Molecular marker for low linolenic acid of cabbage type rape, preparation method and application thereof

Abstract

The invention belongs to the technical field of rape molecular breeding, in particular relates to a preparation method of a codominant SNP (Single Nucleotide Polymorphism) molecular marker closely linked with the linolenic acid content of a cabbage type rape and application thereof as the marker in the auxiliary selection of the low linolenic acid cabbage type rape. The low linolenic acid cabbage type rape strain A254 and high linolenic acid cabbage type rape strain A177 genomes DNA are amplified by using primers to respectively obtain two DNA amplified fragments, then cloning, sequencing and nucleotide sequences comparison are carried out on the four DNA amplified fragments; according to the difference of the sequences, the second bases at 3'-end of a forward primer and a reverse primer are mispaired, primer pairs of YQ-fad31-1, YQ-fad31-2, YQ-fad32-1 and YQ-fad32-2 are designed, the PCR (Polymerase Chain Reaction) amplification is carried out on the primer pairs of YQ-fad31-1, YQ-fad31-2, YQ-fad32-1 and YQ-fad32-2, and the PCR amplification and group detection effects are analyzed to obtain the codominant SNP molecular marker linked with low linolenic acid genes of the cabbage type rape. The invention provides a new marker for the rape molecular breeding. In the invention, a mispairing strategy of the 3'-end of the primer is firstly applied to the development and the detection of the SNP polymorphic marker of the cabbage type rape, and a means of synchronously mispairing the forward primer and the reverse primer is firstly adopted to develop the SNP marker based on the PCR amplification and an agarose gel electrophoresis in the cabbage type rape of an allopolyploid crop. The invention synchronously discloses a method for preparing the molecular marker and the application thereof.

Description

technical field [0001] The invention belongs to the technical field of molecular breeding of rapeseed, and in particular relates to the preparation and application of a molecular marker of low linolenic acid in Brassica napus. The molecular marker can be used as a marker-assisted selection for low-linolenic acid Brassica napus breeding, and provides new molecular markers for cultivating genetically stable low-linolenic acid rapeseed varieties. Background technique [0002] The composition of fatty acids in rapeseed directly determines the quality of rapeseed oil and its nutritional value to humans. Linolenic acid is the fatty acid with the highest oxidation rate among highly unsaturated fatty acids. It is easily oxidized to form odorous oxides after contact with air, light, heat, etc., which makes rapeseed oil deteriorate and cannot be stored. What is more serious is that the oxidation products of linolenic acid are harmful to the human body and can cause cardiovascular ath...

AI Technical Summary

Problems solved by technology

However, the SNP markers obtained by Somers et al. (Daryl Somers, US 2003 / 0150020 A1, Aug.7, 2003; Daryl Somers, US 7081564, July 25, 2006) and Hu et al. (2006) only have one 3' The effect of linolenic acid-assisted selection breeding in Brassica napus is affected by various factors such as DNA template concentration, primer synthesis quality, primer concentration, PCR reaction system and parameters, etc., making its application in actual production subject to great limitations. big limit

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