SNP molecular marker of wheat leaf rust resistant gene Lr42, detection method and application

An anti-leaf rust gene and molecular marker technology, which can be used in the determination/inspection of microorganisms, biochemical equipment and methods, recombinant DNA technology, etc., can solve the problems of inability to meet the needs of modern breeding, complex detection procedures, and poor accuracy. Achieve the effect of easy automation, high detection throughput, and high accuracy

Active Publication Date: 2019-01-08
HENAN ACAD OF AGRI SCI XIAOMAI INST
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0004] Sun et al. (2010) developed three SSR molecular markers linked to Lr42, namely Xwmc432, Xcfd15, and Xgdm33. These three markers have the problems of complicated detection procedures, long time and poor accuracy in the application process, which cannot satisfy The needs of modern breeding

Method used

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  • SNP molecular marker of wheat leaf rust resistant gene Lr42, detection method and application
  • SNP molecular marker of wheat leaf rust resistant gene Lr42, detection method and application
  • SNP molecular marker of wheat leaf rust resistant gene Lr42, detection method and application

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Effect test

Embodiment 1

[0036] The method for obtaining the SNP molecular marker TC113325 of wheat leaf rust resistance gene Lr42 comprises the following steps:

[0037] (1) Construction of high-generation genetic segregation population:

[0038] Using the existing molecular markers of Lr42: Xcfd15 and Xwmc432 to scan the F4 generation population of KS93U50 / Morocco, it was found that the genetically isolated individual plants were heterozygous genotypes at the disease resistance site. Planted as high generation genetic segregants;

[0039] (2) Obtain SNP molecular markers and design amplification primers:

[0040] Using the homologous segment sequences of the wheat chromosome segments of the existing molecular markers Xcfd15 and Xwmc432 of Lr42 in closely related species such as rice, Brachypodium, and barley, to find genes with disease resistance domains, and use the found genes to Primers were designed for its conserved domain, and after verification by the KS93U50 / Morocco high-generation segrega...

Embodiment 2

[0042] The detection method of wheat leaf rust resistance gene Lr42 comprises the following steps:

[0043] (1) Take 93 seeds of an isolated F5 population of KS93U50 / Morocco and plant them in the greenhouse. When the population grows to three leaves and one heart, extract population DNA: use QIAGEN's DNeasy Plant MiniKit to extract according to the instructions; all populations After the sample DNA is extracted, perform DNA integrity electrophoresis detection, and use a concentration meter to measure the sample DNA concentration, and dilute all sample concentrations to 50-100ng / ul according to the measurement results;

[0044] (2) PCR amplification of molecular markers: PCR amplification system is: DNA 2.5ul (50-100ng / ul); mixed primer 0.07ul (SEQ ID NO.1:SEQ ID NO.1:SEQ ID NO.1: H2O=12:12:30:46); MasterMix 3ul (2×); ddH2O 0.493ul; PCR reaction total volume 6.0ul;

[0045] The PCR amplification program was as follows: denaturation at 94°C for 15 min; denaturation at 95°C for ...

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Abstract

The invention discloses an SNP molecular marker TC113325, of a wheat leaf rust resistant gene Lr42, a method of detecting the wheat leaf rust resistant gene Lr42 by using the molecular marker and application thereof. The molecular marker is obtained by amplifying the following primers, wherein the upstream primer sequences are as shown in SEQ ID NO. 1 and SEQ ID NO. 3 and a downstream primer sequence is as shown in SEQ ID NO. 3. The method of detecting the wheat leaf rust resistant gene Lr42 by using the molecular marker TC113325 has the characteristics of being high in detection flux, short in time, high in accuracy, easy to implement automation, low in cost, environmentally friendly and the like, and is applied on a large scale in breeding work favorably.

Description

technical field [0001] The invention relates to the fields of crop molecular biology and molecular breeding, in particular to a SNP molecular marker of wheat leaf rust resistance gene Lr42, a detection method and application thereof. Background technique [0002] As the staple food of 40% of the world's population, wheat is one of the three major staple foods in my country. my country is the most populous country in the world. Food supply security is a strategic task that my country must always adhere to. Leaf rust is a major leaf disease of wheat, caused by the fungus Puccinia triticina Eriks, which can cause a 40% yield loss in severe years (Knott1989). Breeding and popularizing disease-resistant samples is the most effective strategy for disease prevention and control. Aggregating multiple disease-resistant genes into the same variety through molecular markers is the main method of disease-resistant breeding. [0003] Lr42 is a leaf rust resistance gene from A. tauschii ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 李春鑫王会伟赵明忠张建周郭瑞韩留朋杜习军
Owner HENAN ACAD OF AGRI SCI XIAOMAI INST
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