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Primers and their application for screening wheat leaf rust resistance qtl

A technology for leaf rust and wheat, applied in primers for screening wheat leaf rust resistance QTL and its application field, can solve problems such as yield loss, and achieve the effect of prolonging disease resistance

Inactive Publication Date: 2018-08-31
BAODING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the early stage of the disease, chlorotic spots first appeared on the leaves, and then slowly expanded. In the later stage, brown uredia spores piled up on the leaves. This disease is an airborne disease with a wide distribution range. can cause severe yield loss

Method used

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  • Primers and their application for screening wheat leaf rust resistance qtl
  • Primers and their application for screening wheat leaf rust resistance qtl
  • Primers and their application for screening wheat leaf rust resistance qtl

Examples

Experimental program
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Effect test

experiment example

[0034] A new QTL for wheat leaf rust resistance- QLr.hebau-2DS The SSR markers gwm261 and gwm484 are obtained:

[0035] Fundulea 900, Thatcher and 188 Fundulea 900 / ThatcherF 2:3 The populations were planted in Baoding Experimental Field, Hebei during the wheat growing season for three consecutive years in 2010-2011, 2011-2012 and 2012-2013 respectively. A completely randomized block design was adopted for field planting, and a total of 2 repetitions were set up. The row length was 1.2 m and the row spacing 0.25 m, about 50 grains per row. One row of high-sensitivity variety Zhengzhou 5389 was planted in every 10 rows as a control. The induced row (Zhengzhou 5389) was planted vertically with the test material.

[0036] The rows were inoculated with mixed leaf rust strains, and when the leaves of the induced rows were fully infected, the rust resistance of the population was identified, and the field maximum disease severity (MDS) was statistically analyzed.

[0037] The DN...

Embodiment 1

[0043] 1. Extract Thatcher genomic DNA as a template;

[0044] 2. Using wms261 and wms484 as primers and Thatcher genomic DNA as a template, carry out PCR amplification. 10 μL PCR reaction system: 10×buffer 1 μL, 10 mmol L -1 dNTPs 0.2 μL, 4 μmol L -1 Primer 1 μL, 30 ngμL -1 DNA template 1 μL, Taq Enzyme 0.5 U, ddH 2 O 6.7 μL.

[0045] The reaction program was pre-denaturation at 94°C for 5 min; denaturation at 94°C for 45 sec, annealing at 55°C for 45 sec, extension at 72°C for 45 sec, 35 cycles; final extension at 72°C for 10 min, and storage at 10°C.

[0046] 3. Separation and detection of the amplified products: add 2.5 μL 6×Loading buffer to the amplified products, separate them by electrophoresis in a non-denaturing polyacrylamide gel with a concentration of 12% (w / v), and develop the color by silver staining. Such as figure 1 and figure 2 shown ( figure 1 Non-denaturing polyacrylamide gel electrophoresis pictures of wms261 amplification products, figure...

Embodiment 2

[0048] 1. Extract Fundulea 900 genomic DNA as a template;

[0049] 2. Using wms261 and wms484 as primers and Fundulea 900 genomic DNA as a template, carry out PCR amplification. 10 μL PCR reaction system is: 10×buffer 1 μL, 10 mmol L -1 dNTPs 0.2 μL, 4 μmol L -1 Primer 1 μL, 30ng μL -1 DNA template 1 μL, Taq Enzyme 0.5 U, ddH 2 O 6.7 μL.

[0050] The reaction program was pre-denaturation at 94°C for 5 min; denaturation at 94°C for 45 sec, annealing at 55°C for 45 sec, extension at 72°C for 45 sec, 35 cycles; final extension at 72°C for 10 min, and storage at 10°C.

[0051] 3. Separation and detection of the amplified products: add 2.5 μL 6×Loading buffer to the amplified products, separate them by electrophoresis in a non-denaturing polyacrylamide gel with a concentration of 12% (w / v), and develop the color by silver staining. Such as figure 1 and figure 2 shown ( figure 1 Non-denaturing polyacrylamide gel electrophoresis pictures of wms261 amplification product...

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Abstract

The invention discloses a primer used for screening wheat brown leaf rust resisting QTL and application thereof and relates to the technical field of nucleic acid determination or test methods. The QTL is located on a chromosome 2DS, the marker interval is Xgwm261-Xgwm484, and the QTL is from a susceptible parent Thatcher; the primer is a primer pair composed of upstream and downstream nucleotide sequences of a primer wms261 and a primer wms484. The primer is used for screening wheat brown leaf rust resisting QTL-QLr.hebau-2DS, and molecular markers in close linkage with the QTL can be used for marking the QTL for auxiliary selection, so that accumulation of multiple effective anti-disease genes are achieved, and disease resistance of varieties is improved. The discovered novel brown leaf rust resisting QTL and the special primer play an important role in wheat disease resisting breeding process.

Description

technical field [0001] The present invention relates to the technical field of assay or detection methods involving nucleic acids. Background technique [0002] Grass crop wheat is an important food crop widely planted all over the world. Its total output is second only to corn. 35%-40% of the world's population uses wheat as their staple food. In my country, wheat is planted in a wide area, mainly planted in Henan, Shandong, Jiangsu, Hebei, Hubei, Anhui and other provinces, mainly winter wheat, its planting area and total output are second only to rice, ranking first in my country's food crops two. by wheat leaf rust ( Puccinia triticina ) caused by wheat leaf rust is a worldwide disease, which can cause a lot of yield loss when it occurs seriously. It mainly damages wheat leaves and rarely occurs on leaf sheaths and stems. In the early stage of the disease, chlorotic spots first appeared on the leaves, and then slowly expanded. In the later stage, brown uredia spores pil...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 周悦崔彬彬贾晓梅曹柳青靳茜郝征
Owner BAODING UNIV
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