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SNP (single nucleotide polymorphism) site of anti-wheat leaf rust gene Lr13 and application of SNP site

A leaf rust resistance gene and leaf rust technology, applied in the field of biogenetics, can solve problems affecting protein functions and changes, and achieve the effects of improving selection efficiency, high throughput, and accelerating the process of disease-resistant breeding

Active Publication Date: 2020-04-28
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This single nucleotide polymorphism may change the protein sequence encoded by the gene containing this site, thereby affecting the function of the protein

Method used

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  • SNP (single nucleotide polymorphism) site of anti-wheat leaf rust gene Lr13 and application of SNP site
  • SNP (single nucleotide polymorphism) site of anti-wheat leaf rust gene Lr13 and application of SNP site
  • SNP (single nucleotide polymorphism) site of anti-wheat leaf rust gene Lr13 and application of SNP site

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The invention provides a method for screening molecular markers of wheat leaf rust resistance, specifically as follows:

[0033] 1. Genetic analysis

[0034] F 2 Phenotypic identification of individual plants in the population, F 2 The population contained 3057 individuals, of which 2300 were resistant and 757 were susceptible. F 2 The population contained 851 individual plants, including 624 resistant individual plants and 227 susceptible individual plants. Use SPSS to carry out chi-square test (suitability test) to the phenotype results. When the expected ratio is 3:1, the chi-square values ​​are 0.092 and 1.268, P>0.05 (Table 2), and the resistance of Liaochun 10 is obtained. Leaf rust traits are controlled by a dominant single gene. Liaochun 10 and F derived from RL4031(Lr13) 2 All 1395 individual plants in the population were resistant to the disease, and it was concluded that the leaf rust resistance gene in Liaochun 10 was Lr13.

[0035] Table 1 Leaf rust i...

Embodiment 2

[0044] In this example, KASP marker primer Lseq302 was used to screen 33 materials known to contain or not contain Lr13. The specific steps are as follows:

[0045] 1. Using the CTAB method to extract DNA from leaf tissues of 33 wheat materials.

[0046] 2. With the genomic DNA extracted in step 1 as a template, PCR amplification is carried out with a primer pair consisting of a single-stranded DNA molecule (upstream primer) and a reverse single-stranded DNA molecule (downstream primer) shown in the sequences FAM and HEX in Table 3 .

[0047] Table 3 The upstream and downstream primer sequences of KASP marker Lseq302

[0048]

[0049]

[0050] Lseq302 KASP labeled primers were dissolved to make a primer mix (12 mM primer sequence FAM and HEX, 30 mM reverse primer sequence), the total volume of the 384 plate reaction system was 5 μL, and 2×KASP V4.0 Mastermix was 2.5 μL (product of LGC Company) ), the primer mix is ​​0.056μL, the DNA template is about 50-100ng, and ddH ...

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Abstract

The invention relates to an SNP (single nucleotide polymorphism) site of an anti-wheat leaf rust gene Lr13 and application of the SNP site. The SNP site is positioned at a 157695819 locus of a 2B chromosome of wheat, and a KASP (kompetitive allele specific polymerase chain reaction) primer combination capable of amplifying the SNP site comprise three primers of SEQ ID NO.2, SEQ ID NO.3 and SEQ IDNO.4. The SNP site interlinked with the anti-wheat leaf rust gene Lr13 is screened, designed and developed, in addition, the KASP primer combination is designed for the SNP site, the type of the SNP site can be accurately identified through the KASP primer combination, then a wheat strain with the anti-wheat leaf rust gene Lr13 can be screened, the selection efficiency can be improved, the anti-disease breeding progress of wheat can be accelerated, and the target of pyramiding breeding of multiple anti-disease genes can be achieved.

Description

technical field [0001] The invention relates to the field of biogenetics, in particular to the SNP site of wheat leaf rust resistance gene Lr13 and its application. Background technique [0002] Wheat leaf rust is a fungal disease caused by the infection of wheat leaf rust (Puccinia triticina). Infect autumn seedlings and spread to adjacent areas. After germination, the uredospores of the fungus produce germ tubes and invade from the stomata of the leaves, producing uredospore piles and uredospores on the leaf surface, and repeatedly infect. When leaf rust occurs, it seriously affects the production of wheat. Wheat leaf rust will reduce the yield by 3% when the disease is mild, and it will reach 25-30% when the disease is severe, and the severity of the disease is positively correlated with the loss of yield. [0003] At present, it is mainly through planting disease-resistant varieties, supplemented by chemical control and cultivation. Chemical control can be done in th...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 解超杰胡兆荣邱丽娜孙其信马骏王卫东王慧芳刘雨佳穆君怡
Owner CHINA AGRI UNIV
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