SNP Molecular Markers Associated with Corn Ear Row Number and Its Application
A molecular marker, corn ear technology, applied in the field of molecular markers of crop genetics and breeding, can solve the problems of unclear mechanism and quality control, little understanding of cellulose synthesis, etc., and achieve the effect of improving accuracy and speeding up the process.
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Embodiment 1
[0026] 1.1 Construction of GWAS groups
[0027]The maize genome-wide association analysis population includes 431 inbred lines with different kinship. All inbred lines are provided by the School of Plant Science, Jilin University, and are planted in the Teaching and Research Experimental Base of the School of Plant Science, Jilin University (Lvyuan District, Changchun City, Jilin Province). The arrangement of the plots followed a random block design, the block was repeated 3 times, the single-row plot, the row length was 3 m, the row spacing was 65 cm, and the plant spacing was 20 cm. The field management measures were the same as those in the field.
[0028] 1.2 Collection of samples and investigation of phenotypic data
[0029] At the 5-6 leaf stage of the maize seedlings in the plot, 3 seedlings were collected, cleaned and stored in a -20°C refrigerator. After the corn ears were mature, the representative kernel row number (Kernel Row Number, KRN) of 5 ears was investigat...
Embodiment 2
[0041] 2.1 Construction of sequencing library
[0042] (1) 200ng of genomic DNA was digested with the restriction enzyme EcoRI, and after the digestion was complete, it was purified and recovered by magnetic beads.
[0043] (2) The purified DNA after enzyme digestion was connected to Barcode adapters PI with T4 DNA Ligase, the ligation product was purified and recovered by magnetic beads, and the qualified P1 primer label was recorded.
[0044] (3) All samples were mixed in equal proportions into a total DNA mix, and the DNA was fragmented using covaris220. The fragment length of the fragmented DNA was about 200-500 bp.
[0045] (4) After End Repair & dA-tailing, connect to Adaptor P2, and purify and recover the ligated product with magnetic beads.
[0046] (5) Use the KAPA 2G Robust PCR Kit to amplify and enrich the linker ligation products, use magnetic beads to purify and sort the PCR reaction products, and use 2% agarose gel electrophoresis to detect the quality of the co...
Embodiment 3
[0062] Intergroup comparison of different genotypes detected at significant loci
[0063] 3.1 Zm00001d022502 gene Chr7: 179016330 locus comparison between groups
[0064] This locus includes three genotypes, namely G / G, A / A, and A / G. The box plots for the distribution of the number of spike rows in samples of different genotypes are shown in Figure 5 . Group G / G has a median of 14.88 rows, lower quartile Q1 is 13.68 rows, and upper quartile Q3 is 16.29 rows; A / A group has a median of 14.15 rows, lower quartile Q1 is 13.14 rows Row, upper quartile Q3 was 14.37 rows; A / G group median was 14.22 rows, lower quartile Q1 was 13.25 rows, upper quartile Q3 was 15.00 rows.
[0065] The t-test results of the phenotypic mean difference between different genotype groups are shown in Table 2. It can be seen from Table 2 that the difference between G / G and A / A groups is extremely significant (P=0.0007), and the mean difference between groups is 1.163 lines; G / G The difference between G / ...
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