gene tdr (tapetum degradation delay gene) that common Genetic Sterility mutant force is transported in round-grained rice 7 (japonica rice 9522) carries out molecular markers development, obtain and the common closely linked molecular marker S of Genetic Sterility gene tdr, and molecular marker R closely linked with the educated gene tdr ' corresponding to Male sterile gene tdr;
(2) rice strain of plantation force fortune round-grained rice 7 (japonica rice 9522) containing common Genetic Sterility gene tdr, its molecular marker is S/S type or S/R type, with described strain for female parent, with molecular marker type be S/R type or R/R type long-grained nonglutinous rice 9311 for recurrent parent, i.e. male parent, hybridize, it is thus achieved that the seed of first generation of hybrid F1;
(3) plantation F1For hybrid seed, it is thus achieved that F1Markers for Detection is carried out for hybrid plant and to it, selection prospect molecular marker is S/R, its full-length genome molecular marker type is identical with recurrent parent long-grained nonglutinous rice 9311 molecular marker type or more identical strain, in conjunction with traditional breeding way, selecting the strain that field character meets breeding objective in above-mentioned strain is female parent, with recurrent parent long-grained nonglutinous rice 9311 for male parent, backcross, it is thus achieved that first backcross generation first generation of hybrid BC1F1;
(4) plantation BC1F1For hybrid seed, it is thus achieved that BC1F1Markers for Detection is carried out for hybrid plant and to it, selection prospect molecular marker is S/R, its full-length genome molecular marker type is identical with recurrent parent long-grained nonglutinous rice 9311 molecular marker type or more identical strain, in conjunction with traditional breeding way, selecting the strain that field character meets breeding objective in above-mentioned strain is female parent, with recurrent parent long-grained nonglutinous rice 9311 for male parent, backcross, it is thus achieved that second backcross generation first generation of hybrid BC2F1;
(5) plantation BC2F1For hybrid seed, it is thus achieved that BC2F1Markers for Detection is carried out for hybrid plant and to it, selection prospect molecular marker is S/R, its full-length genome molecular marker type is identical with recurrent parent long-grained nonglutinous rice 9311 molecular marker type or more identical strain, in conjunction with traditional breeding way, above-mentioned strain select field character meet the strain selfing of breeding objective, it is thus achieved that BC2F2Seed;
(6) plantation BC2F2For hybrid seed, it is thus achieved that BC2F2Markers for Detection is carried out for hybrid plant and to it, selection prospect molecular marker is S/S, and its full-length genome molecular marker type is entirely pure and mild type or homozygous more strain, in conjunction with traditional breeding way, above-mentioned strain select field character meet the strain selfing of breeding objective, it is thus achieved that BC2F3Seed;
(7) to the BC obtained in step (6)2F3Carry out sterile line qualification, it is thus achieved that meet the common line with genic sterile of sterile line standard of perfection.
Described step (1) Middle molecule marker development adopts the exploitation of third generation SNP marker.
1, the present embodiment utilizes the Male sterile gene tdr gene of wide material sources to originate as sterile gene, can expand and heterotic utilize scope, and comprehensive raising is heterotic utilizes level.
2, the Male sterile gene tdr gene in the present embodiment is controlled its fertility by Recessive genes, and molecule mechanism is relatively easy, therefore, can simplify the process of molecular markers development.
3, the present embodiment adopts the exploitation of third generation SNP marker, can improve automation mechanized operation degree and the property again of molecule exploitation.
4, the sterile line breeding method of the present embodiment is to hand over selection-breeding sterile line either directly through Xian round-grained rice, rather than the maintenance by Xian round-grained rice friendship selection-breeding Cheng Keyu is that transformation becomes sterile line again, the procedure of breeding is simple, breeding cycle can be shortened greatly, needing the sterile line that the Xian round-grained rice that could realize in more than ten years (even more than 20 generation) is handed over before this, the present invention had only to for 5 generations and can realize.
Embodiment 2
The present embodiment is in that from the different of embodiment 1:
Step (3) is: to F1Markers for Detection is carried out for seed, selection prospect molecular marker is S/R, its full-length genome molecular marker type is identical with recurrent parent molecular marker type or more identical seed plantation, obtain F1 generation hybridization strain, in conjunction with traditional breeding way, selecting the strain that field character meets breeding objective in above-mentioned strain is female parent, with recurrent parent long-grained nonglutinous rice 9311 for male parent, backcross, it is thus achieved that first backcross generation first generation of hybrid BC1F1;
Step (4) is: to BC1F1Carrying out Markers for Detection for seed, selecting prospect molecular marker is S/R, and its full-length genome molecular marker type is identical with recurrent parent molecular marker type or more identical seed is planted, it is thus achieved that BC1F1Generation hybridization strain, in conjunction with traditional breeding way, selecting the strain that field character meets breeding objective in above-mentioned strain is female parent, with recurrent parent long-grained nonglutinous rice 9311 for male parent, backcrosses, it is thus achieved that second backcross generation first generation of hybrid BC2F1;
Step (5) is: to BC2F1Carrying out Markers for Detection for seed, selecting prospect molecular marker is S/R, and its full-length genome molecular marker type is identical with recurrent parent molecular marker type or more identical seed is planted, it is thus achieved that BC2F1Generation hybridization strain, in conjunction with traditional breeding way, selects field character to meet the strain selfing of breeding objective, it is thus achieved that BC in above-mentioned strain2F2Seed;
Step (6) is: to BC2F1Carrying out Markers for Detection for seed, selecting prospect molecular marker is S/S, and its full-length genome molecular marker type is entirely pure and mild type or homozygous more seed plantation, it is thus achieved that BC2F1Generation hybridization strain, in conjunction with traditional breeding way, selects field character to meet the strain selfing of breeding objective, it is thus achieved that BC in above-mentioned strain2F2Seed.
Embodiment 3
The present embodiment and embodiment 1 are distinctive in that:
Step (3) is:
Plantation F1For hybrid seed, it is thus achieved that F1For hybrid plant, fertile plant system therein is selected to carry out Markers for Detection, selection prospect molecular marker is S/R, its full-length genome molecular marker type is identical with recurrent parent molecular marker type or more identical strain, in conjunction with traditional breeding way, selecting the strain that field character meets breeding objective in above-mentioned strain is female parent, with recurrent parent long-grained nonglutinous rice 9311 for male parent, backcross, it is thus achieved that first backcross generation first generation of hybrid BC1F1;
Step (4) is:
Plantation BC1F1For hybrid seed, it is thus achieved that BC1F1For hybrid plant, fertile plant system therein is selected to carry out Markers for Detection, selection prospect molecular marker is S/R, its full-length genome molecular marker type is identical with recurrent parent molecular marker type or more identical strain, in conjunction with traditional breeding way, selecting the strain that field character meets breeding objective in above-mentioned strain is female parent, with recurrent parent long-grained nonglutinous rice 9311 for male parent, backcross, it is thus achieved that second backcross generation first generation of hybrid BC2F1;
Step (5) is:
Plantation BC2F1For hybrid seed, it is thus achieved that BC2F1For hybrid plant, fertile plant system therein is selected to carry out Markers for Detection, selection prospect molecular marker is S/R, the strain that its full-length genome molecular marker type is complete or more identical with recurrent parent molecular marker type, in conjunction with traditional breeding way, above-mentioned strain select field character meet the strain selfing of breeding objective, it is thus achieved that BC2F2;
Step (6) is:
Plantation BC2F1For hybrid seed, it is thus achieved that BC2F1For hybrid plant, fertile plant system therein is selected to carry out Markers for Detection, selection prospect molecular marker is S/S, its full-length genome molecular marker type is entirely pure and mild type or homozygous more strain, in conjunction with traditional breeding way, above-mentioned strain select field character meet the strain selfing of breeding objective, it is thus achieved that BC2F2。
Embodiment 4:
The present embodiment and embodiment 1 are distinctive in that:
Described Male sterile gene A is OsADF gene (the F-box albumen that anther development is relevant), and OsADF, at fringe portion specifically expressing, belongs to F-box protein family, and female parent is OsADF gene mutation body.
OsADF mutant, RNAi transgenic line tapetum degradation is abnormal, and microspore development stops, and ultimately results in pollen fertility and reduces.
Embodiment 5:
The present embodiment and embodiment 1 are distinctive in that:
Described Male sterile gene A is cytochrome P450 gene family member: cyp703a3 gene, and female parent is cyp703a3 gene mutation body.
Cyp703a3-2 mutant flower pesticide surface horny layer and exposore developmental defect, the content of cutin monomer and wax component significantly reduces, and causes that anther development is abnormal, and flower pesticide diminishes as white yellow, male sterility mature flower powder can not be formed, thus can not gather in the crops mature seed.
Embodiment 6:
The present embodiment and embodiment 1 are distinctive in that:
Described Male sterile gene A is cytochrome P450 gene family member, i.e. cyp704b2 gene.
ω-the hydroxylating of cyp704b2 catalysis fatty acid, it is required to be that Rice Anther cutin biosynthesis and exposore form institute. The tapetum of cyp704b2 mutant sporinite expands, and pollen grain causes sterile owing to being formed without outer wall and flower pesticide horny layer ateliosis, it addition, be difficult in mutant flower pesticide cutin monomer be detected.
Embodiment 7:
The present embodiment and embodiment 1 are distinctive in that:
Described rice strain's female parent containing Male sterile gene A is that Japan is fine;
Described recurrent parent, i.e. male parent, for Peiai 64;
Described Male sterile gene A is cytochrome P450 gene family member: cyp703a3 gene.
Embodiment 8:
The present embodiment and embodiment 1 are distinctive in that:
Described rice strain's female parent containing Male sterile gene A is long-grained nonglutinous rice 9311;
Described recurrent parent, i.e. male parent, for java rice lemont;
Described Male sterile gene A is cytochrome P450 gene family member, i.e. cyp704b2 gene.
Above-described embodiment is further illustrating the present invention, rather than restriction the scope of the present invention. Without departing from the whole technical scope of the present invention, various amendment and change can be carried out.

Claims (10)

1. the selection of the common line with genic sterile of Oryza sativa L., it is characterised in that specifically include following steps:
(1) the Male sterile gene A in common Genetic Sterility mutant is carried out molecular markers development, obtain with the closely linked molecular marker S of common Genetic Sterility Gene A, and with the educated Gene A corresponding to Male sterile gene A ' closely linked molecular marker R;
(2) with the rice strain containing Male sterile gene A for female parent, its molecular marker is S/S type or S/R type; With molecular marker type be S/R type or R/R type Oryza sativa L. for recurrent parent, i.e. male parent, hybridize to obtain F1;
(3) carrying out Markers for Detection in F1, selection prospect is S/R genotype, and full-length genome labelling selects the strain that deflection wheel returns parent molecules type to be female parent, carries out many for backcrossing with recurrent parent, obtains BCnF1, wherein: n >=2;
(4) carrying out Markers for Detection in BCnF1, selection prospect is S/R genotype, and full-length genome labelling selects deflection wheel to return the strain of parent molecules type, and selfing obtains BCnF2;
(5) carrying out Markers for Detection in BCnF2, selection prospect is S/S genotype, and full-length genome labelling selects partially homozygous strain, and inbreeding of more generation obtains BCnFm, wherein: m >=3;
(6) BCnFm obtained in step (5) is carried out sterile line qualification, it is thus achieved that meet the common line with genic sterile of sterile line standard of perfection.
2. the selection of the common line with genic sterile of a kind of Oryza sativa L. according to claim 1, it is characterised in that
Described step (3) particularly as follows:
Plantation F1For hybrid seed, it is thus achieved that F1Markers for Detection is carried out for hybrid plant and to it, selection prospect molecular marker is S/R, its full-length genome molecular marker type is identical with recurrent parent molecular marker type or more identical strain, in conjunction with traditional breeding way, selecting the strain that field character meets breeding objective in above-mentioned strain is female parent, with recurrent parent for male parent, repeatedly backcrosses, obtain to obtain BCnF1, wherein: n >=2;
Or it is:
To F1Markers for Detection is carried out for seed, selection prospect molecular marker is S/R, its full-length genome molecular marker type is identical with recurrent parent molecular marker type or more identical seed plantation, obtaining F1 generation hybridization strain, in conjunction with traditional breeding way, selecting the strain that field character meets breeding objective in above-mentioned strain is female parent, with recurrent parent for male parent, repeatedly backcross, it is thus achieved that obtain BCnF1, wherein: n >=2;
Or it is:
Plantation F1For hybrid seed, it is thus achieved that F1For hybrid plant, fertile plant system therein is selected to carry out Markers for Detection, selection prospect molecular marker is S/R, its full-length genome molecular marker type is identical with recurrent parent molecular marker type or more identical strain, and in conjunction with traditional breeding way, selecting the strain that field character meets breeding objective in above-mentioned strain is female parent, with recurrent parent for male parent, repeatedly backcross, it is thus achieved that obtain BCnF1, wherein: n >=2.
3. the selection of the common line with genic sterile of a kind of Oryza sativa L. according to claim 1 and 2, it is characterised in that
Described step (4) particularly as follows:
Plantation BCnF1For hybrid seed, it is thus achieved that BCnF1Markers for Detection is carried out for hybrid plant and to it, selection prospect molecular marker is S/R, its full-length genome molecular marker type is identical with recurrent parent molecular marker type or more identical strain, in conjunction with traditional breeding way, above-mentioned strain select field character meet the strain selfing of breeding objective, it is thus achieved that BCnF2Seed;
Or it is:
To BCnF1Carrying out Markers for Detection for seed, selecting prospect molecular marker is S/R, and its full-length genome molecular marker type is identical with recurrent parent molecular marker type or more identical seed is planted, it is thus achieved that BCnF1Generation hybridization strain, in conjunction with traditional breeding way, selects field character to meet the strain selfing of breeding objective, it is thus achieved that BC in above-mentioned strainnF2Seed;
Or it is:
Plantation BCnF1For hybrid seed, it is thus achieved that BCnF1For hybrid plant, fertile plant system therein is selected to carry out Markers for Detection, selection prospect molecular marker is S/R, the strain that its full-length genome molecular marker type is complete or more identical with recurrent parent molecular marker type, in conjunction with traditional breeding way, above-mentioned strain select field character meet the strain selfing of breeding objective, it is thus achieved that BCnF2。
4. the selection of the common line with genic sterile of a kind of Oryza sativa L. according to claim 1 and 2, it is characterised in that
Described step (5) particularly as follows:
Plantation BCnF2For hybrid seed, it is thus achieved that BCnF2Markers for Detection is carried out for hybrid plant and to it, selection prospect molecular marker is S/S, its full-length genome molecular marker type is entirely pure and mild type or homozygous more strain, in conjunction with traditional breeding way, above-mentioned strain select field character meet the strain selfing of breeding objective, it is thus achieved that BCnFm seed;
Or it is:
To BCnF1Carrying out Markers for Detection for seed, selecting prospect molecular marker is S/S, and its full-length genome molecular marker type is entirely pure and mild type or homozygous more seed plantation, it is thus achieved that BCnF1Generation hybridization strain, in conjunction with traditional breeding way, selects field character to meet the strain selfing of breeding objective, it is thus achieved that BCnFm seed in above-mentioned strain;
Or it is:
Plantation BCnF1For hybrid seed, it is thus achieved that BCnF1For hybrid plant, fertile plant system therein is selected to carry out Markers for Detection, selection prospect molecular marker is S/S, its full-length genome molecular marker type is entirely pure and mild type or homozygous more strain, in conjunction with traditional breeding way, above-mentioned strain select field character meet the strain selfing of breeding objective, it is thus achieved that BCnFm.
5. the selection of the common line with genic sterile of a kind of Oryza sativa L. according to claim 1 and 2, it is characterised in that
N=2～7 described in described step (3), it may be assumed that n is 2 or 3 or 4 or 5 or 6 or 7, is specially 2～7 generations that backcrossed.
6. the selection of the common line with genic sterile of a kind of Oryza sativa L. according to claim 5, it is characterised in that
N=2 in described step (3), namely backcrossed 2 generations.
7. the selection of the common line with genic sterile of a kind of Oryza sativa L. according to claim 1-2 or 6 any one, it is characterised in that
M=2 in described step (5), i.e. selfing 2 generation.
8. the selection of the common line with genic sterile of a kind of Oryza sativa L. according to claim 1-2 or 6 any one, it is characterized in that, described step (1) Middle molecule is labeled as and can reflect between bion or population the DNA fragment specific of certain species diversity in genome.
9. the selection of the common line with genic sterile of a kind of Oryza sativa L. according to claim 1-2 or 6 any one, it is characterized in that, described step (1) Middle molecule labelling is specially restriction fragment length polymorphism molecular marker, simple repeated sequence molecular marker, single nucleotide polymorphism molecular marker.
10. the selection of the common line with genic sterile of a kind of Oryza sativa L. described in any one according to Claim 8, it is characterised in that described step (1) Middle molecule marker development is single nucleotide polymorphism molecular markers development.
CN201610012557.6A 2016-01-11 2016-01-11 Breeding method of rice common genic male sterile line Pending CN105660361A (en)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102876711A (en) * 2012-10-31 2013-01-16 湖南杂交水稻研究中心 Cultivation method of rice engineering maintainer line and application thereof to breeding of rice genic male sterile line
CN104894144A (en) * 2015-07-06 2015-09-09 海南波莲水稻基因科技有限公司 Rice CYP704B2 gene mutant, as well as molecule identification method and applications thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102876711A (en) * 2012-10-31 2013-01-16 湖南杂交水稻研究中心 Cultivation method of rice engineering maintainer line and application thereof to breeding of rice genic male sterile line
CN104894144A (en) * 2015-07-06 2015-09-09 海南波莲水稻基因科技有限公司 Rice CYP704B2 gene mutant, as well as molecule identification method and applications thereof

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