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TaqMan probe and real-time fluorescent primers for detecting Fusarium oxysporum f.sp.cubense race1 (FOC1) and application thereof

A Cuban-specialized, Fusarium oxysporum technology, applied in the biological field, which can solve the problems of low content and high interference

Inactive Publication Date: 2014-05-14
中华人民共和国中山出入境检验检疫局
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It is hoped that the establishment of this method can solve the problem of low content and many interferences; at the same time, a method for the quantitative detection of Fusarium oxysporum Cuban specialization No. 1 physiological race has been established to be applied to the comparative study of bacterial content

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  • TaqMan probe and real-time fluorescent primers for detecting Fusarium oxysporum f.sp.cubense race1 (FOC1) and application thereof
  • TaqMan probe and real-time fluorescent primers for detecting Fusarium oxysporum f.sp.cubense race1 (FOC1) and application thereof
  • TaqMan probe and real-time fluorescent primers for detecting Fusarium oxysporum f.sp.cubense race1 (FOC1) and application thereof

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Experimental program
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Effect test

Embodiment Construction

[0059] 1. Use FOC1 and FOC4 conidia 5×10 6 The spore liquid was used to inoculate 20 plants of Guangfen No. 1 Fenjiao respectively, and the symptoms were observed 14 days after the inoculation. The vascular bundles of the two groups of inoculated plants turned black and the leaves turned yellow. From the point of view of the symptoms, the degree of yellowing of leaves was heavier in those inoculated with FOC1 than in those inoculated with FOC4. Bulb tissues with symptoms were taken for single-tube dual real-time fluorescent PCR detection with TaqMan probe method, and the content of FOC1 bacteria in the bulbs was compared with the copy number difference of FOC4 bacteria. The results showed that the average Ct value of FOC1 detected in the pseudostem inoculated with FOC1 was 28.74, and the copy number reached 1130934; the average Ct value of FOC4 detected in the pseudostem inoculated with FOC4 was 34.22, and the copy number reached 31653. Therefore, 14 days after the inoculatio...

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Abstract

The invention provides a real-time fluorescent PCR (polymerase chain reaction) qualitative detection method based on specific conservative primers and a TaqMan probe of a pathogen Fusarium oxysporum f.sp.cubense race1 (FOC1). The real-time fluorescent PCR qualitative detection method can be used for simultaneously carrying out accurate and efficient qualitative and quantitative detection on the FOC1 respectively within 1.5 hours, has high specificity and high sensitivity, improves the detection sensitivity to 1-2 orders of magnitude compared with previous common PCR detection and research, can be used for quantitative detection, breaks through the limitation that an SYBRGREEN fluorescent dye method can not be used for double or multiple detection, is accurate and efficient, and can be used for reinforcing prevention and control detection and quarantine of the disease.

Description

technical field [0001] The invention belongs to the field of biological technology, in particular to a TaqMan probe real-time fluorescent primer for rapid identification and quantitative detection of the physiological race No. Background technique [0002] Fusarium wilt of bananas caused by Fusarium oxysporum f.sp.cubense race1 (FOC1) and race 4 (F.oxysporum f.sp.cubense race4, FOC4) is the world's largest banana wilt disease. The devastating disease has brought huge harm to the banana industry in the world. Banana is mainly concentrated in Africa, South America and Asia, with an annual output of nearly 100 million tons. It is the second largest fruit crop in the world and one of the main grains for nearly 500 million people. It is the fourth largest after rice, wheat and corn. food crops. A total of 120 countries and regions currently produce bananas. In recent years, my country's banana production has developed rapidly, ranking second in the world. The banana industry ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/77
CPCC12Q1/04C12Q1/686C12Q2561/101C12Q2561/113
Inventor 杨雷亮孙丽霞邱德义陈定虎李华平管维王章根
Owner 中华人民共和国中山出入境检验检疫局