Highland barley stem rot fusarium avenaceum loop-mediated isothermal amplification detecting method and application thereof

A technology for oat Fusarium and stem rot, which is applied in the field of temperature amplification detection, can solve the problems of different judgment standards, long cultivation period, large-scale disasters, etc., and achieve the effect of high sensitivity and high accuracy

Inactive Publication Date: 2018-08-10
INST OF PLANT PROTECTION GANSU ACAD OF AGRI SCI +1
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Problems solved by technology

Although the above culture method can detect Fusarium avenatum, it requires staff to observe the shape change in real time and record it before making a judgment
The above method has a long cultivation cycle and different judgment standa...

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  • Highland barley stem rot fusarium avenaceum loop-mediated isothermal amplification detecting method and application thereof
  • Highland barley stem rot fusarium avenaceum loop-mediated isothermal amplification detecting method and application thereof
  • Highland barley stem rot fusarium avenaceum loop-mediated isothermal amplification detecting method and application thereof

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Embodiment Construction

[0063] A ring-mediated isothermal amplification detection method for highland barley stem rot disease Fusarium oats, comprising the following steps:

[0064] A. Collection of disease samples

[0065] Collect the diseased plants of highland barley stalk rot and the soil samples around the diseased plants, and isolate the pathogenic bacteria from the diseased plants and soil samples in an indoor environment by conventional methods, and store the pathogenic bacteria at 4°C;

[0066] B. Strain culture

[0067] The pathogenic bacteria strains were cultured on PDA medium, and the PDA medium formula was 200 g of potatoes, 20 g of glucose, 15-20 g of agar, 1000 mL of tap water, and cultured at room temperature 25°C under light conditions;

[0068] C. Strain genomic DNA extraction

[0069] Use E.Z.N.A TM For the HP Fungal DNA Kit kit, take the mycelium cultured in step B and grind it into a powder under the protection of liquid nitrogen. Add 600 µL of CPL buffer and 10 µL of β-merc...

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Abstract

The invention relates to the technical field of plant disease pathogenic bacteria detection, in particular to a highland barley stem rot fusarium avenaceum loop-mediated isothermal amplification detecting method and application thereof. In a LAMP system, four primers specifically recognize six regions of a target gene, but in a PCR system, only two primers recognize two regions. The four primers are designed by a LAMP design software, and comprise two outer primers F3 and B3 and two inner primers FIP and BIP, highland barley stem rot fusarium avenaceum of seven different geographical origins is yellow-green positive through LAMP detection, gradient bands appear in 2% agarose gel electrophoresis detection, but a contrast group and other pathogenic bacteria are orange negative, and bands donot appear in electrophoresis detection. The four primers are designed by LAMP, and the effects can be implemented. Meanwhile, comparison judgment is carried out by two methods of electrophoresis anddeveloping, and the detection accuracy is greatly improved.

Description

technical field [0001] The invention relates to the technical field of detection of plant disease pathogenic bacteria, in particular to a method for detection of Fusarium avenatum ring-mediated isothermal amplification of stem base rot of highland barley and its application. Background technique [0002] Symptoms of stem rot of highland barley: There are obvious disease centers in the field at the seedling stage, and the plants in the diseased area grow weakly, appear yellow or wilt, and in severe cases, symptoms of lack of ridges, broken ridges and bald spots will occur; After the rot, most of them will wilt or turn yellow. In severe cases, the whole plant will die. It is found that the roots and stem bases are browned and constricted or rotted. After adult stage highland barley plants are infected with stalk rot, they usually show as white ears, shriveled grains, brown or black red stems, constricted roots and stem bases, dry or rotten. [0003] Existing studies have foun...

Claims

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Application Information

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IPC IPC(8): C12Q1/6844C12Q1/6895C12Q1/04C12R1/77
CPCC12Q1/6844C12Q1/6895C12Q2531/119C12Q2563/107C12Q2565/125
Inventor 漆永红李敏权曹素芳李雪萍李潇郭炜
Owner INST OF PLANT PROTECTION GANSU ACAD OF AGRI SCI
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