CN118222747A - Wheat leaf rust resistance molecular marker and application thereof - Google Patents
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Abstract
The invention discloses a wheat leaf rust resistance molecular marker and application thereof. The wheat leaf rust resistance molecular marker disclosed by the invention is a nucleotide corresponding to the 71 st position of SEQ ID No.4 in a sequence table on a wheat 5B chromosome, and is C or T. Experiments prove that the wheat leaf rust resistance molecular marker is related to wheat leaf rust resistance, the wheat homozygous with the C nucleotide corresponding to the 71 st position of SEQ ID No.4 in the genome DNA is higher than the wheat homozygous with the T nucleotide corresponding to the 71 st position of SEQ ID No.4 in the genome DNA, and the wheat leaf rust resistance molecular marker can be used for detecting the wheat leaf rust resistance and further used for molecular marker assisted breeding, and has important significance for cultivating the wheat with enhanced leaf rust resistance.
Description
Technical Field
The invention relates to a wheat leaf rust resistance molecular marker and application thereof in the field of biotechnology.
Background
Wheat leaf rust is caused by wheat leaf rust bacteria (Puccinia tirticina), is a fungal disease, mainly damages wheat leaves, and can infect wheat stems and ears when serious. Wheat leaf rust has the characteristics of air transmission, wide popularity, strong destructiveness, parasitic specificity, cyclic infection and the like, and is frequently generated in wheat main production areas in China. Wheat leaf rust is mainly harmful to wheat leaves, reduces the photosynthetic rate of plants, further influences grain grouting, leads to thousand grain weight reduction, can generally reduce the yield by 5% -15%, and can lead the yield loss of susceptible varieties to be up to 40%. The mode of preventing and treating leaf rust by the medicament not only increases the production cost, but also is easy to cause environmental pollution. Cultivation and planting of wheat leaf rust resistant varieties is the most economical, efficient and environmentally friendly control method.
To date, more than 82 leaf rust resistance genes or alleles have been found in common wheat, durum wheat and diploid wheat varieties using a variety of different genetic research approaches, including Lr1 to Lr82, lrAlt and LrTt, among others. However, due to the constant evolution and mutation of Pt races, most initially effective rust resistant genes typically have a very short life span, and genes that remain effectively resistant to production today are only Lr9, lr19, lr24, lr38, lr47, lr51 and Lr53. The Lr genes found to have adult stage resistance are Lr12, lr13, lr22 (a and b), lr34, lr35, lr37, lr44, lr46, lr48, lr49, lr67, lr68, lr74, lr75, lr77 and Lr78, which Lr12, lr13 and Lr22b have race specialization and are involved in cell allergic death, while other genes are non-race specialization genes capable of conferring resistance to a variety of physiological races. Currently, 8 wheat leaf rust resistance genes have been cloned, respectively: lr1, lr10, lr13, lr14a, lr21, lr22a, lr34 and Lr67. The number of reported wheat leaf rust QTLs is 80, and the positions of the QTLs are almost distributed on 21 chromosomes of wheat, except five chromosomes of 1A, 1D, 3D, 6D and 7A, QTL sites are probably not present on the five chromosomes, and QTLs on the five chromosomes are probably still undetected. The markers selected during the positioning of the QTL locus are different, but the markers are the same in the general position of the chromosome, and the QTL locus which can be positioned in different disease-resistant materials has higher stability and wider distribution and is more worthy of deep research by researchers. Because of the rapid mutation speed of wheat leaf rust bacteria, some genes lose resistance, and part of genes are linked with bad shapes, the genes truly capable of being used for improving the leaf rust resistance in wheat breeding are very limited. Therefore, it is important to dig a new leaf rust resistance gene (QTL) and develop molecular markers closely linked thereto for breeding against diseases.
Molecular marker assisted breeding accelerates the traditional breeding process. The single nucleotide polymorphism (single nucleotide polymorphisms, SNP) is a polymorphism in a DNA sequence due to the substitution, deletion or addition of a single base pair in the genome, thereby changing the base sequence. SNP is widely distributed in genome and has high stability, and with the rapid development of molecular biology technology, the SNP has higher use value and larger development space, and people can use the SNP for molecular marker assisted selection. Thus, the competitive allele-specific PCR (kompetitive ALLELE SPECIFIC PCR, KASP) technique for detecting SNPs shows good development prospects, mainly due to the advantages of high throughput, high stability and high accuracy of KASP markers. At present, the KASP labeling technology is very mature and is very widely applied to wheat genetic breeding.
The Jimai 22 is a high-yield, multi-resistant and high-quality medium-gluten wheat variety, is inspected by national Huang-Huai-Bei-pian in 2006, and respectively completes the introduction and recording work of two provinces of Anhui and Henan in 2010 and 2011, and is suitable for planting in suitable areas of Henan and Anhui of the North-Huang-Huai winter wheat and the south-Huang-Huai-winter wheat. The middle wheat 578 is a high-yield, multi-resistant and high-quality strong-gluten wheat variety, is approved by the country in month 6 of 2021, and is suitable for being planted in the northern water of Huang-Huai winter wheat.
Disclosure of Invention
The invention aims to solve the technical problem of how to detect wheat leaf rust resistance.
In order to solve the technical problems, the invention firstly provides application of a substance for detecting wheat leaf rust resistance molecular markers in detecting or assisting in detecting wheat leaf rust resistance;
the wheat leaf rust resistance molecular marker is a nucleotide corresponding to the 71 st position of SEQ ID No.4 in a sequence table on a wheat 5B chromosome, and is C or T.
In the above application, the substance for detecting the wheat leaf rust resistance molecular marker may include: a primer set capable of amplifying a DNA fragment shown at position 71 of SEQ ID No.4 of the sequence Listing.
In the above application, the primer set may be composed of three single-stranded DNAs named upstream primer F1, upstream primer F2 and downstream primer R, respectively, wherein the upstream primer F1 contains the single-stranded DNA shown in positions 22-41 of SEQ ID No.1, the upstream primer F2 contains the single-stranded DNA shown in positions 22-41 of SEQ ID No.2, and the downstream primer R is the single-stranded DNA shown in SEQ ID No. 3.
Specifically, the upstream primer F1 may be a single-stranded DNA shown in SEQ ID No.1, and the upstream primer F2 may be a single-stranded DNA shown in SEQ ID No. 2.
The present invention also provides a method of detecting wheat leaf rust resistance, the method comprising: detecting the leaf rust resistance of the homozygous wheat to be detected, of which the nucleotide corresponding to the 71 st position of SEQ ID No.4 in the sequence table is C, in the genomic DNA, of which the nucleotide corresponding to the 71 st position of SEQ ID No.4 in the sequence table is higher than or is higher than the homozygous wheat to be detected, of which the nucleotide corresponding to the 71 st position of SEQ ID No.4 in the sequence table is T, in the genomic DNA.
In the method, the detection of the nucleotide at position 71 in the genomic DNA of the wheat to be tested, which corresponds to SEQ ID No.4 in the sequence Listing, can be performed by using the substance for detecting the wheat leaf rust resistance molecular marker.
In the method, the detection of the 71 st nucleotide corresponding to SEQ ID No.4 in the genome DNA of the wheat to be detected can be finished by direct sequencing, or can be finished by amplifying three single-stranded DNAs shown as SEQ ID No.1, SEQ ID No.2 and SEQ ID No.3 in a reaction system of KASP primers and then detecting fluorescent signals in the reaction system.
In one embodiment of the present invention, the reaction system may contain KASP 2x Master Mix (LGC, cat# 13448166). The fluorescent signal in the reaction system is shown as FAM, and the nucleotide at the 71 st position in the genomic DNA of the wheat to be detected, which corresponds to SEQ ID No.4 in the sequence table, is C (namely, the wheat to be detected is homozygous for C); the fluorescent signal in the reaction system is shown as HEX, and the nucleotide at the 71 st position in the genomic DNA of the wheat to be detected, which corresponds to SEQ ID No.4 in the sequence table, is T (namely, the wheat to be detected is homozygous for T); the fluorescence signals in the reaction system are shown as FMA and HEX, and the nucleotides corresponding to the 71 st position of SEQ ID No.4 in the sequence table in the genome DNA of the wheat to be detected are C and T (namely the wheat to be detected is heterozygous of C and T).
The detection of the wheat leaf rust resistance molecular marker substance also belongs to the protection scope of the invention.
The substance for detecting the wheat stripe rust resistance molecular marker can be a kit.
The DNA molecule shown in SEQ ID No.4 also belongs to the protection scope of the present invention.
The invention also provides any one of the following applications of the wheat leaf rust resistance molecular marker or the DNA molecule shown in SEQ ID No. 4:
x1) detecting or aiding in the detection of wheat leaf rust resistance;
X2) comparing the leaf rust resistance of different wheat;
x3) breeding strong leaf rust resistant wheat;
x4) screening or rejecting wheat with weak leaf rust resistance;
x5) wheat breeding.
The invention also provides any one of the following applications of the substance for detecting the wheat leaf rust resistance molecular marker:
Y1) preparing a product for detecting or assisting in detecting wheat leaf rust resistance;
Y2) comparing the leaf rust resistance of different wheat;
Y3) preparing products with strong and weak leaf rust resistance of different wheat;
Y4) breeding wheat with strong leaf rust resistance;
y5) preparing a product for breeding wheat with strong leaf rust resistance;
y6) screening or rejecting wheat with weak leaf rust resistance;
y7) preparing a product of wheat selected or knocked out for weak leaf rust resistance;
Y8) wheat breeding.
The invention also provides a wheat breeding method, which comprises the following steps: detecting the nucleotide corresponding to the 71 st position of SEQ ID No.4 in a sequence table in the genome DNA of wheat, selecting homozygous wheat with the 71 st position of C corresponding to the SEQ ID No.4 in the sequence table in the genome DNA as a parent for breeding, and screening homozygous wheat with the 71 st position of C corresponding to the SEQ ID No.4 in the sequence table in the genome DNA of offspring to obtain wheat with leaf rust resistance.
In the present invention, the wheat may be selected from the filial generation of the middle wheat 578 and the ji wheat 22, or 115 parts of wheat material in table 2 or its progeny.
In a specific embodiment of the invention, the leaf rust resistance is manifested by the MDS level of the leaf. A high MDS in the leaf indicates weak leaf rust resistance, and a low MDS in the leaf indicates strong leaf rust resistance.
The wheat leaf rust resistance molecular marker is related to wheat leaf rust resistance, and the wheat homozygous for which the nucleotide corresponding to the 71 st position of SEQ ID No.4 in the sequence table is C in the genomic DNA is higher than the wheat homozygous for which the nucleotide corresponding to the 71 st position of SEQ ID No.4 in the sequence table is T in the genomic DNA. The wheat leaf rust resistance can be detected by using the wheat leaf rust resistance molecular marker, and the wheat leaf rust resistance can be further used for molecular marker assisted breeding, and the wheat leaf rust resistance detection method has important significance for cultivating wheat with enhanced leaf rust resistance.
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.
Drawings
FIG. 1 is a graph of QLr.caas-5BL located in the middle wheat 578 XJi wheat 22RIL population.
FIG. 2 is the genotyping results of KASP marker Kasp _5B_LR on 115 parts of wheat material. Blue is the middle wheat 578 genotype and red is the ji wheat 22 genotype.
Detailed Description
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, instruments and the like used in the examples described below are commercially available unless otherwise specified. In the following examples, unless otherwise specified, the 1 st position of each nucleotide sequence in the sequence listing is the 5 'terminal nucleotide of the corresponding DNA/RNA, and the last position is the 3' terminal nucleotide of the corresponding DNA/RNA.
Example 1 discovery of leaf rust resistance QTL in the middle wheat 578 XJi wheat 22RIL population and acquisition and use of KASP markers
1. Phenotype data acquisition
The F5 RIL group containing 262 families is constructed by taking Zhongmai 578 as a female parent and Jimai 22 as a male parent through a single grain transmission method, 2021-2022 is planted in Henan Xinxiang, 2022-2023 is planted in Henan Xinxiang, henan Zhengzhou and Hebei guarantying test points, a completely random block design is adopted, three times of repetition are adopted, 6 rows of regions are adopted, the row length is 4m, the row width is 0.25m, and 270 seeds are sowed per square meter. In the whole wheat growing process, the field management is carried out according to a local conventional cultivation technology. The field disease investigation was performed on the population and its parents when the leaf rust onset was the most severe (i.e., when the leaves of the disease control Zhengzhou 5389 were nearly full of leaf rust mass). The investigation was maximum severity (MDS), i.e., the percentage of leaf rust spore pile area on the leaf as a percentage of the total leaf area.
Extracting 262 family young leaf genome DNA by adopting a CTAB method, measuring the DNA concentration by using a Nanodrop2000c spectrophotometer, regulating the DNA sample to a standard concentration of 50 ng/. Mu.l, detecting the DNA quality by using 0.8% agarose gel, and carrying out SNP typing on the DNA with qualified quality. SNP analysis was performed using a 50K SNP chip completed by the cooperation of crop science institute and Affymetrix Axiom company of China national academy of agricultural science.
2. Linkage map construction
RILs and two parents were genotyped using CapitalBio Corporation (http:// www.capitalbio.com) wheat 50K SNP chip, containing 55,224 SNPs in total. Genotype data filtering criteria was screening for non-polymorphic markers, families with a marker deletion rate greater than 20% and markers with a minimum allele frequency of less than 30%, the remaining 9354 high quality polymorphic markers were used for subsequent analysis. The filtered polymorphic markers were treated with BIN functions in ICIMAPPING V4.2 (http:// www.isbreeding.net/; meng et al 2015) to separate the same genotyping markers into one BIN, creating 34 linkage clusters, including 1501 BINs. Genetic linkage maps were mapped using JoinMap v4.0 and MAPCHART V2.32 software (https:// www.wur.nl/en/show/Mapchart.htm; voorrips, 2002).
3. QTL analysis
QTL was detected using QTL Cartographer v2.5 complete interval mapping and LOD threshold was chosen to be 2.5. The 5B chromosome was mapped to 1 more stable QTL, which was designated QLr.caas-5BL (FIG. 1). The flanking markers are AX-1105294353 and AX-95660479, and the physical interval is 669.1-702.9Mb. Under different environmental conditions, 3.42-10.55% of the phenotypic variation can be explained (see Table 1, FIG. 1). Its flanking marker AX-95660479 was converted to Kasp _5b_lr and the genotype of 115 parts of wheat material was detected.
TABLE 1 QLr.caas-5BL of the wheat 578 XJimai 22RIL population detected by composite interval mapping method
| Genetic location (cM) | Marking section | Physical location (Mb) | LOD value | PVE(％) | Add |
| 55.51 | AX-1105294353-AX-95660479 | 669.1-702.9 | 2.67-5.82 | 3.42-10.55 | -2.36--5.31 |
4. Design and utilization of KASP primer
1. Design of KASP primer
Corresponding to the Kasp B_LR marker, SNP site AX-95660479 is located at 702.9Mb (website https:// ugi. Versailles. Inra. Fr/blast_ iwgsc /) of the wheat reference genome (5B chromosome) CHINESE SPRING REFSEQ V1.0.0. The SNP variation between the middle wheat 578 and the Jimai 22 for the marker AX-95660479 (sense strand) closely linked to QLr.caas-5BL and its surrounding nucleotides are shown in SEQ ID No.4 (Y stands for C or T).
Designing a KASP (KASP sequence) marker primer according to the antisense strand of the SNP locus marker AX-95660479, wherein the sequence is as follows:
Upstream primer F1:5'-GAAGGTGACCAAGTTCATGCT ACCCCTCAAGTCTGAGCTTG-3' (SEQ ID No.1, underlined is the specific fluorescent tag sequence FAM);
the upstream primer F2:5'-GAAGGTCGGAGTCAACGGATT ACCCCTCAAGTCTGAGCTTA-3' (SEQ ID No.2, underlined is the specific fluorescent tag sequence HEX);
the downstream primer R:5'-GCTGACAAGGACCTCGACG-3' (SEQ ID No. 3).
The last base at the 3' -end of the two upstream primers corresponds to SNP site AX-95660479 (antisense strand). SNP site AX-95660479 is C or T (represented by Y in SEQ ID No. 4) at position 71 of wheat 5B chromosome corresponding to SEQ ID No.4 (sense strand).
The upstream primer F1 is used for amplifying the condition that the nucleotide at the SNP site AX-95660479 (antisense strand) on the wheat 5B chromosome is G (corresponding to the sense strand, the nucleotide at the SNP site is C), and the upstream primer F2 is used for amplifying the condition that the nucleotide at the SNP site AX-95660479 (antisense strand) on the wheat 5B chromosome is A (corresponding to the sense strand, the nucleotide at the SNP site is T); the downstream primer R is a universal primer.
The single-stranded DNA molecule shown in SEQ ID No.1 and the single-stranded DNA molecule shown in SEQ ID No.3 amplify a fragment in which the nucleotide at the SNP site AX-95660479 (antisense strand) on the wheat 5B chromosome is homozygous for G (corresponding to the sense strand, the genotype of the SNP site is C: C homozygote).
The single-stranded DNA molecule shown in SEQ ID No.2 and the single-stranded DNA molecule shown in SEQ ID No.3 amplify a fragment in which the nucleotide at the SNP site AX-95660479 (antisense strand) on the wheat 5B chromosome is homozygous for A (corresponding to the sense strand, the genotype of the SNP site is homozygous for T: T).
The single-stranded DNA molecule shown in SEQ ID No.1, the single-stranded DNA molecule shown in SEQ ID No.2 and the single-stranded DNA molecule shown in SEQ ID No.3 amplify fragments with the nucleotides G and A heterozygous (corresponding to the sense strand, the genotype of the SNP locus is T: C heterozygous) at the SNP locus AX-95660479 (antisense strand) on the wheat 5B chromosome.
2. Establishment of KASP detection method
Principle of KASP: two forward competitive primers (primer 5 'end has base sequence complementary pairing with fluorophore HEX and FAM, other sequences only have difference at SNP of 3' end) and one reverse common primer; the PCR reaction system contains a fluorescent group and a quenching group modified universal sequence (Master Mix is provided by LGC company), two forward primers can emit light with two different colors, if the locus is homozygous, a single fluorescence is emitted, and if the locus is heterozygous, two kinds of fluorescence are emitted simultaneously.
The KASP marker PCR amplification system was as follows: 2.0. Mu.l of KASP 2 XMaster Mix (LGC, cat# 13448166), 0.048. Mu.l of KASP primer (3 primers mixed at a total concentration of 50. Mu.M, with a molar ratio of two upstream primers to one downstream primer of 2:2:5), 1.952. Mu.l of template DNA (50 ng/. Mu.l). Amplification was performed using a 384-well PCR apparatus (BIO-RAD, S1000TMTHERMAL CYCLER) as follows: 94 ℃ for 15min;94 ℃ for 20s, 63-55 ℃ for 1min (1 ℃ drop per cycle), 10 cycles; 94℃for 20s, 55℃for 60s,32 cycles. The PCR amplified products were placed in an autofocus fluorescence multifunctional microplate reader (PHERAstarplus SNP, BMG LABTECH) to read the final fluorescence data, which was then imported into Klustercaller v3.4 (LGC, hoddesdon, UK) for genotyping.
For Kasp _5b_lr marker (SNP site AX-95660479): if the fluorescence signal data of the amplified product is analyzed by genotyping software KlusterCaller to be close to the Y axis (FAM fluorescence signal), the genotype representing the locus (sense strand) is CC homozygous; if the fluorescence signal data of the amplified product is analyzed by genotyping software KlusterCaller to be close to the X axis (HEX fluorescence signal), the genotype representing the locus (sense strand) is TT homozygosity; if the fluorescence signal data of the amplified product is located in the middle between the X-axis and the Y-axis (both FAM and HEX signals are present) by genotyping software KlusterCaller, the genotype representing this site (sense strand) is TC heterozygous.
3. Kasp detection
The experimental materials were 115 parts Huang Huai wheat district materials ((Liu JD,He ZH,Rasheed A,Wen WE,Yan J,Zhang PZ,Wan YX,Zhang Y,Xie CJ,Xia XC(2017)Genome wide association mapping of black point reaction in common wheat(Triticum aestivum L.).BMC Plant Biology,https://doi.org/10.1186/s12870-017-1167-3.), specifically shown in Table 2.
115 Parts of Huang-Huai wheat material are planted in Henan Zhengzhou (year 2020-2021) and Hebei baoding (year 2021-2022). All experiments were performed using a random block design, 3 replicates, 3 rows of blocks, 2m rows long, 25cm row spacing, 50 grains/row. And carrying out field management according to a local conventional management rule, and identifying leaf rust resistance. In the wheat jointing stage, the mixed inoculation (THTT, PHTN, THKT, THDL, THGS) in the field is adopted, and the specific reference is made to the strain and inoculation method Li Z F,Xia X C,He Z H,Li X,Zhang L J,Wang H Y,Meng Q F,Yang W X,Li G Q,Liu D Q.Seedling and slow rusting resistanceto leaf rust in Chinese wheat cultivars.Plant Dis,2010,94:45–53.
And when the leaf rust disease is most serious, the group and the parents thereof are subjected to field disease investigation. The investigation was maximum severity (MDS), i.e., the percentage of leaf rust spore pile area on the leaf as a percentage of the total leaf area. The average values in both environments are shown in table 2.
And (3) respectively extracting genome DNA of all experimental materials, taking the genome DNA as a template, and detecting by using the KASP primer designed in the step (2), wherein the specific operation is shown in the step (2).
The results are shown in Table 2 and FIG. 2. Of 115 parts of wheat material, 92 parts of material was CC homozygous (middle wheat 578 genotype), 23 parts of material was TT homozygous (ji wheat 22 genotype), and the average value of MDS of the CC homozygous wheat material was 8.46% lower than that of the TT homozygous wheat material, with a significant difference at the 0.05 level (table 3). Illustratively, the Kasp _5b_lr markers of the present invention are associated with wheat leaf rust resistance.
Table 2, genotype test results of 115 parts of wheat Material
Note that: CC: the middle wheat 578 genotype; TT: genotype of ji mai 22.
TABLE 3 QLr.caas-5B115 Natural population MDS Effect
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.
Claims (10)
1. The application of a substance for detecting wheat leaf rust resistance molecular markers in detecting or assisting in detecting wheat leaf rust resistance;
the wheat leaf rust resistance molecular marker is a nucleotide corresponding to the 71 st position of SEQ ID No.4 in a sequence table on a wheat 5B chromosome, and is C or T.
2. The use according to claim 1, characterized in that: the substance for detecting the wheat leaf rust resistance molecular marker comprises the following components: a primer set capable of amplifying a DNA fragment shown at position 71 of SEQ ID No.4 of the sequence Listing.
3. The use according to claim 2, characterized in that: the primer group consists of three single-stranded DNAs named as an upstream primer F1, an upstream primer F2 and a downstream primer R, wherein the upstream primer F1 contains single-stranded DNAs shown in 22-41 positions of SEQ ID No.1, the upstream primer F2 contains single-stranded DNAs shown in 22-41 positions of SEQ ID No.2, and the downstream primer R is single-stranded DNA shown in SEQ ID No. 3.
4. A use according to claim 3, characterized in that: the upstream primer F1 is single-stranded DNA shown in SEQ ID No.1, and the upstream primer F2 is single-stranded DNA shown in SEQ ID No. 2.
5. A method of detecting wheat leaf rust resistance comprising: detecting the leaf rust resistance of the homozygous wheat to be detected, of which the nucleotide corresponding to the 71 st position of SEQ ID No.4 in the sequence table is C, in the genomic DNA, of which the nucleotide corresponding to the 71 st position of SEQ ID No.4 in the sequence table is higher than or is higher than the homozygous wheat to be detected, of which the nucleotide corresponding to the 71 st position of SEQ ID No.4 in the sequence table is T, in the genomic DNA.
6. The method according to claim 5, wherein: the detection of the nucleotide at position 71 in the genomic DNA of wheat to be tested, which corresponds to SEQ ID No.4 of the sequence Listing, is carried out using the substance for detecting the molecular marker for leaf rust resistance of wheat as described in any one of claims 1 to 4.
7. A substance according to any one of claims 1 to 4 for detecting a marker of said wheat leaf rust resistance molecule, or a DNA molecule represented by SEQ ID No.4.
8. Use of the wheat leaf rust resistance molecular marker as set forth in claim 1 or any one of the following DNA molecules as set forth in SEQ ID No. 4:
x1) detecting or aiding in the detection of wheat leaf rust resistance;
X2) comparing the leaf rust resistance of different wheat;
x3) breeding strong leaf rust resistant wheat;
x4) screening or rejecting wheat with weak leaf rust resistance;
x5) wheat breeding.
9. Use of any one of the following substances according to any one of claims 1-4 for detecting a molecular marker for wheat leaf rust resistance:
Y1) preparing a product for detecting or assisting in detecting wheat leaf rust resistance;
Y2) comparing the leaf rust resistance of different wheat;
Y3) preparing products with strong and weak leaf rust resistance of different wheat;
Y4) breeding wheat with strong leaf rust resistance;
y5) preparing a product for breeding wheat with strong leaf rust resistance;
y6) screening or rejecting wheat with weak leaf rust resistance;
y7) preparing a product of wheat selected or knocked out for weak leaf rust resistance;
Y8) wheat breeding.
10. A wheat breeding method comprising: detecting the nucleotide corresponding to the 71 st position of SEQ ID No.4 in a sequence table in the genome DNA of wheat, selecting homozygous wheat with the 71 st position of C corresponding to the SEQ ID No.4 in the sequence table in the genome DNA as a parent for breeding, and screening homozygous wheat with the 71 st position of C corresponding to the SEQ ID No.4 in the sequence table in the genome DNA of offspring to obtain wheat with leaf rust resistance.
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| CN202410470457.2A CN118222747A (en) | 2024-04-18 | 2024-04-18 | Wheat leaf rust resistance molecular marker and application thereof |
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