Molecular marker s5.7 co-segregated with spinach sex gene y and its application
A technology of molecular markers and co-segregation, applied in the field of plant molecular genetics and breeding, can solve the problem of insufficient stability of V20A amplification effect, and achieve the effect of improving breeding efficiency
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Embodiment 1
[0036] Example 1 Acquisition of Molecular Marker S5.7 Co-segregated with Spinach Sex Gene Y
[0037] 1. Construction of spinach genetic population
[0038] The F2 population was constructed by using female plants×male plants of the first-generation commercial spinach hybrid variety Donglu 1 (F1). The traits of the F2 population were investigated at the flowering stage ( figure 1 , figure 2 ), there were 343 plants in the F2 population, including 189 female plants and 154 male plants, which were 1:1 by chi-square test, and the sex of spinach was controlled by sex gene X / Y.
[0039] 2. Genomic DNA extraction and construction of male and female pools
[0040] Genomic DNA of 343 F2 individual plants was extracted by CTAB method.
[0041] Take 10 female plants and 10 male plants in F2 to construct female and male pools for Bulk Segregating Analysis (BSA), and measure the concentration of DNA with a DNA detector (NanoDrop Spectrophotometer). Dilute to the same concentration of...
Embodiment 2
[0051] Example 2 Application of Molecular Marker S5.7
[0052] Using the successfully transformed SCAR marker S5.7, construct F2 populations in the other 7 populations of spinach [Description of populations: Female plants × male plants in the first-generation spinach hybrid commercial variety Spinach (F1), and the female plants in the F2 population × Male plants were used to construct F3 populations, and 4 F3 populations were selected for molecular marker gender identification (that is, the experimental population numbers were 48, 50, 52, and 53 respectively); F2 population was constructed by plant x male plant, and F3 population was constructed by female plant x male plant in F2 population, and three F3 populations were selected for molecular marker sex identification (that is, the experimental population numbers were 54, 58, 59)]. Gender identification, molecular marker identification results are consistent with phenotype identification results, see image 3 , so it can be ...
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