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A pair of cabbage turnip mosaic virus EST-SSR markers and application thereof

A technology of turnip mosaic virus and Chinese cabbage, which is applied in the direction of DNA/RNA fragments, measurement/inspection of microorganisms, recombinant DNA technology, etc., can solve problems such as difficult to distinguish homozygous and heterozygous, achieve high accuracy and speed up The effect of selective breeding and efficient selection

Inactive Publication Date: 2010-09-22
VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Zhang Xiaowei et al. (2009) screened three AFLP markers with linkage distances of 2.9cM, 2.4cM and 0.5cM for QTLs related to TuMV resistance in Chinese cabbage, but they were all dominant markers, and it was difficult to distinguish homozygous and Heterozygote

Method used

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  • A pair of cabbage turnip mosaic virus EST-SSR markers and application thereof
  • A pair of cabbage turnip mosaic virus EST-SSR markers and application thereof
  • A pair of cabbage turnip mosaic virus EST-SSR markers and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] (1) Using the improved CTAB method to extract the genomic DNA of the single plant to be tested

[0028]In this example, the Chinese cabbage inbred line material "73" with high resistance to TuMV and the inbred line material "71-36-2" with high TuMV susceptibility were selected as parent materials, and the national-level resistant source of TuMV "8407" was selected. The core germplasm material "Guan 291" sensitive to TuMV was used as the control material. Take 0.1 g of the young leaves of the above materials, grind them into powder in liquid nitrogen, and put them into a 1.5 mL centrifuge tube. Add 700 μL of 65°C preheated CTAB extraction buffer (2.0% CTAB (g / ml); 1.4mol / L NaCl; 1mol / L Tris-HCl, pH=8.0; 0.5mol / L EDTA; 1.0% β-mercaptoethanol (V / V)), mix well, bathe in water at 65°C for 45min-1h, and shake once every 5-10min. Cool to room temperature, centrifuge to take the supernatant, extract with an equal volume of Tris-saturated phenol:chloroform:isoamyl alcohol (25:...

Embodiment 2

[0035] Such as figure 1 As shown, using the primer HCC259, a 157bp band was amplified in the resistant parental material "73" and the disease-resistant control material "8407", named ATA157, and in the susceptible parental material "71-36-2" A 154bp band was amplified in the control material "Crown 291" and the susceptible control material, named ATA154.

Embodiment 3

[0037] Such as figure 2 As shown, using primer HCC259 to detect 10 individual plants (A-J) of F2 generation isolated populations, wherein, some individual plants (such as No. C, E, and No. , J individual plants) the susceptible band ATA154 was expanded, and some (A, B, D, G, H individual plants) expanded the heterozygous band.

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Abstract

The invention discloses a pair of EST-SSR markers ATA157 and ATA154 which are closely interlocked with the disease resistant and susceptible genes of cabbage turnip mosaic virus, and belong to the technical field of biology. The markers are a pair of allelic markers, and the interlocking distance thereof with cabbage TuMV resistance locis is 3.8cM. Wherein the fragment length of the marker ATA157 is 157bp, and the fragment length of the marker ATA154 is 154bp. The invention also discloses a pair of EST-SSR labeled primers HCC259 which are provided with sequences in sequence tables SEQ ID No.3 and SEQ ID No. 4. Through the primers, the genome DNA of a single plant to be tested is adopted as the template and the markers are amplified so as to realize the high-efficiency selection of cabbage TuMV resistance, speed up the breeding of novel TuMV resistant breed strains of cabbage, and establish a novel Chinese cabbage hybrid parent selection system.

Description

technical field [0001] The invention relates to a pair of EST-SSR markers, especially the EST-SSR markers closely linked with Chinese cabbage turnip mosaic virus (TuMV) disease-resistant and susceptible genes, and belongs to the field of biotechnology. Background technique [0002] Chinese cabbage (Brassica rapa L. ssp. pekinensis) originates in my country and is an important vegetable in China, Japan, North Korea, South Korea and other countries. Virus disease is one of the three major diseases of Chinese cabbage in my country, which causes an average annual output loss of 5%, and in some years, the output is reduced by more than 10%. It is difficult to prevent and control the disease. At present, there is no universally applicable antiviral agent or other effective control technology. Breeding disease-resistant varieties is still the most effective and sustainable control measure. However, the conventional breeding procedure is cumbersome, and it is difficult to observe t...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 李巧云佟海申张志刚宋希云赵智中刘栓桃
Owner VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
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