Molecular marker closely linked with QTL resistant to wheat crown rot and application of molecular marker

A technology for stem rot and molecular marker, which is applied in the field of wheat breeding and molecular biology to achieve the effects of stable PCR amplification, simple operation and improved breeding efficiency

Active Publication Date: 2019-02-22
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, the QTL markers that have been reported so far are obtained from foreign disease-resistant germplasm, and the QTL markers used in domestic wheat breeding have not been reported yet.

Method used

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  • Molecular marker closely linked with QTL resistant to wheat crown rot and application of molecular marker
  • Molecular marker closely linked with QTL resistant to wheat crown rot and application of molecular marker
  • Molecular marker closely linked with QTL resistant to wheat crown rot and application of molecular marker

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] F 2 plant, F 2 The offspring of the plant adopt the method of single grain propagation, and finally obtain F 6 Recombinant inbred line populations; use SSR and other markers to analyze the genotypes of the obtained recombinant inbred line populations, and use mapping software such as JoinMap to construct genetic linkage maps; Sex identification; comprehensive genetic linkage map and stem rot resistance identification data were analyzed by QTL analysis software such as MapQTL, and the disease resistance QTL was located and the molecular markers closely related to the disease resistance QTL were obtained. Xwmc388-205 shown in the present invention is a molecular marker closely linked with the 7A chromosome disease resistance QTL of CI12633. This tag can be obtained by:

[0024] CI12633 leaf DNA was extracted by CTAB method (Proc Natl Acad Sci USA, 1984, 81(24):8014-8018).

[0025]PCR amplification was performed using the following reaction system and operating procedu...

Embodiment 2

[0032] 1. Predict the strain resistance of C01~C67

[0033] The strains defined as C01~C67 in the examples are based on CI12633 as the male parent and Yangmai 158 as the female parent. 8 Generation of high-generation strains.

[0034] (1) Extract DNA from leaves of C01-C67 lines by CTAB method.

[0035] (2) Using the wheat DNA obtained in step (1) as a template and using SEQ NO:1 and SEQ NO:2 as primers to perform PCR amplification.

[0036] PCR reaction system: total volume 20μl, including 10×buffer 2μl, 1.5mM MgCl 2 , 0.2mM dNTPs, 0.25μM each of SEQ NO:1 and SEQ NO:2, 50ng of template DNA.

[0037] PCR operation program: pre-denaturation at 94°C for 5 minutes; then denaturation at 94°C for 30 sec, annealing at 55°C for 30 sec, extension at 72°C for 30 sec, a total of 40 cycles; and finally extension at 72°C for 5 min.

[0038] (3) After the amplified product is separated by electrophoresis on a 12% polyacrylamide gel, check whether it contains the Xwmc388-205 molecular m...

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Abstract

The invention discloses a molecular marker Xwmc388-205 closely linked with QTL resistant to wheat crown rot and application of the molecular marker Xwmc388-205. The molecular marker is a DNA segment with the size of 205 bp, wherein the DNA segment is obtained in the mode that the a wheat DNA is adopted as a template, PCR amplification is conducted through primer pairs of with nucleotide sequencesshown as SEQ No.1 and SEQ No.2, and , and then electrophoretic separation is conducted through 12% polyacrylamide gel of 12%. The molecular marker Xwmc388-205 can conduct forecasting and screening indoors by detecting resistance of the molecular marker to wheat crown rot, susceptible plants are weeded out, and thus the breeding efficiency is improved.

Description

technical field [0001] The invention relates to the fields of wheat breeding and molecular biology, in particular to a molecular marker closely linked with wheat stalk rot resistance QTL and its application. Background technique [0002] Wheat crown rot, also known as Fusarium crown rot, is an important worldwide wheat disease that occurs in arid and semi-arid regions. Occurs in the southern Pacific coast, Canada, Italy, the Middle East, North Africa and China (Molecular Plant Pathology, 2018, 19(7): 1547-1562). The disease is mainly caused by Fusarium graminearum in Australia (Plant Breeding, 2015,134:365-372), and in China by Fusarium graminearum and Fusarium asiaticum, However, F. graminearum has been on the rise in recent years (Journal of Wheat Crops, 2016, 36(11):1547-1552). The pathogen begins to infect the base of the wheat stem, and the leaf sheaths and stems at the base of the stem become brown at the seedling stage, sometimes causing browning and rot at the root...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12N15/11C12Q1/6895C12Q2600/13C12Q2600/156
Inventor 周淼平张鹏陈达姚金保马鸿翔
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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