CN114774573B - KASP markers associated with southern rust resistance in corn and uses thereof - Google Patents
KASP markers associated with southern rust resistance in corn and uses thereof Download PDFInfo
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Abstract
The invention discloses a KASP marker related to southern rust resistance of corn and application thereof. The invention also discloses a method for identifying or assisting in identifying the southern rust resistance of corn, which comprises the steps of detecting whether genotypes of 1608911 th deoxyribonucleotide of chromosome 10 of corn to be detected are GG, GA or AA, and determining the southern rust resistance according to the genotypes of the corn to be detected: the corn to be tested of the GG genotype and the GA genotype appears to be resistant to southern rust; the corn to be tested of the AA genotype appears to feel southern rust. Experiments prove that: the KASP marker provided by the invention can effectively identify the southern rust resistance of corn, is simple to operate, has accurate and reliable detection results, can be used for identifying the southern rust resistance of corn and assisting in selective breeding, and lays a theoretical foundation for breeding southern rust disease-resistant corn varieties.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a KASP marker related to southern rust resistance of corn and application thereof.
Background
Corn (Zea mays L.) is the grain and cash crop with the largest planting area and the highest total yield in China, and plays an important role in national economy. Southern rust is a serious air-borne fungal disease caused by rust of the multiple piles (Puccinia polysora. Underworkings) and has a wide disease scope. The disease can cause yield reduction and grain quality deterioration, and forms a great threat to the safe production of corn. In the 70 s of our country, corn southern rust was first found in the southwest (le dong, south shore, cliff etc.). In the 90 s, the disease is outbreak in Zhejiang Chunan, and the area of the diseased corns accounts for about 20% of the area of autumn corns. In recent years, southern rust has been epidemic and spreading in yellow-Huai-Hai-Xia corn production areas (Henan, anhui, shandong, etc.) in northern China. If the southern rust is happened in the summer corn area, the leaves are covered by a yellow orange summer spore pile of the germ for about one week, so that the leaves quickly dry up and die, and the production loss is 20% -40%; when in outbreak, the quality of the seeds is reduced, the yield can be reduced by more than 80 percent, and even the seeds are in harvest. Because the resistance level of the corn popularized variety to southern rust is generally not ideal, the control of the disease in production mainly depends on chemical control. However, the prevention and control effect is generally poor due to a plurality of factors such as untimely medication or improper method, pathogen resistance and the like. Therefore, the cultivation and popularization of disease-resistant varieties are the only way to fundamentally solve the danger of southern rust, and also become urgent in our country for corn production.
At present, the cultivation of the disease-resistant corn variety aiming at southern rust is mainly improved by experience and phenotype selection, and the problems of long breeding period, low efficiency, poor predictability and the like exist. Based on the characteristics of the southern rust pathogenic bacteria, the resistance identification mainly depends on natural disease in the field, so that the phenotype identification cannot be normally performed for the low disease area of the southern rust of corn or the year which is difficult to disease due to environmental factors, and the strain breeding speed is greatly reduced. The molecular markers are developed aiming at the disease-resistant genes, so that germplasm resources can be identified, disease-resistant germplasm is screened, and a resistance source is provided for variety breeding; the method can also carry out prospect selection on the improved inbred line, quickly and efficiently improve target characters, obviously improve the breeding efficiency and predictability of the inbred line with excellent disease resistance, and has important promotion effect on accelerating the breeding process of new disease-resistant varieties.
Disclosure of Invention
The invention aims to rapidly and accurately identify the southern rust resistance of corn.
In order to solve the technical problems, the invention firstly provides a method for identifying or assisting in identifying the southern rust resistance of corn.
The method for identifying or assisting in identifying the southern rust resistance of the corn provided by the invention is used for detecting whether genotypes of 1608911 th deoxyribonucleotide of chromosome 10 of the corn to be detected are GG, GA or AA, and determining the southern rust resistance according to the genotypes of the corn to be detected: the corn to be tested for the GG genotype and the GA genotype appears or is candidate to appear to be resistant to southern rust; the corn to be tested of AA genotype is expressed or candidate is expressed as feeling south rust;
The GG genotype is a homozygote of the 1608911 th deoxyribonucleotide of chromosome 10 of a corn genome;
The GA genotype is a heterozygote of G and A of the 1608911 th deoxyribonucleotide of chromosome 10 of the corn genome;
the AA genotype is the homozygote of which the 1608911 th deoxyribonucleotide of chromosome 10 of the corn genome is A.
Further, the method for detecting whether the genotype of the 1608911 th deoxyribonucleotide of the chromosome 10 of the corn to be detected is GG, GA or AA can comprise the following steps: taking genome DNA of corn to be detected as a template, adopting a KASP primer group to carry out PCR amplification, carrying out fluorescent signal scanning on the obtained amplification product, and judging whether the genotype of the 1608911 th deoxyribonucleotide of chromosome 10 of the corn to be detected is GG, GA or AA according to the fluorescent signal.
The KASP primer group consists of an upstream primer F1, an upstream primer F2 and a downstream primer R;
The upstream primer F1 is single-stranded DNA shown in a sequence 1;
The upstream primer F2 is single-stranded DNA shown in a sequence 2;
The downstream primer R is single-stranded DNA shown in a sequence 3.
Further, the PCR amplification system is as follows: 30ng of template DNA, 0.5. Mu.L of 2X KASP MASTER Mix, 0.486. Mu.L of deionized water or ultrapure water, and 0.014. Mu.L of primer working solution.
The PCR amplification procedure was as follows: 95 ℃ for 15min;94 ℃ for 20s; 1min 10 cycles at 61-55 ℃;94 ℃ for 20s;68 ℃ for 1min; 1min 32 cycles at 55 ℃.
Reading a fluorescence signal value by using PHERA starplus SNP fluorescence detector after the PCR amplification is finished, and deriving a genotyping result by using Kraken software to judge whether the genotype of 1608911-deoxyribonucleotide of chromosome 10 of the corn to be detected is GG, GA or AA: if fluorescence signal data of the amplification product of the corn to be detected is blue through Kraken software analysis, the genotype of the corn to be detected is GG; if fluorescence signal data of the amplification product of the corn to be detected is red through Kraken software analysis, the genotype of the corn to be detected is AA; if fluorescence signal data of the amplification product of the corn to be detected is green through Kraken software analysis, the genotype of the corn to be detected is GA.
In order to solve the technical problems, the invention also provides a novel application of the substance for detecting the genotype of the 1608911 th deoxyribonucleotide of the 10 th chromosome of the corn to be detected.
The invention provides application of a substance for detecting genotype of a 1608911 th deoxyribonucleotide of a 10 th chromosome of corn to be detected in any one of the following (a 1) to (a 8):
(a1) Identifying or assisting in identifying southern rust resistance of the corn to be tested;
(a2) Preparing a product for identifying or assisting in identifying the southern rust resistance of the corn to be tested;
(a3) Screening or assisting in screening of southern rust resistant corn varieties or southern rust susceptible corn varieties;
(a4) Preparing and screening or assisting in screening products of southern rust resistant corn varieties or southern rust resistant corn varieties;
(a5) Improving corn varieties;
(a6) Preparing a corn variety improved product;
(a7) Breeding corn;
(a8) And (5) preparing a corn breeding product.
In order to solve the technical problems, the invention also provides a product as described in any one of the following (b 1) - (b 3):
(b1) The above-mentioned KASP primer set;
(b2) A PCR reagent comprising the KASP primer set of (b 1);
(b3) A kit comprising the KASP primer set of (b 1) or the PCR reagent of (b 2).
The use of the above-mentioned products in any of the following (c 1) - (c 4) is also within the scope of the present invention:
(c1) Identifying or assisting in identifying southern rust resistance of the corn to be tested;
(c2) Screening or assisting in screening of southern rust resistant corn varieties or southern rust susceptible corn varieties;
(c3) Improving corn varieties;
(c4) And (5) breeding corn.
In order to solve the technical problems, the invention also provides a method for screening or assisting in screening of the southern rust resistant corn varieties or the southern rust susceptible corn varieties.
The method for screening or assisting in screening the southern rust resistant corn varieties comprises the step of selecting the corn varieties with GG genotype or GA genotype;
The GG genotype is a homozygote of the 1608911 th deoxyribonucleotide of chromosome 10 of a corn genome;
The GA genotype is a heterozygote of G and A of the 1608911 th deoxyribonucleotide of chromosome 10 of the corn genome.
The method for screening or assisting in screening the southern rust-infected corn varieties comprises the step of selecting the corn varieties with AA genotype;
the AA genotype is the homozygote of which the 1608911 th deoxyribonucleotide of chromosome 10 of the corn genome is A.
In order to solve the technical problems, the invention finally provides a method for improving corn varieties.
The method for improving the corn variety provided by the invention comprises the step of breeding the southern rust-resistant corn variety screened by the method as a breeding material. The improvement is southern rust resistance property improvement.
Further, the breeding method may include the steps of: and backcrossing the southern rust-resistant corn variety serving as a donor parent and the southern rust-sensitive corn variety serving as a recurrent parent. The number of backcrosses may be three.
Further, the donor parent is Beijing 2416K, and the recurrent parent is Beijing 92H, beijing X005, beijing 2416C92 or Beijing 2416B92.
The application of any of the above methods in maize breeding is also within the scope of the present invention. The purpose of the breeding is to cultivate southern rust resistant corn varieties.
The method, application, or product of any of the above, wherein the maize reference genome version number is b73_refgen_v4.
In any of the methods, applications or products described above, the corn may specifically be a corn inbred line jing 2416K, a corn inbred line jing 2416, a corn inbred line jing 724, a corn inbred line jing MC01, an F 1 population individual of corn inbred line jing 2416 and corn inbred line jing 2416K, an F 2 segregating population individual of corn inbred line jing 2416 and corn inbred line jing 2416K, and a corn inbred line jing 2416K as recipient parents, a corn inbred line jing 92H, a corn inbred line jing X005, a corn inbred line jing 2416C92, or a corn inbred line 2416B92 as recurrent parents, a BC 3F2 population individual obtained by 3 backcross and 1 inbred, respectively.
According to the invention, firstly, by comparing the difference sites of the southern rust disease-resistant inbred line and the disease-susceptible inbred line of corn, a SNP site related to the southern rust disease resistance of corn is found, which is positioned at 1608911 th site of chromosome 10 of corn genome, the polymorphism of the site is G/A, and then KASP marker KM23 is obtained according to the development design of the SNP site. Experiments prove that: the KASP marker KM23 provided by the invention can effectively identify the southern rust resistance of corn, is simple to operate, has accurate and reliable detection results, can be used for identifying the southern rust resistance of corn and assisting in selective breeding, and lays a theoretical foundation for breeding southern rust disease-resistant corn varieties.
Drawings
FIG. 1 is a SNP site associated with southern rust resistance of maize.
FIG. 2 is a graph of KM23 markers associated with southern rust resistance in maize versus KASP typing of different inbred lines. Red: a (AA genotype); blue: g: G (GG genotype); green: g: A (GA genotype); black: NTC blank.
FIG. 3 is a graph showing the F 2 population individual phenotype and genotyping of KM23 markers. 1-10: isolating disease resistant individuals in the population; 11-15: and isolating the infected individuals in the population.
FIG. 4 shows the use of KM23 marker in backcross transformation. A: backcross transformation flow chart. B: number of plants of each genotype in the BC 3F2 population. Wherein, B1: backcross populations of Beijing 2416K and Beijing 92H; b2: backcross populations of Beijing 2416K and Beijing X005; b3: a backcross population of jing 2416K and jing 2416C 92; b4: a backcross population of jing 2416K and jing 2416B 92. C-F: phenotype and genotype of individual plants of 4 backcross populations. RR stands for homozygous disease resistance; rr represents heterozygous disease resistance; rr represents a disease.
Detailed description of the preferred embodiments
The following detailed description of the invention is provided in connection with the accompanying drawings that are presented to illustrate the invention and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the invention in any way.
The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are carried out according to techniques or conditions described in the literature in the field or according to the product specifications. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
Corn inbred Jing 2416: the maize research center of the academy of agriculture and forestry science in Beijing city breeds, new variety application number: 20080695.5, grant bulletin number: CNA004319G. The maize inbred line jing 2416 is a high-sensitivity southern rust phenotype.
Maize inbred line jing 2416K: the maize research center of the academy of agriculture and forestry science in Beijing city breeds, new variety application number: 20170479.9, application bulletin number: CNA017957E. The maize inbred line Jing 2416K is a phenotype with high resistance to southern rust.
Maize inbred line jing 92H: the maize research center of the academy of agriculture and forestry science in Beijing city breeds, new variety application number: 20162099.6, application bulletin number: CNA016888E. The maize inbred line Beijing 92H is a high-sensitivity southern rust phenotype.
Maize inbred line jing X005: the maize research center of the academy of agriculture and forestry science in Beijing city breeds, new variety application number: 20160549.6, grant bulletin number: CNA014512G. The maize inbred line Beijing X005 is a high-susceptibility southern rust phenotype.
Maize inbred line jing 2416C92: the maize research center of the academy of agriculture and forestry science in Beijing city breeds, new variety application number: 20184308.7, application bulletin number: CNA025328E. The maize inbred line jing 2416C92 is a high-susceptibility southern rust phenotype.
Maize inbred line jing 2416B92: the maize research center of the academy of agriculture and forestry science in Beijing city breeds, new variety application number: 20184313.0, application bulletin number: CNA025333E. The maize inbred line Beijing 2416B92 is a high-susceptibility southern rust phenotype.
Maize inbred line jing 724: the maize research center of the academy of agriculture and forestry science in Beijing city breeds, new variety application number: 20110278.8, grant bulletin number: CNA005913G. The maize inbred line Beijing 724 is a high-sensitivity southern rust phenotype.
Maize inbred line jingmc 01: the maize research center of the academy of agriculture and forestry science in Beijing city breeds, new variety application number: 20151547.7, grant bulletin number: CNA011488G. The maize inbred line Beijing MC01 is a high-sensitivity southern rust phenotype.
EXAMPLE 1 development of KASP markers associated with southern rust resistance in corn
1. Discovery of SNP markers related to southern rust resistance in corn
By comparing the differential sites of maize inbred lines Beijing 2416K (disease resistance), beijing 2416 (disease sensitivity), beijing 724 (disease sensitivity) and Beijing MC01 (disease sensitivity), a SNP site related to the southern rust resistance of maize was found, which was located at position 1608911 of chromosome 10 of the maize genome (reference genome version number: B73_RefGen_v4) (FIG. 1), and the polymorphism at this site was G/A.
2. Development of KASP markers associated with southern rust resistance in maize
Based on the SNP site found in the first step, a KASP marker KM23 was designed, and the KM23 marker information is specifically shown in Table 1.
Table 1 KM23 flag information
Note that: the underlined sequence in the upstream primer F1 is FAM fluorescent sequence, and the underlined sequence in the upstream primer F2 is HEX fluorescent sequence; y is a degenerate base and represents bases C and T.
3. Verification of the KASP marker
Test material: jing 2416K (disease resistance), jing 2416 (disease susceptibility), jing 724 (disease susceptibility), jing MC01 (disease susceptibility), and the F 1 population of Jing 2416 and Jing 2416K.
1. Identification of southern rust resistance of corn
Identification of southern rust resistance in maize is based on natural morbidity in three parts of the southwest. Investigation was conducted from the maize milk maturation stage to the wax maturation stage (about 15 days after pollination). The key points of investigation are 3 leaves above and below the corn ear. And adopting 1, 3, 5, 7 and 9 grade grading standards, recording the disease grade by parts of materials according to disease symptom description, and carrying out disease resistance comprehensive evaluation according to the disease grade. The disease resistance grade classification standard of the southern rust of the corn is shown in table 2.
TABLE 2 disease resistance classification of southern rust in corn
| Grade of illness | Description of symptoms | Resistance to |
| 1 | Allergic reactions or lesions with or without only sporopiles on leaves | High Resistance (HR) |
| 3 | The spore pile on the leaf is small, the area of the leaf is less than 25% | Anti (R) |
| 5 | The spore pile on the leaf is medium and occupies 26 to 50 percent of the leaf area | Middle Resistance (MR) |
| 7 | The spore pile on the leaf is a large number and occupies 51 to 75 percent of the leaf area | Disease (S) |
| 9 | The spore pile on the leaf is a large number, which occupies 76-100% of the leaf area, and the leaf is dead | High Sense (HS) |
2. Genotyping
Genotyping was performed on the F 1 individuals of maize inbred lines Jing 2416K (disease resistance), jing 2416 (disease sensitivity), jing 724 (disease sensitivity), jing MC01 (disease sensitivity) and Jing 2416K by using KM23 markers. The specific method comprises the following steps: KASP high-throughput SNP genotyping was performed according to the standard experimental procedures of Laboratory of the Government Chemist (LGC) company, which were mainly: extracting corn leaf genome DNA by using plant DNA extraction kit, and then passing through NanoDetecting the quality and measuring the concentration by a spectrophotometer, and finally diluting to about 30 ng/mu L for later use; opening Kraken a corresponding project of software, inputting the name and the bar code number of a sample plate, and setting the positions of a DNA sample, a control sample and an NTC blank control; sample information is imported, and the sample information is filled in according to a template, wherein the sample information comprises a sample bar code number, a sample plate number, a hole site number, a sample plate type and a sample plate bar code number; transferring 1.5 mu L of each DNA sample into a corresponding plate by using a Replikator pore plate replicator, and drying at 55 ℃; preparing reaction liquid according to KASP genotyping standard reaction system of LGC company, wherein the total volume of each reaction is 1 μl, each reaction liquid comprises 0.5 μl of 2× KASP MASTER Mix,0.486 μl of deionized water or ultrapure water, and 0.014 μl of primer working solution, separating liquid by Merdian micropipette after preparation, centrifuging, checking whether there is a defect in the hole by using electricity, and sealing with Fusion laser sealing film instrument; PCR amplification was performed in a high throughput water bath thermal cycler, PCR amplification procedure: 95 ℃ for 15min;94 ℃ for 20s; 1min 10 cycles at 61-55 ℃;94 ℃ for 20s;68 ℃ for 1min; 1min 32 cycles at 55deg.C; after amplification, a PHERA starplus SNP fluorescence detector is used for reading the fluorescence signal value, and Kraken software is used for deriving the genotyping result.
The results show that: the genotype of Jing 2416K is GG (homozygous disease resistance, blue); the genotypes of the F 1 single plants of Jing 2416 and Jing 2416K are GA (heterozygous disease resistance, green); genotypes of jing 2416, jing 724, and jing MC01 were AA (disease-causing, red) (fig. 2).
Example 2 use of the KASP marker KM23 associated with southern rust resistance in maize
1. Identification of southern rust resistance of maize under test
Test material: and separating 15 single plants of the colony from F 2 assembled by Jing 2416 and Jing 2416K.
The test materials were subjected to southern rust resistance identification and genotyping according to the method in example 1.
The results show that 10 disease-resistant individuals and 5 disease-sensitive individuals are all obtained in 15 individuals, 4 genotypes in the 10 disease-resistant individuals are GG, and 6 genotypes are GA; the genotypes of the 5 susceptible individuals were all AA (FIG. 3).
2. Improvement of corn variety
The maize inbred lines Beijing 92H, beijing X005, beijing 2416C92 and Beijing 2416B92 are excellent inbred lines bred by the institute of corn at the national academy of sciences of agriculture and forestry in Beijing city, have high mating strength and excellent comprehensive agronomic characters, but are sensitive to southern rust. The invention takes the inbred line Jing 2416K with high resistance to the southern rust disease of corn as a resistance source, and improves the southern rust disease resistance of the susceptible inbred lines Jing 92H, jing X005, jing 2416C92 and Jing 2416B92, thereby obtaining the anti-rust Jing 92H, anti-rust Jing X005, anti-rust Jing 2416C92 and anti-rust Jing 2416B92. The method comprises the following specific steps:
Beijing 2416K is used as donor parent, beijing 92H, beijing X005, beijing 2416C92 and Beijing 2416B92 are respectively used as recurrent parent, BC 3 is obtained through 3 backcrosses and resistance identification, and BC 3F2 group is obtained through 1 selfing (FIG. 4A). The BC 3F2 population of jing 2416K and of jing 92H yielded 208 individuals, the BC 3F2 population of jing 2416K and of jing X005 yielded 237 individuals, the BC 3F2 population of jing 2416K and of jing 2416C92 yielded 252 individuals, and the BC 3F2 population of jing 2416K and of jing 2416B92 yielded 208 individuals (fig. 4B).
Genotyping and resistance identification was performed on all BC 3F2 individuals using the marker KM23 according to the procedure in example 1. The result shows that 52 of the backcross offspring of Beijing 92H are homozygous for disease resistance, and the genotype is GG;101 are heterozygous disease-resistant, and the genotype is GA;55 were susceptible and were of genotype AA (fig. 4B, C). In the backcross offspring of Beijing X005, 57 are homozygous disease resistance, and the genotype is GG;128 are heterozygous disease-resistant, and the genotype is GA;52 were susceptible and were AA in genotype (fig. 4B, D). In the backcross offspring of Jing 2416C92, 63 are homozygous disease-resistant, and the genotype is GG;131 are heterozygous disease resistance, and the genotype is GA;58 were susceptible and were of genotype AA (fig. 4B, E). In the backcross offspring of Jing 2416B92, 56 are homozygous disease-resistant, and the genotype is GG;103 are heterozygous disease-resistant, and the genotype is GA;49 were susceptible and were of genotype AA (fig. 4B, F). The field resistance phenotype of all BC 3F2 individuals was consistent with the marker KM23 genotype.
The result shows that the genotyping result of the KASP marker KM23 on the offspring of the segregating population and the backcross population is consistent with the field resistance phenotype, and the molecular marker is proved to be applicable to the resistance identification of the southern rust of corn and the molecular marker assisted selective breeding.
The present application is described in detail above. It will be apparent to those skilled in the art that the present application can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the application and without undue experimentation. While the application has been described with respect to specific embodiments, it will be appreciated that the application may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the application following, in general, the principles of the application and including such departures from the present disclosure as come within known or customary practice within the art to which the application pertains. The application of some of the basic features may be done in accordance with the scope of the claims that follow.
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Claims (9)
1. A method for identifying or assisting in identifying southern rust resistance of corn, which is to detect whether the genotype of 1608911 th deoxyribonucleotide of chromosome 10 of corn to be detected is GG, GA or AA, and determine southern rust resistance according to the genotype of the corn to be detected: the corn to be tested for the GG genotype and the GA genotype appears or is candidate to appear to be resistant to southern rust; the corn to be tested of AA genotype is expressed or candidate is expressed as feeling south rust;
The GG genotype is a homozygote of the 1608911 th deoxyribonucleotide of chromosome 10 of a corn genome;
The GA genotype is a heterozygote of G and A of the 1608911 th deoxyribonucleotide of chromosome 10 of the corn genome;
the AA genotype is a homozygote of which the 1608911 th deoxyribonucleotide of chromosome 10 of a corn genome is A;
The maize genome has a reference genome version number B73_RefGen_v4.
2. The method according to claim 1, characterized in that: the method for detecting whether the genotype of the 1608911 th deoxyribonucleotide of the chromosome 10 of the corn to be detected is GG, GA or AA comprises the following steps: taking genome DNA of corn to be detected as a template, adopting a KASP primer group to carry out PCR amplification, carrying out fluorescent signal scanning on the obtained amplification product, and judging whether the genotype of 1608911 th deoxyribonucleotide of chromosome 10 of the corn to be detected is GG, GA or AA according to the fluorescent signal;
The KASP primer group consists of an upstream primer F1, an upstream primer F2 and a downstream primer R;
The upstream primer F1 is single-stranded DNA shown in a sequence 1;
The upstream primer F2 is single-stranded DNA shown in a sequence 2;
The downstream primer R is single-stranded DNA shown in a sequence 3.
3. Use of a substance for detecting the genotype of deoxyribonucleotide 1608911 on chromosome 10 of a maize to be tested in any one of the following (a 1) to (a 8):
(a1) Identifying or assisting in identifying southern rust resistance of the corn to be tested;
(a2) Preparing a product for identifying or assisting in identifying the southern rust resistance of the corn to be tested;
(a3) Screening or assisting in screening of southern rust resistant corn varieties or southern rust susceptible corn varieties;
(a4) Preparing and screening or assisting in screening products of southern rust resistant corn varieties or southern rust resistant corn varieties;
(a5) Improving corn varieties;
(a6) Preparing a corn variety improved product;
(a7) Breeding corn;
(a8) Preparing a corn breeding product;
The maize genome has a reference genome version number B73_RefGen_v4.
4. A product of any one of the following (b 1) to (b 3):
(b1) A KASP primer set; the KASP primer group consists of an upstream primer F1, an upstream primer F2 and a downstream primer R; the upstream primer F1 is single-stranded DNA shown in a sequence 1; the upstream primer F2 is single-stranded DNA shown in a sequence 2; the downstream primer R is single-stranded DNA shown in a sequence 3;
(b2) A PCR reagent comprising the KASP primer set of (b 1);
(b3) A kit comprising (b 1) the KASP primer set or (b 2) the PCR reagent.
5. Use of the product of claim 4 in any one of the following (c 1) - (c 4):
(c1) Identifying or assisting in identifying southern rust resistance of the corn to be tested;
(c2) Screening or assisting in screening of southern rust resistant corn varieties or southern rust susceptible corn varieties;
(c3) Improving corn varieties;
(c4) And (5) breeding corn.
6. A method of screening or aiding in screening for a southern rust resistant maize variety comprising the step of selecting a maize variety of the GG genotype or the GA genotype;
The GG genotype is a homozygote of the 1608911 th deoxyribonucleotide of chromosome 10 of a corn genome;
The GA genotype is a heterozygote of G and A of the 1608911 th deoxyribonucleotide of chromosome 10 of the corn genome;
The maize genome has a reference genome version number B73_RefGen_v4.
7. A method of screening or aiding in screening a southern rust-susceptible maize variety comprising the step of selecting a maize variety of AA genotype; the AA genotype is a homozygote of which the 1608911 th deoxyribonucleotide of chromosome 10 of a corn genome is A; the maize genome has a reference genome version number B73_RefGen_v4.
8. A method for improving maize varieties, comprising the step of breeding a southern rust-resistant maize variety selected by the method of claim 6 as a breeding material.
9. Use of the method of any one of claims 6-8 in maize breeding.
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| CN105861647A (en) * | 2016-01-26 | 2016-08-17 | 河南农业大学 | Molecular marker combination realizing close linkage with southern corn rust gene and applications of molecular marker combination |
| CN106282394A (en) * | 2016-10-28 | 2017-01-04 | 中玉金标记（北京）生物技术股份有限公司 | The method of high throughput testing Semen Maydis south rust resistance gene type and test kit thereof |
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| BR112021013923A2 (en) * | 2019-03-11 | 2021-09-21 | Pioneer Hi-Bred International, Inc. | METHODS OF IDENTIFICATION, SELECTION AND PRODUCTION OF RUST RESISTANT HARVEST OF SOUTHERN CORN |
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| CN106282394A (en) * | 2016-10-28 | 2017-01-04 | 中玉金标记（北京）生物技术股份有限公司 | The method of high throughput testing Semen Maydis south rust resistance gene type and test kit thereof |
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