The indel marker of the bra013400 frameshift mutation in Chinese cabbage and its application in breeding practice

A Chinese cabbage, 140bp technology, applied in the fields of application, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problems that have not yet been reported on the cloning of the receptor class TNL-WRKY gene, so as to improve breeding efficiency, simplify screening methods, The effect of shortening the breeding period

Inactive Publication Date: 2021-09-03
VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there is no report on the cloning of receptor-like TNL-WRKY gene in Chinese cabbage

Method used

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  • The indel marker of the bra013400 frameshift mutation in Chinese cabbage and its application in breeding practice
  • The indel marker of the bra013400 frameshift mutation in Chinese cabbage and its application in breeding practice
  • The indel marker of the bra013400 frameshift mutation in Chinese cabbage and its application in breeding practice

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1: Cloning of the Bra013400 target sequence in two typical Chinese cabbage inbred lines 06-247 and Guan 291

[0053] 1.1 Chinese cabbage genomic DNA extraction

[0054] (1) Chinese cabbage seedling leaves are put into a liquid nitrogen precooled mortar, fully ground into powder in liquid nitrogen;

[0055] (2) After the liquid nitrogen evaporates to dryness, transfer it to a 2ml centrifuge tube immediately, and add about 0.6ml of CTAB extract solution preheated to 65°C for every 100mg of material. 60 minutes to lyse the cells;

[0056] (3) After the lysis is completed, take out the sample and allow it to cool down to room temperature completely. Add an equal volume of chloroform (chloroform), gently invert to mix, and place at room temperature for 10 minutes;

[0057] (4) room temperature, 12000rpm centrifugation for 15 minutes;

[0058] (5) Use a pipette to carefully suck out the upper aqueous phase, add it to a new 1.5ml centrifuge tube, add 500 μl of isop...

Embodiment 2

[0072] Application of Embodiment 2 Marks

[0073] In order to verify the practicability of the markers, we used the Bra013400F / Bra013400R primer combination to detect some of the germplasm resources created by the inventors of the present application. Some individual plants of the F2 population were detected in combination, which further verified the practicability of the markers. The specific implementation rules are as follows:

[0074] (1) Part of the germplasm resources and each individual plant genomic DNA of the F2 population were extracted as described in "1.1" in Example 1.

[0075] (2) PCR amplification: The preparation and amplification conditions of the PCR reaction solution are as described in item (2) of "1.2" in Example 1.

[0076] (3) Detection of PCR products is as described in item (3) of "1.2" in the examples. Test results such as Figure 5 As shown, A is the detection result of some germplasm resources, in which CK1 is the inbred line 06-247, CK2 is the ...

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Abstract

The invention discloses a co-dominant marker for identifying the wild type and mutant of the coding gene Bra013400 of Chinese cabbage TNL-WRKY, the co-dominant marker includes: a marker related to the wild type Bra013400W, with a fragment size of 140bp, such as SEQ ID Shown in No.1; the marker Bra013400m related to the mutant, with a size of 135bp, is shown in SEQ ID No.2. The present invention obtains a wild type and a mutant of a gene Bra013400 possibly encoding TNL-WRKY located on the A01 chromosome, and develops a co-dominant marker for identifying the wild type and the mutant. The marker can be used to screen natural populations of Chinese cabbage, providing efficient detection means of Chinese cabbage disease-resistant R gene loci, and the materials carrying wild-type genes detected by the co-dominant marker of the present invention can be used as disease-resistant breeding It is an important material for breeding new varieties of Chinese cabbage with disease resistance.

Description

technical field [0001] The invention relates to the technical field of vegetable breeding, in particular to an InDel marker for a Chinese cabbage Bra013400 frameshift mutation and its application in breeding practice. Background technique [0002] There are abundant disease resistance genes (R genes) in plant genomes, which ensure that they can resist a wide variety of pathogenic microorganisms throughout the growth and development cycle. So far, more than 100 R genes have been cloned. Studies have found that a large class of R gene coding products generally include Toll / Interleukin-1 receptor-like structures (Toll / Interleukin-1 receptor, TIR), nucleic acid binding sites (Nucleotide binding site, NBS) and Leucine-rich repeats (Leucine-rich repeats, LRRs) and so on. As far as a specific R gene is concerned, it is often one or a combination of several motifs mentioned above. According to the motifs contained in R genes, R genes can be roughly divided into eight categories (G...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11A01H1/04
CPCA01H1/04C12Q1/6895C12Q2600/13C12Q2600/156
Inventor 赵智中刘栓桃张志刚李巧云王立华卢金东
Owner VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
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