Molecular marker for detecting wheat backbone germplasm weekly 8425B specific chromosome segment and application of molecular marker

A technology of chromosome fragments and molecular markers, which is applied in the field of molecular markers for the detection of 8425B specific chromosome fragments of wheat backbone germplasm, can solve the problems of lack of specific detection molecular markers, and achieve the effect of improving detection efficiency

Active Publication Date: 2021-12-24
HENAN AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0008] The technical problem to be solved by the present invention is: aiming at the lack of molecular markers for the specific detection of chromosome 1B of wheat backbone germplasm week 8425B, the present invention provides a molecular marker for specific detection of chromosome 1B of wheat backbone germplasm week 8425B, which can be judged Whether the wheat plants have inherited the 1B chromosome of Zhou 8425B, and then predict the resistance of wheat plants to leaf rust, stripe rust, and powdery mildew, and accelerate the breeding process of high-yield and multi-resistant wheat varieties

Method used

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  • Molecular marker for detecting wheat backbone germplasm weekly 8425B specific chromosome segment and application of molecular marker
  • Molecular marker for detecting wheat backbone germplasm weekly 8425B specific chromosome segment and application of molecular marker
  • Molecular marker for detecting wheat backbone germplasm weekly 8425B specific chromosome segment and application of molecular marker

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Experimental program
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Effect test

Embodiment 1

[0034] Specific detection of germplasm resources: The parents of wheat backbone germplasm Zhou 8425B, Guangmai 74, Lianfeng 1, Shanqianmai and Anong 7959, collected other well-known 1BL / 1RS wheat germplasms at home and abroad, Aimengniu, Luomengniu Fulin No. 10, Lovelin No. 13, Lankao 906, Niuzhut, Aurora, Caucasus Mai, Bainong 207, Zhoumai 36, Zhoumai 22, Zhengmai 366, Xinong 388, Pingan 518, Saidmai 5, Xinmai 36, Aikang 58 and other wheat varieties widely popularized in the production of Huanghuai wheat region and the germplasm China Spring commonly used in scientific research.

[0035] Wheat backbone germplasm Zhou 8425B is a new type of 1BL / 1RS translocation wheat germplasm created by the distant hybridization of triticale and common wheat varieties through stepwise compound hybridization, radiation mutagenesis and chromosome detection. Firstly, the hexaploid triticale Guangmai 74 was crossed with the common wheat Lianfeng 1, and its F 1 Further crossing with the common 1...

Embodiment 2

[0055] In March 2020, 74 derivative varieties of Zhou 8425B were molecularly detected by the method of the present invention, and their resistance to stripe rust and leaf rust was predicted. The detailed steps of detection are as follows:

[0056] a. Sample collection: At the jointing stage of wheat, take 184 weekly 8425B derivative varieties with 1.5cm long leaves, put them into sterilized 1.5mL centrifuge tubes, put them in the ice box and bring them back to the laboratory for DNA extraction;

[0057] b. DNA extraction: use the 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,

[0058] ① After 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 for 1 hour, and shake well every 15 minutes;

[0059] ②Take out the sample and cool it on ice, add 600 μL chloroform / isoamyl alco...

Embodiment 3

[0077] Example 3: The present invention can predict the incidence rate in advance, do hybridization one year in advance, and improve breeding efficiency. The detection material is BC 1 Population of 45 individuals, BC 1 The population was obtained by crossing the high-yielding variety Zhoumai 36 (female parent) with the main parent germplasm Zhou 8425B (male parent) and backcrossing once. 45 BC 1 Individual plants are defined as ZM36 / Z84-1~ZM36 / Z84-45.

[0078] The basic detection steps are basically the same as in Example 2, except that:

[0079] In step b: ⑥Dry the DNA precipitate, add 100 μl of ultrapure water, let stand at room temperature for 60 minutes, and store at -20°C for later use.

[0080] 45 BC planted in March 2021 1 Molecular testing was carried out on individual plants, and the test results are shown in Table 2. In April 2021, 14 individual plants carrying the Z84-1B-1 marker were screened for backcrossing with the high-yielding reincarnation parent Zhoumai...

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Abstract

The invention relates to the field of wheat breeding and molecular biology, in particular to a molecular marker for detecting a wheat backbone germplasm weekly 8425B specific chromosome segment and application of the molecular marker. The molecular marker is characterized in that wheat DNA is used as a template, a primer pair as shown in nucleotide sequences Z84-1B-1F and Z84-1B-1R is used for PCR amplification, a DNA fragment with the size of 207bp is obtained after electrophoretic separation is performed by polyacrylamide gel with the mass fraction of 8%, and the marker is named as ZH84-1B-1 by the applicant. The nucleotide sequence of the molecular marker ZH84-1B-1 is as shown in SEQ ID NO: 1. The molecular marker can be used for detecting whether a wheat plant has a specific 1BL / 1RS translocation fragment of the week-8425B. The molecular marker is specific to Zhou 8425B, is linked with three disease genes, can be simultaneously used for molecular marker-assisted selection of the three diseases, and can simultaneously detect whether three disease-resistant genes are introduced into a breeding material or not after one-time hybridization, so that the detection efficiency of disease-resistant breeding is improved by two times.

Description

technical field [0001] The invention relates to the fields of wheat breeding and molecular biology, in particular to a molecular marker for the detection of 8425B specific chromosome fragments of wheat backbone germplasm and its application. Background technique [0002] Wheat is one of the most important ration crops in my country. The sown area of ​​wheat in the Huanghuai wheat region, the total output and the transfer rate of commercial grain account for more than 60%, 70% and 80% of the country respectively, which plays a decisive role in the national food security and the effective supply of agricultural products. Impact. After the 1970s, the wheat yield in this area quickly transitioned from long-term low yield to medium-high yield. However, at that time, most of the wheat varieties had high stalks and lodging and frequent occurrence of diseases, which seriously affected the further improvement of wheat yield. In wheat breeding, it is urgent to create short-stalked, hig...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/13C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 殷贵鸿李巧云唐建卫李俣佳高艳许豪高闯
Owner HENAN AGRICULTURAL UNIVERSITY
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