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Method for Quantitative Detection of Banana Fusarium wilt Race 4 from Soil

A technology for quantitative detection of Fusarium wilt of banana, which is applied in the direction of determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of poor reaction stability, low specificity and low sensitivity, and achieve stable reaction, high sensitivity, low The effect of pollution

Inactive Publication Date: 2016-07-13
ENVIRONMENT & PLANT PROTECTION INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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Problems solved by technology

[0014] The purpose of the embodiments of the present invention is to provide a method for quantitatively detecting the No. 4 race of Fusarium wilt of banana from soil, aiming to solve the problems of low sensitivity, low specificity, serious pollution, and poor reaction stability of the previous first-generation LAMP detection method. question

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  • Method for Quantitative Detection of Banana Fusarium wilt Race 4 from Soil
  • Method for Quantitative Detection of Banana Fusarium wilt Race 4 from Soil
  • Method for Quantitative Detection of Banana Fusarium wilt Race 4 from Soil

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Embodiment Construction

[0060] In order to make the object, technical solution and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0061] figure 1 Shown is the flow of the method for the quantitative detection of Fusarium wilt race 4 from soil provided by the present invention. For ease of illustration, only the parts relevant to the present invention are shown.

[0062] The application principle of the present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.

[0063] Step 1. Pretreat the sample to be tested and quickly extract the DNA of the sample to be tested

[0064] DNA extraction step: add SPCB buffer (100mmol / LTris-HCl, 100mmol / LEDTA, 120mM Na 3PO4, 2%CTAB, 1.5MNaCl, pH8.0, 2% ...

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Abstract

The invention discloses a quantitative detection method for FOC (Fusarium oxysporum f. sp. cubense (E.F. Smith) Snyd. & Hans.) race 4 from the soil. The method comprises the following steps: pretreating a sample to be detected and extracting DNA of the sample to be detected; preparing specific primers; conducting fluorescent nucleic acid isothermal amplification detection technique RealAmp reaction; constructing a RealAmp standard curve; and determining the result. The real-time fluorescent nucleic acid isothermal amplification detection technology provided by the invention is a novel nucleic acid detection technology combining a new generation of isothermal nucleic acid amplification technology with the real-time fluorescent detection technology, and has the advantages of high sensitivity, high specificity, low pollution and stable reaction. RealAmp can quantitatively detect pathogens; because of the characteristics of LAMP, Bst DNA polymerase has chain exchange capacity and good tolerance of inhibiting substances contained in the soil DNA extracts; the method carries out detection work on the soil, avoids cover opening and influence by contaminants and conducts on-site soil sample detection; the method for analysis and determination of reaction products is very simple; therefore, the method is suitable for wide application.

Description

technical field [0001] The invention belongs to the field of banana fusarium wilt detection, in particular to a method for quantitatively detecting race 4 of banana fusarium wilt from soil. Background technique [0002] Banana is a plant of the genus Musa of the Musaceae family, and is an important economic crop and food crop in tropical and subtropical regions. Banana Fusarium Wilt is a devastating disease of the banana industry, which seriously restricts the healthy development of my country's banana industry. [0003] The pathogen of banana wilt is FusariumoxysporumSchlevht.f.sp.cubense(E.F.Smith)Snyd.&Hans.(Foc), formerly known as FusariumcubenseE.F.Smith, belongs to half-knowledge fungi, Tumoraceae, Fusarium genus, Chinese The name is Fusarium oxysporum Cuban specialization. The fungus is divided into 3 races. In my country, there are mainly race 1 (race1, R1) and race 4 (race4, R4). Race No. 1 has a long history and mainly infects bananas, etc. The harm of race No....

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/06C12Q1/04
CPCC12Q1/6851C12Q2531/119C12Q2561/113C12Q2563/107
Inventor 张欣彭军张贺蒲金基谢艺贤漆艳香喻群芳谢培兰
Owner ENVIRONMENT & PLANT PROTECTION INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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