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A SNP molecular marker for detecting clubroot resistance in non-heading Chinese cabbage and its application

A technology for head cabbage and clubroot resistance, which is applied in recombinant DNA technology, microbial measurement/inspection, DNA/RNA fragments, etc., can solve the economic loss of growers, time-consuming and laborious field and indoor identification, etc., to eliminate Aerosol pollution, conducive to high-throughput rapid detection, and the effect of eliminating EB pollution to the environment

Active Publication Date: 2021-06-01
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, the incidence of clubroot of non-heading cabbage has become more and more serious, causing huge economic losses to growers
The onset of clubroot is greatly affected by environmental factors, and field and indoor identification is time-consuming and laborious. The substrate treatment after identification also brings problems to the environment and the secondary transmission of clubroot. Therefore, the development is suitable for high-throughput seedlings. Identification of SNP molecular markers closely linked to clubroot resistance traits in non-heading Chinese cabbage plants will help to quickly and effectively detect the resistance of non-heading Chinese cabbage to clubroot and facilitate large-scale commercial molecular breeding. Has a good application prospect and economic value

Method used

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  • A SNP molecular marker for detecting clubroot resistance in non-heading Chinese cabbage and its application
  • A SNP molecular marker for detecting clubroot resistance in non-heading Chinese cabbage and its application
  • A SNP molecular marker for detecting clubroot resistance in non-heading Chinese cabbage and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035]Example 1 Development of SNP molecular markers of anti-rooted caucasi anti-root lanes

[0036]The non-gauge material self-cultivation of the 7th physiological small root bacteria obtained by indoor identification is a mother. The patient's self-communications CS22 is the father. By hybrid, self-adding 285 orders from the F2 separation population Stage, through indoor artificial identification, according to the size of the root of the plant, the morbidity level is divided into 0 (no pathogenesis), level 1, 2 and 3. The incidence of 285 monosone of the F2 group is: 0 grade 80 strains, 1 stage 143 strain, 2 levels of 40 strains and 3 grades, 22 monographs of F2 population of 0 (unchanging plants) Constructing Extreme Actresso Gene Chine CR-Pool, the 12th single strain of the pathogenesis constructs the relegation of the patients CS-POool in the patient, and then performs BSA positioning analysis, positioning 1 QTL named QBrCR38-07 The region of 18m-21m in chromosomal region is colle...

Embodiment 2

[0044]Example 2 Development of the SNP molecular marker of the KASP reaction detection

[0045]1. In response to the molecular labeling of the present invention, the detection of high flux is detected by a high-flux of the non-tappable cabbage, and the primer combination is designed in Example 1.

[0046]2, using a simplified CTAB method, genomic DNA is extracted from non-comrades.

[0047](1) Take a proper amount of the cabbage blade in a fresh or freeze-like, add 2 ml of centrifuge tube, and add two steel beads to ground on the tissue grinding instrument;

[0048](2) Add 750 μl of CTAB solution, oscillating stellate, 65 ° C shocking temperature bath 0.5-1 h;

[0049](3) Cooling to room temperature, add 750 μL of chloroform in the fume hood: isoethane (24: 1) electric knife 3-4 times mix;

[0050](4) 12000RMP centrifuge for 10 min, take 500 μL to the new 1.5 ml of centrifuge tube;

[0051](5) Add an equal volume isopropanol solution gently shaken, and precipitate for 1 hour -20 ° C, 12000 RMP centrifug...

Embodiment 3

[0058]Example 3: F2 separation population resistance identification and analysis of molecular markers and phenotype correlation analysis

[0059]The label in the present invention is verified, and the specific steps are as follows:

[0060]1, will be mixed with a sump and sterilization matrix containing 7th physiological and small species, and the concentration of root lactimosis is 5 × 106Spore / g, seed seeding into the mixed matrix, indoor temperature 25 / 20 ° C (day / night), optical cycle 14h / 10h (day / night), according to the size of the root infection of roots, each single The case of strain is investigated, and the degree of preciseness of the disease without jaw cabbage is:

[0061]0 - level - no pathogenesis;

[0062]Level 1 - The main root did not have a significant swelling, the root, and the side roots have individual tumors

[0063]Level 2 - The main root is clearly swollen, 2 to 3 times the cross-cut area of ​​the tumor size of the stem.

[0064]Level 3 - The main root is obviously s...

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Abstract

The invention discloses a SNP molecular marker for detecting clubroot resistance of non-heading Chinese cabbage and its application. The SNP molecule is marked as Br_K_070103, and the Br_K_070103 is located at base 201813113 of chromosome 7 of non-heading Chinese cabbage. The non-heading Chinese cabbage with the base of G at this place is disease-resistant, and the non-heading Chinese cabbage with the base of T at this place is not Heading cabbage is susceptible. The invention applies the KASP technology to genotype the found SNP molecular markers, can quickly and accurately detect the resistance of the non-heading Chinese cabbage to clubroot, and greatly improves the genetic transfer efficiency. And the detection process does not require enzyme digestion, electrophoresis, and sequencing, etc., which is easy to operate, facilitates high-throughput rapid detection, and completely eliminates aerosol pollution of PCR products, environmental pollution by EB, and harm to the human body.

Description

Technical field[0001]The present invention belongs to the field of SNP molecular markers, and more particularly to SNP molecules and their applications for detecting anti-rootable diseases of non-commented cabbage.Background technique[0002]Traditional system-spectrum breeding methods are universally used breeding methods that do not commented on breeders, through breeding their many years of hard work, cultivating and creating a large number of high-quality, high-yield and disease-free nonsense cabbage. However, the traditional breeding model identified by the phenotype is not accurate, the amount of genetic group is large, high human cost, and the length of breeding cycle, it is difficult to achieve large-scale commercial integrated breeding or material genetic breeding. . With the development of molecular biology and bioinformatics, Molecular Assisted Selection, MAS has shown a huge technical advantage, achieving effective combination of genetic foundations and target status, and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 朱红芳李晓锋朱玉英翟文
Owner SHANGHAI ACAD OF AGRI SCI