CN103858747A - Molecule designing and breeding method of plant backcross line population - Google Patents
Molecule designing and breeding method of plant backcross line population Download PDFInfo
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Abstract
The invention relates to a molecule designing and breeding method of plant backcross line population, which is characterized in that a donor plant containing different exogenous target genes and different individuals of an acceptor plant to be improved are subjected to hybridization and backcross with an acceptor parent; auxiliary selection is carried out according to a molecular mark tightly linked with the target gene, and the stable backcross lines containing different target genes can be bred; the backcross lines constitute the backcross line population containing several different target gene; the backcross line population seed production is carried out according to ratio while matching and designing population, and thereby the backcross line population product for commercialization can be generated. The molecule designing and breeding method changes a highly difficult technology which is characterized in that many target genes enable polymerization towards to one individual to a technology which is characterized in that different genes or less gene enables polymerization to different individuals of one same acceptor, and mark selective introduction of multiple individuals and less gene can be simultaneously carried out, so that purpose of acceptor colony having many excellent target genes can be realized; different individuals are used for carrying with diversity population of different target genes to reach an effect of polymerization of many excellent genes by colony level, and the molecule designing and breeding method has better continuous production prospect.
Description
Technical field:
The present invention relates to that a Plants backcrosses is the Molecular design breeding method of population, belongs to plant molecular marker assistant breeding and Molecular design breeding technical field.
Background technology:
Molecular mark is the major technique that modern molecular biology combines with thremmatology, can improve efficiency and the purpose of traditional breeding method, especially improves the efficiency of the complicated economical character improvement to being difficult to directly observation.Current molecular mark refers to be designed and the closely linked molecular labeling of different target genes, the method of selecting by assistant breeding, all types of target gene is imported in the particular individual of acceptor, make this receptor contain numerous target genes that molecular marker assisted selection imports, i.e. the auxiliary pyramiding breeding of common genetic marker.But because the complexity of organism is affected by linkage relationship and the position effect of target recombinant gene etc. between gene especially, cause the target gene of importing more, difficulty and expression effect are just more difficult to expect.Therefore, current molecular breeding is mainly emphasized certain excellent genes of donor by the molecular labeling locating information having obtained, synthetic molecules mark, in donor and acceptor crossover process, with molecular mark, select and contain donor target gene, and other background proterties and original receptor do not have the kind of significant difference, be mainly used in the backcross improvement plan of single or minority gene.In fact improve a proterties and at least after hybridization, will carry out backcrossing of 3-7 generation, and carry out the screening of molecular labeling simultaneously, could acquired character obtain the individuality of improveing, improve next proterties and restart again hybridization and backcross process.Therefore, the difficulty that realizes more than 3 gene pyramiding is large, efficiency is low, and the time of improvement acceptor at least needs 9-21 generation.In the time that improvement completes, this receptor kind may be out-of-date, or replaced by kind of new generation.Therefore, numerous target genes are aggregated to cycle of an acceptor individual plant is long, difficulty is large, efficiency is low, and its breeding efficiency has limitation.
Summary of the invention:
In order to overcome the deficiencies in the prior art, the object of the present invention is to provide a Plants to backcross is the Molecular design breeding method of population.
It is the Molecular design breeding method of population that plant provided by the invention backcrosses, and comprises the steps:
(1) the donor plant that contains different external source target genes and the Different Individual of recipient plant to be improved are hybridized;
(2) will carry out rotational crossing with recipient plant parent again through the recipient plant individuality of hybridization;
(3), according to carrying out MOLECULE DESIGN assisted Selection with the closely linked molecular labeling of target gene, stable the backcrossing that incubation contains respectively different target gene is;
(4) according to breeding objective collocation design, by 2-100 backcross be or backcross be between hybridization first generation of hybrid composition to contain backcrossing of multiple different target genes be population;
(5) it is the seed production of population that the proportioning during according to collocation design population backcrosses, and is population product thereby produce business-like backcrossing.
Described recipient plant is self-pollinated plant, cross-pollinatd plant, Constantly allogamous plant or asexually propagated plant.
Described donor plant is the plant that can hybridize with recipient plant.
Described external source target gene be can increase the gene of recipient plant to biological stress resistance, can increase the gene of recipient plant to abiotic stress defensive ability/resistance ability, can improve recipient plant output gene, can improve recipient plant quality characteristic gene, can increase the gene of recipient plant adaptive capacity.
After the hybridization of donor plant and recipient plant, backcross and carry out 1-15 time, formation Other Main Agronomic Characters is similar to recurrent parent or from stable the backcrossing of donor plant target gene be better.
The hybridization of donor plant and recipient plant and backcross after, backcross between being and carry out again phase mutual cross, form the first generation of hybrid; Or backcross, stable the backcrossing that forms 1-10 target gene of polymerization is again.
Described MOLECULE DESIGN be using stable the 2-100 of acquisition backcross be and or can propagating materials and or backcross be the first generation of hybrid by phase mutual cross as component, then by population arrange in pairs or groups that forming backcrosses and is pure lines population with or first generation of hybrid population or vegetative propagation population.
Described population collocation is the collocation of the each component in population being carried out to different proportion, identify each component by molecular labeling, and detect the proportioning of the each component of population by statistical test, thus guarantee to carry out accurate seed production, realize that to backcross be the commercialization of population seed.
The present invention has following beneficial effect compared with prior art:
1, the present invention by highly difficult to the technology of the numerous target genes of body polymerization one by one, change respectively the different individuality to same acceptor by different genes or minority gene pyramiding into, the Marker selection that simultaneously carries out multiple bodies, minority gene imports, again by collocation and Combination Design between same acceptor different target gene individuality, realize acceptor colony (backcross is population) and possess the object of numerous good target genes; Utilize Different Individual to carry the diversity of different target gene (polygene type) population, reach the effect of the numerous excellent genes of population level polymerization, especially provide horizontal resistance for the biological adverse circumstance of opposing, also reduce the selection of damage by disease and insect is pressed, reach the effect of alleviating damage by disease and insect sudden change, possess better sustainable prospect of production.
2, the present invention is take multiple individualities of recipient plant as unit, simultaneously from the donor plant hybridization that contains different marker gene, then carrying out molecular marker assisted selection and background selects, select respectively multiple the backcrossing of containing multiple donor target genes and be and/or backcross to be that a phase mutual cross backcrosses again, the target gene of polymerization moderate amount, improvement when realizing multiple proterties, reduce the difficulty and the time cycle that numerous target genes are imported to a single receptor, forming 2-100 backcrosses and to be or the first generation of hybrid of the line cross that backcrosses, backcross with these again and be or the first generation of hybrid is arranged in pairs or groups and design object gene colony product according to a certain percentage, realization is set up and accurately backcrossed is the target of population kind.In commercialization population product, each component and population ratio can sample by multiple individual plants, carry out Markers for Detection and statistical test and determine, the molecular labeling of component presents homozygote or first-filial generation or specific clone state.And in the product of the auxiliary pyramiding breeding of traditional molecular labeling, just only have a kind of homozygote genotype, between individuality without any difference.
3, carry out in the process of background selection backcrossing, except the target gene difference of mark, all characteristics are all identical with original receptor or better than original receptor, then according to the requirement of breeding objective, be collocation backcrossing of different marker gene, being designed to specifically backcrosses is population, can obtain the combination resistance to different damage by disease and insect and different biological strains, and keeps that specifically to backcross be accurate population.The different marker gene strain of same acceptor phenotype is basic identical, it is only marker gene object difference, therefore, the strain that contains different marker gene can be arranged in pairs or groups according to a certain percentage and form accurate business population, enter production link, thereby slowing down kind selects to press to the specificity of damage by disease and insect, delay the mutagenesis of varietal resistance gene pairs damage by disease and insect, extend kind service life, improve the efficiency of molecular mark, realize in population Molecular design breeding between individuality, many (genes) to backcross be population Molecular design breeding.
Donor can possess the phenotypic material of target gene from any, and possess and the molecular labeling of this gene linkage, or can by molecular labeling localization method find this gene linkage molecule mark material or from the transgenic progeny material of target gene.
Embodiment:
Below in conjunction with specific embodiment, the invention will be further described.
Embodiment 1:
Take self pollination crop paddy rice as example, kind ' Guangdong show accounts for ' is acceptor, with 6 acceptor individual plants respectively with contain blast resistant gene Pi-b, Pi-55(t), anti-brown back rice plant hopper gene Bph14, Bph15 and bacterial leaf spot resistant ospc gene Xa-7, the donor hybridization of Xa-23, then use respectively ' Guangdong show accounts for ' to backcross, each individuality of selecting to import target-marking gene in backcross progeny, and other proterties individuality the most similar to ' Guangdong show accounts for ', and repeated multiple times backcrossing, finally obtain respectively containing Pi-b, Pi-55(t), Bph14, Bph15 and Xa-7, 6 target genes of Xa-23, other comprehensive agronomy proterties 6 stable objects gene strains similar or better to ' Guangdong show accounts for ', it is bred separately.
Will be containing Pi-b and Pi-55(t) the strain phase mutual cross of gene, then backcross, select the dual anti-pest strain containing 2 blast resistant genes; By the strain phase mutual cross containing Bph14 and Bph15 gene, then backcross, select the dual anti-worm strain containing 2 anti insect genes; By the strain phase mutual cross containing Xa-7 and Xa-23 gene, then backcross, select the dual anti-bacterial leaf spot strain containing 2 bacterial leaf spot resistant genes; By the strain phase mutual cross containing Bph14 and Pi-b gene, then backcross, select the Resistant strain containing blast resisting and anti insect gene.
The molecular labeling of above-mentioned each gene can select to be less than from open source information and target gene the molecular labeling of 10 Morgans (cM) distance.
Giving an example in one or more sources that table 1 provides wherein, but is not limited to these sources.
Table 1, genetic donor material and mark source thereof are for example
| Gene | Donor material for example | Linked marker (cM) for example | Chromosome |
| Pi-b | IR24,BL1 | C379～C2782B(is included) | 2 |
| Pi-55(t) | No. 2, Yuejingsimiao | RM1345-RM3452(is included) | 8 |
| Bph14 | O.officinalis | G1318-R1925(is included) | 3 |
| Bph15 | O.officinalis | C820-S11182(is included) | 4 |
| Xa-7 | DV85,DV86,DZ78 | G1091(6.0cM),AFLP31-10(3cM) | 6 |
| Xa-23 | Common wild-rice | OSR6(5.4cM),RM206(1.9cM) | 11 |
1-1：
Will be respectively containing Pi-b, Pi-55(t), 6 target genes of Bph14, Bph15 and Xa-7, Xa-23 seed that is that backcrosses breeds separately, according to each be seed mixed in equal amounts, it is population that the multiple gene of composition blast resisting, brown back rice plant hopper and bacterial leaf-blight equilibrium backcrosses, and enters production link.
1-2：
Will be respectively containing Pi-b, Pi-55(t), 6 target genes of Bph14, Bph15 and Xa-7, Xa-23 seed that is that backcrosses breeds separately, then by seed according to 2: 2: 2: the ratio of 2: 1: 1 is mixed, it is population that the multiple gene that composition is laid equal stress on blast resisting and brown back rice plant hopper backcrosses, and enters production link.
1-3：
Will be respectively containing Pi-b, Pi-55(t), the seed that is that backcrosses of 6 target genes of Bph14, Bph15 and Xa-7, Xa-23 breeds separately, then by seed according to 1: 1: 1: the ratio of 1: 2: 2 is mixed, it is population that composition backcrosses with the disease-resistant multiple gene of taking as the leading factor of bacterial leaf spot resistant, enters production link.
1-4：
Will be respectively containing Pi-b and Pi-55(t), the seed of 3 dual anti-strains of Bph14 and Bph15 and Xa-7 and Xa-23 breeds separately, then seed is mixed according to the ratio of 1:1:1, forming that the comprehensive multiple gene of dual anti-target gene backcrosses is population, enters production link.
1-5：
Will be respectively containing Pi-b and Pi-55(t), the seed of 4 dual anti-strains of Bph14 and Bph15, Xa-7 and Xa-23 and Bph14 and Pi-b breeds separately, then seed is mixed according to the ratio of 1:1:1:2, it is population that the comprehensive multiple gene of the two Resistant Genes of composition backcrosses, and enters production link.
In commercialization seed, sample by individual plant, carry out Markers for Detection and statistical test, can detect accurately component and the proportioning thereof of each population, so that seed law enforcement.
Embodiment 2:
Take cross pollinated plant corn single cross hybrid, ' in Zheng Dan 958 ' combination, ' Zheng 58 ' is as example for inbred line female parent, with 6 ' Zheng 58 ' acceptor individual plant respectively with the inbred line ' PI186208 ' that contains anti-southern type rust Rpp9 gene, containing the inbred line ' neat 319 ' of RppQ gene, contain the inbred line ' P25 ' of Rpp3 gene and the inbred line ' Pa405 ' containing anti-short mosaic disease Mdm1 gene, containing the inbred line ' 41 ' of Mdm2 gene, containing inbred line ' FAP1360A ' the donor hybridization of anti-short mosaic disease SCMV1 gene, then use respectively that ' Zheng 58 ' backcrosses, each individuality of selecting to import target gene in backcross progeny with molecular labeling, and other proterties and the ' individuality that Zheng 58 ' is the most similar, and repeated multiple times backcrossing, finally obtain respectively containing Rpp9, RppQ, Rpp3, Mdm1 and Mdm2, 6 target genes of SCMV1, other comprehensive agronomy proterties and ' similar or better 6 the stable objects gene strains of Zheng 58 ', it is bred separately.
The molecular labeling selection of above-mentioned each gene is less than the molecular labeling of 15 Morgans (cM) distance from open source information and genes of interest.
Giving an example in one or more sources that table 2 provides wherein, but is not limited to these sources.
Table 2, genetic donor material and mark source thereof are for example
2-1：
The seed that contains respectively the inbred line of 6 target genes of Rpp9, RppQ, Rpp3, Mdm1, Mdm2 and SCMV1 is bred separately, then seed is mixed according to the ratio of 1:1:1:1:1:1, the maternal population of multiple gene inbred line that composition is laid equal stress on anti-southern type rust and maize dwarf mosaic, enter crossbreed ' the maternal group's of system of Zheng Dan 958 ' production link, and then with former ' Zheng Dan 958 ' male parent ' prosperous 72 ' assembly first generation of hybrid population, form polygene type MOLECULE DESIGN first-filial generation commercialization population.
2-2：
The seed that contains respectively the inbred line of 6 target genes of Rpp9, RppQ, Rpp3, Mdm1, Mdm2 and SCMV1 is bred separately, then seed is mixed according to the ratio of 2:2:2:1:1:1, the multiple gene inbred line population that composition is taken as the leading factor with anti-southern type rust, enter crossbreed ' the maternal group's of system of Zheng Dan 958 ' production link, and then with former ' Zheng Dan 958 ' male parent ' prosperous 72 ' assembly first generation of hybrid population, form polygene type MOLECULE DESIGN first-filial generation commercialization population.
2-3：
The seed that contains respectively the inbred line of 6 target genes of Rpp9, RppQ, Rpp3, Mdm1, Mdm2 and SCMV1 is bred separately, then seed is mixed according to the ratio of 1:1:1:1:2:2:2, the multiple gene inbred line population that composition is taken as the leading factor with anti-short mosaic disease, enter crossbreed ' the maternal group's of system of Zheng Dan 958 ' production link, and then with former ' Zheng Dan 958 ' male parent ' prosperous 72 ' assembly first generation of hybrid population, form polygene type MOLECULE DESIGN first-filial generation commercialization population.
In commercialization seed, sample by individual plant, carry out Markers for Detection and statistical test, can detect accurately component and the proportioning thereof of each population, so that seed law enforcement.
Embodiment 3:
Take Chang Yihua crop cotton as example, ' middle cotton 49 ' is acceptor, with 6 acceptor individual plants respectively with contain resisting verticillium gene GBVe1, GBVe2, VBR1; Anti-blight gene FW
r; The donor hybridization of anti insect gene BT, Sck, then use respectively that ' middle cotton 49 ' backcrosses, each individuality of selecting to import target gene in backcross progeny, and other proterties is to ' middle cotton 49 ' similar or better individual, and repeated multiple times backcrossing, finally obtain respectively containing GBVe1, GBVe2, VBR1, FW
r, BT and Sck 6 target genes, other comprehensive agronomy proterties are with ' middle cotton 49 ' consistent or better 6 stable objects gene strains, breed separately it.
The molecular labeling of above-mentioned each gene or external source marker gene can be selected to be less than 15cM distance from the document of publishing and target gene.
Giving an example in the one source that table 3 provides wherein, but is not limited to these sources.
Table 3: genetic donor material and mark source thereof are for example
3-1：
GBVe1, GBVe2, VBR1, FW will be contained respectively
r, BT and Sck the seed that is that backcrosses of 6 target genes breed separately, according to each be seed mixed in equal amounts, it is population that the multiple gene of composition bollworm resisting, verticillium wilt and fusarium wilt equilibrium backcrosses, and enters production link.
3-2：
GBVe1, GBVe2, VBR1, FW will be contained respectively
r, BT and Sck the seed that is that backcrosses of 6 target genes breed separately, according to each be seed, 2:2:2:2:1:1 ratio is mixed, forming that multiple gene withered to resist, that verticillium wilt is taken as the leading factor backcrosses is population, enters production link.
3-3：
GBVe1, GBVe2, VBR1, FW will be contained respectively
r, BT and Sck backcrossing of 6 target genes be that seed is bred separately, according to each be that seed 1:1:1:1:2:2 ratio is mixed, it is population that the multiple gene that composition bollworm resisting is taken as the leading factor backcrosses, and enters production link.
3-4: 2 strain phase mutual crosses that contain GBVe1, GBVe2 gene are formed to the first generations of hybrid, with first generation of hybrid seed and contain respectively VBR1, FW
r, BT and Sck 4 strains form pure lines and the mixed population of taking of the first generation of hybrid according to the ratio collocation of 1:3:3:3:3.
3-5: by containing the strain of GBVe1, GBVe2 gene, respectively at the strain cross of VBR1 gene, then backcross and select the stable individuality that contains 3 resisting verticillium genes, 3 valency resisting verticillium genes backcross and are, then with contain respectively FW
r, BT and Sck 3 strains to form new backcrossing according to the ratio collocation of 1:1:1:1 be population.
In commercialization seed, sample by individual plant, carry out Markers for Detection and statistical test, can detect accurately component and the proportioning thereof of each population, so that seed law enforcement.
Embodiment 4:
Take asexually propagated crop potato as example, kind ' No. 2, middle potato ' is acceptor, with 5 acceptor individual plants respectively with contain anti-late blight gene (Rpil, R10); The donor hybridization of resistance to bacterial wilt gene (1,2,3), then use respectively ' No. 2, middle potato ' to backcross, each individuality of selecting to import target gene in backcross progeny with molecular labeling, and other proterties is similar to ' No. 2, middle potato ' or better individual, and carry out Comprehensive Traits selection, obtain respectively containing anti-late blight gene (Rpil and R10) and resistance to bacterial wilt gene, other comprehensive agronomy proterties and ' No. 2, middle potato ' similar or better 5 target gene strains, carry out separately vegetative propagation to it.
The molecular labeling of above-mentioned each gene or external source marker gene can be selected to be less than 15cM distance from the document of publishing and target gene.
Giving an example in the one source that table 4 provides wherein, but is not limited to these sources.
Table 4: genetic donor material and mark source thereof are for example
4-1：
To contain respectively 2 target genes of Rpi1, R10 and the clone of disease-resistant gene 1, disease-resistant gene 2 and disease-resistant gene 3 is bred separately, according to each clone potato seed 1:1:1:1:1:1 collocation, the multiple gene that forms anti-late blight and the bacterial wilt equilibrium clone potato seed group that backcrosses, enters production link.
4-2：
The clone of 5 target genes that contain respectively Rpi1, R10, disease-resistant gene 1, disease-resistant gene 2 and disease-resistant gene 3 is bred separately, according to each clone potato seed 2:2:1:1:1 ratio collocation, the composition multiple gene of taking as the leading factor with the anti-late blight clone potato seed group that backcrosses, enters production link.
4-3：
The clone of 5 marker gene that contain respectively Rpi1, R10, disease-resistant gene 1, disease-resistant gene 2 and disease-resistant gene 3 is bred separately, according to each clone potato seed 1:1:2:2:2 ratio collocation, the composition resistance to bacterial wilt multiple gene of taking as the leading factor the clone potato seed group that backcrosses, enters production link.
In commercialization seed, sample by individual plant, carry out Markers for Detection and statistical test, can detect accurately component and the proportioning thereof of each population, so that seed law enforcement.
Claims (8)
- Plant to backcross be the Molecular design breeding method of population, it is characterized in that comprising the steps:(1) the donor plant that contains different external source target genes and the Different Individual of recipient plant to be improved are hybridized;(2) will carry out rotational crossing with recipient plant parent again through the recipient plant individuality of hybridization;(3), according to carrying out MOLECULE DESIGN assisted Selection with the closely linked molecular labeling of target gene, stable the backcrossing that incubation contains respectively different target gene is;(4) according to breeding objective collocation design, by 2-100 backcross be or backcross be between hybridization first generation of hybrid composition to contain backcrossing of multiple different target genes be population;(5) it is the seed production of population that the proportioning during according to collocation design population backcrosses, and is population product thereby produce business-like backcrossing.
- Plant according to claim 1 to backcross be the Molecular design breeding method of population, it is characterized in that described recipient plant is self-pollinated plant, cross-pollinatd plant, Constantly allogamous plant or asexually propagated plant.
- Plant according to claim 1 to backcross be the Molecular design breeding method of population, it is characterized in that described donor plant is the plant that can hybridize with recipient plant.
- Plant according to claim 1 to backcross be the Molecular design breeding method of population, it is characterized in that described external source target gene be can increase the gene of recipient plant to biological stress resistance, can increase the gene of recipient plant to abiotic stress defensive ability/resistance ability, can improve recipient plant output gene, can improve recipient plant quality characteristic gene, can increase the gene of recipient plant adaptive capacity.
- Plant according to claim 1 to backcross be the Molecular design breeding method of population, it is characterized in that after donor plant and recipient plant hybridization, backcross and carry out 1-15 time, form Other Main Agronomic Characters similar to recurrent parent or from stable the backcrossing of donor plant target gene be better.
- Plant according to claim 5 to backcross be the Molecular design breeding method of population, after it is characterized in that donor plant and recipient plant hybridization and backcrossing, backcross between being and carry out again phase mutual cross, form the first generation of hybrid; Or backcross, stable the backcrossing that forms 1-10 target gene of polymerization is again.
- Plant according to claim 6 to backcross be the Molecular design breeding method of population, it is characterized in that described MOLECULE DESIGN be using the 2-100 of acquisition stable backcross be or and can propagating materials and or backcross be the first generation of hybrid by phase mutual cross as component, then by population arrange in pairs or groups that forming backcrosses and is pure lines population with or first generation of hybrid population or vegetative propagation population.
- Plant according to claim 7 to backcross be the Molecular design breeding method of population, it is characterized in that described population collocation is the collocation of the each component in population being carried out to different proportion, identify each component by molecular labeling, and detect the proportioning of the each component of population by statistical test, thereby guarantee to carry out accurate seed production, it is the commercialization of population seed that realization backcrosses.
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