Breeding method and application of a kind of high oleic acid rapeseed male sterile line
A technology of nuclear sterile line and high oleic acid, which is applied in the field of rapeseed breeding, to achieve the effect of increasing market value and supply, reducing production costs, and improving economic benefits and market competitiveness of enterprises
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Embodiment 1
[0025] Example 1 Development and Verification of Molecular Markers
[0026] 1. Development and verification of Ms3 / ms3 markers
[0027] In this example, the gene sequence at the Bnms3 locus and the sequence between the corresponding wild-type BnC9.Tic40 and the homologous gene BnA10.Tic40 were analyzed. See figure 1 shown, where Indicates the start position and end position of the primer; → indicates the sequence of m3c9-4F / 2R; → indicates the sequence of M3C9-1F / 1R. Aiming at the difference of Bnms3 at 1200 to 1222 and the difference of BnC9.Tic40 at 1630 to 2050, an ARMS-PCR allele-specific marker M3C9-1F / 1R and m3c9-4F / 2R dual PCR system was developed, and the primer sequences are as follows :
[0028] M3C9-1F: 5'-CTGAAGCTTCTTTAACTGTATGATGC-3', see SEQ ID NO: 1;
[0029] M3C9-1R: 5'-TAGAGGGAACAATCTAGATGAG-3', see SEQ ID NO:2;
[0030] m3c9-4F: 5'-GGACATGTTTTTGTTTTTTGG-3', SEQ ID NO: 3;
[0031] m3c9-2R: 5'-AGGTTGTGGCTTAGACACAGGAG-3', SEQ ID NO:4.
[0032] The PCR sy...
Embodiment 2
[0053] Example 2 Breeding of GMS Lines of High Oleic Rapeseed
[0054] 1. See the breeding route of the present invention Figure 5 . The target genotype identified by prospect selection in the breeding roadmap of the present invention is ms3ms3ms4 with high oleic acid b ms4 b (homozygous sterile strain A) and Ms3ms3ms4 b ms4 b (maintaining strain B) and ms3ms3ms4 c ms4 c (temporary maintenance strain C). The background selection is based on the results of the foreground selection. The selected individual plants with consistent growth and shape are analyzed using the 6K breeding chip (produced by Zhongyujin Company) markers. The recurrent parents are used as a reference, and the genetic similarity of the three ABC lines is compared at the same time. Individual plants with better similarity enter into the next generation. In the present invention, the non-transgenic high oleic acid sterility material with a stable oleic acid content greater than 75% is finally bred thro...
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