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Group of cabbage mustard InDel molecular markers as well as development method and application thereof

A molecular marker, kale technology, applied in recombinant DNA technology, biochemical equipment and methods, DNA/RNA fragments, etc., to achieve the effects of high polymorphism, good stability, and improved work efficiency

Active Publication Date: 2020-02-14
INST OF VEGETABLES GUANGDONG PROV ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is to establish a technical system for the development of Chinese kale InDel markers, for the location of important trait genes of Chinese kale, the construction of genetic maps, the analysis of genetic diversity and Molecular marker-assisted selection breeding provides more new InDel markers to make up for the current lack of InDel markers in kale

Method used

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  • Group of cabbage mustard InDel molecular markers as well as development method and application thereof
  • Group of cabbage mustard InDel molecular markers as well as development method and application thereof
  • Group of cabbage mustard InDel molecular markers as well as development method and application thereof

Examples

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Embodiment 1

[0033] Example 1 Development of a group of kale InDel molecular markers

[0034] In order to improve the success rate of the development of kale InDel molecular markers, the present invention carried out whole-genome resequencing on the representative varieties of kale, purple kale "HJJL" and green kale "612F", and compared with the reference genome Brassica oleracea (B. oleracea) comparison, screening of InDel loci with differences between two kale varieties for marker development.

[0035] (1) Extraction of kale DNA:

[0036] ①Take about 2 g of young kale leaves, grind them with liquid nitrogen, and quickly add 1000 μL of 2% CTAB extraction buffer when the liquid nitrogen quickly evaporates to dryness, mix well, place in a water bath at 65°C for 50 minutes, and shake every 10 minutes;

[0037] ② After standing at room temperature, centrifuge at 12000rpm for 10min at 4°C, and transfer about 800μL of the supernatant to a new 2mL centrifuge tube;

[0038] ③ Add an equal volum...

Embodiment 2

[0056] Example 2 Application of a group of kale InDel molecular markers in the analysis of kale genetic diversity

[0057] Genomic DNA was extracted from young leaves of 33 Chinese kale materials of different varieties (refer to the method in Example 1), and the names and corresponding DNA sequence numbers of the 33 kinds of Chinese kale materials are shown in Table 2 below. With the genomic DNA of 33 kale materials as a template, 33 parts of kale DNA samples were carried out to PCR amplification (reaction conditions in reference example 1) with 60 pairs of InDel labeled primers further selected in Example 1, using 8% The amplification products were separated by polyacrylamide gel electrophoresis, and the electrophoresis results of InDel primers J252, J117, and J813 were as follows: figure 2 , 3 , 4 shown. Each band in the electrophoretic pattern is regarded as a site, and the presence or absence of the amplified fragment is assigned "1" and "0" respectively, and a 0-1 matr...

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Abstract

The invention provides a group of cabbage mustard InDel molecular markers as well as a development method and an application thereof, which relate to the technical field of molecular markers. The cabbage mustard InDel molecular marker provided by the invention comprises primer pairs corresponding to 60 sites, and the nucleotide sequences of the primer pairs are shown as SEQ ID NO. 1 to SEQ ID NO.120. The marker developed by the invention has the characteristics of simple operation, good stability and high polymorphism; the primer pairs are good in stability, are uniformly distributed in 9 linkage groups, can be used for positioning of important agronomic trait genes of cabbage mustard, genetic diversity analysis, fingerprint construction, genome-wide association analysis and genetic linkage map construction or molecular marker-assisted selective breeding, and can improve the working efficiency.

Description

technical field [0001] The invention relates to the technical field of molecular markers, in particular to a group of kale InDel molecular markers and a development method and application thereof. Background technique [0002] Chinese kale (Brassica alboglabra) is a subspecies of Brassica oleracea in the family Brassicaceae. It is mainly cultivated in Guangdong, Guangxi, Fujian and other places. It is a characteristic vegetable in southern China and feeds on flower sprouts and leaves. The traditional breeding method of kale has a long cycle and low efficiency. Combining molecular marker-assisted selection with traditional breeding will speed up the process of new breed selection and genetic improvement. The development of molecular markers will lay the foundation for molecular marker-assisted breeding. [0003] Molecular markers are genetic markers based on nucleotide sequence variation in genetic material between individuals, and are a direct reflection of genetic polymor...

Claims

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Application Information

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IPC IPC(8): C12Q1/6809C12Q1/6895C12N15/11
CPCC12Q1/6809C12Q1/6895C12Q2600/13C12Q2600/156C12Q2535/122C12Q2537/165
Inventor 张艳洪晓如陈汉才黎庭耀沈卓杨易
Owner INST OF VEGETABLES GUANGDONG PROV ACAD OF AGRI SCI
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