CN113234852A - Molecular marker and primer group for identifying wheat powdery mildew resistance and application - Google Patents
Molecular marker and primer group for identifying wheat powdery mildew resistance and application Download PDFInfo
- Publication number
- CN113234852A CN113234852A CN202110737370.3A CN202110737370A CN113234852A CN 113234852 A CN113234852 A CN 113234852A CN 202110737370 A CN202110737370 A CN 202110737370A CN 113234852 A CN113234852 A CN 113234852A
- Authority
- CN
- China
- Prior art keywords
- kasp
- powdery mildew
- mildew resistance
- wheat
- molecular marker
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/13—Plant traits
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Analytical Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Botany (AREA)
- Mycology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a molecular marker for identifying wheat powdery mildew resistance, a primer group and application. The molecular marker is a KASP specific linkage marker which is developed by utilizing two pairs of KASP specific linkage markers of a QPm, saas-4AS flanking SNP probe AX-110406692 and AX-109926787, wherein the linkage markers are a KASP marker of KASP-AX-110406692 and a KASP marker of KASP-AX-109926787. The molecular marker, the primer set and the application for identifying the wheat powdery mildew resistance provided by the invention develop the molecular marker which can be used for the auxiliary selection of the wheat powdery mildew resistance, can quickly screen the wheat variety or the line with high powdery mildew resistance containing the gene for the disease-resistant breeding utilization of wheat, and obtains the wheat variety containing the wheat powdery mildew resistance gene, thereby not only saving the production cost, but also greatly improving the selection efficiency of wheat materials with powdery mildew resistance and greatly shortening the breeding cycle of the wheat disease-resistant variety; and the detection is convenient and quick and is not influenced by the environment.
Description
Technical Field
The invention relates to the field of molecular biology, in particular to a molecular marker for identifying wheat powdery mildew resistance, a primer group and application.
Background
Wheat powdery mildew is a disease caused by powdery mildew, Buchner burley of the Gramineae family, which occurs on wheat. Mainly harms leaves, and in severe cases, leaf sheaths, stalks and spikes are all infected. Yellow spots are seen in the disease part at the early stage of disease onset, the spots gradually develop into spots with the disease condition being serious, the spots are oval or round, a powdery mildew layer is arranged on the surface, the spots develop to white gray at the middle stage, the spots develop to light brown at the later stage, and a closed capsule shell is generated. When the disease condition is mild, the mildew spots are distributed dispersedly, and gradually expand into pieces as the disease condition worsens, and finally cover the whole leaves; the lower and surrounding tissues of the lesion are faded, the diseased leaves are yellow and withered early, and if the disease affects the stem and leaf sheath, the whole plant can be fallen down; the plants are short, small and weak, the ears are small, the thousand kernel weight is obviously reduced, and the wheat yield is finally influenced. Therefore, the discovery and the utilization of the new wheat powdery mildew resistance gene have important significance for wheat powdery mildew resistance breeding.
The research on wheat powdery mildew resistance genes in the prior art is not deep; no molecular marker for identifying wheat powdery mildew resistance is reported, and no breeding selection for wheat powdery mildew resistance by using the molecular marker is reported.
Disclosure of Invention
The invention aims to provide a molecular marker for identifying wheat powdery mildew resistance, a primer group and application, and aims to solve the technical problem that the prior art lacks of breeding selection for wheat powdery mildew resistance by utilizing the molecular marker.
In order to achieve the purpose, the invention provides the following technical scheme:
the invention provides a molecular marker for identifying wheat powdery mildew resistance based on QPm.saas-4AS gene, wherein QPm.saas-4AS is positioned in the interval of 1.06Mb on the end of chromosome 4AS, the molecular marker is two pairs of KASP specific linkage markers developed by utilizing QPm.saas-4AS gene flanking SNP probes AX-110406692 and AX-109926787, and the linkage markers are KASP markers of KASP-AX-110406692 and KASP markers of KASP-AX-109926787; the nucleotide sequences of the primer sets of KASP-AX-110406692 and KASP-AX-109926787 are respectively,
first, the KASP-AX-110406692 primer sequence is as follows:
FAM：GAAGGTGACCAAGTTCATGCTACTGAATAGTCTTTTTGGTGACAAA，
HEX：GAAGGTCGGAGTCAACGGATTCTGAATAGTCTTTTTGGTGACAAG，
COM：TTACATGTACCCATCCCATGAACC；
the second set of KASP-AX-109926787 primer sequences are as follows:
FAM：GAAGGTGACCAAGTTCATGCTTCCCCTTCTCCTTTTGTTTTCCTC，
HEX：GAAGGTCGGAGTCAACGGATTTCCCCTTCTCCTTTTGTTTTCCTT，
COM：AAAATCAGAGGAAATGAAAACATC。
further, said KASP-AX-110406692 and KASP-AX-109926787 are designed according to the probe sequences of AX-110406692 and AX-109926787, respectively, and said probe sequences of AX-110406692 and AX-109926787 are:
AX-110406692：
CTGTAGTTGTAACTGAATAGTCTTTTTGGTGACAA[A/G]GGTTCATGGGATGGGTACATGTAATTGATCACACG；
AX-109926787：
TATCAGTTTTTATCCCCTTCTCCTTTTGTTTTCCT[C/T]GATGTTTTCATTTCCTCTGATTTTTTGGTTTTGTT。
the preparation method of the molecular marker for identifying the wheat powdery mildew resistance provided by the invention comprises the following steps:
(1) construction of high-generation recombinant inbred line genetic population
Constructing a high-generation recombinant inbred line genetic population F8 by adopting a common cultivated wheat variety Sichuan wheat 104 with high powdery mildew resistance and a wheat variety Baimao with high susceptibility to powdery mildew;
(2) construction of genetic linkage maps
Performing whole genome scanning on the population by using a 50K high-density SNP chip, constructing a genetic linkage map, performing multi-point powdery mildew resistance identification on the genetic population and parents for many years, and positioning a powdery mildew resistance gene QPm.saas-4 AS;
(3) genetic mapping
Genetic localization is carried out on powdery mildew resistant gene QPm.saas-4AS, and the probe intervals are AX-110406692 and AX-109926787;
(4) obtaining molecular marker for identifying wheat powdery mildew resistance gene
Using qpm. saas-4AS site flanking probes AX-110406692 and AX-109926787 to develop two pairs of KASP specific linkage markers, KASP-AX-110406692 and KASP-AX-109926787;
the probe sequences of the AX-110406692 and AX-109926787 are respectively as follows:
AX-110406692：
CTGTAGTTGTAACTGAATAGTCTTTTTGGTGACAA[A/G]GGTTCATGGGATGGGTACATGTAATTGATCACACG；
AX-109926787：
TATCAGTTTTTATCCCCTTCTCCTTTTGTTTTCCT[C/T]GATGTTTTCATTTCCTCTGATTTTTTGGTTTTGTT。
further, in the step (3), the method for carrying out genetic localization on the powdery mildew resistance gene QPm.saas-4AS is a QTL localization method.
The primer group provided by the invention for identifying the molecular marker has the nucleotide sequences as follows:
first, the KASP-AX-110406692 primer sequence is as follows:
FAM：GAAGGTGACCAAGTTCATGCTACTGAATAGTCTTTTTGGTGACAAA，
HEX：GAAGGTCGGAGTCAACGGATTCTGAATAGTCTTTTTGGTGACAAG，
COM：TTACATGTACCCATCCCATGAACC；
the second set of KASP-AX-109926787 primer sequences are as follows:
FAM：GAAGGTGACCAAGTTCATGCTTCCCCTTCTCCTTTTGTTTTCCTC，
HEX：GAAGGTCGGAGTCAACGGATTTCCCCTTCTCCTTTTGTTTTCCTT，
COM：AAAATCAGAGGAAATGAAAACATC。
the invention provides a method for identifying wheat powdery mildew resistance genes, which comprises the following steps:
(1) extracting wheat genome DNA to be identified;
(2) carrying out quantitative PCR amplification on the DNA extracted in the step (1) by adopting a primer group of KASP-AX-86175290 and KASP-AX-86172908;
(3) and identifying the amplification product.
Further, in the step (2), when the quantitative PCR amplification is performed:
and (3) PCR reaction system: 2 μ l of DNA; KASP Master mix 5. mu.l; 3 mul of KASP primer group, and 3 primers of the primer group are mixed according to the volume ratio of FAM to HEX to COM =1 to 2; total reaction volume 10. mu.l;
PCR reaction procedure: pre-denaturation at 94 ℃ for 15 min; denaturation at 94 ℃ for 20sec, gradient renaturation/elongation at 61-55 ℃ for 60sec, reduction per cycle of 0.6 ℃, for 10 cycles; denaturation at 94 ℃ for 20sec, renaturation/elongation at 55 ℃ for 60sec, for 26 cycles; the KaSP-typed fluorescence data were read at 40 ℃ or lower.
The molecular marker provided by the invention is applied to identification of wheat powdery mildew resistance.
Based on the technical scheme, the embodiment of the invention can at least produce the following technical effects:
the molecular marker, the primer set and the application for identifying the wheat powdery mildew resistance provided by the invention develop the molecular marker which can be used for auxiliary selection of the wheat powdery mildew resistance, greatly improve the utilization efficiency of an important gene resource, and further can quickly screen the wheat variety or the strain with high powdery mildew resistance containing the gene for disease-resistant breeding utilization of wheat, so as to obtain the wheat variety containing the wheat powdery mildew resistance gene, thereby not only saving the production cost, but also greatly improving the selection efficiency of wheat materials with powdery mildew resistance and greatly shortening the breeding cycle of the wheat disease-resistant variety; and the detection is convenient and quick and is not influenced by the environment.
Drawings
FIG. 1 is a technical roadmap for an embodiment of the invention;
FIG. 2 shows the resistance reaction of the highly susceptible powdery mildew material barley mallow selected in example 1 and the highly powdery mildew resistant variety Chuan wheat 104 against wheat powdery mildew;
FIG. 3 is a typing chart of the detection of the specific molecular marker KASP-AX-110406692 by the fluorescent quantitative PCR instrument in example 2 of the present invention;
FIG. 4 is a typing chart of KASP-AX-109926787 as a specific molecular marker in example 2 of the present invention by using a fluorescence quantitative PCR instrument.
Detailed Description
As shown in fig. 1-4:
the powdery mildew resistance gene QPm.saas-4AS of the present invention is disclosed in Liu Z, Wang Q, Wan H, et al, QTL mapping for adult-plant resistance to powder family in Chinese elite common wheat Chuanmai104[ J ]. real Research Communications, 2020, 49(12): 1-10.
Example 1:
the preparation method of the molecular marker for identifying the wheat powdery mildew resistance provided by the invention comprises the following steps:
(1) construction of high-generation recombinant inbred line genetic population
Constructing a high-generation recombinant inbred line genetic population F8 by adopting a common cultivated wheat variety Sichuan wheat 104 with high powdery mildew resistance and a wheat variety Baimao with high susceptibility to powdery mildew;
(2) construction of genetic linkage maps
Performing whole genome scanning on the population by using a 50K high-density SNP chip, constructing a genetic linkage map, performing multi-point powdery mildew resistance identification on the genetic population and parents for many years, and positioning a powdery mildew resistance gene QPm.saas-4 AS;
(3) genetic mapping
Genetic localization is carried out on powdery mildew resistance gene QPm.saas-4AS through a QTL localization method, and the probe intervals are AX-110406692 and AX-109926787;
(4) obtaining molecular marker for identifying wheat powdery mildew resistance gene
Two pairs of KASP-specific linkage markers were developed using QPm. saas-4AS site flanking probes AX-110406692 and AX-109926787, which linkage markers are KASP-AX-110406692 and KASP-AX-109926787.
Example 2:
to verify KASP-AX-110406692 and KASP-AX-109926787 in example 1, mix reagents from Eigy technologies, Inc. were used as KASP-labeled primer detection reagents, and CFX384 Real-Time System from BIO-RAD was used as a quantitative PCR instrument; the phenotype identification of the powdery mildew resistance is to carry out grading identification on the infection degree of each material leaf by pathogenic bacteria; the detection of the specific marker is the typing detection through a fluorescent quantitative PCR instrument; the method specifically comprises the following steps:
(1) extracting the wheat genome DNA to be identified and subpackaging the wheat genome DNA to a quantitative PCR reaction plate;
(2) carrying out quantitative PCR amplification on the DNA extracted in the step (1) by adopting a primer group of KASP-AX-86175290 and KASP-AX-86172908;
the nucleotide sequences of the primer groups are respectively as follows:
first, the KASP-AX-110406692 primer sequence is as follows:
FAM：GAAGGTGACCAAGTTCATGCTACTGAATAGTCTTTTTGGTGACAAA，
HEX：GAAGGTCGGAGTCAACGGATTCTGAATAGTCTTTTTGGTGACAAG，
COM：TTACATGTACCCATCCCATGAACC；
the second set of KASP-AX-109926787 primer sequences are as follows:
FAM：GAAGGTGACCAAGTTCATGCTTCCCCTTCTCCTTTTGTTTTCCTC，
HEX：GAAGGTCGGAGTCAACGGATTTCCCCTTCTCCTTTTGTTTTCCTT，
COM：AAAATCAGAGGAAATGAAAACATC；
the quantitative PCR amplification method comprises the following steps: PCR amplification reaction and data observation and analysis were carried out using a fluorescence quantitative PCR instrument manufactured by BIO-RAD, and PCR amplification was carried out using Master mix reaction solution for KASP marker detection provided by LGC Biotechnology.
And (3) PCR reaction system: 2 μ l of DNA; KASP Master mix 5. mu.l; 3 mul of KASP primer group, and 3 primers of the primer group are mixed according to the volume ratio of FAM to HEX to COM =1 to 2; total reaction volume 10. mu.l;
PCR reaction procedure: pre-denaturation at 94 ℃ for 15 min; denaturation at 94 ℃ for 20sec, gradient renaturation/elongation at 61-55 ℃ for 60sec, reduction per cycle of 0.6 ℃, for 10 cycles; denaturation at 94 ℃ for 20sec, renaturation/elongation at 55 ℃ for 60sec, for 26 cycles; the KaSP-typed fluorescence data were read at 40 ℃ or lower.
(3) And identifying the amplification product.
According to the wheat powdery mildew resistance grading identification standard, the concrete grading standard is 0 grade and 0 grade; stage, stage 1, stage 2, stage 3, stage 4; wherein, the 0 grade is that the leaves have no infection trace and are represented as immunity, 0; the grade is that the leaves have no fresh spores but a small amount of necrotic spots, the grade 1 is that the leaves have a small amount of fresh spores but show high powdery mildew resistance, the grade 2 is that the leaves have a certain proportion of fresh spores but show medium powdery mildew resistance, the grade 3 is that the leaves have more fresh spores, show medium powdery mildew resistance, and the grade 4 is that the leaves are completely infected, show high powdery mildew resistance. The results of phenotype grading identification and detection of the infected degree of the parental leaves are shown in fig. 2, and show that the results of KASP marker detection are stable and reliable according to phenotype analysis and positioning results, and can be used for molecular detection of gene locus QPm.
Claims (8)
1. A molecular marker for identifying wheat powdery mildew resistance based on the gene locus qpm saas-4AS, which is located within the interval of 1.06Mb on the end of chromosome 4AS, characterized in that: the molecular marker is a KASP specific linkage marker which is developed by utilizing QPm, saas-4AS gene flanking SNP probes AX-110406692 and AX-109926787 and is a KASP marker of KASP-AX-110406692 and a KASP marker of KASP-AX-109926787; the nucleotide sequences of the primer sets of KASP-AX-110406692 and KASP-AX-109926787 are respectively,
first, the KASP-AX-110406692 primer sequence is as follows:
FAM：GAAGGTGACCAAGTTCATGCTACTGAATAGTCTTTTTGGTGACAAA，
HEX：GAAGGTCGGAGTCAACGGATTCTGAATAGTCTTTTTGGTGACAAG，
COM：TTACATGTACCCATCCCATGAACC；
the second set of KASP-AX-109926787 primer sequences are as follows:
FAM：GAAGGTGACCAAGTTCATGCTTCCCCTTCTCCTTTTGTTTTCCTC，
HEX：GAAGGTCGGAGTCAACGGATTTCCCCTTCTCCTTTTGTTTTCCTT，
COM：AAAATCAGAGGAAATGAAAACATC。
2. the molecular marker for identifying wheat powdery mildew resistance of claim 1, wherein: the KASP-AX-110406692 and KASP-AX-109926787 are designed according to the probe sequences of AX-110406692 and AX-109926787 respectively, and the probe sequences of AX-110406692 and AX-109926787 are respectively as follows:
AX-110406692：
CTGTAGTTGTAACTGAATAGTCTTTTTGGTGACAA[A/G]GGTTCATGGGATGGGTACATGTAATTGATCACACG；
AX-109926787：
TATCAGTTTTTATCCCCTTCTCCTTTTGTTTTCCT[C/T]GATGTTTTCATTTCCTCTGATTTTTTGGTTTTGTT。
3. the method for preparing a molecular marker for identifying wheat powdery mildew resistance according to claim 1 or 2, wherein: the method comprises the following steps:
(1) construction of high-generation recombinant inbred line genetic population
Constructing a high-generation recombinant inbred line genetic population F8 by adopting a common cultivated wheat variety Sichuan wheat 104 with high powdery mildew resistance and a wheat variety Baimao with high susceptibility to powdery mildew;
(2) construction of genetic linkage maps
Performing whole genome scanning on the population by using a 50K high-density SNP chip, constructing a genetic linkage map, performing multi-point powdery mildew resistance identification on the genetic population and parents for many years, and positioning a powdery mildew resistance gene QPm.saas-4 AS;
(3) genetic mapping
Genetic localization is carried out on powdery mildew resistant gene QPm.saas-4AS, and the probe intervals are AX-110406692 and AX-109926787;
(4) obtaining molecular marker for identifying wheat powdery mildew resistance gene
Two pairs of KASP-specific linkage markers were developed using QPm. saas-4AS site flanking probes AX-110406692 and AX-109926787, which linkage markers are KASP-AX-110406692 and KASP-AX-109926787.
4. The method for preparing a molecular marker for identifying wheat powdery mildew resistance according to claim 3, wherein the molecular marker comprises: in the step (3), the method for carrying out genetic positioning on the powdery mildew resistance gene QPm.saas-4AS is a QTL positioning method.
5. Primer set for identifying a molecular marker according to claim 1 or 2, characterized in that: the nucleotide sequences of the primer groups are respectively as follows:
first, the KASP-AX-110406692 primer sequence is as follows:
FAM：GAAGGTGACCAAGTTCATGCTACTGAATAGTCTTTTTGGTGACAAA，
HEX：GAAGGTCGGAGTCAACGGATTCTGAATAGTCTTTTTGGTGACAAG，
COM：TTACATGTACCCATCCCATGAACC；
the second set of KASP-AX-109926787 primer sequences are as follows:
FAM：GAAGGTGACCAAGTTCATGCTTCCCCTTCTCCTTTTGTTTTCCTC，
HEX：GAAGGTCGGAGTCAACGGATTTCCCCTTCTCCTTTTGTTTTCCTT，
COM：AAAATCAGAGGAAATGAAAACATC。
6. a method for identifying a wheat powdery mildew resistance gene is characterized in that: the method comprises the following steps:
(1) extracting wheat genome DNA to be identified;
(2) carrying out quantitative PCR amplification on the DNA extracted in the step (1) by adopting a primer group of KASP-AX-86175290 and KASP-AX-86172908;
(3) and identifying the amplification product.
7. The method for identifying wheat powdery mildew resistance gene of claim 6, which comprises: in the step (2), when quantitative PCR amplification is performed:
and (3) PCR reaction system: 2 μ l of DNA; KASP Master mix 5. mu.l; 3 mul of KASP primer group, and 3 primers of the primer group are mixed according to the volume ratio of FAM to HEX to COM =1 to 2; total reaction volume 10. mu.l;
PCR reaction procedure: pre-denaturation at 94 ℃ for 15 min; denaturation at 94 ℃ for 20sec, gradient renaturation/elongation at 61-55 ℃ for 60sec, reduction per cycle of 0.6 ℃, for 10 cycles; denaturation at 94 ℃ for 20sec, renaturation/elongation at 55 ℃ for 60sec, for 26 cycles; the KaSP-typed fluorescence data were read at 40 ℃ or lower.
8. Use of a molecular marker according to claim 1 or 2 for identifying wheat powdery mildew resistance.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110737370.3A CN113234852B (en) | 2021-06-30 | 2021-06-30 | Molecular marker and primer group for identifying wheat powdery mildew resistance and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110737370.3A CN113234852B (en) | 2021-06-30 | 2021-06-30 | Molecular marker and primer group for identifying wheat powdery mildew resistance and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113234852A true CN113234852A (en) | 2021-08-10 |
| CN113234852B CN113234852B (en) | 2022-11-04 |
Family
ID=77141169
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110737370.3A Active CN113234852B (en) | 2021-06-30 | 2021-06-30 | Molecular marker and primer group for identifying wheat powdery mildew resistance and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113234852B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113278724A (en) * | 2021-06-01 | 2021-08-20 | 四川省农业科学院作物研究所 | Molecular marker, primer group and application for identifying wheat leaf villus gene |
| CN116121433A (en) * | 2022-09-09 | 2023-05-16 | 河北省农林科学院粮油作物研究所 | KASP molecular marker for detecting wheat powdery mildew resistance and application thereof |
| CN117660691A (en) * | 2024-01-03 | 2024-03-08 | 四川省农业科学院作物研究所 | Molecular marker for identifying wheat spike length gene, primer group and application |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101736076A (en) * | 2008-11-19 | 2010-06-16 | 朱玉丽 | Research progress in molecular marker positioning of wheat powdery mildew resistance gene |
| US20140020128A1 (en) * | 2012-07-13 | 2014-01-16 | E.I. Du Pont De Nemours And Company | Molecular markers for various traits in wheat and methods of use |
| CN108165656A (en) * | 2017-11-08 | 2018-06-15 | 河南丰德康种业有限公司 | Wheat molecular marker and its application in wheat powdery mildew resistance is identified |
| CN108300799A (en) * | 2018-04-19 | 2018-07-20 | 山东农业大学 | The high-throughput detection label of wheat powdery mildew resistant gene Pm 5 e and its application in breeding |
| CN108977572A (en) * | 2018-09-05 | 2018-12-11 | 中国农业科学院作物科学研究所 | Mildew-resistance gene label and application based on 895 genetic background of wheat in wheat breed |
| CN110923352A (en) * | 2019-12-02 | 2020-03-27 | 河南大学 | KASP marker of wheat powdery mildew resistance gene PmDTM and application thereof |
| CN111719008A (en) * | 2019-03-19 | 2020-09-29 | 中国科学院遗传与发育生物学研究所 | SNP coseparated with wheat powdery mildew disease-resistant gene Pm5e and application thereof |
| CN112501345A (en) * | 2020-12-23 | 2021-03-16 | 山东农业大学 | SNP loci significantly associated with wheat powdery mildew resistance and application thereof in genetic breeding |
-
2021
- 2021-06-30 CN CN202110737370.3A patent/CN113234852B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101736076A (en) * | 2008-11-19 | 2010-06-16 | 朱玉丽 | Research progress in molecular marker positioning of wheat powdery mildew resistance gene |
| US20140020128A1 (en) * | 2012-07-13 | 2014-01-16 | E.I. Du Pont De Nemours And Company | Molecular markers for various traits in wheat and methods of use |
| CN108165656A (en) * | 2017-11-08 | 2018-06-15 | 河南丰德康种业有限公司 | Wheat molecular marker and its application in wheat powdery mildew resistance is identified |
| CN108300799A (en) * | 2018-04-19 | 2018-07-20 | 山东农业大学 | The high-throughput detection label of wheat powdery mildew resistant gene Pm 5 e and its application in breeding |
| CN108977572A (en) * | 2018-09-05 | 2018-12-11 | 中国农业科学院作物科学研究所 | Mildew-resistance gene label and application based on 895 genetic background of wheat in wheat breed |
| CN111719008A (en) * | 2019-03-19 | 2020-09-29 | 中国科学院遗传与发育生物学研究所 | SNP coseparated with wheat powdery mildew disease-resistant gene Pm5e and application thereof |
| CN110923352A (en) * | 2019-12-02 | 2020-03-27 | 河南大学 | KASP marker of wheat powdery mildew resistance gene PmDTM and application thereof |
| CN112501345A (en) * | 2020-12-23 | 2021-03-16 | 山东农业大学 | SNP loci significantly associated with wheat powdery mildew resistance and application thereof in genetic breeding |
Non-Patent Citations (4)
| Title |
|---|
| JINGHUANG HU等: "development of SNP，KASP，and SSR markers by BSR-Seq technology for saturation of genetic linkage map and efficient detection of wheat powdery mildew resistance gene pm61", 《INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES》 * |
| XU ZHONGYANG等: "expression analysis of wheat transcription factorTaTGA1 gene responding to infection of powdery mildew", 《ACTA PHYTOPATHOLOGICA SINICA》 * |
| Z LIU等: ""QTL mapping for adult-plant resistance to powdery mildew in Chinese elite common wheat Chuanmai104", 《CEREAL RESEARCH COMMUNICATIONS》 * |
| 徐晓丹: "5份小麦农家品种的抗白粉病基因分析及定位", 《中国博士学位论文全文数据库农业科技辑》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113278724A (en) * | 2021-06-01 | 2021-08-20 | 四川省农业科学院作物研究所 | Molecular marker, primer group and application for identifying wheat leaf villus gene |
| CN116121433A (en) * | 2022-09-09 | 2023-05-16 | 河北省农林科学院粮油作物研究所 | KASP molecular marker for detecting wheat powdery mildew resistance and application thereof |
| CN116121433B (en) * | 2022-09-09 | 2023-08-29 | 河北省农林科学院粮油作物研究所 | KASP molecular marker for detecting wheat powdery mildew resistance and application thereof |
| CN117660691A (en) * | 2024-01-03 | 2024-03-08 | 四川省农业科学院作物研究所 | Molecular marker for identifying wheat spike length gene, primer group and application |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113234852B (en) | 2022-11-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113234852B (en) | Molecular marker and primer group for identifying wheat powdery mildew resistance and application | |
| CN105803071B (en) | SNP marker relevant to melon powdery mildew resistance and its application | |
| CN112725506B (en) | KASP primer group for detecting gibberellic disease resistance of Yangmai series wheat and application thereof | |
| CN113584216B (en) | Development and application of KASP marker of wheat grain weight gene TaCYP78A16 | |
| CN1079114C (en) | Nucleic acid markers for rice blast resistance genes and rice blast resistance genes isolated by the use of these markers | |
| CN109735648A (en) | A kind of method and its dedicated kit for screening different mass of 1000 kernel wheats | |
| CN117683927A (en) | Functional KASP molecular marker of rice blast resistance gene and application thereof | |
| CN109628628A (en) | The development and application of the SNP marker of rice blast resistant gene Pi2 | |
| CN106755465B (en) | Molecular marker closely linked with wheat flag leaf length QTL QFLL | |
| CN111926104A (en) | SSR molecular marker and method for identifying authenticity of filial generation of sugarcane and saccharum arundinaceum | |
| CN106929585A (en) | The detection method of blast resistant gene Pigm and its application | |
| CN104946630B (en) | Disease-resistant linkage molecular marker for cucumber target spot disease and special primer and application thereof | |
| CN113278724B (en) | Molecular marker for identifying wheat leaf villus gene, primer set and application | |
| CN115927718A (en) | KASP molecular marker for identifying wheat grain weight and grain width and application thereof | |
| KR102429219B1 (en) | Marker composition for discrimination of soybean cultivar resistant or susceptible to Phytophthora sojae and uses thereof | |
| CN111607664B (en) | Application of SNP molecular marker on 1DS chromosome related to wheat stripe rust | |
| CN111334597B (en) | SNP (Single nucleotide polymorphism) site and KASP (Kaempferi protein) marker for detecting powdery mildew resistance of watermelon and application thereof | |
| CN113278723A (en) | Composition for analyzing genetic diversity of Chinese cabbage genome segment or genetic diversity introduced in synthetic mustard and application | |
| KR100984169B1 (en) | Primer set, method and kit for selecting TMV-resistant pepper cultivar | |
| CN109797235A (en) | With the molecular labeling R112823 of resistance gene of rice blast Pi1 close linkage | |
| JP3999254B2 (en) | Genetic markers linked to loci involved in resistance to barley stripe dwarf disease and their utilization | |
| CN109825619A (en) | With the molecular labeling R060939 of resistance gene of rice blast Pigm close linkage | |
| CN112752843A (en) | Smut resistance-related marker of Saccharum plant and use thereof | |
| CN109797234A (en) | With the molecular labeling R060939-2 of resistance gene of rice blast Pi2 close linkage | |
| CN108546778A (en) | A kind of SNP marker and its application for detecting anti-cucumber powdery mildew character |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |