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Molecular marker for detecting wheat leaf rust resistance gene Lr47, detection method and application of molecular marker

A technology of leaf rust resistance gene and molecular marker, which is applied in the field of molecular genetic breeding, can solve the problem of the difficulty of wheat rust resistance gene, and achieve the effect of low detection difficulty

Active Publication Date: 2022-05-13
PEKING UNIV INST OF ADVANCED AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The main purpose of the present invention is to provide a molecular marker for detecting wheat leaf rust resistance gene Lr47, a detection method and its application, so as to solve the difficult problem of detecting wheat rust resistance gene in the prior art

Method used

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  • Molecular marker for detecting wheat leaf rust resistance gene Lr47, detection method and application of molecular marker
  • Molecular marker for detecting wheat leaf rust resistance gene Lr47, detection method and application of molecular marker
  • Molecular marker for detecting wheat leaf rust resistance gene Lr47, detection method and application of molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1. Indoor seedling stage leaf rust resistance identification

[0052] The disease-resistant parent Kern Lr47 and the susceptible parent CSph1b were planted in the artificial climate chamber. The conditions of the artificial climate chamber were: 16 hours of light, 8 hours of darkness, 25°C during the day, 22°C at night, and 80-90% humidity. When the seedlings grow to the stage of two leaves and one heart, they are inoculated with the highly toxic physiological race PHQS of leaf rust by manual sweeping. Dark and moisturizing treatment for 24 hours after inoculation. After 12 days of inoculation, the materials were identified for leaf rust resistance, and the identification method was carried out according to the grading standard of 0-4 (Note: 0 is immune; 0 is near immune; 1 is high resistance; 2 is moderate resistance; 3 Grade 4 is medium sense; grade 4 is high sense). The results showed that: Kern Lr47 showed near immune resistance, while CSph1b showed hig...

Embodiment 2

[0054] Example 2. Creation of a short-segment translocation line of Pseudospeltia spp.

[0055] The lack of high-quality reference genomes and the cross-pollination characteristics of this species lead to large sequence differences among different materials, which has caused certain difficulties in the development of specific molecular markers. The transcriptomes and The exome sequencing data, combined with the reference genome sequence and bioinformatics analysis of all sequenced wheat, found the specific SNPs (single nucleotide polymorphism, single nucleotide polymorphism) of the 7S chromosome of Alpinia spelta, and obtained SNPs data also indicated that Kern Lr47 carried about 150Mb of 7S chromosome fragment. This application uses the method of the Chinese spring ph1b mutant to narrow down the chromosome interval where the foreign gene is located. After knowing the range of the gene position, it is possible to further develop tightly linked molecular markers. Multiple gen...

Embodiment 3

[0060] Example 3. Short fragments of Goatwort pseudospelts resistant to leaf rust are transferred to the main cultivars of common wheat

[0061] To transfer the reduced 7S chromosome segment to common wheat cultivars, individual plants containing homozygous short 7S chromosome segments identified from recombinant F284 progeny (eg, image 3 The individual plants corresponding to lanes 4, 6, and 11) were crossed with the common wheat variety Yangmai 21 to obtain F 1 .

[0062] Using the molecular marker Pku1176 to perform PCR amplification on the genome DNA of a single plant of the progeny population of the recombinant F284 x Yangmai 21, the specific reaction system is as follows:

[0063] DNA template (100ng / μL) 0.5μL

[0064] 2×Taq Plus PCR MasterMix 6.25 μL (Ruibo Xingke Company)

[0065] 0.5 μL each of 10 μM primers

[0066] ddH 2 O make up to 12.5 μL

[0067] PCR amplification reaction program: pre-denaturation at 94°C for 3min; denaturation at 94°C for 30s, annealing...

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Abstract

The invention provides a molecular marker for detecting a wheat leaf rust resistance gene Lr47, a detection method and application of the molecular marker. The invention develops a molecular marker Pku1176 closely linked with a leaf rust resistant gene Lr47 by means of an induced wheat-Aegilops spelltuifolia leaf rust resistant short fragment translocation line and in combination with reference sequences of wheat and Aegilops spelltuifolia. By utilizing the molecular marker, translocated short fragment 7S chromosomes under different wheat backgrounds can be accurately and quickly detected, the early molecular assisted selection of the leaf rust resistant gene Lr47 is realized, and the wheat disease-resistant breeding efficiency is improved. The method solves the problem of high difficulty in detecting the wheat rust-resistant gene in the prior art, and is suitable for the field of molecular genetic breeding.

Description

technical field [0001] The invention relates to the field of molecular genetic breeding, in particular to a molecular marker for detecting wheat leaf rust resistance gene Lr47, a detection method and an application thereof. Background technique [0002] Wheat leaf rust (Leaf rust) is an airborne fungal disease caused by the infection of wheat leaf rust (Puccinia triticina), which has the characteristics of wide distribution, fast transmission speed and large damage loss. In recent years, with climate change, changes in farming systems and the emergence of new physiological races of leaf rust, the damage of wheat leaf rust has become increasingly serious, and it has become an important disease that seriously threatens the safe production of wheat in my country. Therefore, controlling wheat leaf rust has become an important task in wheat production. [0003] Breeding and using disease-resistant varieties is the most economical, safe and effective way to control wheat leaf rus...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11C12Q1/6844C12Q1/6827
CPCC12Q1/6895C12Q1/6844C12Q1/6827C12Q2600/13C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 陈时盛李洪娜王桂平华蕾沈涛李洪雨姜登基罗静白升升
Owner PEKING UNIV INST OF ADVANCED AGRI SCI
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