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SSCP marker closely linked with major wheat scab resistance QTL and application thereof

A technology for wheat scab and scab, applied in the fields of wheat breeding and molecular biology, to achieve the effects of low cost, easy operation and improved breeding efficiency

Inactive Publication Date: 2010-11-24
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Molecular marker analysis by ABI DNA sequence analyzer can distinguish head blight resistant varieties, but most laboratories engaged in wheat breeding in my country cannot apply this technology for marker analysis

Method used

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  • SSCP marker closely linked with major wheat scab resistance QTL and application thereof
  • SSCP marker closely linked with major wheat scab resistance QTL and application thereof
  • SSCP marker closely linked with major wheat scab resistance QTL and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Chromosomal location of markers

[0025] DNA was extracted from the leaves of wheat varieties Ning 7840 (high resistance to scab), DL 2000Marker (susceptible to scab), Chinese spring, tetrasomy N3BT3A, N3AT3D N3DT3A of Chinese spring, and telosome DT3BL and DT3BS of Chinese spring.

[0026] According to the design and synthesis of PCR amplification primers (LIUSX, Cereal Research Communications, 2008, 36: 195-200.) according to the second gene sequence reported by Liu et al. (2008): Forward primer 5'-CGTGGTTCCACGTCTTCTTA-3' ; The reverse primer 5'-TGAAGTTCATGCCACGCATA-3' (primers were synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd., the same below) was used to amplify the DNA of each leaf by PCR. Electrophoresis was carried out on 2.5% agarose gel, and the Chinese Spring, N3AT3D, N3DT3A, and DT3BS with 3BS regions could amplify a band of about 240 bp, while the N3BT3A and DT3BL lacking 3BS could not amplify this band. Determine that...

Embodiment 2

[0028] Example 2 Sequence alignment of PCR amplification products of 9 wheat varieties

[0029] DNA from leaves of 8 wheat varieties Sumai 3, Wangshuibai, Ning 894037, Ning 7840, Annong 8455, Alondra, Clark and Frotana were extracted. The 8 wheat varieties were amplified by PCR with the same primers and separated by agarose gel electrophoresis. Among these cultivars, Sumai 3, Wangshuibai, Ning 894037, and Ning 7840 are four wheat cultivars with main QTLs for scab 3BS resistance; Anong 8455, Alondra and Clark are susceptible cultivars; Frotana is American There is no major QTL for 3BS resistance in varieties resistant to head blight. Amplification result shows, the amplified band of these 8 kinds all can amplify the band of about 240bp in 2.5% agarose gel electrophoresis (such as figure 2 ). ,

[0030] exist figure 2 Middle, 1: Sumai 3; 2: Wangshuibai; 3: Ning 894037; 4: Ning 7840; 5: Frotana; 6: Annong 8455; 7: Alondra; 8: Clark; 9: DL 2000Marker.

[0031] Depend on ...

Embodiment 3

[0033] Embodiment 3, the preparation of denaturing sample solution and non-denaturing polyacrylamide gel

[0034] 1. Preparation of denatured sample solution: deionized formamide with a volume ratio of 98%, EDTA with a pH of 10mmol / L = 8.0, xylene cyanol FF with a mass ratio of 0.025%, and bromophenol blue with a mass ratio of 0.025% .

[0035] Preparation method:

[0036] The first is to prepare 10 times xylene cyanol FF and bromophenol blue mother liquor, weigh 0.125g xylene cyanol FF and 0.125g bromophenol blue and dissolve in 50ml formamide.

[0037] Next is the preparation of the denatured sample solution. The method is to take 5ml of 10 times xylene cyanol FF and bromophenol blue mother solution, 1ml of 0.5M EDTA solution with pH 8.0, and add 44ml of formamide to a total volume of 50ml.

[0038] 2. Preparation of 12% non-denaturing polyacrylamide gel: 100 ml of polyacrylamide gel solution contains 11.6 g of acrylamide and 0.4 g of methylenebisacrylamide.

[0039] Prep...

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Abstract

The invention relates to a single strand conformation polymorphism (SSCP) marker closely linked with a major wheat scab resistance quantitative trait locus (QTL). The SSCP marker is characterized in that: scab resistance candidate genes of a scab resistance QTL in a 3BS area of a wheat variety or strain resisting scab is PCR amplified by using a primer; after denaturalization, a PCR-amplified product has different single strand conformation; and the SSCP marker closely linked with the major wheat scab resistance QTL is established for detecting a genotype of the wheat variety or strain during breeding. The SSCP marker has the advantages of: overcoming the disadvantages that the wheat scab resistance screening can only be authenticated in a flowering period and is easily influenced by the environment in conventional breeding, predicting and screening wheat plants with the scab resistance by detecting a molecular marker at a seedling stage, eliminating disease plants, reducing waste of labor and materials and improving the breeding efficiency. Compared with an ABI DNA sequence analysis meter-based marker, the SSCP marker closely linked with the major wheat scab resistance QTL has the advantages of simple and convenient operation, low cost and same sensitivity.

Description

A technical field [0001] The invention relates to the fields of wheat breeding and molecular biology, in particular, selecting and utilizing molecular markers closely linked to main effect QTLs for resistance to scab resistance of wheat for assisted breeding, so as to improve the efficiency of wheat scab resistance breeding. Background technique [0002] Wheat head blight is an important disease in warm and humid wheat regions, mainly caused by Fusarium graminearum. Wheat head blight not only causes serious yield loss, but also mycotoxins such as deoxynivalenol (DON) remaining in the diseased wheat grains seriously threaten the health of humans and animals. Breeding disease-resistant varieties is the most economical way to control this disease. and effective means. A lot of previous work (Bai Guihua, Shanghai Agricultural Journal, 1989, 5(4): 17-23; Sinijders CHA, Euphytica, 1990, 50: 11-18; Singh RP, PlantDisease, 1995, 79: 238-240; van Ginkel M, Plant Disease, 1996, 80:8...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11C12R1/77
Inventor 余桂红杜军凯马鸿翔张旭周淼平任丽娟张鹏孙晓波
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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