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Detection primers and molecular detection method for pathogenic fungi of corn common rust

A technology for corn rust and pathogenic fungi is applied in the field of molecular biology to achieve the effects of low test cost, simple and fast operation, and good practicability

Inactive Publication Date: 2011-08-10
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the research on molecular detection of corn rust fungus has not been reported at home and abroad.

Method used

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  • Detection primers and molecular detection method for pathogenic fungi of corn common rust
  • Detection primers and molecular detection method for pathogenic fungi of corn common rust
  • Detection primers and molecular detection method for pathogenic fungi of corn common rust

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Field corn rust monitoring comparison test

[0027] (1) Sample collection

[0028] On February 17, 2009, Chongpo Town, Sanya City, Hainan Province, on August 26, Changchun City, Jilin Province, and on September 24, Qianjiang Xinhua City, Chongqing City, respectively randomly took 10, 7, and 4 corn leaf samples,- Store at 80°C for later use.

[0029] (2) Synthesize common rust detection primers according to the following nucleotide sequence (the same below):

[0030] Upstream primer: 5'-TTTAGTAGTCTCTATCTCCAACAACA-3',

[0031] Downstream primer: 5'-AAGACTCTTTTTGCATGGTTT-3'.

[0032] (3) Detection of corn rust

[0033] ① Extract genomic DNA from maize leaves;

[0034] ②PCR amplification, 25μL reaction system was placed in the PCR thin-walled tube in turn:

[0035] 10′Buffer 2.5μL, 25mmol / LMgCL 2 2.0μL, 2.5mmol / LdNTP 2.0μL, 2.5μL each of the above common corn rust upstream primer and 2mmol / L downstream primer, 10-50ng of corn leaf genomic DNA, 1.0U of Taq ...

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Abstract

The invention relates to specific primers and a molecular detection method for pathogenic fungi of corn common rust. The specific primers comprise an upstream primer: 5'-TTTAGTAGTCTCTATCTCAACAACA-3', and a reverse primer: 5'-AAGACTCTTTTGCATGGTTT-3'. PCR (Polymerase Chain Reaction) amplification and agarose gel electrophoresis are performed on the specific primers, wherein the amplification parameters are as follows: pre-denaturating for 5 minutes at 95 DEG C, then denaturating for 45 seconds at 94 DEG C, annealing for 45 seconds at 55 DEG C, extending for 90 seconds at 72 DEG C, circulating for 30 times, and extending for 10 minutes at 72 DEG C, and the germ carrying condition of field corn is detected by the amplified product (with the fragment length of about 480bp). The primer provided by the invention has strong specificity on corn strip rust fungi and can be used for detection of the corn rust with the sensitivity of 0.001ng / mu L, and the detection method provided by the invention has good practicability and is simple and quick to operate.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a specific primer and a molecular detection method for common corn rust pathogenic fungi. Background technique [0002] Corn rust is an increasingly serious fungal disease of corn worldwide. There are three kinds of corn rusts that have been reported so far, namely, the common corn rust caused by Puccinia sorghhi Schw., the southern corn rust caused by Puccinia polysora Underw. and the corn rust caused by corn hulls. Tropical corn rust caused by the fungus [Physopella zeae(Mains) Cummins & Ramachar]. Common corn rust is the most common type of corn rust. [0003] Corn rust occurs in a wide range in my country, covering the main corn producing areas in the north and south. There are two types of common rust and southern rust. Common rust mostly occurs in the northeast and northwest corn areas with low temperature and high humidity; southern rust occurs in Taiwan , Hainan, and Yu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 郑文明邢国珍蒋士君许君袁红霞王明凤杨文博张磊
Owner HENAN AGRICULTURAL UNIVERSITY
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