Safflower mosaic disease pathogen detection and identification method

An identification method, mosaic disease technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as safflower yield and quality loss

Pending Publication Date: 2022-03-11
SHANXI AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] According to the survey, during the growth process of safflower, more than a dozen kinds of diseases and insect pests may suffer, such as safflower finger aphid, safflower rust, etc., which will cause great losses to the yield and quality of safflower.

Method used

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  • Safflower mosaic disease pathogen detection and identification method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] dsRNA was obtained from safflower-susceptible plants showing typical mosaic leaves and necrotic spots in the safflower cultivation test field, and healthy safflower plants were used as controls.

[0037] The following steps are used for pathogen detection and identification:

[0038] (1) amplification, using primers to obtain the amplified product of recombinase polymerase amplification (RPA) from the safflower sample; said primers include a first set of primers and a second set of primers, and the first set of primers includes The forward primer that NO:1 nucleotide sequence is made up and the reverse primer that is made up of SEQ ID NO:2 nucleotide sequence; The second group of primers includes the forward primer that is made up of SEQ ID NO:3 nucleotide sequence and A reverse primer consisting of the nucleotide sequence of SEQ ID NO:4;

[0039] (2) analyze the amplification product of RPA;

[0040] (3) Judgment is made on the analyzed amplification results. The occ...

Embodiment 2

[0062] The difference between this embodiment and embodiment 1 is:

[0063] In the step (2), after the obtained RPA amplification product is recovered and purified by electrophoresis and gel slicing, healthy safflower is used as a negative control, and DL 1000 Marker is used as a standard molecular weight; electrophoresis at 120V / 90mA for 45min, and observed with an imaging system And take pictures.

[0064] When extracting dsRNA,

[0065] In step b, take the supernatant, add an equal volume of the above phenol / chloroform, centrifuge at 10,000 r / min at 4°C for 10 min, and repeat 3 times;

[0066] In step f, discard the supernatant, add 3mL binding solution, vortex mix, centrifuge at 4800r / min for 3min at 4°C; discard the supernatant and repeat 3 times.

[0067] The invention can scientifically and targetedly detect and identify pathogens, is fast and accurate, has good detection stability and strong specificity, can provide effective monitoring means for the health detection...

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Abstract

The invention relates to a safflower mosaic disease pathogen detection and identification method which comprises the following steps: (1) amplification: obtaining an amplification product of recombinase polymerase amplification (RPA) from a safflower sample by using a primer; (2) analyzing an amplification product of RPA; (3) performing result judgment on the analyzed amplification result, and indicating that the safflower sample is infected with the corresponding virus if a corresponding bp characteristic strip appears; if the corresponding bp characteristic strip does not appear, the sample is not infected with the mosaic virus. According to the method, pathogen detection and identification can be scientifically and specifically carried out, the method is rapid and accurate, the detection stability is good, the specificity is strong, an effective monitoring means can be provided for health detection of safflower, the production safety is ensured, and the method has a certain reference value for detection of other pathogens.

Description

technical field [0001] The invention relates to the technical field of plant virus detection, in particular to a method for detecting and identifying the pathogen of safflower mosaic disease. Background technique [0002] Safflower, also known as grass safflower, belongs to the genus Safflower of Compositae. It is an annual or overwintering herb plant. It is widely planted in all countries in the world. It is a traditional medicinal plant and a high-quality edible oil source plant. Viral diseases are common diseases. Occurs in all production areas. [0003] The dried flowers of safflower are used as medicine, which has the effects of promoting blood circulation, promoting menstrual flow, dispelling blood stasis and relieving pain, clearing toxins and clearing rashes. Studies have found that a class of lignan active compounds isolated from safflower has the ability to effectively inhibit HIV-1 reverse transcriptase, integrase and its own transcription in the early stage of H...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11
CPCC12Q1/701C12Q1/6844C12Q2600/166C12Q2531/119C12Q2521/507C12Q2537/1376C12Q2545/113C12Q2565/125
Inventor 王德富牛颜冰张学治杨智明贾彬良崔丽艳杜江
Owner SHANXI AGRI UNIV
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