Molecule detection method for puccinia striiformis f.sp.tritici No.17 physiological race

A technology for wheat stripe rust and physiological races, which can be used in the determination/inspection of microorganisms, biochemical equipment and methods, etc., and can solve the problems of misjudgment of changes in environmental conditions, time-consuming, and heavy workload.

Inactive Publication Date: 2014-02-05
NORTHWEST A & F UNIV
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  • Claims
  • Application Information

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Problems solved by technology

[0003] The traditional identification of physiological races adopts the method of identifying the host, whi...

Method used

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  • Molecule detection method for puccinia striiformis f.sp.tritici No.17 physiological race
  • Molecule detection method for puccinia striiformis f.sp.tritici No.17 physiological race
  • Molecule detection method for puccinia striiformis f.sp.tritici No.17 physiological race

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Experimental program
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Embodiment 1

[0018] Embodiment 1: detection of diseased leaves

[0019] The susceptible cultivar 'Mingxian 169' was planted in a nutrient pot of 13×12 cm, 10 seeds were planted in each pot, and 6 plants were kept in each pot after emergence. When the first leaf of wheat seedlings was fully unfolded, the saturated inoculation method was used to inoculate uredia spores, and sterile water was used as a control. The inoculated wheat seedlings were kept moist for 36 hours under dark conditions (temperature 10°C, relative humidity 100%) and then moved to an artificial climate incubator (temperature 14-16°C, light 16h a day, relative humidity 60-80%, light intensity 8000 ~10000Lx) culture. On the 3rd day, the 5th day, the 7th day, the 9th day, and the 12th day after inoculation respectively (production of mottling). On the 14th day (spore generation), wheat leaves were collected, total DNA was extracted, PCR amplification was performed, and agarose gel electrophoresis was used for detection. Th...

Embodiment 2

[0020] Example 2: Detection of prevalence frequency of wheat stripe rust No. 17 physiological race in the field

[0021] From June 3rd to 5th, 2012, 50 fields were randomly collected in Tianshui, Longnan, and Pingliang, Gansu Province, and 20 leaves of typical wheat stripe rust were randomly collected from each field, and brought back to the laboratory. CTAB (Cetyltrimethyl Ammonium Bromide ) method to extract the total DNA of morbidity wheat blade, adopt the specificity primer designed by the present invention and PCR reaction system to carry out the detection of standard sample, the result shows, No. 17 physiological races of wheat stripe rust are popular in Tianshui, Longnan, Pingliang, Gansu The frequency was 11.2% (Table 1).

[0022] Table 1 Detection results of prevalence frequency of wheat stripe rust No. 17 in Tianshui, Gansu in 2012

[0023]

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Abstract

The invention discloses a molecule detection method for puccinia striiformis f.sp.tritici No.17 physiological race. By employing a method for converting a RAPD molecular marker into a SCAR molecular marker, the specific primers (upstream primer 5' -TTCCCCCGCTGTCT and downstream primer 5'- GTTGAGCGGGGGAA) of No.17 physiological race are obtained, and are subjected to PCR amplified reaction (reaction conditions: 94 DEG C, 5 min, 1 cycle; 94 DEG C, 30 s, 60 DEG C, 1 min, 72 DEG C, 90 s, 35 cycles; 72 DEG C, 7 min, 1 cycle) and 2% agarose gel electrophoresis, so that an specific strip with a length of about 1000 bp is obtained. Therefore, the molecule detection method helps to rapidly discriminate and distinguish puccinia striiformis f.sp.tritici No.17 physiological race in molecular level, and establishes a base for developing research on wheat stripe rust molecular epidemiology.

Description

technical field [0001] The invention relates to the conversion of RAPD (Random Ampiified Polymorphic DNA) molecular markers into SCAR (Sequence Characterized Ampiified Region) molecular markers in biotechnology, and designs specific primers that can be used for monitoring and identifying physiological race 17 of wheat stripe rust. technical background [0002] my country is one of the countries with the largest wheat planting area and the largest consumption in the world, and the abundance and shortage of wheat are directly related to the national economy and people's livelihood. Wheat stripe rust is a worldwide wheat disease that occurs in most wheat producing areas around the world. Wheat stripe rust (Puccinia striiformis f.sp.tritici) can be spread in high altitude and long distance by means of airflow, causing the disease to spread in different wheat production areas, which is a major biological disaster affecting food safety production. Wheat stripe rust has frequently...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/04C12Q1/686C12Q2535/139C12Q2531/113
Inventor 胡小平鲁传强马丽杰
Owner NORTHWEST A & F UNIV
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