Molecular marker closely linked with major QTL (Quantitative Trait Locus) RtA07-2 of lateral root number of rape and application of molecular marker

A molecular marker, rapeseed technology, applied in the fields of molecular biology and genetic breeding, to reduce workload, speed up the breeding process, and shorten the breeding cycle.

Active Publication Date: 2018-05-18
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the cloning of QTL for root morp...

Method used

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  • Molecular marker closely linked with major QTL (Quantitative Trait Locus) RtA07-2 of lateral root number of rape and application of molecular marker

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Experimental program
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Effect test

Embodiment 1

[0025] Obtaining the main QTL Rt.A07-2 for the traits of lateral root number in rape:

[0026] (1) The segregated population adopted by the present invention is a recombinant inbred population formed by the single-seed propagation method for seven consecutive generations after the hybridization of Shuang 11 and No. 73290 in the parent, including 175 strains, named BnaZNRIL.

[0027] (2) Collect the leaves of 175 strains in the BnaZNRIL population and the individual leaves of Shuang 11 and No. 73290 in the parent, extract the total DNA using the CTAB method, and use the rape 60K SNP chip to analyze the genotype of each sample, and count the polymorphic positions Point on the genetic types in the population, and use Joinmap 4.0 analysis software to construct a genetic map of rapeseed based on SNP markers.

[0028] (3) The 175 strains and parents of the BnaZNRIL population were cultivated by hydroponics for Shuang11 and No.73290, and the seedlings developed to the 3-leaf stage, 5-leaf s...

Embodiment 2

[0034] Obtaining a molecularly labeled primer closely linked to Rt.A07-2 of the major QTL site on the number of side roots of rape

[0035] (1) Analyzing the double 11 and No.73290 resequencing data of the two parents, it was found that there were multiple indels in the initial location interval of Rt.A07-2 and its left and right sides, and 58 of them were extracted upstream and downstream. Each 150bp sequence, in accordance with the principle of primer design, develop and design Indel labeled primers.

[0036] (2) Using the DNA of 175 individual strains of the BnaZNRIL population and Shuang 11 and No. 73290 in the two parents as templates, using the above 78 molecular markers and Shuang 11 and No. 73290 in the two parents as templates, According to acrylamide gel electrophoresis, 44 primers had specific polymorphisms between the two parents. Using these 44 markers to import into the Joinmap4.0 software, the results showed that 32 of the Indel markers are closely linked.

[0037] (...

Embodiment 3

[0039] The application of a molecular marker primer closely linked to the major QTL site Rt.A07-2 of rape side root number in rape root configuration breeding, the application process includes:

[0040] The F1 and Zhongshuang 11 obtained from the hybridization of the parent Zhongshuang 11 and No.73290 were used for continuous backcrossing, and the BC was marked on the left and right sides of the main QTL site Rt.A07-2. 1 F 1 , BC 2 F 1 And BC 3 F 1 The materials were analyzed, and the main QTL site Rt.A07-2 heterozygous single plant was selected for backcrossing in each generation, and finally in BC 4 F 1 Select heterozygous single plants for bagging and selfing to obtain BC 4 F 2 seed. BC under hydroponic conditions 4 F 2 Identification of the number of lateral roots per plant; simultaneous use of P323 and I42 for BC 4 F 2 The genotype of each individual plant is determined, and the genotype that is consistent with the Zhongshuang 11 genotype is marked as "A", and the genotype tha...

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Abstract

The invention discloses a molecular marker closely linked with a major QTL (Quantitative Trait Locus) Rt.A07-2 of lateral root number of rape and application of the molecular marker. The invention locates a major gene locus Rt.A07-2 for controlling the lateral root number of the rape on a rape A07 linkage group for the first time, and 28.7% phenotypic variance can be explained. The locus is closely linked with insertion-deletion (Indel) molecular markers P323 and I42. The lateral root number of the rape can be predicted by checking the molecular markers P323 and I42 related to the traits of the lateral root number of the rape, thereby quickly and accurately screening fine individual plants with more lateral roots.

Description

Technical field [0001] The invention belongs to the technical field of molecular biology and genetic breeding, and specifically relates to a major gene locus of a side root number trait in Brassica napus and two molecular markers closely linked to it, and also relates to gene locus and molecular markers in the root structure of rapeseed. Application in type breeding. Background technique [0002] In recent years, the self-sufficiency rate of domestic vegetable oil in my country is only about 35%, which seriously threatens the safety of edible oil supply in my country (Hua Wei et al., 2016). Rapeseed is the largest oil crop in my country (Hu et al., 2016), and rapeseed oil is the largest source of domestic edible vegetable oil. However, compared with rapeseed exporting countries such as Canada, my country's rapeseed industry has the problem of low oil production, which restricts its international competitiveness. As the scale of urbanization continues to expand and the area of ​...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11A01H1/04
CPCA01H1/04C12Q1/6895C12Q2600/13C12Q2600/156
Inventor 王汉中顿小玲匡列琼刘贵华王新发
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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