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Molecular marker of brassica napus dominant nucleic sterility restoring line and preparation method and application thereof

Brassica napus and molecular marker technology, applied in biochemical equipment and methods, microbe measurement/inspection, DNA/RNA fragments, etc., to achieve the effect of reducing breeding workload, shortening breeding period, and speeding up the process

Inactive Publication Date: 2010-05-05
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there have been reports on the localization of sterility genes and restorer genes, so far there has been no report on the application of molecular markers of dominant sterility restorer lines in Brassica napus to the identification and selection of restorer genes

Method used

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  • Molecular marker of brassica napus dominant nucleic sterility restoring line and preparation method and application thereof
  • Molecular marker of brassica napus dominant nucleic sterility restoring line and preparation method and application thereof
  • Molecular marker of brassica napus dominant nucleic sterility restoring line and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Analysis of Genetic Characteristics of Rapeseed Rape Dominant Genic Sterile Materials

[0042] One of the materials used in the present invention (i.e., one of the materials used to extract genomic DNA from the male sterile line of Brassica napus, i.e. the parent 4185A is from the homozygous two-type line GMS3 of the Brassica napus dominant sterility material reported by Zhou Yongming (see Zhou Yongming, Brassica oleracea Brassica napus recurrent selection research, Acta Crops, 1993, 1 (19): 70-75), the plants in this line were fertile and sterile plants were segregated at 1:1 (due to the homozygous genic sterile material of Brassica napus It is impossible to harvest seeds, and each generation needs to maintain a sterile line through sibling crossing, in other words, to preserve the homozygous two-type line GMS3, which can be obtained every year through a commonly used aforementioned hybridization and simple selection method for preparing genomes DNA sterile ...

Embodiment 2

[0071] Example 2: Co-dominant SCAR marker development of dominant sterility restorer lines in Brassica napus

[0072] 1. Extraction of Brassica napus genomic DNA

[0073] The three isolated populations obtained from the above-mentioned Example 1 and the parents (i.e. the dominant genic male sterile line 4185A of Brassica napus described in the above steps and the dominant genic sterile restorer lines 4185B-2 and F of Brassica napus 1 (4185A × 4185B-2) Genomic DNA is extracted from fresh and tender leaves, the specific preparation method is with reference to Li Jia et al. 521-523), the quality of the extracted DNA sample was detected by 1.2% agarose gel electrophoresis, and the concentration of the DNA sample was detected by an ultraviolet spectrophotometer (model: Pharmacia Biotech, GeneQuant II).

[0074] 2. Using the primer pair YQ-Rf1 and YQ-Rf2 to amplify and analyze the double-affinity F 1 Genomic DNA

[0075] Using the primer pair YQ-Rf1 and YQ-Rf2 designed by the app...

Embodiment 3

[0091] Example 3: Verification of application effect of co-dominant SCAR markers

[0092] The codominant SCAR markers of the dominant genic male sterile restorer line of Brassica napus obtained in the above steps were analyzed for the three segregated populations in the present invention, and the analysis results are shown in Table 7. At (4185A×4185B-2)F 2In the population, all 171 sterile individual plants only had 441bp amplified fragments as the parent sterile line 4185A. In the 4185A×(4185A×4185B-2) backcross population, 71 fertile individuals had the same DNA fragment as 4185B-2, and 77 sterile individuals had the same DNA fragment as 4185A. In the (4185A×4185B-2)×4187 population, 102 fertile individuals had the same DNA fragment as 4185B-2, 100 individuals had the same DNA fragment as 4185A, and 96 sterile individuals The strain showed the same DNA fragment as 4185A. These data indicate that this marker is highly tightly linked to the dominant genic male sterile resto...

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Abstract

The invention belongs to the technical field of breeding of rape molecules, in particular to a preparation method of a co-dominant SCAR molecular marker of a brassica napus dominant nucleic sterility restoring line and application of the molecular marker as marker-assisted selection on selectively breeding the brassica napus dominant nucleic sterility restoring line. The genome DNA of a sterile line 4185A separated by the first generation after a dominant nucleic sterility line brassica napus material homozygous two-type line GMS3 is planted and a brassica napus dominant nucleic sterility restoring line 4185B-2 is amplified by a primer YQ-Rf1 and a primer YQ-Rf2, and four amplified DNA segments are respectively obtained; then the cloning, the sequence test and the nucleotide sequence comparison are carried out on the DNA amplified segments, and the primer YQ-Rfa and the primer YQ-Rfb are designed according to the difference of sequences; and the PCR amplification and selective effect inspection are carried out, thereby obtaining the co-dominant SCAR molecular marker of the brassica napus dominant nucleic sterility restoring line. The invention provides a new marker for the breeding of the rape molecules. The invention also discloses a preparation method and application of the molecular marker.

Description

technical field [0001] The invention belongs to the technical field of molecular breeding of rapeseed, and in particular relates to a molecular marker of a dominant sterility restorer line of Brassica napus, its preparation method and application. Background technique [0002] Brassica napus dominant genic sterility is a type of male sterility formed by nuclear gene mutation, which has the characteristics of complete abortion, stable sterility, less environmental factors affecting fertility, and no cytoplasmic effect ( Zhou Yongming, Study on Recurrent Selection of Brassica napus. Acta Crops, 1993, 1(19): 70-75). There are differences in the genetic basis of the dominant sterility materials of Brassica napus from different sources. Mathias (Mathias R. Anew dominant gene for male sterility in rapeseed (Brassica Napus L.). Z Pflanzenzüchtg, 1985, 94: 170-173) reported a rapeseed dominant sterility material controlled by a single dominant gene . Through the genetic analysis ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12P19/34
Inventor 周永明杨庆勇
Owner HUAZHONG AGRI UNIV
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