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Molecular marker closely linked with wheat bipolaris sorokiniana black point resistant QTL and application of molecular marker

A technology for molecular markers of Helminthosporium umbilicalis, which is applied in the field of molecular markers closely linked to QTL for resistance to black germ disease of wheat root rot, can solve problems such as laborious, time-consuming, and unstable results, and achieve rapid identification, Stable results and time-saving effect

Active Publication Date: 2021-05-04
HENAN AGRICULTURAL UNIVERSITY
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AI Technical Summary

Problems solved by technology

[0008] Aiming at the problems of lack of practical molecular markers in the current breeding of wheat resistance to black germ disease, time-consuming, laborious and unstable results of existing identification methods for resistance to black germ disease, the invention provides a method for detecting whether there is a wheat plant with resistance to black germ disease wheat Molecular markers closely linked to the resistance QTL of the line Yuyou 1 can be used to determine whether wheat plants carry the black germ resistance QTL, and then evaluate the resistance of wheat plants to Bipolaris sorokiniana black germ disease, so as to speed up the selection process of wheat resistant to black germ disease. The specific technical scheme is as follows:

Method used

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  • Molecular marker closely linked with wheat bipolaris sorokiniana black point resistant QTL and application of molecular marker
  • Molecular marker closely linked with wheat bipolaris sorokiniana black point resistant QTL and application of molecular marker
  • Molecular marker closely linked with wheat bipolaris sorokiniana black point resistant QTL and application of molecular marker

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] F 2 plant, F 2 The offspring of the plant adopt the method of single grain propagation, and finally obtain F 7 Recombinant inbred line population, for F 7 The recombinant inbred line population was inoculated and identified, and 30 extremely disease-resistant and extremely susceptible families were selected from each to construct a disease-resistant pool and a disease-susceptible pool. The disease-resistant parents, disease-susceptible parents, and disease-resistant pools were connected to the susceptible pool through a 660K chip. Genotype analysis was performed on the disease pool to detect the SNPs associated with resistance to black embryo disease. The most associated SNPs were detected on chromosome 4B. The SNP markers on chromosome 4B were converted to dCAPS markers, and the genes of each family of the recombinant inbred line were tested. Type detection, using the QTL IciMapping V4.1 mapping software to construct a genetic linkage map, and combining the genetic l...

Embodiment 2

[0050]In March 2020, use the present invention to quickly predict F 5 Resistance of population lines to B. sorokiniana black embryo disease, F 5 The female parent of the population is the high-yielding variety Zhoumai 36, and the male parent is the disease-resistant line Yuyou 1, with 120 F 5 The strains are defined as ZY-BP-1~ZY-BP-120, and the detailed steps of detection are as follows:

[0051] a. Sample collection: at the jointing stage of wheat, take 120 F 5 Put the 1.5cm-long leaves of each family into a sterilized 1.5mL centrifuge tube, put them in an ice box and bring them back to the laboratory for DNA extraction;

[0052] b. DNA extraction: use CTAB method (Wang Guanlin, Fang Hongyun. Principles and Technology of Plant Genetic Engineering. Science Press, 1998: 370-372) to extract the DNA of the sample in step a,

[0053] ①After grinding the leaves with liquid nitrogen, quickly add 600 μL CTAB extract to the centrifuge tube, mix well, place in a water bath at 65°C ...

Embodiment 3

[0075] The detection material is BC 1 Population of 50 individuals, BC 1 The population was obtained by crossing the high-yielding variety Saidemai No. 7 (female parent) with the disease-resistant line Yuyou No. 1 (male parent), and then backcrossing again. 50 BC 1 Individual plants are defined as SY-BP-1~SY-BP-50.

[0076] The detection steps are basically the same as in Example 2, the difference is that:

[0077] In step b: ⑥Dry the DNA precipitate, add 50 μL of ultrapure water, let it stand at room temperature for 30 minutes, and store it at -20°C for later use.

[0078] 50 BC planted in March 2020 1 Molecular testing was carried out on individual plants, and the test results are shown in Table 2. In April 2020, 22 individual plants carrying the BP-4B-c1 marker were screened and the individual plants with excellent agronomic traits were backcrossed with the high-yielding recurrent parent Saidemai No. 7. If the field black germ disease resistance identification is carri...

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Abstract

The invention relates to the field of wheat disease resistance breeding and molecular biology, in particular to a molecular marker closely linked with wheat bipolaris sorokiniana black point resistant QTL and application of molecular marker. The invention provides a molecular marker for detecting whether a wheat plant has a nucleotide sequence which is shown as SEQ ID NO: 1, wherein the molecular marker is tightly linked with a black-point-resistant wheat line Youyou 1 resistance QTL. The molecular marker is a DNA fragment with the size of 249bp, which is obtained by carrying out PCR (polymerase chain reaction) amplification enzyme digestion electrophoresis separation on a primer pair shown in nucleotide sequences SEQ ID NO: 2 and SEQ ID NO: 3 by taking wheat DNA as a template, and the marker is named as BP-4B-c1. According to the method, through molecular marker detection in the seedling stage, resistance prediction and screening of the wheat to the B.sorokiniana black point can be rapidly conducted, precious scientific research time and a large amount of manpower and material resources are saved, and the identification result is stable. Therefore, the method can accurately and efficiently screen the black-point-resistant wheat strain, and the breeding process of the high-yield black-point-resistant wheat is greatly improved.

Description

technical field [0001] The invention relates to the field of wheat disease-resistant breeding and molecular biology, in particular to a molecular marker closely linked with wheat root rot Helminthosporium umbilicalisis resistance QTL and its application. Background technique [0002] Wheat black point (black point) is a common wheat disease, which refers to black and brown lesions of different shapes and degrees on the surface of wheat grains. . Wheat black germ disease not only affects the appearance quality of wheat grains and leads to the reduction of wheat purchase grade, but also reduces the germination rate of wheat and the dry weight and fresh weight of seedlings. What is more serious is that some black germ disease pathogens will produce toxins and seriously affect human health. Therefore, different countries have strict restrictions on the black germ rate of wheat seeds. my country's national standard for wheat (GB 1351-2008) stipulates that the imperfect kernels ...

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 李巧云殷贵鸿唐建卫姜玉梅王丝雨高闯高艳
Owner HENAN AGRICULTURAL UNIVERSITY
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