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Development and application of KASP molecular marker of maize sugarcane mosaic virus resistant gene

A sugarcane mosaic virus and molecular marker technology, applied in the application, plant genetic improvement, recombinant DNA technology and other directions, to achieve the effects of stable labeling, short detection period and low cost

Pending Publication Date: 2022-06-10
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004]However, although the current linkage markers of maize resistance to sugarcane mosaic virus disease related genes can be used for molecular marker-assisted selection, most of them are RFLP markers, SSR markers, and InDel markers Or enzyme-cut markers, the detection efficiency is low, and it may also produce aerosols to pollute the environment, which is not suitable for high-throughput molecular detection platforms (Wang Bangtai, Lu Xiangling, Xi Zhangying, etc. Development of molecular markers for corn resistance to sugarcane mosaic virus [J]. Journal of Plant Genetic Resources, 2009.)

Method used

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  • Development and application of KASP molecular marker of maize sugarcane mosaic virus resistant gene
  • Development and application of KASP molecular marker of maize sugarcane mosaic virus resistant gene
  • Development and application of KASP molecular marker of maize sugarcane mosaic virus resistant gene

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Effect test

Embodiment 1

[0035] Development of a KASP marker closely linked to the gene SCMV2 related to resistance to sugarcane mosaic virus disease in maize

[0036] 1. Design of primers: According to the KASP_scmv site, extract the flanking sequences of 100 bp on the left and right sides, and design three sets of KASP primers preferably using Primer5.0 software. After detection by the ArrayTape platform of Douglas Scientific, a set of KASP primers with good polymorphism was selected for subsequent verification.

[0037] The KASP primers used to detect KASP_scmv are as follows:

[0038] Primer X: 5'-gaaggtcggagtcaacggattGAGAAGGCGTCGCAGC-3' (SEQ ID No.1, the lowercase part is the specific fluorescent tag sequence VIC);

[0039] Primer Y: 5'-gaaggtgaccaagttcatgctGGAGAAGGCGTCGCTGT-3' (SEQ ID No.2, the lowercase part is the specific fluorescent tag sequence FAM);

[0040] Primer R: 5'-AGTAGAACTCCCTGAGGCAACG-3' (SEQ ID No. 3).

[0041] 2. DNA extraction: Genomic DNA was extracted from maize leaves by ...

Embodiment 2

[0063] Application of KASP marker closely linked to maize sugarcane mosaic virus resistance related gene SCMV2 in molecular marker-assisted selection of maize plants resistant to sugarcane mosaic virus disease

[0064] In order to detect the practicability of the KASP_scmv marker of the present invention, the maize variety Zheng 58 (as the female parent) and Shen 137 (as the male parent) described in Table 1 were crossed to prepare F2 segregation populations, and the F2 segregation populations were tested for KASP markers and sugarcane flowers Inoculation identification of leaf virus disease (test results see figure 2 , Table 2), marker detection and virus disease inoculation identification implementation method reference example 1. Through statistics on the disease resistance of the tested varieties, among the 48 isolated populations, only 3 individual plant genotypes did not match the sugarcane mosaic virus disease resistance phenotype, and the sugarcane mosaic virus diseas...

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Abstract

The invention discloses development and application of a KASP molecular marker of a maize sugarcane mosaic virus resistant gene, and belongs to the technical field of biology. The method comprises the following steps: detecting the genotype of to-be-detected corn, and identifying or assisting in identifying the sugarcane mosaic virus resistance of the corn according to the genotype of the to-be-detected corn, the genotype is the genotype of a KASPsmmv site in a corn genome; the KASPsmmv site is an InDel site in a corn genome, and the KASPsmmv site is an InDel site in a corn genome. The molecular marker is a KASP marker KASPscmv closely linked with the maize sugarcane mosaic virus disease resistant gene SCMV2, the marker is developed based on the KASP technology, the 129376861th basic group of the maize genome No.3 chromosome can be detected in a high-throughput mode, the KASP technology is applied to conduct genotype identification on the maize sugarcane mosaic virus disease resistant gene SCMV2, and the molecular marker has the advantages of being high in specificity, high in specificity and high in specificity. The method has the advantages of simplicity and convenience in operation, low cost, short detection period, stability in marking, environment friendliness and the like, can be used for accurately detecting the sugarcane mosaic virus resistant gene SCMV2 of the corn, and is of great significance in promoting sugarcane mosaic virus resistant breeding of the corn.

Description

technical field [0001] The invention relates to the development and application of a KASP molecular marker of a corn anti-sugarcane mosaic virus disease gene, and belongs to the field of biotechnology. Background technique [0002] Maize dwarf mosaic disease is one of the important diseases affecting maize production. At present, except for the occurrence of dwarf mosaic disease in Northeast my country, it is common in all parts of China. In the general occurrence year, dwarf mosaic disease causes yield reduction of 5% to 10%, and serious disease field can cause greater production loss, or even no harvest. Sugarcane mosaic virus (SCMV) is the main pathogen that causes maize dwarf mosaic disease in my country. By cultivating maize varieties resistant to sugarcane mosaic virus disease, we can prevent and control maize dwarf mosaic disease and ensure the most efficient and economical corn production. , Environmentally friendly methods (Yu Weipeng. Screening of corn varieties (l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11A01H1/04
CPCC12Q1/6895A01H1/04C12Q2600/13C12Q2600/156
Inventor 徐扬周恺李成杨文艳焦宇馨薛钱承于广宁杨泽峰徐辰武
Owner YANGZHOU UNIV