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Wheat-powdery-mildew-resistance-associated gene molecular marker primers and use thereof
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A molecular marker, wheat technology, applied in the field of plant biology, can solve the problem of limited resistance genes, and achieve the effect of accelerating the breeding process and improving the breeding efficiency
Inactive Publication Date: 2012-07-04
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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However, the resistance genes used in breeding are very limited
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Embodiment 1
[0036] Example 1: Example of Primer Xcib9
[0037] (1) Using the CTAB method to extract wheat genomic DNA, the extraction steps:
[0038] 1) Take 2g of fresh young leaves, grind them into fine powder with liquid nitrogen, add 2×CTAB extract (2% CTAB; 1.4M NaCl, 0.1M Tris-HCl, pH 8.0, 0.1M EDTA, pH 8.0) 15ml, mix well.
[0039] 2) 30-45min in a water bath at 65°C, during which time shake gently to mix. After cooling to room temperature, add an equal volume of chloroform:isoamyl alcohol (24:1), mix gently until the supernatant becomes milky, and centrifuge at 4000rpm for 10min.
[0040] 3) Take the supernatant, add an equal volume of isopropanol, and place in an ice bath to precipitate DNA.
[0041] 4) The DNA was hooked out, washed twice with 70% ethanol and once with absolute ethanol, air-dried the DNA, and dissolved in an appropriate amount of 1×TE solution with pH 8.0. Add RNase to a final concentration of 100 μg / μl.
[0048] (1) Using the CTAB method to extract wheat genomic DNA, the extraction steps:
[0049] 1) Take 2g of fresh young leaves, grind them into fine powder with liquid nitrogen, add 2×CTAB extract (2% CTAB; 1.4M NaCl, 0.1M Tris-HCl, pH 8.0, 0.1M EDTA, pH 8.0) 15ml, mix well.
[0050] 2) 30-45min in a water bath at 65°C, during which time shake gently to mix. After cooling to room temperature, add an equal volume of chloroform:isoamyl alcohol (24:1), mix gently until the supernatant becomes milky, and centrifuge at 4000rpm for 10min.
[0051] 3) Take the supernatant, add an equal volume of isopropanol, and place in an ice bath to precipitate DNA.
[0052] 4) The DNA was hooked out, washed twice with 70% ethanol and once with absolute ethanol, air-dried the DNA, and dissolved in an appropriate amount of 1×TE solution with pH 8.0. Add RNase to a final concentration of 100 μg / μl.
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Abstract
The invention discloses a wheat-powdery-mildew-resistance-associated genemolecular marker primers and use thereof. The molecular marker primers provided by the invention for tracing a wheat-powdery-mildew-resistance-associated gene in a wheat line 07jian126 are at least one of the molecular marker primer pair Xcib9 or the molecular marker primer pair Xcib5, wherein the molecular marker primer pair Xcib9 is used for performing polymerasechain reaction(PCR) amplification on the disease resistant wheat line 07jian126 with a 145bp specific fragment; and the molecular marker primer pair Xcib5 isused for performing PCR amplification on wheat line 07jian126 with a 385bp specific fragment. When the two molecular marker primers disclosed by the invention or one of the two molecular marker primers is used, the wheat-powdery-mildew-resistance-associated gene in the wheat line 07jian126 can be traced conveniently through PCR amplification detection. When the molecular markers disclosed by the invention are used in a wheat breeding process, the molecular marker assisted selection of the wheat-powdery-mildew-resistance-associated gene in wheat breeding materials can be performed before wheatpowdery mildew occurs in a seedling stage, and therefore the breeding efficiency is increased and the breeding process is accelerated.
Description
technical field [0001] The invention relates to marker primers for wheat powdery mildew resistance gene molecules, belonging to the field of plantbiotechnology, in particular to a molecular marker primer that can be used to trace the powdery mildew resistance gene in wheat strain 07jian126 and its application. Background technique [0002] Wheat is an important food crop in the world, and it is planted on a large scale in all major crop producing areas around the world. In my country, especially in the north, winter wheat production accounts for about 56% of the total national wheat production. The high and stable yield of wheat is of great significance to solving the world food crisis. Wheat powdery mildew is a major worldwide disease caused by obligate parasites (Blumeria graminis DC.), which can lead to more than 50% reduction in wheat yield in severe years. Introducing resistance genes into wheat is the most economical, effective and safe control method. There are 57...
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