CN113373256A - Development and application of KASP molecular marker of sugarcane mosaic virus disease resistance gene of corn - Google Patents
Development and application of KASP molecular marker of sugarcane mosaic virus disease resistance gene of corn Download PDFInfo
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Abstract
The invention discloses development and application of KASP molecular markers of sugarcane mosaic virus disease resistance genes of corn, and belongs to the technical field of biology. Detecting the genotype of the corn to be detected, and identifying or assisting in identifying the sugarcane mosaic virus disease resistance of the corn according to the genotype of the corn to be detected; the genotype is the genotype of KASP _ scmv locus in the corn genome; the KASP _ scmv site is an InDel site in the maize genome. The molecular marker is KASP _ SCMV which is closely linked with a corn sugarcane mosaic virus disease resistance gene SCMV2, the marker is developed based on KASP technology and can detect 129376861 th base of a corn genome chromosome 3 in a high-throughput manner, the method disclosed by the invention is used for carrying out genotype identification on the corn sugarcane mosaic virus disease resistance gene SCMV2 by applying the KASP technology, and the method has the advantages of simplicity and convenience in operation, low cost, short detection period, stable marker, environmental friendliness and the like, can accurately detect the corn sugarcane mosaic virus disease resistance gene SCMV2, and has important significance in promoting the breeding of the corn sugarcane mosaic virus disease resistance gene SCMV.
Description
Technical Field
The invention relates to development and application of KASP molecular markers of sugarcane mosaic virus disease resistant genes of corn, belonging to the technical field of biology.
Background
The maize dwarf mosaic disease is one of important diseases affecting maize production, the dwarf mosaic disease is common in various places in China except for the light occurrence of dwarf mosaic disease in northeast, and the local incidence of the dwarf mosaic disease is serious in northern China, northwest east spring maize regions and southwest regions. In the common years, the dwarf mosaic disease causes the yield reduction of 5 to 10 percent, and the retransmission of the dwarf mosaic disease causes large production loss and even no harvest. Sugarcane mosaic virus (SCMV) is a main pathogen causing maize dwarf mosaic disease in China, and the cultivation of Sugarcane mosaic virus disease resistant maize varieties is the most efficient, economic and environment-friendly method for preventing and treating maize dwarf mosaic disease and ensuring maize production (Weipeng Sugarcane mosaic virus (SCMV) resistant maize variety (line) screening and SCMV weed host identification [ D ] Shandong agricultural university, 2020.).
Currently, some maize sugarcane mosaic virus resistance genes have been explored, such as SMMV2 (schingying, zhanghou, lisnhai, etc.. discovery and initial localization of a new maize dwarf mosaic virus resistance gene [ J ] crop press, 2008.; Qu meeting. Meanwhile, molecular markers linked with genes are also developed, and the corn disease resistance improvement breeding is carried out by a Marker-assisted Selection (MAS) technology. The molecular marker can select a target plant from DNA level at any stage of plant development (Tanksley et al, RFLP mapping in plant breeding: New tools for an old science.1989, Biotechnology,7:257-263), thereby making up for many disadvantages in traditional breeding and becoming an effective way for solving the problem of difficult variety breeding.
However, although the linked markers of the genes related to sugarcane mosaic virus resistance of corn at present can be used for molecular marker-assisted selection, most of the linked markers are RFLP markers, SSR markers, InDel markers or enzyme cutting markers, the detection efficiency is low, aerosol can be generated to pollute the environment, and the linked markers are not suitable for a high-flux molecular detection platform (Wangbai, Luxianglingying, Sheying, etc.. the molecular markers of sugarcane mosaic virus resistance of corn are developed [ J ]. plant genetic resources academic, 2009.). Therefore, the development of low-cost molecular markers capable of being detected at high throughput is an urgent need to promote the identification of sugarcane mosaic virus disease resistant genes of corn, increase the breeding accuracy of sugarcane mosaic virus disease resistance and improve the breeding efficiency.
KASP (competitive Allele-Specific PCR) realizes the genotyping effect by specifically identifying gene sites through fluorescent probes, and can be used for detecting SNP sites and InDel sites. Compared with molecular markers such as SSR, RFLP, InDel and the like, the KASP marker has the characteristics of high detection speed, low cost, easiness in large-scale application and the like, does not need to be typed according to the size of a DNA fragment, can get rid of the detection method of the traditional gel electrophoresis, which has relatively complicated steps, low throughput and higher price, and is more suitable for a high-throughput molecular detection platform which is rapidly developed at the present stage. Therefore, the development of the corn sugarcane mosaic virus disease resistance gene KASP molecular marker which is low in cost and suitable for a high-throughput molecular detection platform has important significance for popularizing the application of a molecular marker technology and improving the disease resistance breeding efficiency and the breeding level of corn in China.
Disclosure of Invention
The present invention is implemented by the following technical solutions in view of the technical problems mentioned in the background art:
the development of the KASP molecular marker of the sugarcane mosaic virus disease resistance gene of the corn comprises the steps of detecting the genotype of the corn to be detected, and identifying or assisting in identifying the sugarcane mosaic virus disease resistance gene of the corn according to the genotype of the corn to be detected; the genotype is the genotype of KASP _ scmv locus in the corn genome; the KASP _ scmv site is an InDel site in the maize genome, is positioned at 129376861 th chromosome 3 of maize and is nucleotides 51 to 59 th in SEQ ID No.4, and the nucleotide type of the KASP _ scmv site is CGTCGCAGC (hereinafter, all indicated by "+") or deletion (hereinafter, all indicated by "-");
the sugarcane mosaic virus resistance of the corn to be detected with the KASP _ scmv locus genotype as a + + genotype or a + -genotype is higher than or is candidate to be higher than that of the corn to be detected with the KASP _ scmv locus genotype as a- -genotype; the corn to be detected with the KASP _ scmv locus genotype of + -genotype and the corn to be detected with the KASP _ scmv locus genotype of + + genotype have no significant difference in sugarcane mosaic virus resistance; wherein said + + genotype represents a homozygous for nucleotide species + at said KASP _ scmv locus in the maize genome; said-genotype represents homozygous for the nucleotide species of said KASP _ scmv locus in the maize genome; the + -genotype represents a heterozygote of the nucleotide species of the KASP _ scmv locus in the maize genome, which is + and-.
As a preferred example, the molecular marker is used in corn breeding.
As a preferred example, the molecular marker is used for detecting the polymorphism or genotype of KASP _ scmv locus in claim 1 and is used for identifying or assisting in identifying the sugarcane mosaic virus disease resistance of corn.
As a preferred example, the molecular marker is used for detecting the polymorphism or genotype of KASP _ scmv locus in claim 1 and preparing products for identifying or assisting in identifying sugarcane mosaic virus disease resistance of corn.
As a preferred example, the molecular marker is used for detecting the polymorphism or genotype of KASP _ scmv locus as defined in claim 1 in the auxiliary breeding of corn or the preparation of auxiliary breeding products of corn.
The breeding method of the corn by applying the KASP molecular marker of the sugarcane mosaic virus disease resistance gene to the corn breeding comprises the following steps,
comprising selecting as a parent maize having as- -the genotype of said KASP _ scmv site of claim 1, which- -genotype represents homozygous for the nucleotide species of said KASP _ scmv site in the maize genome; the corn breeding can be the cultivation of the sugarcane mosaic virus disease resistant corn.
As a preferred example, the molecular marker also applies to the product for detecting the polymorphism or genotype of KASP _ scmv locus as defined in claim 1, which includes any one of the following products C1) -C3);
C1) detecting products of InDel polymorphism or genotype related to sugarcane mosaic virus resistance of corn;
C2) identifying or assisting in identifying a sugarcane mosaic virus disease resistant product of corn;
C3) the product is used for auxiliary breeding of corn.
As a preferred example, the substance for detecting the polymorphism or genotype of KASP _ scmv site according to claim 1 is D1), D2) or D3) as follows;
D1) said means for detecting polymorphisms or genotypes at KASP _ scmv sites of claim 1 comprising PCR primers for amplifying maize genomic DNA fragments comprising said KASP _ scmv sites;
D2) the substance for detecting the polymorphism or genotype of KASP _ scmv locus in claim 1 is PCR reagent containing the PCR primer;
D3) a kit containing the PCR primer described in D1) or the PCR reagent described in D2).
As a preferable example, the PCR primer is a primer set consisting of a single-stranded DNA whose nucleotide sequence is the 22 nd to 37 th positions of SEQ ID No.1 of the sequence Listing, a single-stranded DNA whose nucleotide sequence is the 22 nd to 38 th positions of SEQ ID No.2 of the sequence Listing, and a single-stranded DNA whose nucleotide sequence is SEQ ID No.3 of the sequence Listing.
As a preferable example, the PCR primer is a primer set of a single-stranded DNA represented by SEQ ID No.1 of the sequence Listing, a single-stranded DNA represented by SEQ ID No.2 of the sequence Listing, and a single-stranded DNA represented by SEQ ID No.3 of the sequence Listing.
The invention has the beneficial effects that: the molecular marker is KASP _ SCMV which is closely linked with a corn sugarcane mosaic virus disease resistance gene SCMV2, the marker is developed based on KASP technology and can detect 129376861 th base of a corn genome chromosome 3 in a high-throughput manner, the method disclosed by the invention is used for carrying out genotype identification on the corn sugarcane mosaic virus disease resistance gene SCMV2 by applying the KASP technology, and the method has the advantages of simplicity and convenience in operation, low cost, short detection period, stable marker, environmental friendliness and the like, can accurately detect the corn sugarcane mosaic virus disease resistance gene SCMV2, and has important significance in promoting the breeding of the corn sugarcane mosaic virus disease resistance gene SCMV.
Drawings
FIG. 1 is a chart showing the typing of 4 test maize varieties of materials tested using the molecular marker KASP _ scmv in example 1, in which FAM indicates genotype- -, VIC indicates genotype + +, and H indicates genotype + -.
FIG. 2 is a graph of example 2 typing 48F 2 test populations using the molecular marker KASP _ scmv, in which FAM indicates genotype- - -, VIC indicates genotype + +, and H indicates genotype + -.
Detailed Description
In order to make the technical means, the original characteristics, the achieved purposes and the effects of the invention easy to understand, the invention is further explained by combining the specific drawings and the embodiments.
The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention. The examples provided below serve as a guide for further modifications by a person skilled in the art and do not constitute a limitation of the invention in any way.
The experimental procedures in the following examples, unless otherwise indicated, are conventional and are carried out according to the techniques or conditions described in the literature in the field or according to the instructions of the products. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
The corn varieties used in the examples described below are all commercially available.
The inventor of the invention found an InDel site related to the resistance of corn to sugarcane mosaic virus through literature (Qu meeting, discovery of a candidate gene of corn to sugarcane mosaic virus (SCMV2), cloning and differential site analysis [ D ]. Shenyang agriculture university, 2016.), the site is anchored at position 129376861 of chromosome 3 of corn (https:// www.ncbi.nlm.nih.gov/assembly/GCF _902167145.1/, genome version Zm-B73-REFERENCE-NAM-5.0), and the InDel site is named KASP _ SCMV, and the flanking sequence of the InDel site is shown as SEQ ID No. 4. KASP _ scmv is located at positions 51-59 of SEQ ID No.4 where the nucleotide sequence is + or-.
The + + genotype below represents the homozygous type with the nucleotide sequence of KASP _ scmv as +; - -genotype means homozygous for the nucleotide sequence of KASP _ scmv; the + -genotype represents the heterozygote type of the nucleotide species of KASP _ scmv as + and-.
Example one
Development of KASP marker closely linked with corn sugarcane mosaic virus disease resistance related gene SCMV2
1. Designing a primer: flanking sequences of 100bp each left and right were extracted based on the KASP _ scmv sites, and 3 sets of KASP primers were preferably designed using Primer5.0 software. After detection using the ArrayTape platform from Douglas Scientific, 1 set of KASP primers with good polymorphism was selected for subsequent validation.
The KASP primers used to detect KASP _ scmv were as follows:
primer X: 5'-gaaggtcggagtcaacggattGAGAAGGCGTCGCAGC-3' (SEQ ID No.1, lower case letter portion is specific fluorescent tag sequence VIC);
primer Y: 5'-gaaggtgaccaagttcatgctGGAGAAGGCGTCGCTGT-3' (SEQ ID No.2, lower case letter portion is specific fluorescent tag sequence FAM);
Primer R：5’-AGTAGAACTCCCTGAGGCAACG-3’(SEQ ID No.3)。
DNA extraction: the genome DNA is extracted from the corn leaves by the conventional CTAB method.
KASP reaction test
KASP marker amplification and reaction system:
(1) and (3) detecting by using a fluorescent quantitative PCR instrument AB-Q6 Flex:
the detection reaction system of the 5-mu-L PCR fluorescence quantitative instrument comprises: 50ng of genomic DNA, and 0.07. mu.L of a Primer mixture (preferably, the ratio of the Primer mixture: 12. mu.L each of forward Primer X and Primer Y100 pmol. L-1, reverse Primer R100 pmol. L-130. mu.L, ddH2O 46. mu.L, and the same detection object can be achieved by using other reasonable ratios of the Primer mixture), 2.5. mu.L of LGC company 2 XKASP Mix (Low Rox). Editing a sample table, executing a running program and storing data according to an AB-Q6 instrument operation manual of the fluorescent quantitative PCR instrument.
The above reaction system is the preferred reaction system of AB-Q6 Flex, and other reasonable reaction systems can also achieve the same detection purpose.
(2) The selection of the ArrayTape platform from Douglas Scientific
1.6 u L PCR ArrayTape platform detection reaction system includes: genomic DNA 50 ng/. mu.L 0.8. mu.L, Primer Mix 0.03. mu.L (preferred Primer Mix ratios: 12. mu.L for each of forward Primer X and Primer Y100 pmol. L-1, reverse Primer R100 pmol. L-130. mu.L, ddH2O 46. mu.L, the same detection object can be achieved by using other reasonable Primer Mix ratios), LGC 2 XKASP Mix (Std Rox) 0.8. mu.L. According to an ArrayTape platform instrument operation manual, a sample table is compiled, a program is operated, and data are read.
The above reaction system is the preferable reaction system of the ArrayTape platform of Douglas Scientific company, and other reasonable reaction systems can achieve the same detection purpose.
Note: the above is a recommended detection method, and other detection methods capable of achieving the same detection purpose can also be applied to the molecular marker-assisted breeding process of the marker.
Wherein, the 2 XKASP Mix consists of a fluorescent probe A, a fluorescent probe B, a quenching probe A, a quenching probe B, high fidelity Taq enzyme, dNTP, Mg2+, and the like. The nucleotide sequence of the fluorescent probe A is as follows: 5'-GAAGGTCGGAGTCAACGGATT-3', connecting a VIC fluorescent group at the 5 ' end; the nucleotide sequence of the fluorescent probe B is as follows: 5'-GAAGGTGACCAAGTTCATGCT-3', wherein the 5 ' end is connected with a FAM fluorescent group; the nucleotide sequence of the quenching probe A is as follows: 5'-AATCCGTTGACTCCGACCTTC-3', wherein the 3 ' end is connected with a quenching group BHQ; the nucleotide sequence of the quenching probe B is as follows: 5'-AGCATGAACTTGGTCACCTTC-3', wherein the 3 ' end is connected with a quenching group BHQ.
And (3) amplification procedure: pre-denaturation at 95 ℃ for 10min for 1 cycle; denaturation at 95 ℃ for 20s, annealing at 55-62 ℃ (preferably 55 ℃) for 60s, setting 40 cycles.
The experiment was carried out while setting a blank control (NTC) without adding template DNA to the reaction system, and 1 or more blank controls were set for each plate.
Analyzing the scanning data, and then determining the genotype of the KASP _ scmv locus in the corn genome to be detected (namely, detecting whether the 129376861 th base of the chromosome 3 in the corn genome is + or-), wherein if the fluorescence signal data of the amplification product of the corn to be detected is analyzed to be close to the X axis (VIC signal) by a Douglas genotyping software, the genotype of the KASP _ scmv locus in the corn genome to be detected is + + homozygote (namely, the 129376861 th base of the chromosome 3 in the corn genome is + + homozygote); if the fluorescence signal data of the amplified product of the corn to be detected is analyzed to be close to a Y axis (FAM signal) by Douglas genotyping software, the genotype of the KASP _ scmv locus in the corn genome to be detected is-homozygous (i.e. the 129376861 th basic group of the No.3 chromosome of the corn genome is-homozygous); if the fluorescence signal data of the amplified product of the corn to be detected is positioned between an X axis and a Y axis (VIC and FAM signals) through analysis of Douglas genotyping software, the genotype of the KASP _ scmv site in the corn genome to be detected is a + -heterozygote (namely, the 129376861 th base of the No.3 chromosome of the corn genome is a + -heterozygote). The samples shown black in the lower left corner are blank controls.
5. Label typing data analysis
To verify the reliability of the KASP _ SCMV marker, 4 maize variety materials were inoculated and identified to obtain disease-resistant phenotype data of the material, sugarcane mosaic virus inoculation identification methods and evaluation criteria are described in the literature (Qu part. discovery of maize sugarcane mosaic virus resistant candidate gene (SCMV2), cloning and differential site analysis [ D ] shenyang agriculture university, 2016). The genotype of the material was detected using the ArrayTape platform from Douglas Scientific, and the detection method, reaction system and amplification procedure were carried out according to the preferred protocol described above.
The amplification results showed that the KASP _ scmv marker was able to obtain stable PCR products in 4 material and both + and-allelic sites could be detected (fig. 1). The genotypes of the two disease-resistant corns are + + genotypes, and the genotypes of the two susceptible corns are-genotypes. Therefore, the KASP _ scmv marker can be used for the breeding work of the sugarcane mosaic virus disease resistance of the corn.
Therefore, the KASP _ SCMV marker provided by the invention can be used for molecular marker assisted breeding of a corn sugarcane mosaic virus disease resistance related gene SCMV 2.
TABLE 1.4 sugarcane mosaic virus resistance and genotype information for test maize varieties
| Serial number | Name of breed | Genotype(s) | Disease resistance | Serial number | Name of breed | Genotype(s) | Type (B) |
| No.01 | Zheng 58 | -- | HS | No.03 | Qi 319 | ++ | HR |
| No.02 | Shen 137 | ++ | HR | No.04 | Sinker 118 | -- | HS |
Note: HR, high resistance; r, disease resistance; MR, medium resistance; s, infection; HS, high feeling; +, genotype is homozygous for +; - -the genotype is-homozygous in spring; + -, genotype is heterozygote of + and-; ' indicates no detection signal.
Example 2
Application of KASP marker closely linked with corn sugarcane mosaic virus resistance related gene SCMV2 in molecular marker-assisted selection of corn sugarcane mosaic virus resistance plants
To examine the utility of the KASP _ scmv marker of the present invention, corn variety Zheng 58 (as female parent) and Shen 137 (as male parent) described in Table 1 were hybridized to formulate F2 isolate, KASP marker detection and sugarcane mosaic virus inoculation identification were performed on the F2 isolate (see FIG. 2, Table 2 for detection results), marker detection and virus inoculation identification were performed as in example 1. By counting the disease resistance of the test varieties, only 3 individuals in 48 segregating population individuals did not match the sugarcane mosaic virus disease resistant phenotype, and the consistency of the sugarcane mosaic virus disease resistant phenotype and the genotype was found to be P93.75% (consistency P is the number of consistent population individuals/total number of population). The result shows that the marker KASP _ scmv has higher practicability in screening sugarcane mosaic virus resistant plants of corn.
TABLE 2 phenotypic and genotypic information for isolated populations of maize
Note: HR, high resistance; r, disease resistance; MR, medium resistance; s, infection; HS, high feeling; +, genotype is homozygous for +; - -the genotype is-homozygous in spring; + -, genotype is heterozygote of + and-; ' indicates no detection signal.
The foregoing shows and describes the general principles and broad features of the present invention and advantages thereof. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, and that various changes and modifications may be made without departing from the spirit and scope of the invention as defined in the appended claims. The scope of the invention is defined by the appended claims and equivalents thereof.
Sequence listing
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Claims (10)
1. The development of KASP molecular marker of sugarcane mosaic virus disease resistance gene of corn is characterized in that: detecting the genotype of the corn to be detected, and identifying or assisting in identifying the sugarcane mosaic virus disease resistance of the corn according to the genotype of the corn to be detected; the genotype is the genotype of KASP _ scmv locus in the corn genome; the KASP _ scmv site is an InDel site in the maize genome, is positioned at 129376861 th chromosome 3 of maize and is nucleotides 51 to 59 th in SEQ ID No.4, and the nucleotide type of the KASP _ scmv site is CGTCGCAGC (hereinafter, all indicated by "+") or deletion (hereinafter, all indicated by "-");
the sugarcane mosaic virus resistance of the corn to be detected with the KASP _ scmv locus genotype as a + + genotype or a + -genotype is higher than or is candidate to be higher than that of the corn to be detected with the KASP _ scmv locus genotype as a- -genotype; the corn to be detected with the KASP _ scmv locus genotype of + -genotype and the corn to be detected with the KASP _ scmv locus genotype of + + genotype have no significant difference in sugarcane mosaic virus resistance; wherein said + + genotype represents a homozygous for nucleotide species + at said KASP _ scmv locus in the maize genome; said-genotype represents homozygous for the nucleotide species of said KASP _ scmv locus in the maize genome; the + -genotype represents a heterozygote of the nucleotide species of the KASP _ scmv locus in the maize genome, which is + and-.
2. The KASP molecular marker of a maize sugarcane mosaic virus disease resistance gene of claim 1, which is characterized in that: the molecular marker is applied to corn breeding.
3. The KASP molecular marker of a maize sugarcane mosaic virus disease resistance gene of claim 1, which is characterized in that: the application of the molecular marker in detecting the polymorphism or genotype of KASP _ scmv locus in claim 1 in identifying or assisting in identifying sugarcane mosaic virus disease resistance of corn.
4. The KASP molecular marker of a maize sugarcane mosaic virus disease resistance gene of claim 1, which is characterized in that: the application of the molecular marker in detecting the polymorphism or genotype of KASP _ scmv locus in claim 1 in preparing and identifying or assisting in identifying sugarcane mosaic virus disease resistant products.
5. The KASP molecular marker of a maize sugarcane mosaic virus disease resistance gene of claim 1, which is characterized in that: the application of the molecular marker in detecting the polymorphism or genotype of KASP _ scmv locus in claim 1 in the auxiliary breeding of corn or the preparation of auxiliary breeding products of corn.
6. The method for breeding corn by applying the KASP molecular marker of the sugarcane mosaic virus disease resistance gene to corn breeding according to claim 2, which is characterized in that: comprises the following steps of (a) carrying out,
comprising selecting as a parent maize having as- -the genotype of said KASP _ scmv site of claim 1, which- -genotype represents homozygous for the nucleotide species of said KASP _ scmv site in the maize genome; the corn breeding can be the cultivation of the sugarcane mosaic virus disease resistant corn.
7. The KASP molecular marker of a maize sugarcane mosaic virus disease resistance gene of claim 1, which is characterized in that: the molecular marker is also applied to a product for detecting the polymorphism or genotype of KASP _ scmv locus as defined in claim 1, wherein the product comprises any one of the following products C1-C3);
C1) detecting products of InDel polymorphism or genotype related to sugarcane mosaic virus resistance of corn;
C2) identifying or assisting in identifying a sugarcane mosaic virus disease resistant product of corn;
C3) the product is used for auxiliary breeding of corn.
8. The use of KASP molecular markers for sugarcane mosaic virus disease resistance genes in maize according to any one of claims 3-5 or 7, wherein: the substance for detecting the polymorphism or genotype of KASP _ scmv locus according to claim 1 is D1), D2) or D3);
D1) said means for detecting polymorphisms or genotypes at KASP _ scmv sites of claim 1 comprising PCR primers for amplifying maize genomic DNA fragments comprising said KASP _ scmv sites;
D2) the substance for detecting the polymorphism or genotype of KASP _ scmv locus in claim 1 is PCR reagent containing the PCR primer;
D3) a kit containing the PCR primer described in D1) or the PCR reagent described in D2).
9. The use of KASP molecular markers for sugarcane mosaic virus disease resistance genes in maize as claimed in claim 8, wherein: the PCR primer is a primer group consisting of single-stranded DNA with a nucleotide sequence of 22 th to 37 th positions of SEQ ID No.1 in a sequence table, single-stranded DNA with a nucleotide sequence of 22 nd to 38 th positions of SEQ ID No.2 in the sequence table and single-stranded DNA with a nucleotide sequence of SEQ ID No.3 in the sequence table.
10. The use of KASP molecular markers for sugarcane mosaic virus disease resistance genes in maize according to any of claim 9, wherein: the PCR primer is a primer group consisting of a single-stranded DNA shown by SEQ ID No.1 in a sequence table, a single-stranded DNA shown by SEQ ID No.2 in the sequence table and a single-stranded DNA shown by SEQ ID No.3 in the sequence table.
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| CN202110690760.XA CN113373256A (en) | 2021-06-22 | 2021-06-22 | Development and application of KASP molecular marker of sugarcane mosaic virus disease resistance gene of corn |
| CN202210282536.1A CN114606335A (en) | 2021-06-22 | 2022-03-22 | Development and application of KASP molecular marker of sugarcane mosaic virus disease resistance gene of corn |
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