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Primer for screening wheat leaf rust resistance QTL and application of primer

A technology for leaf rust and wheat, which is applied to primers for screening wheat leaf rust resistance QTL and its application field, can solve the problems of loss, affect disease prevention effect and quality, and single-gene resistance is not persistent, and achieves long-term extension. The effect of disease resistance

Inactive Publication Date: 2015-09-16
BAODING UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the continuous evolution of new virulence genes of pathogenic bacteria and the increasing selection pressure, this kind of single-gene resistance is generally not durable, and with the widespread planting of single-gene-resistant crop varieties, this resistance will disappear in a short period of time. will be lost, seriously affecting the effect and quality of disease prevention

Method used

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  • Primer for screening wheat leaf rust resistance QTL and application of primer
  • Primer for screening wheat leaf rust resistance QTL and application of primer
  • Primer for screening wheat leaf rust resistance QTL and application of primer

Examples

Experimental program
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Effect test

experiment example

[0033] A new QTL for wheat leaf rust resistance- QLr.zh-4B The SSR markers wmc692 and wmc657 are obtained:

[0034] During the three consecutive growth years of 2011-2012, 2012-2013 and 2013-2014, Zhoumai 22, Chinese Spring and their hybrids were planted in the experimental field of Hebei Agricultural University in Baoding by using a completely randomized block design method and self-crossed year by year. f 2:3 family lineage. Two replicates were planted each year, and the first two rows of each replicate were planted with the parents Zhoumai 22 and China Spring respectively, so as to facilitate the observation of the disease status of the parents, and then the 215 families planted were randomly arranged in sequence. Plant 50 grains in each family, plant in a single row, the row length is 1.5m, and the row spacing is 0.25m. Zhengzhou 5389 is planted in every ten rows, which is convenient for counting and used as a susceptible control. Zhengzhou 5389 is planted on both sides...

Embodiment 1

[0041] 1. Extract Chinese spring genomic DNA as a template;

[0042] 2. Using wmc692 and wmc657 as primers and Chinese spring genome DNA as template, carry out PCR amplification. 10μL PCR reaction system: 10×buffer 1 μL, 10 mmol L -1 dNTPs 0.2 μL, 4 μmol L -1 Primer 1 μL, 40 ng μL -1 DNA template 1 μL, Taq Enzyme 1 U, ddH 2 O 6.7 μL.

[0043] The PCR reaction program was pre-denaturation at 94°C for 5 min; denaturation at 94°C for 1 min; the annealing temperatures of primers wmc692 and wmc657 were 51°C and 61°C respectively, the annealing time was 1 min, and extension at 72°C for 1 min; 35 cycles; Extend for 10 min at ℃ and store at 4℃.

[0044] 3. Separation and detection of the amplified products: add 2.5 μL 6×Loading buffer to the amplified products, take 5 μL for each amplified sample, and run it on a non-denaturing polyacrylamide gel with a concentration of 12% (w / v). Separated by electrophoresis and visualized by silver staining. Such as figure 1 and figur...

Embodiment 2

[0046] 1. Extract Zhoumai 22 genomic DNA as a template;

[0047] 2. Using wmc692 and wmc657 as primers and Zhoumai 22 genomic DNA as template, carry out PCR amplification. 10μL PCR reaction system: 10×buffer 1 μL, 10 mmol L -1 dNTPs 0.2 μL, 4 μmol L -1 Primer 1 μL, 40 ng μL -1 DNA template 1 μL, Taq Enzyme 1 U, ddH 2 O 6.7 μL.

[0048] The PCR reaction program was pre-denaturation at 94°C for 5 min; denaturation at 94°C for 1 min; the annealing temperatures of primers wmc692 and wmc657 were 51°C and 61°C respectively, the annealing time was 1 min, and extension at 72°C for 1 min; 35 cycles; Extend for 10 min at ℃ and store at 4℃.

[0049] 3. Separation and detection of the amplified products: add 2.5 μL of 6×Loading buffer to the amplified products, separate 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 polyacryl...

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Abstract

The invention discloses a primer for screening wheat leaf rust resistance QTL and an application of the primer, and relates to the technical field of determination or detection method of nucleic acid. The primer is a primer pair consisting of upstream and downstream nucleotide sequences of primers wmc692 and wmca657. The invention provides novel SSR markers wmc692 and wmc657 of the wheat leaf rust resistance QTL-(i) QLr.zh-4B. A molecular marker which is tightly linked to the QTL can be utilized to perform the assistant marker selection for the QTL, so that the accumulation of a plurality of active disease-resistant genes can be realized, and the disease resistance of a species can be prolonged. The novel leaf rust resistance QTL and the special primer can play an important role in the long-term disease resistance breeding of wheat.

Description

technical field [0001] The present invention relates to the technical field of assay or detection methods involving nucleic acids. Background technique [0002] Wheat is the food crop with the largest planting area, the largest total output and total trade volume in the world, and has high nutritional value. China ranks first among the major wheat-producing countries in the world, and wheat is one of the three major staple food crops in my country. The planting area is only Next to rice and corn, wheat is the main food for about half of the country's population. Proper production of wheat plays an important role in my country's social stability and the sustainable and healthy development of the national economy. by wheat leaf rust ( Puccinia triticina ) caused by wheat leaf rust is a worldwide disease, which can cause great yield loss when it occurs seriously. In my country, in the past, wheat leaf rust mainly occurred in the southwest and parts of the Yangtze River Basin, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 周悦靳茜秦金燕贾晓梅崔彬彬曹柳青郝征
Owner BAODING UNIV
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