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FLC1 specific SNP molecular marker based on competitive allele specificity PCR technology and application

An allele-specific, DNA molecular technology, applied in the field of FLC1-specific SNP molecular markers and applications, can solve problems such as misselection or missed selection, loss of close linkage relationship, etc., and achieve high accuracy, high specificity, and good application value Effect

Inactive Publication Date: 2016-01-13
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the application of molecular markers that are closely linked to the target gene may be limited by the genetic background, and the polymorphisms of the parents need to be detected in different populations; in addition, in the process of meiosis, this type of molecular markers There is still the possibility that the close linkage relationship with the target gene will be lost due to the exchange of chromosomes, which will cause misselection or missed selection during the identification of the target gene (Hayashi et al., 2006; Ingvardsene et al., 2008)

Method used

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  • FLC1 specific SNP molecular marker based on competitive allele specificity PCR technology and application
  • FLC1 specific SNP molecular marker based on competitive allele specificity PCR technology and application
  • FLC1 specific SNP molecular marker based on competitive allele specificity PCR technology and application

Examples

Experimental program
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Effect test

Embodiment 1

[0041] Example 1, the acquisition of molecular markers

[0042] In order to screen and obtain bolting-resistant resources better and faster, and to promote the practice of molecular marker-assisted selection breeding, through the analysis of resequencing data of 5 bolting-resistant Chinese cabbage materials and 5 bolting-easy Chinese cabbage materials, it was found that the 3763th The base at position has a G / A mutation SNP site, which is located at base 13859895 on chromosome A10, and the SNP site is named FLC1-13859895G / A site.

[0043] According to the FLC1-13859895G / A site, the following FLC1-specific primers that can be used in KASP technology were designed as molecular markers: upstream primer FLC1-FF, upstream primer FLC1-FV and downstream primer FLC1-R.

[0044] The above-mentioned FLC1-FF, FLC1-FV and FLC1-R primers were entrusted to LGC (Laboratory of the Government Chemist Government Chemist Laboratory) Ltd., UK.

[0045] The sequence characteristics of FLC1-FF, FL...

Embodiment 2

[0050] Example 2. Application of molecular markers of FLC1-13859895G / A loci in identifying early-bolting and late-bolting materials

[0051] 1. Molecular markers are used in the identification of early bolting and late bolting materials

[0052] 1) DNA extraction

[0053] Genomic DNA of the 105 cabbage materials to be tested in Table 1 were extracted by conventional CTAB method.

[0054] The quality of the extracted DNA was detected by agarose electrophoresis and Nanodrop2100 respectively, and it was found that the extracted genomic DNA met the relevant quality requirements, that is, the agarose electrophoresis showed a single DNA band without obvious dispersion; the A260 / 280 detected by Nanodrop2100 was between 1.8- Between 2.0 (DNA sample without protein contamination); A260 / 230 between 1.8-2.0 (DNA sample with low salt ion concentration); no obvious light absorption at 270nm (DNA sample without phenol contamination); for competitive allele The amount of DNA detected by ge...

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Abstract

The invention discloses an FLC1 specific SNP molecular marker based on the competitive allele specificity PCR technology and an application, and provides an application of polymorphism or genotype matter for detecting FLC1-13859895G / ASNP locuses in a Chinese cabbage genome in authenticating or assisting in authenticating the bolting state of Chinese cabbages. The experiment proves that a developed SNP marker specific primer can well classify bolting characters, high application value is achieved, and preselection and assistant breeding of bolting resisting genetic materials can be achieved.

Description

technical field [0001] The invention belongs to the technical field of biotechnology, and relates to FLC1 specific SNP molecular markers and applications based on competitive allele specific PCR technology. Background technique [0002] Cabbage crops are the vegetable crops with the largest cultivated area in my country, and play an important role in the vegetable basket project in China. In recent years, with the improvement of people's living standards, the production of cabbage has shifted from ensuring quantity supply to improving quality, meeting annual production supply and high-end demand. Special Chinese cabbage varieties with good edible quality and meeting diversified needs, such as spring sowing cabbage, baby cabbage and fast cabbage, are becoming more and more popular in the market, making the cultivation area of ​​spring cabbage and spring and summer cabbage in alpine regions expand year by year. Early bolting is the main limiting factor restricting the product...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 苏同兵于拴仓张凤兰余阳俊赵岫云张德双卢桂香隋光磊
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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