Detection primer, detection kit and detection method for watermelon wilt disease fungi

A watermelon fusarium wilt, detection kit technology, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, and microorganism determination/inspection, can solve the problems of watermelon fusarium wilt inaccuracy and inconvenient operation, and achieves rapid Reliable detection and identification, easy operation, high precision results

Inactive Publication Date: 2015-04-29
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In order to solve the defects of inaccurate and inconvenient operation in the detection of Fusarium wilt of watermelon through morphological characteristics in the prior art, the present invention discloses a detection kit for Fusarium wilt of watermelon and a detection

Method used

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  • Detection primer, detection kit and detection method for watermelon wilt disease fungi
  • Detection primer, detection kit and detection method for watermelon wilt disease fungi
  • Detection primer, detection kit and detection method for watermelon wilt disease fungi

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Embodiment 1

[0032] Embodiment 1: The present invention discloses detection primers for detecting Fusarium wilt of watermelon, a double detection primer set composed of general primers for Fusarium wilt and primers that can only specifically amplify Fusarium wilt of watermelon, wherein, the general primer for Fusarium wilt, its upstream The primer sequence is Fos-R: 5'-GCAGTCGTACGTCATCGACC-3', the downstream primer sequence is Fos-S: 5'-CCATGGCAGATGGCGAGTCA-3'; the primer pair for specific amplification of Fusarium wilt of watermelon, and its upstream primer sequence is Fow- R: 5'-CCCGCTGGCTACTAATCTTTGA-3', the downstream primer sequence is: Fow-S: 5'-ATGTAGTGGCGATACTTCCTTG-3'.

[0033] When designing the primers, the sequence of the Fos primer pair is a publicly published general primer pair for Fusarium wilt, and the Fow primer is designed according to the specific sequence of the genome of Fusarium wilt of watermelon, which can be obtained at ftp: / / foc1:foc1linjianbrassicadb.org. The sp...

Embodiment 2

[0036] Embodiment two: In addition to the primers and detection kits for the detection of Fusarium wilt of watermelon, the present invention also discloses a detection method, which includes the following steps, (1) extracting plant tissue DNA infected by Fusarium watermelon;

[0037] (2) Using a detection kit to perform PCR amplification on the DNA isolated in step (1);

[0038] (3) 6 μ l of the PCR amplification product of step (2) is separated by 1.5% agarose gel electrophoresis with a mass concentration, and then stained with ethidium bromide under an ultraviolet lamp, and the result is judged according to the size of the amplification product, When the 491bp product is specifically amplified, it can be judged that there is Fusarium wilt of watermelon in the plant tissue.

[0039] The extraction method of described step (1) comprises the following steps: 1. get 250mg of watermelon wilt pathogen thalline and liquid nitrogen, quartz sand and fully grind into white powder, pa...

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Abstract

The invention discloses a detection primer and a detection kit for watermelon wilt disease fungi. The detection primer is a dual-detection primer set consisting of a wilt disease fungus universal primer and a primer which only can amplify a watermelon wilt disease fungi. A reagent of the detection kit comprises the detection primer, a buffer solution, dNTPs, Taq polymerase, watermelon wilt disease fungus positive control DNA and ultrapure water. The invention further discloses a detection method. The detection method comprises the following steps: (1) extracting DNA of a plant tissue infected by the watermelon wilt disease fungi; (2) carrying out PCR amplification on DNA which is separated in the step (1) by virtue of the detection kit; and (3) separating PCR amplification products in the step (2) by virtue of sepharose gel electrophoresis, dyeing the products with ethidium bromide under an ultraviolet lamp after the separation, and determining a result according to the sizes of the amplified products. According to the invention, a method for detecting watermelon wilt disease fungi at a biological molecular level is provided and has the characteristics of high precision and convenient operation.

Description

technical field [0001] The invention belongs to the fields of crop disease detection and identification and plant quarantine, and specifically relates to a detection primer, a detection kit and a detection method for watermelon Fusarium wilt. Background technique [0002] Fusarium wilt of watermelon, commonly known as vine wilt or wilt, is a worldwide soil-borne disease of melons caused by Fusarium oxysporum f.sp.niveum. Its occurrence is very common and poses a serious threat to the yield and quality of watermelon. It is an important disease in watermelon production. At present, the effective control method for this kind of vascular diseases mainly relies on the breeding of disease-resistant varieties, but the resistance source of the disease-resistant varieties available for breeding is single, and the efficiency is not high. Some researchers have screened out actinomycete strains XF9, XF18 and XF22 that have good control effects on watermelon Fusarium wilt and survive we...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/77
CPCC12Q1/686C12Q1/6895C12Q2600/16C12Q2537/143
Inventor 凌键曹月霞杨宇红谢丙炎
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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