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Rapid detection method for millet smut fungi

A detection method, millet technology, applied in the detection of millet smut infection and the field of rapid molecular detection of millet smut, can solve the problems of unfavorable disease transmission, time-consuming, labor-intensive, time-consuming and other problems, and achieve timely control of diseases Spread, simple operation, high specific effect

Inactive Publication Date: 2016-09-07
SHANXI UNIV
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Problems solved by technology

These are the basic methods for diagnosing the presence of plant pathogens, however, they rely on time-consuming laboratory techniques and skilled taxonomic knowledge, which are time-consuming and thus not conducive to timely control of the spread of diseases

Method used

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  • Rapid detection method for millet smut fungi
  • Rapid detection method for millet smut fungi
  • Rapid detection method for millet smut fungi

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Embodiment 1

[0018] Method for detection of millet smut pathogen by PCR technique:

[0019] 1. Design of PCR-specific primers: According to the difference between the ITS sequence of the pathogen of millet smut and the ITS sequence of other smut species on NCBI, PCR-specific primers were designed using the software Primer Premier 5.0, and the upstream primer F1 sequence was 5'-AGACGGGTTTACCACTCAAC -3', the downstream primer R1 sequence is 5'-AAGCCACGGTGAATGGCAAAG-3', and was synthesized by Bioengineering (Shanghai) Co., Ltd.

[0020] 2. Detection of primer specificity: the genomic DNA of the smut spores and the tested pathogenic bacteria and the millet leaves of the control group were subjected to primer specificity detection. The pathogenic bacteria tested included spores of white pathogen of millet, spores of corn smut, and hyphae of blast fungus.

[0021] The reaction system is 20 μL, including: 2 μL of 10×Buffer, 1.6 μL of dNTP, 0.4 μL of each primer, 0.4-1 μL of DNA template, 0.2 μL ...

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Abstract

The invention provides a rapid detection method for millet smut fungi and belongs to the technical field of biological detection. Whether smut fungi exists in a sample or not is detected through extracting millet smut fungi spore genome DNA (Deoxyribonucleic Acid), designing a PCR (Polymerase Chain Reaction) specific primer according to the difference between a millet smut fungi pathogenic bacteria ITS sequence and an ITS sequence of other ustilago on NCBI, carrying out PCR amplification by applying the designed specific primer of the millet smut fungi, carrying out electrophoresis on an amplification product with 1 percent agarose gel, photographing with a gel imaging system and comparing with a standard molecular weight. The detection verifies that a detection system provided by the invention has the advantages of simplicity in operation, rapidness and accurate detection result.

Description

Technical field: [0001] The invention belongs to the field of crop disease prevention and plant quarantine, and in particular relates to a molecular rapid detection method and application of the pathogen of grain smut, that is, to detect the pathogen of grain smut with high sensitivity by using polymerase chain reaction (PCR) molecular biology technology Rapid test for the detection of smut infestation in millet. Background technique: [0002] The pathogenic bacterium of millet smut is Ustilago crameri, which belongs to the smut genus of the Basidiomycotina family Ustilagoceae. The pathogen is mainly infected by seeds, and it is not obvious during the vegetative growth process of the plant, and it begins to appear in the late stage of grain filling. When the millet is harvested and threshed, it will pollute other millet and cause disease in the next year. After sowing, the teliospores of the pathogenic bacteria will germinate and spread in the plant with the growth point, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/645
CPCC12Q1/6895C12Q1/686C12Q2565/125
Inventor 刘丽青仪慧兰
Owner SHANXI UNIV
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