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Molecular marker linked with wheat mildew-resistance gene

A powdery mildew resistance gene and molecular marker technology, applied in the field of agricultural biotechnology engineering, can solve the problem of resistance gene loss of function

Inactive Publication Date: 2006-04-19
TIANJIN NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Some of the molecular markers that have been obtained so far are invalid due to the variation of the physiological race of powdery mildew, and the resistance gene loses its effect

Method used

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  • Molecular marker linked with wheat mildew-resistance gene
  • Molecular marker linked with wheat mildew-resistance gene
  • Molecular marker linked with wheat mildew-resistance gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Using the RAPD molecular marker method, a new molecular marker OPP15 linked to the wheat powdery mildew resistance gene was obtained 900 .

[0042] The specific method is: the parent of the disease-resistant gene donor is from the British common wheat variety Brock, and it is crossed with the susceptible common wheat parent 015 and the susceptible parent Jing 411. The hybrid combination is "Brock / 015 / / Jing 411", and the resistant list 92 BC strains were obtained from high-generation lines with stable resistance to powdery mildew through crossing, backcrossing and selfing 2 f 5 ;

[0043] 1. Extract DNA:

[0044] Take 0.2 g of wheat leaves, grind them into powder in liquid nitrogen, quickly add DNA extraction buffer (50mM Tris-HCl, pH8.0; 20mM EDTA, pH8.0; 50mM NaCl), incubate at 65°C for 15 minutes, and mix them by inversion 2-3 times, 0 ℃ ice bath for 10 minutes. Centrifuge at 3,000 rpm for 10 minutes, take the supernatant, add an equal volume of Tris-H...

Embodiment 2

[0058] Example 2: Using the SSR molecular marker method, a new molecular marker Xgwm114 linked to the wheat powdery mildew resistance gene was obtained 120 .

[0059] Plant material is with embodiment 1, and concrete way is as follows:

[0060] 1. Extract DNA:

[0061] Take 0.2 g of wheat leaves, grind them into powder in liquid nitrogen, quickly add DNA extraction buffer (50 mM Tris-HCl, pH 8.0; 20 mM EDTA, pH 8.0; 50 mM NaCl, incubate at 65 ° C for 15 minutes, and mix them by inversion 2-3 times, ice bath at 0°C for 10 minutes. Centrifuge at 3000 rpm for 10 minutes, take the supernatant, add an equal volume of Tris-HCl (pH8.0) saturated phenol, invert and mix well, and centrifuge at 3000 rpm for 10 minutes Take the supernatant, add an equal volume of phenol, chloroform and isoamyl alcohol (25:24:1) to extract, mix upside down, and centrifuge at 3000 rpm for 10 minutes. Take the supernatant, add an equal volume of chloroform, mix upside down Evenly, centrifuge at 3000 rpm ...

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Abstract

The present invention relates to a molecular marker linked with wheat anticalcino gene and its obtaining method. Said invention adopts two molecular marking methods of RAPD and SSR to make screening of resistance marker for wheat parent and nosotropic filial generation "Brock / 015 / / Jing 411", in which RAPD marker uses 10bp oligonucleotide developed by American OPERON co. as random primer, and SSR marker uses the primer synthesized by SSR primer sequence reported by R & der in 1998, after screening, the molecular marker OPP15900 and Xgwm 114120 linked with wheat anticalcino gene can be obtained. These markers can be used for auxiliary selection breeding of anticalcino wheat, at the same time can lay the foundation of cloning new anticalcino gene.

Description

[0001] Technical field: The present invention belongs to agricultural biotechnology engineering, and particularly relates to molecular markers linked with powdery mildew resistance genes and methods for obtaining them. technical background: [0002] Wheat powdery mildew caused by the obligate parasitic fungus Erysiphe graminis f.sp.tritici is one of the main diseases that endanger wheat production. In recent years, with the promotion of dwarf and semi-dwarf wheat varieties, the increase of field planting density and the improvement of water and fertilizer conditions, the disease has become more and more serious in the northern and southern wheat regions of my country, and has become one of the main obstacles to high and stable wheat yields. one. In 1990, wheat powdery mildew was prevalent in 39% of the country's wheat growing areas, resulting in an output loss of 400 million kilograms. At present, wheat powdery mildew has become one of the common diseases that threaten wheat p...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12P19/34
Inventor 王振英陈宏彭永康
Owner TIANJIN NORMAL UNIVERSITY
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