CN-114582426-B - Corn breeding method based on MAS technology

CN114582426BCN 114582426 BCN114582426 BCN 114582426BCN-114582426-B

Abstract

The application provides a corn breeding method based on a MAS technology, which comprises the steps of marking existing corn low temperature resistant quantitative trait locus information to a reference map, obtaining corn low temperature resistant consistent quantitative trait loci through analyzing the marked reference map by using a Meta, obtaining corn germination low temperature resistant quantitative trait locus information based on a strongest low temperature resistant corn inbred line and a weakest low temperature resistant corn inbred line, determining candidate region excavation response low temperature significant difference expression candidate genes of the corn germination low temperature resistant quantitative trait loci based on the consistent quantitative trait locus information and the corn germination low temperature resistant quantitative trait locus information, and carrying out corn breeding according to the candidate genes. The method integrates the low temperature resistant quantitative trait locus information of the preset quantity of corns, evaluates the relationship between the low temperature resistant related quantitative trait locus effect and the position of the corns, provides powerful help for the molecular marker technology which can be applied to the MAS technology in the development process of the corresponding section, and improves the auxiliary breeding efficiency of the MAS.

Inventors

YU TAO
LI SINAN
WANG WEI
ZHAO WEI
WANG MINGQUAN
YANG JIANFEI
LI BO
ZHANG LIGUO
WU ERNA
MENG XIANGYU
ZHANG JIANGUO
CAO JINGSHENG
CAO SHILIANG
HU GUANGHUI
WANG CHENGBO
LI WENYUE
YANG GENGBIN
LI XIN

Assignees

黑龙江省农业科学院玉米研究所
黑龙江省农业科学院玉米研究所

Dates

Publication Date: 20260421
Application Date: 20220111
Priority Date: 20220111

Claims (10)

1. A MAS technology-based maize breeding method comprising: marking the existing corn low temperature resistant quantitative trait locus information to a reference map, and obtaining corn low temperature resistant consistent quantitative trait loci through analyzing the marked reference map by using a Meta; Selecting the strongest low temperature resistant corn inbred line and the weakest low temperature resistant corn inbred line from the corn inbred lines with preset quantity, and obtaining low temperature resistant quantitative character site information of corn germination period based on the strongest low temperature resistant corn inbred line and the weakest low temperature resistant corn inbred line; obtaining two ribonucleic acid data before and after the strongest low temperature resistant corn inbred line and the weakest low temperature resistant corn inbred line are subjected to low temperature stress treatment in a germination period, and obtaining change data before and after the strongest low temperature resistant corn inbred line and the weakest low temperature resistant corn inbred line are subjected to low temperature stress treatment in the germination period; Determining a candidate region of low temperature resistance in the corn germination period based on the consistent quantitative trait locus information and the low temperature resistance quantitative trait locus information in the corn germination period, and mining response low temperature significant difference expression candidate genes according to the change data; And (5) carrying out corn breeding according to the candidate genes.
2. The MAS technology-based maize breeding method according to claim 1, wherein the step of labeling the existing maize low temperature resistant quantitative trait locus information to a reference map and obtaining the maize low temperature resistant consistent quantitative trait locus from the reference map labeled by Meta analysis comprises: Acquiring existing low temperature resistant quantitative trait locus information of corn, wherein the quantitative trait locus information comprises a quantitative trait locus information name, a quantitative trait locus information chromosome, a quantitative trait locus information LOD value, a quantitative trait locus information population type and a quantitative trait locus information size; Constructing a quantitative trait locus information original map based on the existing low temperature resistant quantitative trait locus information of the corn; comparing the original quantitative trait locus information spectrum with the reference spectrum, and adjusting the original quantitative trait locus information spectrum; marking the original quantitative trait locus information to the reference map in proportion by using a ranking function; and obtaining the corn low temperature resistant consistency quantitative character site by analyzing the marked reference map through Meta.
3. The MAS technology-based maize breeding method according to claim 2, wherein the step of obtaining a maize low temperature resistant consistent quantitative trait locus from a Meta-analysis annotated reference map comprises: Determining the position of the quantitative trait locus on a chromosome by analyzing the marked reference map through Meta; and taking the quantitative trait locus with the determined position as the consistent quantitative trait locus.
4. The MAS technology-based maize breeding method according to claim 1, wherein the step of selecting the strongest low temperature resistant maize inbred line and the weakest low temperature resistant maize inbred line among a predetermined number of maize inbred lines comprises: Obtaining experimental data of the preset number of corn inbred lines, wherein the experimental data are continuous field and indoor low temperature resistance identification data of the preset number of corn inbred lines for at least two years; counting the growth data of the preset number of corn inbred lines based on the low temperature resistant identification data, wherein the growth data comprise relative emergence indexes, relative germination rates and relative germination indexes; and determining the strongest low temperature resistant corn inbred line and the weakest low temperature resistant corn inbred line based on the relative emergence index, the relative germination rate and the relative germination index.
5. The MAS technology-based maize breeding method according to claim 1, wherein the step of obtaining low temperature resistant quantitative trait locus information for maize germination based on the strongest low temperature resistant maize inbred line and the weakest low temperature resistant maize inbred line comprises: Constructing a segregation population by taking the strongest low temperature resistant corn inbred line and the weakest low temperature resistant corn inbred line as basic data; Screening the extreme offspring of the segregating population through indoor low temperature resistance identification to obtain extreme offspring data; And carrying out genome resequencing on the extreme offspring data by a cluster separation analysis method to obtain low temperature resistant quantitative trait locus information of the corn in the germination period.
6. The MAS technology-based maize breeding method according to claim 1, wherein the step of obtaining two pieces of ribonucleic acid data of the strongest low temperature resistant maize inbred line and the weakest low temperature resistant maize inbred line before and after germination period low temperature stress treatment comprises: acquiring initial ribonucleic acid data of the strongest low temperature resistant corn inbred line and the weakest low temperature resistant corn inbred line before low temperature stress treatment in germination period; and carrying out germination period low-temperature stress treatment on the strongest low-temperature resistant corn inbred line and the weakest low-temperature resistant corn inbred line to obtain target ribonucleic acid data after germination period low-temperature stress treatment on the strongest low-temperature resistant corn inbred line and the weakest low-temperature resistant corn inbred line.
7. The MAS technology-based maize breeding method according to claim 6, wherein the step of obtaining variation data of the strongest low temperature resistant maize inbred line and the weakest low temperature resistant maize inbred line before and after germination period low temperature stress treatment comprises: comparing the initial ribonucleic acid data with the target ribonucleic acid data to obtain differential expression ribonucleic acid data and remarkable enrichment metabolic pathway data before and after low temperature stress treatment in germination period of the strongest low temperature resistant corn inbred line and the weakest low temperature resistant corn inbred line; The differential expression ribonucleic acid data and the significantly enriched metabolic pathway data are used as the change data.
8. The MAS technology-based maize breeding method according to claim 1, wherein the step of determining a candidate region of low temperature resistance in maize germination based on the consistent quantitative trait locus information and the maize germination low temperature resistance quantitative trait locus information, mining a candidate gene in response to significant differential expression at low temperature according to the variation data comprises: comparing and screening transcriptome significant differential expression genes positioned in the consistent quantitative trait locus information and the low temperature resistant quantitative trait locus information of the corn germination period; the transcriptome significantly differentially expressed genes are taken as candidate genes for low temperature significantly differential expression in response.
9. The MAS technology-based maize breeding method according to claim 1, further comprising, prior to the step of maize breeding according to the candidate gene: and verifying the candidate gene of the response low-temperature significant differential expression.
10. The MAS technology based maize breeding method according to claim 1, where the reference profile is the maize IBM 2 20088 neighbors reference profile.

Description

Corn breeding method based on MAS technology Technical Field The application relates to the technical field of breeding, in particular to a corn breeding method based on MAS technology. Background The optimal growth temperature of the corn is 28-32 ℃, and the germination, growth and development of seeds and the accumulation of dry matters are very sensitive to the temperature. After corn sowing, if the average temperature is reduced to below 8 ℃, the seeds can be subjected to powder planting, rot or even difficult seedling formation after the average temperature is continued for 3-4 days, so that serious yield reduction is caused. The low temperature cold damage of corn in germination period is a serious problem in middle and high latitude corn producing areas in the world. In the northeast China, the low-temperature cold injury is serious, and the corn is even reduced by more than 20 percent. The average occurrence rate of the low-temperature cold injury in the Heilongjiang province is highest and reaches 38 percent from the aspect of the occurrence frequency of the low-temperature cold injury in 1961 to 2007. The breeding and creating of corn germplasm with strong low temperature resistance in germination period is the most economical and effective method for solving low temperature hazard. The low temperature resistance of corn is a complex quantitative character controlled by micro-effect multiple genes, and the difficulty of breeding and creating low temperature resistant germplasm by a conventional method is high. With the rapid development of biotechnology, students at home and abroad have developed some beneficial exploration on the research of low-temperature resistant molecular design breeding of corn, but the main effect QTL or key genes are not positioned at present. In recent years, the genetic improvement process of crop stress resistance traits is accelerated by means of MAS molecular marker assisted breeding technology, but because most genes for controlling crop stress resistance traits are quantitative trait inheritance, the MAS technology is limited by a plurality of factors of QTL positioning and has not been widely applied to corn breeding practice. Disclosure of Invention In view of the above, the present application aims to provide a MAS technology-based corn breeding method, which is characterized in that IBM2 2008Neighbors is used as a reference map, and through a meta-analysis method, 295 corn low temperature resistant related QTLs are integrated, so as to obtain a 'consistent QTL' with larger effect values, which can be detected under different conditions, and evaluate the relationship between the corn low temperature resistant related QTL effect and the position, thereby providing a powerful help for developing molecular markers applicable to MAS technology in the corresponding sections in the future, and greatly improving the auxiliary breeding efficiency of MAS. The method has the advantages that the method is limited by various factors, all target QTLs are difficult to obtain by one-time QTL positioning, the QTL phenotype contribution rate is poor in reliability, the method is used for researching and adopting Meta to integrate the pre-human-corn low-temperature-resistant related QTL to provide more reliable molecular markers for developing MAS, the problem of low marker density exists in a genetic map constructed by the conventional QTL positioning, the reference value for fine positioning is limited, the MAS efficiency is low, the research is carried out by integrating the pre-human results, the corn low-temperature-resistant related QTL integration map is constructed, the molecular markers of the map are abundant, MQTL which can be detected in different QTL positioning can be obtained by virtue of the Meta technology, and the MAS efficiency is improved by developing molecular markers closely linked with MQTL. The embodiment of the application provides a corn breeding method based on MAS technology, which comprises the following steps: marking the existing corn low temperature resistant quantitative trait locus information to a reference map, and obtaining corn low temperature resistant consistent quantitative trait loci through analyzing the marked reference map by using a Meta; Selecting the strongest low temperature resistant corn inbred line and the weakest low temperature resistant corn inbred line from the corn inbred lines with preset quantity, and obtaining low temperature resistant quantitative character site information of corn germination period based on the strongest low temperature resistant corn inbred line and the weakest low temperature resistant corn inbred line; obtaining two ribonucleic acid data before and after the strongest low temperature resistant corn inbred line and the weakest low temperature resistant corn inbred line are subjected to low temperature stress treatment in a germination period, and obtaining change data before and after the strongest