A Molecular Marker Closely Linked to Vitality Traits of Sweet Corn Seeds and Its Application
A technology of molecular markers and sweet corn, applied in the field of molecular biology, can solve the problems of hindered starch synthesis, negative correlation of sugar content, low emergence rate of sweet corn, etc., and achieve the effect of improving seed vigor and shortening the breeding process
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Embodiment 1
[0036] 1. Experimental materials: 295 Temperate Tropical Sweet Corn Self-tone from China, the United States, Thailand, constitutes the associated group of all genome association analysis (provided by the Research Institute of Crop Body, Guangdong Academy of Agricultural Sciences), which has rich Genetic diversity.
[0037] Sample source is shown in Table 1.
[0038] Table 1
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[0048] 2. Field Test Design: In 2017, it was planted in the Guangzhou Dafeng Base. It adopts a complete random area design, two repetitions, 0.25 meters per line, 10 plants, fertilization, watering, weeding and other field management measures in accordance with local Normal levels are managed, physiological mature, harvest, naturally dry.
[0049] 3. Associated group Phenotype Identification: After the dried, each self-container is selected from 3-5 sizes uniform cutting hair, take 30 capsules, uniform seeds, sod...
Embodiment 2
[0056] Example 2, SNP site screening and target gene positioning
[0057] All genome association analysis: By reducing the associated group, a total of 9.8 million high-quality SNP sites, combined with 9.8 million SNP, group structure, intra-affinity and seed vitality related traits of associated groups, using Tassel The MLM model performs all genome association analysis, and 1 candidate gene (ZM00001D027504) (ZM00001D027504) (ZM00001D027504) is found on the 1st chromosome (P ≤ 1 × 10-6), and the nucleotide sequence is derived from the Gramene website. At 1: 6907596-6912220), the gene encodes a triphosphate nucleoside, and there is three unsecured SNP sites on the sixth exon of the gene, one of which SNP sites (named mark S1_6908324) And seed sprouts significantly correlated (p = 5.19216e-08), can explain 11.9% of phenotypic variation, other two SNP sites (named marked S1_6908332 and mark S1_6908437 respectively) also associated with seed germination, but not significant Embodimen...
Embodiment 3
[0059] Example 3, ZM00001D027504 gene of molecular marker primer design
[0060] For marking S1_6908324, marking S1_6908332, marking S1_6908437, and marking S1_6907721, an allele-specific KASP primer is used to amplify and detect, and 19-21 bases labeled F1 and F2 primers are selected on the left side of the SNP position, and take the SNP A base of the variation site is a F1 primer, and the other base is a F2 primer, and the upper two joints are connected, and 19-22 bases is selected from the right 40-100 bp, respectively, thereby passing through the fluorescence detection platform. The purpose of detecting different SNP genotypes.
[0061] The primer sequences of the label S1_6908324 are as follows:
[0062] Ph1f1: gaaggtgaccaagttcatgcttgtgcctccccccccccgtacatgcc (SEQ ID NO: 1)
[0063] PH1F2: gaaggtcggggggtcaacgatttgtgcctcccccccgtacatgct (SEQ ID NO: 2)
[0064] PH1R: CatgaagtccttcgtcagaGc (SEQ ID NO: 3)
[0065] The primer sequences of the label S1_6908332 are as follows:
[0066...
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