CN109536629B - Method for rapidly improving corn breeding population - Google Patents
Method for rapidly improving corn breeding population Download PDFInfo
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- CN109536629B CN109536629B CN201811268759.2A CN201811268759A CN109536629B CN 109536629 B CN109536629 B CN 109536629B CN 201811268759 A CN201811268759 A CN 201811268759A CN 109536629 B CN109536629 B CN 109536629B
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- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
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- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
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- C12Q2600/00—Oligonucleotides characterized by their use
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Abstract
The invention provides a method for rapidly improving a corn breeding population, which comprises the following steps: carrying out genotype identification on a basic population by using a high-density SNP molecular marker, selecting an excellent single plant population by combining with field phenotype identification, and rapidly producing a large number of DH lines by using a dihaploid technology; performing multipoint field phenotype identification and combining ability measurement on DH groups, selecting single plants with excellent performance to perform molecular marker identification, analyzing and excavating target character association marker sites; establishing a whole genome selection model; screening a plurality of top-quality DH lines by using the established whole genome selection model, and performing mixed pollination to form a new round of basic population; secondly, a double haploid technology is utilized to enable the basic population to rapidly produce a large number of DH lines, a whole genome selection model is utilized to carry out phenotype prediction screening, the screened excellent plants are subjected to multipoint field phenotype identification, and excellent inbred lines are screened; and (5) circularly improving the basic population and breeding more excellent inbred lines.
Description
Technical Field
The invention relates to a breeding technology, in particular to a method for rapidly improving a corn breeding population.
Background
Maize breeding is one of the important directions for agricultural research, and cross breeding is an important mode of maize breeding. In corn cross breeding, creation of excellent breeding basic materials is the key of breeding success and failure. The common population creating methods comprise population improvement, pedigree selection, backcross breeding and the like, but the breeding time of the methods is long, 6-8 generations (3-5 years) are often needed, the genetic gain is small, and the breeding efficiency is low. There are specific problems as follows: 1. low generation population material (e.g. S) 1 -S 3 ) Is in a heterozygous state, and the phenotype identification is inaccurate due to the existence of recessive and gene interaction, so that the phenotype result of the final homozygous inbred line cannot be completely represented, and the prior art usually identifies and screens high-generation materials. 2. The improvement period is long, because the low-generation heterozygous families or single plants are identified and improved, the selected single plants and families can not be directly used for breeding hybrids, and multiple generations of selfing are needed to be carried out, and the hybrid mating group is carried out after pure lines are formed. 3. The prior art does not effectively combine a molecular identification method with a modern biological statistics technology, has insufficient precision on the molecular genetic background identification and genetic diversity control of a group family or a single plant, has low efficiency on the screening and identification of target characters, and can not effectively utilize breeding data. In this regard, there is a pressing need to improve breeding efficiency using modern biotechnology.
The Double Haploid (DH) technology is a typical representative of the modern biological breeding technology, 100% homozygous inbred line can be obtained through 2 generations in a parthenogenesis mode, and the creation of the maize inbred line by utilizing the DH technology is an important technical means of a variety company at present. The whole Genome Selection (GS) technology is a molecular breeding selection technology newly developed after 2000 years, is a breeding prediction method developed on the basis of modern statistics and high-throughput molecular marker rapid development, has the capability of predicting phenotypes according to individual genotypes, such as the capability of predicting the phenotypes of DH phenotypes and filial generations, and can further rapidly perform phenotype identification. It can be seen that DH technology solves the problem of rapid creation of maize inbred line, GS solves the problem of large-scale screening and phenotype identification of inbred line. A DH + GS technology is integrated, a DH + GS-based breeding technology system is established, breeding materials can be created quickly, and breeding research and development can be carried out quickly. The molecular marker assisted technology can identify and mine target character sites more accurately, and ensures that the target characters of selected strains are excellent in performance.
Disclosure of Invention
Aiming at the following problems existing in the existing population improvement technology: 1) The cycle is long, and 5-8 generations are usually passed; 2) Phenotype identification is difficult, the prior art usually identifies heterozygous populations, and the trait identification is inaccurate due to the existence of the obvious and implicit effects; 3) The genetic gain is low, and the genetic progress is low because the heterozygous population is selected and identified; 4) The direct breeding of hybrid seeds is difficult to carry out, and the improved offspring is difficult to directly combine with the hybrid seeds because the heterozygous population is improved. The method for rapidly improving the breeding population of the corn is used for rapidly improving the breeding population by combining a double haploid technology, a molecular marker auxiliary technology and a whole genome selection technology.
The invention relates to a method for rapidly improving a corn breeding population, which comprises the following steps:
a) Firstly, the high-density SNP molecular marker is utilized to pair the basic population P n Performing identification, combining with field phenotype identification, selecting excellent single plant population, and rapidly producing numerous DH lines by using dihaploid technology to obtain D n A population; to D n Carrying out multipoint field phenotype identification and combining ability identification on the colony, and selecting a plurality of excellent single plants to carry out identification by using high-density SNP molecular markers; analysis D n Phenotype identification results and molecular marker identification results of the population are obtained, so as to mine marker sites related to the target characters; establishing a whole genome selection model by combining a phenotype identification result, a combining ability identification result, a target character associated marker locus and a molecular marker identification result;
b) Using the established whole genome selection model in D n Selecting a part of DH lines with the best target characters from the population, mixing and pollinating to form a new round of basic population P n+1 ；
C) Grouping of bases using doubled haploid technologyBody P n+1 Rapid production of large numbers of DH lines to obtain D n+1 A population; identification of D by high-Density SNP molecular marker n+1 The group, utilize the whole genome to select the model to carry on the phenotype prediction to screen against the goal property, carry on the multipoint field phenotype identification to M excellent plants screened out, confirm the excellent DH line bred finally after the phenotype is identified is the inbred line bred, this line can be used in the matched stack hybrid directly;
d) Repeating the steps B-C in a circulating manner, further improving the basic population and breeding more excellent inbred lines;
n and M are both natural numbers.
Wherein, the basal population P n The basal population obtained according to the prior art method or the basal population obtained according to the method of the invention.
The basic population obtained according to the prior art method is a known corn population, or a corn population obtained by selecting an inbred line, a hybrid and the like according to an expected breeding target through inbreeding, hybridization, backcrossing or a combination thereof.
Wherein, the number of the SNP molecular markers used in the high-density SNP molecular markers is more than 10000, preferably 55000.
Wherein the number is numerous, preferably 150 and more, more preferably 500 and more.
The target traits are breeding target traits which are common in the prior art, such as plant height, high panicle position, yield, grain size, grain weight, combining ability and the like.
Wherein, the most elegant part is the part with 5-10% ranking.
The invention also provides application of the method for rapidly improving the corn breeding population in corn variety breeding.
The invention has the beneficial effects that:
firstly, identifying a basic population by using a molecular marker, selecting an excellent single plant population by combining with field phenotype identification, quickly producing a 100% homozygous inbred line (a DH line can be obtained by 2 generations) from the excellent single plant population by using a double haploid technology, carrying out phenotype identification (including a combining ability test) and molecular marker identification on the homozygous inbred line, establishing a whole genome selection model by combining phenotype data and molecular data, then hybridizing the identified excellent inbred line to form a new round of basic population and induce a new DH line, directly carrying out genotype identification on the DH line, planting the excellent DH line, carrying out phenotype verification and assembling hybrid seeds. The method directly identifies the phenotype and the molecule of the homozygous inbred line, the phenotype is accurate, and the genetic diversity is controllable; the identified excellent inbred line can establish a new group on one hand, and can directly assemble hybrid seeds on the other hand for commercial development, thereby improving the breeding efficiency. Meanwhile, the whole genome selection model established by the method can be used for early-stage (seedling stage) identification and screening, so that the time is remarkably saved, and the efficiency is improved.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
In the varieties related in the examples, the variety 909 is a commercially available public variety.
Example 1
1. Constructing a basic comprehensive population according to breeding targets, breeding backgrounds and heterosis population types, wherein 2-5 maize parent inbred lines can be used for mixed pollination construction, or hybrid inbred separation construction can be directly used, the latter is adopted in the embodiment, and particularly F909 in the hybrid is used 2 Generation segregation population construction of the basic population (P) 0 )。
2. The parental material was labeled using 55K high density (preferably over 10000, this practice uses 55K) SNP molecular markers derived from 55K corn chips (Xu c., ren y., jian y., guo z., zhang y., xie c., fu j., wang h., wang g g., xu y., li p., zuo c.development of a mail 55K SNP array with improved genome for molecular breeding,2017, 37).
3. For the basic group P 0 Haploid induction was performed and 150 DH lines were generated in duplicate (D) 0 A population).
4. To D 0 And performing multipoint field phenotype identification and combining ability identification on the colony.
5. Screening D 0 Extracting NDA from individual plant with excellent field performance by using 55K high-density (same as above) SNP molecular marker is used for carrying out marker analysis on the material.
6. Binding of D 0 And performing linkage analysis on the group phenotype and molecular marker data, and mining target character associated marker loci. In this example, 80 marker sites for controlling plant height were excavated, specifically, see table 1, and the markers were derived from a corn 55K chip (Xu c., ren y., jian y., guo z., zhang y., xie c., fu j., wang h., wang g., xu y., li p., zuo c. Development of a ze 55K SNP array with improved gene library for molecular breeding,2017, 37.
TABLE 1 control of plant height marker loci
7. And establishing a whole genome selection model by combining phenotype identification data, character linkage site information and high-density molecular detection data for phenotype prediction of the offspring population. In the embodiment, the plant height expression of 150 DH-line groups derived from a single 909 progeny is predicted, and the prediction accuracy reaches 70.81%.
8. Comprehensive multipoint phenotypic identification at D 0 Screening 10% optimal DH lines from the population, mixing and pollinating to form a new round of basic population (P) 1 )。P 1 The population is selected, is a new improved population, and has target character more than P 0 Is excellent.
9. For comprehensive basic group P 1 Haploid induction and doubling are carried outGeneration of 500 DH lines (D) 1 A population).
10. Extraction of D 1 And (3) carrying out marker analysis on the material by using high-density SNP molecular markers for each pure line DNA of the population, and carrying out phenotype prediction screening by using a whole genome selection model so as to avoid large-scale phenotype determination.
11. For screened D 1 And performing multipoint field phenotype identification on excellent plants in the population, testing and matching, and breeding to obtain 58 excellent inbred lines which are all characterized by moderate plant height, spike position and excellent agronomic characters.
12. Integration of the multipoint phenotypic identification and Whole genome selection predictive screening results, at D 1 Screening 10% optimal DH line in the colony, mixing and pollinating to form a new round of basic colony (P) 2 )。
13. By analogy, new breeding populations and DH lines are continuously created by improvement.
Example 2
The test was carried out in the same manner as in steps 1 to 5 of example 1.
6. A linkage analysis method is used for excavating 80 marked sites for controlling seed weight, and concretely, the marks are derived from a corn 55K chip (Xu C., ren Y., jian Y., guo Z., zhang Y., xie C., fu J., wang H., wang G., xu Y., li P., zou C. Development of a mail 55K SNP array with improved gene coverage for molecular organization, 2017, 37).
TABLE 2 control of seed weight marker sites
7. And establishing a whole genome selection model by combining phenotype identification, character linkage site information and high-density molecular detection data for phenotype prediction of the offspring population. In the embodiment, the established whole genome selection model is used for predicting seed phenotype of 150 DH line populations derived from a single cell 909, and the prediction accuracy reaches 77.71%.
8. Integration of the multipoint phenotype and binding Capacity identification at D 0 Screening 10-15% optimal DH line from the population, mixing and pollinating to form a new round of basic population (P) 1 )。P 1 The population is selected, is a new improved population, and has target character more than P 0 Is excellent.
9. For comprehensive basic group P 1 Haploid induction was performed and 500 DH lines were generated in duplicate (D) 1 A population).
10. Extraction of D 1 And (3) carrying out marker analysis on the material by using high-density SNP molecular markers for each pure line DNA of the population, and carrying out phenotype prediction screening by using a whole genome selection model, so that large-scale phenotype and combining ability determination is omitted.
11. For screened D 1 And performing multipoint field phenotype identification on excellent plants in the population, and breeding to obtain 32 excellent inbred lines which are all represented by high seed weight and the seed weight is increased by 12.3%.
12. Integration of the multipoint phenotypic identification and Whole genome selection predictive screening results, at D 1 Screening 10% optimal DH lines from the population, mixing and pollinating to form a new round of basic population (P) 2 )。P 2 The population is selected, is a new improved population, and has target character more than P 0 、P 1 Is excellent.
13. By analogy, new breeding groups and DH lines are continuously created by improvement.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, some modifications and improvements can be made without departing from the technical principle of the present invention, and these modifications and improvements should also be considered as the protection scope of the present invention.
Claims (2)
1. A method for rapidly improving a corn breeding population, which is characterized by comprising the following steps:
a) Constructing a basic population P by using a single 909F 2 generation self-segregation population n (ii) a Firstly, a 55K high-density SNP molecular marker is utilized to carry out the pair of the basic groups P n Identifying, combining with field phenotype identification, selecting excellent individual plant population, and rapidly producing DH lines of 500 or more by using dihaploid technique to obtain D n A group;
to D n Performing multipoint field phenotype identification and combining ability identification on a group, and selecting a plurality of excellent single plants to be identified by 55K high-density SNP molecular markers;
analysis D n The phenotype identification result and the molecular marker identification result of the population are used for excavating the marker locus associated with the target character;
the target character correlated marker locus is a marker locus for controlling plant height:
or, the target character associated marker site is a control seed weight marker site:
establishing a whole genome selection model by combining a phenotype identification result, a combining ability identification result, a target character associated marker locus and a molecular marker identification result;
b) Using the established whole genome selection model in D n Selecting DH lines with target characters ranking 5-10% from the population, mixing and pollinating to form a new round of basic population P n+1 ；
C) Making the basic population P by using a doubled haploid technique n+1 Rapidly producing more than 150 DH lines to obtain D n+1 A population;
identification of D by 55K high-density SNP molecular marker n+1 The group, utilize the whole genome to select the model to carry on the phenotype prediction to screen against the goal property, carry on the multipoint field phenotype identification to M excellent plants screened out, confirm the excellent DH line bred finally after the phenotype is identified is the inbred line bred, this line can be used in the matched stack hybrid directly;
d) Repeating the steps B-C in a circulating manner, further improving the basic population and breeding more excellent inbred lines;
n and M are both natural numbers.
2. Use of the method of rapid corn breeding population improvement as claimed in claim 1 in corn variety breeding.
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Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110459265B (en) * | 2019-08-14 | 2022-07-05 | 中国农业科学院作物科学研究所 | Method for improving prediction accuracy of whole genome |
| CN111528087A (en) * | 2020-04-17 | 2020-08-14 | 中国农业科学院作物科学研究所 | Method for breeding high-combining ability and excellent germplasm of corn |
| CN111771716B (en) * | 2020-07-24 | 2022-03-04 | 江西省农业科学院作物研究所 | Crop genetic breeding method for efficiently utilizing heterosis |
| CN112365926B (en) * | 2020-11-10 | 2024-02-13 | 沈阳农业大学 | Whole genome prediction method for general and special mating forces of maize inbred line and application thereof |
| CN112687340A (en) * | 2020-12-17 | 2021-04-20 | 河南省农业科学院粮食作物研究所 | Method for breeding corn high-yield material based on whole genome association analysis and whole genome selection |
| CN114793886B (en) * | 2022-06-01 | 2023-07-28 | 新疆农垦科学院 | Method for breeding drought-tolerant male parent germplasm based on maize PB group |
| CN115316266B (en) * | 2022-09-05 | 2023-09-08 | 新疆农垦科学院 | Crop population improvement and variety breeding method |
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