Low-linolenic-acid molecular marker for brassica napus, and preparation method and application of low-linolenic-acid molecular marker

A cabbage rape, molecular marker technology, applied in biochemical equipment and methods, DNA/RNA fragments, microbial determination/inspection, etc., can solve problems such as application limitations, shorten the breeding period, speed up the process, and reduce the breeding work. amount of effect

Inactive Publication Date: 2012-05-02
HUAZHONG AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the SNP markers obtained by Somers et al. (Daryl Somers, US 2003 / 0150020A1, Aug.7, 2003; Daryl Somers, US 7081564, July 25, 2006) and Hu et al. The effect in linolenic acid-assisted selection breeding of 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. limit

Method used

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  • Low-linolenic-acid molecular marker for brassica napus, and preparation method and application of low-linolenic-acid molecular marker
  • Low-linolenic-acid molecular marker for brassica napus, and preparation method and application of low-linolenic-acid molecular marker
  • Low-linolenic-acid molecular marker for brassica napus, and preparation method and application of low-linolenic-acid molecular marker

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Embodiment 1

[0047] 1. Preparation F 1 Generation of hybrids and establishment of double haploid (DH) populations:

[0048]In this example, the Brassica napus strain A A254 with low linolenic acid content (linolenic acid content is 2.28%) bred by this unit (the seeds of this material have been sent to Wuhan City, Hubei Province on November 20, 2009 Preserved by the China Center for Typical Culture Collection in Wuhan University, its preservation number is CCTCC-P200909, the biological material deposited has been disclosed in its parent application, the application number of the parent application is 200910273437.1, and its publication number is: CN101818196A, its The public date is September 01, 2010) as the female parent, and the rapeseed high-generation breeding line A A177 with common linolenic acid content (linolenic acid content is 7.78%) (the seeds of this material have been delivered on November 20, 2009 Preserved by the Chinese Type Culture Collection Center in Wuhan University, W...

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Abstract

The invention belongs to the technical field of molecular breeding of rape, and particularly relates to a preparation method for a codominant single nucleotide polymorphism (SNP) molecular marker which is closely linked with the content of linolenic acid of brassica napus, and application of the marker to the auxiliary selection of low-linolenic-acid brassica napus. The preparation method comprises the following steps of: amplifying genomic deoxyribonucleic acid (DNA) of a low-linolenic-acid brassica napus line AA254 and a high-linolenic-acid brassica napus line AA177 by using primers to obtain two DNA-amplified fragments respectively, cloning the four DNA-amplified fragments, sequencing, and comparing nucleotide sequences; simultaneously mismatching 2nd basic groups of 3' terminals of a forward primer and a reverse primer according to the difference of the sequences so as to design a primer pair, namely YQ-fad32-1 and YQ-fad32-2; and performing polymerase chain reaction (PCR) amplification on the primer pair, namely the YQ-fad32-1 and the YQ-fad32-2 to obtain the codominant SNP molecular marker which is linked with a low linolenic acid gene of the brassica napus through PCR amplification and analysis of a population detection effect. The invention provides a novel marker for the molecular breeding of the rape. Nucleotide sequences of the novel marker are shown as SEQ ID NO:7 and SEQ ID NO:8.

Description

technical field [0001] The present invention is a divisional application of application number 200910273437.1. [0002] 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 [0003] 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 o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 周永明傅廷栋杨庆勇
Owner HUAZHONG AGRI UNIV
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