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Codominance marker primer tightly linked with tomato spotted wilt virus resisting sites RTSW of nicotiana alata, identifying method and application of codominance marker primer

A co-dominant marker, spotted wilt technology, applied in the field of molecular biology, can solve the problems of inability to distinguish homozygous/heterozygous genotypes of resistance loci, false positives of SCAR markers, and large-scale application of restriction markers, etc. The effect of obvious difference in belt type, high detection efficiency and lower detection cost

Pending Publication Date: 2019-11-26
YUNNAN ACAD OF TOBACCO AGRI SCI
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Problems solved by technology

So far, only H. Moon and J.S. Nicholson (2007) have developed the corresponding molecular markers in the world, but the genetic distance between the AFLP marker and the resistance gene is far away, and the linkage is not tight. The developed SCAR markers exist in practical applications. prone to false positives
Moreover, neither marker can distinguish homozygous / heterozygous genotypes at the resistance locus, limiting the large-scale application of these markers

Method used

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  • Codominance marker primer tightly linked with tomato spotted wilt virus resisting sites RTSW of nicotiana alata, identifying method and application of codominance marker primer
  • Codominance marker primer tightly linked with tomato spotted wilt virus resisting sites RTSW of nicotiana alata, identifying method and application of codominance marker primer
  • Codominance marker primer tightly linked with tomato spotted wilt virus resisting sites RTSW of nicotiana alata, identifying method and application of codominance marker primer

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Embodiment Construction

[0033]The present invention will be further described in detail below in conjunction with the examples.

[0034] A co-dominant marker primer closely linked to tobacco spotted wilt locus RTSW, the primer is composed of two single-stranded DNAs of primer 1 and primer 2;

[0035] The primer 1 sequence is Seq ID No.1:

[0036] RTSW_Marker3_F 5'-CCTATGAAGCAACGAAGCGATA-3';

[0037] The primer 2 sequence is Seq ID No.2:

[0038] RTSW_Marker3_R 5'-GTTGACTGTTGACTGTTGATTAGAG-3'.

[0039] The method for using the primers to identify the RTSW allele type of the tobacco spot wilt resistance site, the specific steps are as follows:

[0040] (1) Using the genomic DNA of tobacco to be identified, tobacco spotted wilt resistant Polalta and spotted wilt susceptible variety K326 as templates, PCR primers composed of two single-stranded DNAs of primer 1 and primer 2 were used to pair genomic DNA Carry out PCR amplification, and use PCR primers composed of two single-stranded DNAs of primer 1 ...

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Abstract

The invention relates to a codominance marker primer tightly linked with tomato spotted wilt virus resisting sites RTSW of nicotiana alata, an identifying method and an application of the codominancemarker primer. The molecular marker primer consists of two single-stranded DNAs of a primer 1 and a primer 2. The codominance marker primer can specially perform marking and identification on nicotiana alata genomic DNAs, detect the type of allelic genes of RTSW in the genomic DNAs of nicotiana alata materials, can identify whether tomato spotted wilt virus resisting sites RTSW exist or not, and can identify the type of the allelic genes of the tomato spotted wilt virus resisting sites RTSW of nicotiana alata, and the codominance marker primer is reliable and convenient to use. Compared with aconventional tomato spotted wilt virus resisting marker, the codominance marker primer has the advantages of being tightly linked with target gene RTSW, high in accuracy, low in cost, high in detection efficiency and the like, the screening efficiency of the varieties of the tomato spotted wilt virus resisting genes RTSW of nicotiana alata can be greatly improved, the breeding period of the tomato spotted wilt virus resisting nicotiana alata varieties can be greatly shortened, and the breeding efficiency can be improved.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a molecular marker capable of identifying a tobacco spot wilt resistance gene locus RTSW and its application in positioning, cloning or breeding tobacco spot wilt resistance gene loci. Background technique [0002] Tobacco spotted wilt is a disease of tobacco caused by the infection of Orthotospovirus. This genus is a plant virus with the widest range of hosts and the most serious occurrence. Its representative American species, Tomato Spotted Wilt Virus (TSWV), has posed a greater threat to tobacco leaf production in Yunnan tobacco-growing areas in my country. . According to the test results of samples collected from various places (states, cities) and counties in Yunnan in recent years, TSWV is distributed on tobacco in Yunnan province and tends to expand and intensify. What is even more worrying is that none of the main flue-cured tobacco varieties in Yu...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 黄昌军刘勇
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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