Primer combination assisting in screening stripe-rust-resistant wheat and application of primer combination

A stripe rust and primer combination technology, applied in the direction of recombinant DNA technology, microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problems of wheat stripe rust epidemic, loss of resistance, yield loss, etc.

Active Publication Date: 2015-12-09
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Historically, the known stripe rust resistance genes Yr9, Yr24/Yr26, Yr10, etc. have lost their resistance due to the successive em...

Method used

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  • Primer combination assisting in screening stripe-rust-resistant wheat and application of primer combination
  • Primer combination assisting in screening stripe-rust-resistant wheat and application of primer combination
  • Primer combination assisting in screening stripe-rust-resistant wheat and application of primer combination

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1. Molecular markers Xwmc382 and Xgwm636 of stripe rust resistance genes and the acquisition of their dedicated primers

[0048] Using my country's stripe rust race CYR29 on wheat materials Zhongmai 175 (resistant variety) and Luoxuan 987 (susceptible variety) and their F 1 , F 2 and F 3 The seedling stage resistance identification was carried out in the generation population, and the results are shown in Table 2.

[0049] Table 2 Genetic analysis of resistance of Zhongmai 175, Lunxuan 987 and their progeny

[0050]

[0051] The parent Zhongmai 175 is all disease-resistant, and the parent round selection 987 is all susceptible, F 1 All disease resistant, F 2 There were 252 resistant strains and 92 susceptible strains in the population, which conformed to the ratio of 3:1 (χ 2 =0.56, P 1df = 0.46). 147 F 3 Among the families, 29 families showed homozygous disease resistance, 78 families showed heterozygous disease resistance, 40 families showed homozyg...

Embodiment 2

[0061] Example 2. Source and disease resistance identification of new wheat stripe rust resistance genes

[0062] The pedigree of Zhongmai 175 is BPM27 / Jing 411.

[0063] The disease resistance of Jing 411 and Zhongmai 175 were identified respectively, Jing 411 was highly susceptible (infection type IT=4), and Zhongmai 175 was highly resistant (infection type IT=0;).

[0064] The following molecular identifications were carried out for each tested wheat (Zhongmai 175, Jing 411 or Lunxuan 987): the genomic DNA of the wheat leaves to be tested was extracted; the genomic DNA was used as a template, and the Xwmc382 primer pair and the Xgwm636 primer pair were used for PCR amplification. The PCR amplification product was subjected to 6% denaturing polyacrylamide gel electrophoresis, and then silver stained for color development.

[0065] The band pattern of Jing 411 is consistent with that of round selection 987 (amplified with Xwmc382 primer pair, shown as band pattern A2; amplif...

Embodiment 3

[0067] Example 3. Screening of stripe rust resistant wheat with Xwmc382 primer pair and Xgwm636 primer pair

[0068] The wheat varieties (materials) to be tested are: Zhongmai 175, Jing 411, Lunxuan 987, China Chun, Mingxian 169, AvocetS, Yonggang 2, Zhoumai 26, Jimai 22, Jimai 23, Yangmai 21, Yang Mai 14, Zheng Mai 366, 92 Hui 94-1-12, 99-19-10, CA12107, CA13152 and Lantian 21.

[0069] 1. Extract the genomic DNA of the wheat leaves to be tested.

[0070] 2. Using the genomic DNA extracted in step 1 as a template, PCR amplification was performed using the Xwmc382 primer pair and the Xgwm636 primer pair respectively.

[0071] PCR reaction system: Genomic DNA 1.5μl (50-100ng / μl), upstream and downstream primers (4μmol L -1 ), 7.5 μl of 2×TaqPCRMix (Beijing Huitian Oriental Technology Co., Ltd., Cat: HT201), supplemented to 15 μl with sterile ultrapure water.

[0072] PCR amplification program using Xwmc382 primer pair: 94°C pre-denaturation for 5 min; 94°C denaturation for 1 m...

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Abstract

The invention discloses a primer combination assisting in screening stripe-rust-resistant wheat and an application of the primer combination. The primer combination comprises an Xwmc382 primer pair and an Xgwm636 primer pair, wherein the Xwmc382 primer pair comprises single-chain DNA (deoxyribose nucleic acid) molecules shown in a sequence 1 in a sequence table and single-chain DNA molecules shown in a sequence 2 in the sequence table; the Xgwm636 primer pair comprises single-chain DNA molecules shown in a sequence 3 in the sequence table and single-chain DNA molecules se shown in a quence 4 in the sequence table. The primer pair and molecular marks can be used for breeding of stripe-rust-resistant wheat molecules and cloning of stripe-rust-resistant genes. The special primers and the molecular marks play an important role in breeding of disease-resistant wheat.

Description

technical field [0001] The invention relates to a primer set for auxiliary screening of stripe rust-resistant wheat and its application. Background technique [0002] Wheat stripe rust is a worldwide foliar fungal disease that seriously affects wheat yield and quality. Wheat stripe rust is caused by Puccinia striiformisf.sp.tritici, which likes shade and coolness. It occurs in almost all wheat growing areas in the world, with a loss of 10-70% in average years, and even in severe years. In my country, wheat stripe rust mainly occurs in the northwest and southwest regions (such as Gansu, Sichuan and other provinces), and has also increased in Shaanxi, Hubei and other provinces in recent years. In 1950, 1964, 1980 and 2002, the disease was once widespread in my country, causing huge losses to wheat production. In the past 10 years, stripe rust occurred on an area of ​​about 60 million mu each year. In recent years, agricultural measures and chemical agents have effectively re...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 夏先春卢家玲何中虎陈璨
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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