Single nucleotide polymorphisms (SNP) locus linked to sporisorium-reiliana-resistance-associated gene, molecular marker LSdCAP4 based on same and use of same
A technology of resistance to head smut and molecular marker, applied in the field of genetic engineering, can solve the problems of few application reports of the molecular marker of disease resistance in maize, and achieve the effect of accelerating the process of breeding
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Embodiment 1
[0041] Example 1. Acquisition of SNP loci significantly related to main head smut resistance loci in maize
[0042] 1. Establishment of maize near-isogenic lines containing major head smut resistance loci
[0043] 1 parent material
[0044] Mo17 was introduced from the United States in 1974 by the Institute of Crop Breeding and Cultivation of the Chinese Academy of Agricultural Sciences. It is a bicyclic system of 187-2×C103. This line has outstanding advantages such as rich genetic basis, high combining ability, wide adaptability, strong stalks, high seed yield rate and resistance to head smut. Flat head group, improved Reid group and Lvdahongbonizi all have strong heterosis. Because Mo17 is resistant to head smut and its hybrid (F1) also has excellent disease resistance, it has played a huge role in the production of northern spring corn.
[0045] Qi 319 is an excellent PN group germplasm that was bred by the Maize Research Institute of Shandong Academy of Agricultural Sc...
Embodiment 2
[0060] Example 2. Conversion of SNP markers significantly associated with major head smut resistance loci in maize into dCAPS molecular markers
[0061] 1. SNP-based dCAPs marker development method
[0062] Based on the SNP-4 locus ID information significantly related to maize head smut resistance, the online tool (http: / / helix.wustl.edu / dcaps / dcaps.html) was used to analyze the optimal restriction sites and Choose a restriction enzyme. Through the BLAST function of the MAIZEGDB website, it is necessary to find the highly homologous BAC sequence and download it to the computer, and then use the DNAMAN software to compare and analyze the SNP-4 site ID information with the downloaded BAC sequence to find the SNP site The region of the ID information extends the base sequence from the center of the mutation site to both sides, with a total length of about 500bp. Then use the primer5.0 software to fix the primer at one end of the SNP site (restriction site with changed bases), s...
Embodiment 3
[0143] Example 3. dCAPS molecular marker chromosome physical position base sequence position information search
[0144] For the effectively amplified sequence information of the dCAPS marker, use the B73RefGen_v1 sequence as the reference sequence to find the specific physical position of the corresponding marker on the maize chromosome in the BLAST and Locus Lookup function items of the MAIZEGDB website, which will be compatible with the main anti-head smut in maize The 12 molecular markers related to the site were respectively located in the bin2.09 region of the maize chromosome. The specific search results are as follows:
[0145] 1. Legend analysis of LSdCAP4-marked PCR products after BLAST search
[0146] The result is as Figure 5-7 , shown in Table 9. Figure 5 A graph showing the retrieved distribution of LSdCAP4 markers on 10 chromosomes, Figure 6 A graph showing the retrieved distribution of the LSdCAP4 marker on chromosome 2, Figure 7 A map showing the distr...
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