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Molecular marker 6VS-SPB with specific dasypyrum villosum 6VS chromosome and application of molecular marker

A 6VS-SPB and molecular marker technology, applied in the fields of molecular biology and genetic breeding science, can solve the problems of poor repeatability, instability, harsh requirements for PCR reaction system and reaction conditions, etc., to eliminate false negatives and strong specificity Effect

Inactive Publication Date: 2012-12-05
JIANGSU UNIV
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Problems solved by technology

However, in general, these markers have the disadvantage of instability in marker-assisted selection breeding, one reason is determined by the type of marker, such as RAPD marker OPT17 1400 and OPT17 1900 ; Secondly, due to the harsh requirements of the PCR reaction system and reaction conditions, the poor reproducibility of the PCR results caused
These shortcomings are not conducive to the application of marker-assisted selection technology in wheat breeding practice

Method used

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  • Molecular marker 6VS-SPB with specific dasypyrum villosum 6VS chromosome and application of molecular marker

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Embodiment

[0024] 1. Design and synthesis of primers

[0025] According to the information of the United States Department of Agriculture website http: / / wheat.pw.usda.gov, the EST sequence located in the 6th homology group of common wheat was searched, and the Primer Premier 5.0 software was used to design primers. The primers were provided by Shanghai Jierui Bioengineering Ltd Synthetics.

[0026] 2. Screening of 6VS chromosome-specific EST-STS markers in T. villosa

[0027] (1) PCR amplification: Genome extraction was performed using A. villosa, common wheat, and the translocation line T6AL.6VS of W. villosa (all known and public germplasm materials, provided by the Institute of Cytogenetics, Nanjing Agricultural University). After the DNA is amplified by PCR, the 10mL PCR reaction system contains about 10~20ng template DNA, 1×PCR buffer, 1.5mmol L-1 MgCl2, 200mmol L-1 dNTP, and the final concentration of the two primers is 0.2mmol L-1 , 0.5U Taq DNA polymerase, supplement the reacti...

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Abstract

The invention discloses a molecular marker 6VS-SPB with a specific dasypyrum villosum 6VS chromosome and the application of the molecular marker, and relates to the technical field of molecular biology and genetic breeding science. A marker primer is designed according to an EST (Expressed Sequence Tag) sequence BE499423 of a short arm of a homoeologous group of the 6th part of wheat; the specific sequences comprise 6VS-SPBF of 5'-TCATGCTTTAGCTGAGTTTGACCAGA-3', and 6VS-SPBR of 5'-GAACTTCCACAGCTTCTCATTTGAACAT-3'; a specific DNA strip of 900 bp can be obtained when the marker primer is subjected to PCR (Polymerase Chain Reaction); and the dasypyrum villosum 6VS chromosome and the wheat powdery mildew resistant character controlled by the dasypyrum villosum 6VS chromosome can be effectively traced. Therefore, the molecular marker 6VS-SPB is of potential application value in wheat powdery mildew resistant marker-assisted selection.

Description

technical field [0001] The invention relates to the fields of molecular biology and genetic breeding science and technology, in particular to a molecular marker for specifically tracking the 6VS chromosome of T. villosa and its usage. Background technique [0002] Wheat is one of the three major food crops in the world, and it is also the first food crop in my country's total output. It is one of the important determinants of ensuring my country's food security. Wheat is often affected by biotic or abiotic stresses during its growth process. Wheat powdery mildew is a worldwide wheat disease. In recent years, with the improvement of farmland water conservancy facilities and water and fertilizer conditions in my country, its harm has been increasing year by year. The wheat-T6AL.6VS translocation line bred by distant hybridization and chromosome engineering technology shows high resistance or immunity to the powdery mildew physiological races found so far, and its powdery mild...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 何华纲别同德朱姗颖洪好粱莹孙卫红
Owner JIANGSU UNIV
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