Molecular marker 6VS-LEM with specificity for haynaldia villosa 6VS chromosomes and application thereof

A 6VS-LEM and molecular marker technology, applied in the fields of molecular biology and genetic breeding science, can solve the problems of long marker sequence, poor repeatability, harsh requirements of PCR reaction system and reaction conditions, etc., and achieve good stability and specificity strong effect

Inactive Publication Date: 2013-01-09
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 marker 6VS-LEM with specificity for haynaldia villosa 6VS chromosomes and application thereof

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Embodiment

[0025] 1. Design and synthesis of primers

[0026] Select the conserved genes on the short arm of Brachypodium chromosome 3, perform BLAST analysis in the NCBI or Graingenes database, retrieve the EST sequences of wheat family crops, and use Primer Premier 5.0 software to design primers. The primers were provided by Shanghai Jierui Biological Engineering Co., Ltd. synthesis.

[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×PCRbuffer, 1.5mmol L-1MgCl2, 200mmol L-1dNTP, the final concentration of the two primers is 0.2μmolL-1, 0.5U Taq For 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; denaturation at ...

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Abstract

The invention discloses a molecular marker 6VS-LEM for specifically tracking haynaldia villosa 6VS chromosomes and application of the molecular marker 6VS-LEM, and relates to the technical fields of molecular biology and genetic breeding science. A marking primer is designed according to wheat EST sequence BE404563 capable of being electronically located at slender false brome grass No.3 chromosome short arms, a particular sequence is 6VS-LEMF: 5'-CCAGGAGGGCCTCCAGGA-3' and 6VS-LEMR: 5'-CTGGGAGCTCCTGCTGCGT-3', a 350bp specific deoxyribonucleic acid (DNA) stripe can be amplified when the marking primer is used for polymerase chain reaction (PCR), the haynaldia villosa 6VS chromosomes and wheat powdery mildew resisting characters controlled by the haynaldia villosa 6VS chromosomes can be rapidly and effectively tracked, and the molecular marker 6VS-LEM has stronger practical value on wheat powdery mildew resisting marker-assisted selection breeding.

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 currently discovered physiological races of powdery mildew, and its p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 朱姗颖何华纲别同德蔡阳洋梁莹
Owner JIANGSU UNIV
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