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Molecular markers having haynaldia villosa 6VS chromosome specificity and use thereof

A technology of molecular markers and villous wheat, which is applied in the fields of molecular biology and genetic breeding science, can solve the problems of long and unstable marker sequences, harsh requirements for PCR reaction system and reaction conditions, etc., and achieve strong specificity and good stability Effect

Inactive Publication Date: 2013-02-27
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 long marker sequence (mostly close to or exceeding 1000bp), the harsh requirements of the PCR reaction system and reaction conditions, and the poor reproducibility of PCR results
These shortcomings are not conducive to the application of marker-assisted selection technology in wheat breeding practice

Method used

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  • Molecular markers having haynaldia villosa 6VS chromosome specificity and use thereof

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Embodiment

[0025] 1. Design and synthesis of primers

[0026] 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.

[0027] 2. Screening of 6VS chromosome-specific EST-STS markers in T. villosa

[0028] (1) PCR amplification: Genome extraction was performed using A. villosa, common wheat, and the translocation line T6AL.6VS of wheat-T. Perform PCR amplification after DNA, 10μL PCR reaction system contains about 10-20ng template DNA, 1×PCR buffer, 1.5mmol L -1 MgCl2, 200mmolL -1 dNTP, the final concentration of the two primers is 0.2 μmol L -1 , 0.5U Taq DNA polymerase, supplement the reaction system with sterile distilled water to 10 μL; the PCR reaction program is: pre-denaturation at 94°C for 3 minutes; den...

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Abstract

The invention discloses molecular markers having haynaldia villosa 6VS chromosome specificity and a use thereof, and relates to the technical field of molecular biology and genetic breeding science. The molecular markers can specifically track a haynaldia villosa 6VS chromosome. The molecular markers as primers are designed according to an EST sequence BE585684 of a short arm of a wheat sixth-part homologous group and have sequences of 6VS-SSDF: 5'-CCAACTCTAGCTGACCGCAGACTA-3' and 6VS-SSDR: 5'-ATTCCATGGTGTAATAGCTCCAACTAC-3'. Through PCR, the molecular markers are amplified into a specific DNA band having the length of 350bp. The molecular markers can fast and effectively track a haynaldia villosa 6VS chromosome and wheat powdery mildew resistance controlled by the haynaldia villosa 6VS chromosome, and has strong practical values in wheat powdery mildew-resistant molecular marker-assistant selective breeding of wheat.

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...

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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胡向均李晶晶
Owner JIANGSU UNIV
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