CN107926699A - A kind of method using corn full genome omics technology fast accurate selecting and breeding corn self-mating system - Google Patents
A kind of method using corn full genome omics technology fast accurate selecting and breeding corn self-mating system Download PDFInfo
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- CN107926699A CN107926699A CN201711484640.4A CN201711484640A CN107926699A CN 107926699 A CN107926699 A CN 107926699A CN 201711484640 A CN201711484640 A CN 201711484640A CN 107926699 A CN107926699 A CN 107926699A
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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
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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
- A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
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Abstract
The invention discloses a kind of method using corn full genome omics technology fast accurate selecting and breeding corn self-mating system, traditional breeding method is combined by this method with genome sequencing technology, pass through Agronomic characteristic screening and genotyping, screening is obtained containing parents' merit gene and genetic background is closer to the fine individual plant of target parent, and the stabilization self-mating system for polymerizeing parents' merit is finally bred as by being continuously selfed.Compared with prior art, this method carries out genetic background screening by conventional selection and the genotyping technique of genome sequencing, substantially reduces the Improvement of different generations of self-mating system, just can precisely select the excellent corn inbred line of Comprehensive Traits within only 23 years.
Description
Technical field
It is fast using corn full genome omics technology more particularly to one kind the present invention relates to corn breeding techniques field
The method of fast precisely selecting and breeding corn self-mating system.
Background technology
With deepening continuously for Market Orientation and largely introducing for external fine germplasm resources, the corn that China cultivates
Kind all improves a lot in quality and quantity, and Chinese maize breeder has successively been bred as large quantities of excellent from foreign countries' introduction resource
Good Hybrid, such as tucks in single series, agricultural university's series, Zheng's single series, increases income volume increase for the corn in China and made very big tribute
Offer, but be still short of on the excellent corn breakthrough varieties of Comprehensive Traits.
The breeding practice of corn hybridizating variety proves that the merit of parent is to improve kind Comprehensive Traits and market competition
One of central factor of power, carrying out the selection and breeding of self-mating system using routine techniques, not only the selection and breeding cycle is long, heavy workload, but also due to
Technology is more traditional, and easy leak selects merit single plant to cause to be difficult to determine newly excellent certainly in early generation during early generation selection and breeding
Friendship is monosystem, causes the integrated quality improvement for being applied to the self-mating system of production that breeding new material excellent at present is difficult to be utilized,
The self-mating system with comprehensive excellent character is caused to lack, the development for constraining hybrid maize is broken through.
During breed breeding, preferable germplasm, clearly female parent, male parent self-mating system are selected according to required character requirement, most
Effective method is directly to be made choice according to the genotype of plant, and the fast development of Protocols in Molecular Biology is directly selected to be this
Select and provide possibility.Will rapidly and efficiently selection and breeding polymerize the corn inbred lines of a variety of merits, realize high yield, high-quality, more anti-, wide
The developing goal of suitable, the easy production of hybrid seeds, it is necessary to carry out traditional breeding method and be combined with modern high technology.Modern biotechnology it is fast
The completion of speed development particularly corn genome sequencing, enables corn breeding to be operated from gene level；To be quick, smart
Quasi- selection and breeding polymerize the corn inbred line of excellent trait of multiple resistance and cenospecies has established solid foundation.
The content of the invention
It is an object of the invention to overcome the deficiencies of the prior art and provide a kind of fast using corn full genome omics technology
The method of fast precisely selecting and breeding corn self-mating system, it is intended to which the time limit that solves existing selecting and breeding corn self-mating system is longer, heavy workload, synthesis
The problems such as merit polymerization is inadequate.
The present invention is achieved by the following technical solutions：
The present invention provides a kind of method using corn full genome omics technology fast accurate selecting and breeding corn self-mating system, bag
Include following steps：
(1) using the corn inbred line for treating transformation as female parent, using the corn inbred line containing objective trait as male parent, carry out normal
After advising sexual hybridization, with male parent backcrossing once, in the BC1F1 generations of acquisition, are selfed once again, obtain BC1F2 segregating populations, wherein mesh
Mark character refers to treating some merits lacking in the corn inbred line of transformation；
(2) Agronomic characteristic screening is carried out for segregating population to the BC1F2 of step (1), selection contains male parent Objective
The single plant that shape, optimum selecting contain maternal merit carries out mixed receipts, and plants BC1F2 generations colony of mixed roguing system seed, obtains
BC1F3 colonies；
(3) continue Agronomic characteristic screening to the BC1F3 colonies of step (2), selection containing male parent objective trait,
Optimum selecting contains the single plant of maternal merit；
(4) two generation genome sequencing technologies are utilized, full genome is carried out to the filial generation single plant after parents and step (3) screening
Group sequencing and genotyping, screen containing parents' merit gene and genetic background closer to male parent F3 filial generations single plant into
Row bagging is selfed；
(5) repeat step (4), continuous selfing 1 more than generation are bred as the stabilization self-mating system of polymerization parents' merit.
Further, the objective trait include plant height is shorter, leaf morphology preferably, high yield, stable yields, it is degeneration-resistant, disease-resistant, go out
One or more merits in the characters such as seed rate is high and coordinate force is strong, but not limited to this.
Further, combining ability test is carried out to the single plant after genome sequencing screens, selects the strong list of coordinate force
Strain carries out bagging selfing and sowing.
Further, the selfing process is required for carrying out security isolation.
The present invention has the following advantages compared with prior art：
1) genetic background of corn inbred line is effectively expanded, beneficial to initiative corn new germplasm resource.
2) fast accurate：Genetic background selection is carried out using the genotyping technique based on genome sequencing, so as to contract
The short Improvement of different generations of self-mating system, only 2-3 just can be bred as stable self-mating system, greatly accelerate breeding speed, reduction corn
The problem of genetic recombination single plant that breeding relies on artificial Field Screening and needs to select colony big, good is difficult to find and identifies.
3) it polymerize merit：Agronomic characteristic observation combines genome sequencing, screening polymerization parents' merit
Single plant.
Brief description of the drawings
Fig. 1：The Technology Roadmap of embodiment 1；
Fig. 2：The Technology Roadmap of embodiment 2.
Embodiment
Technical scheme is described in detail below by two groups of specific embodiments, the method being directed to is such as
It is known to those skilled in the art the material such as the conventional method of dawn, reagent used without special instruction, unless otherwise instructed,
It is commercially available products, germ plasm resource used, is commercially available conventional variety, can be directly from Anhui an aromatic plant metioned in ancient books silver high-tech kind industry share
Co., Ltd buys or is directly acquired from national genebank.
Embodiment 1
Present embodiments provide it is a kind of utilize corn full genome omics technology, with plant type is preferable, plant height is shorter, resistance to rich water,
" Zheng 58 " carries out the more excellent jade of fast accurate selection and breeding to drought-enduring, anti-fall, disease-resistant Huang-Huai-Hai core self-mating system to treat transformation parent
The method of rice self-mating system new varieties, its Technology Roadmap is as shown in Figure 1, comprise the following steps：
(1) BC1F2 is obtained for segregating population
With self-mating system " Zheng 58 " is female parent, with Agronomic characteristic is excellent, seed-producing rate is higher, it is resistance to it is close, best in quality from
It is male parent to hand over system " ph6wc ", after the maternal conventional sexual hybridization with male parent progress, with male parent backcrossing once, the BC1F1 generations of acquisition
It is selfed again once, obtains BC1F2 for segregating population, amount to 1031 plants.
(2) BC1F2 is screened for Agronomic characteristic
Agronomic characteristic screening is carried out for segregating population to this 1031 plants of F2, selection plant height is shorter, Agronomic characteristic
Fine individual plant excellent, coordinate force is strong, obtains the preferable fine individual plant of leaf morphology after roguing the weak, amount to 103, mixed
Receive, and plant F2 generations colony of mixed roguing system seed, obtain F3 colonies.
(3) BC1F3 is screened for Agronomic characteristic
Continue Agronomic characteristic screening to BC1F3 colonies, the same step of method (2), obtains 67 fine individual plants.
(4) genome sequencing and screening
Using two generation genome sequencing technologies, to parents, " 67 one's shares of expenses for a joint undertaking of Zheng 58 ", " ph6wc " and step (3) are for excellent
The full-length genome of single plant is sequenced, and step includes：
I) to parents, " Zheng 58 ", " ph6wc " and 67 parts of single plants are numbered, and corresponding plant leaf is taken in seedling stage, utilize
The N96 plant genes groups DNA extraction kit extraction plant leaf genomic DNA of Qiaqen companies；
Ii) genomic DNA to be smashed, connector is connected after being repaired with archaeal dna polymerase, multiple sample mixing carry out electrophoresis, and
The fragment of 300-400bp is recycled, establishes PE400 sequencing libraries；
Iii Illumina Hiseq X10 microarray datasets) are utilized, full-length genome is carried out with the double end sequencing methods of 2 × 150bp
Resurvey sequence, parent be sequenced depth about 50 ×, offspring's single plant sequencing depth about 0.1 ×, can about be completed within one week or so time
Examining order.
Compared and Genetic Variation Analysis identification, acquisition filial generation list by the way that filial generation single plant and parents are carried out whole genome sequence
The fine haplotype map of strain, passes through the fine haplotype map of filial generation single plant, the genotype in analysis favorable genes site, selection
Favorable genes Heterozygosity is low, favorable genes containing parents and genetic background closer to " ph6wc " single plant 16.
This 16 single plants are subjected to combining ability test with ND81-2, B319,7922 self-mating systems at the same time, screening obtains coordinate force
Strong single plant carries out bagging selfing and sowing.
(5) continuous selfing
After repeat step (4), then 1 generation of continuous selfing, stable Elite inbred is obtained, is named as an aromatic plant metioned in ancient books 586C, the selfing
Be shorter plant height, resistance to rich water, it is drought-enduring, resistance to it is close, anti-fall, disease-resistant, seed-producing rate is high and coordinate force is strong, polymerize the excellent spy of parents
Property.
Embodiment 2
Present embodiments provide it is a kind of utilize corn full genome omics technology, with plant type preferably, high temperature resistant, coordinate force height,
" prosperous 7-2 " carries out the more excellent corn inbred line new varieties of fast accurate selection and breeding to wide suitable more anti-self-mating systems to treat transformation parent
Method, its Technology Roadmap is as shown in Fig. 2, comprise the following steps：
(1) acquisitions of the BC1F2 for colony
With self-mating system, " prosperous 7-2 " is female parent, with economical character is excellent, seed-producing rate is high, self-mating system best in quality " ph4cv "
For male parent, after the maternal conventional sexual hybridization with male parent progress, with male parent backcrossing once, in the BC1F1 generations of acquisition, are selfed once again,
BC1F2 is obtained for segregating population, amounts to 958 plants.
(2) BC1F2 is screened for Agronomic characteristic
Agronomic characteristic screening is carried out for segregating population to this 958 plants of BC1F2, selection plant height is shorter, field agronomy
The plant that shape is good, coordinate force is strong, obtains the preferable fine individual plant of leaf morphology after roguing the weak, amount to 91, carries out mixed receipts, and
F2 generations colony of mixed roguing system seed is planted, obtains F3 colonies.
(3) BC1F3 is screened for Agronomic characteristic
Continue Agronomic characteristic screening to BC1F3 colonies, the same step of method (2), obtains 59 fine individual plants.
(4) genome sequencing and screening
Using two generation genome sequencing technologies, to parents, " 59 one's shares of expenses for a joint undertaking of prosperous 7-2 ", " ph4cv " and step (3) are for single plant
Full-length genome be sequenced, step does not do repeated description herein with (4) the step of embodiment 1.By by filial generation single plant and double
Parent carries out whole genome sequence comparison and Genetic Variation Analysis identification, obtains the fine haplotype map of filial generation single plant, and analysis is sub
For the genotype in excellent genes site in the fine haplotype map of single plant, select favorable genes containing parents, Heterozygosity low and
Genetic background obtains BC1F4 closer to 13 progress bagging selfings of single plant of " ph4cv ".
(5) continuous selfing
Repeat step (4), genome sequencing screening is carried out to BC1F4, by the fine individual plant after screening while and ND81-
2nd, B319,7922 self-mating systems carry out combining ability test, and screening obtains the strong single plant of coordinate force and carries out bagging selfing and sowing, to obtain the final product
To stable Elite inbred, an aromatic plant metioned in ancient books 724V is named as, the self-mating system plant type is preferably, high temperature resistant, much wide more suitable anti-, seed-producing rate is high and matches somebody with somebody
It is strong with joint efforts, it has polymerize the good characteristic of parents.
Be above a kind of detailed embodiment of the invention and specific operating process, be using technical solution of the present invention before
Put and implemented, but protection scope of the present invention is not limited to the above embodiments.
Claims (4)
- A kind of 1. method using corn full genome omics technology fast accurate selecting and breeding corn self-mating system, it is characterised in that including Following steps：(1) using the corn inbred line for treating transformation as female parent, using the corn inbred line containing objective trait as male parent, routinely have Property hybridization after, with male parent backcrossing once, in the BC1F1 of acquisition generation, is selfed once again, obtains BC1F2 segregating populations；(2) Agronomic characteristic screening is carried out for segregating population to the BC1F2 of step (1), selection contains male parent objective trait, selects It is preferred that selecting the single plant containing maternal merit carries out mixed receipts, and plants BC1F2 generations colony of mixed roguing system seed, BC1F3 is obtained Colony；(3) Agronomic characteristic screening is continued to the BC1F3 colonies of step (2), selected containing male parent objective trait, preferentially Single plant of the selection containing maternal merit；(4) two generation genome sequencing technologies are utilized, full-length genome survey is carried out to the filial generation single plant after parents and step (3) screening Sequence and genotyping, screen the F3 filial generation single plants containing parents' merit gene and genetic background closer to male parent and are covered Bag selfing；(5) repeat step (4), continuous selfing 1 more than generation are bred as the stabilization self-mating system of polymerization parents' merit.
- A kind of 2. side using corn full genome omics technology fast accurate selecting and breeding corn self-mating system according to claim 1 Method, it is characterised in that the objective trait be selected from plant height is shorter, leaf morphology preferably, high yield, stable yields, it is degeneration-resistant, disease-resistant, go out seed One or more merits of rate height and coordinate force persistent erection.
- A kind of 3. side using corn full genome omics technology fast accurate selecting and breeding corn self-mating system according to claim 1 Method, it is characterised in that combining ability test is carried out to the single plant after genome sequencing screens, select the strong single plant of coordinate force into The selfing of row bagging and sowing.
- A kind of 4. side using corn full genome omics technology fast accurate selecting and breeding corn self-mating system according to claim 1 Method, it is characterised in that the selfing process is required for carrying out security isolation.
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| CN113951134A (en) * | 2021-02-05 | 2022-01-21 | 江苏省农业科学院 | Efficient corn breeding method based on single plant evaluation and whole genome selection technology |
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Effective date of registration: 20210629 Address after: 230000 No.98, Chuangxin Avenue, high tech Zone, Hefei City, Anhui Province Applicant after: ANHUI WIN-ALL HI-TECH SEED Co.,Ltd. Applicant after: Center for excellence and innovation in molecular plant science, Chinese Academy of Sciences Applicant after: SHANGHAI NORMAL University Applicant after: ANHUI KAITAI AGRICULTURE AND ANIMAL HUSBANDRY TECHNOLOGY Co.,Ltd. Address before: 230000 No.98, Chuangxin Avenue, high tech Zone, Hefei City, Anhui Province Applicant before: ANHUI WIN-ALL HI-TECH SEED Co.,Ltd. Applicant before: SHANGHAI INSTITUTES FOR BIOLOGICAL SCIENCES, CHINESE ACADEMY OF SCIENCES Applicant before: SHANGHAI NORMAL University Applicant before: ANHUI KAITAI AGRICULTURE AND ANIMAL HUSBANDRY TECHNOLOGY Co.,Ltd. |