Activity detection method for fusarium virguliforme

A technology for soybean sudden death and syndrome, applied in the preparation of test samples, fluorescence/phosphorescence, material excitation analysis, etc., can solve the problems of large number of spores, long detection time, short detection time, etc., and achieve good repeatability Effect

Inactive Publication Date: 2014-02-19
SHENZHEN AUDAQUE DATA TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a new detection time is short, the required spore amount is very few, even 1 spore can be used for the problems such as long detection time or the large amount of spores that exist in the current North American sudden death syndrome pathogenic bacteria activity detection. A feasible detection method for the activity of North American soybean sudden death syndrome pathogen

Method used

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  • Activity detection method for fusarium virguliforme
  • Activity detection method for fusarium virguliforme
  • Activity detection method for fusarium virguliforme

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0023] Example: North American Soybean Sudden Death Syndrome Bacteria Activity Detection

[0024] 1. Materials and Instruments

[0025] North American soybean sudden death syndrome strain ARG1.1, propidium iodide, LSM5EXCITER laser scanning confocal microscope (ZEISS), DK-S28 electric constant temperature water bath (Shanghai Jinghong Experimental Equipment Co., Ltd.).

[0026] 2. Experimental method

[0027] Taking North American soybean sudden death syndrome pathogen ARG1.1 as the experimental material, freshly prepared 1 mL of spore suspension was divided into two groups: one group was live spores (unkilled, UK) that had not been treated in water bath, and the other group was 50-treated live spores. The dead spores treated in a water bath for 3 minutes at ℃ are expressed as killed (K). Take 1 μL of propidium iodide (dissolved in DMSO) with an initial concentration of 0.5 mmol / L and add 200 μL of spore suspension, mix well, the final concentration of propidium iodide stain...

experiment example 1

[0031] Experimental Example 1: Propidium iodide staining concentration and time screening

[0032] 1. Materials and Instruments

[0033] North American soybean sudden death syndrome strain ARG1.1, propidium iodide, LSM5EXCITER laser scanning confocal microscope (ZEISS), DK-S28 electric heating constant temperature water bath (Shanghai Jinghong Experimental Equipment Co., Ltd.), MLR-350HT biochemical incubator ( SANYO).

[0034] 2. Experimental method

[0035]Using the North American soybean sudden death syndrome pathogen ARG1.1 as material, the dye concentration and dyeing time of propidium iodide were screened. Freshly prepared spore suspensions were divided into three groups; one group was untreated live spores used for germination experiments as a germination test control; one group was untreated live spores used for staining experiments (UK group); The group was dead spores treated in a water bath at 50°C for 3 minutes (group K). Design 4 concentration gradient experim...

experiment example 2

[0043] Experimental example 2: Quantitative analysis of activity detection of North American soybean sudden death syndrome pathogen

[0044] 1. Materials and Instruments

[0045] Eight strains of North American soybean sudden death syndrome pathogen strains from the United States, Argentina and other countries, propidium iodide, LSM5EXCITER laser scanning confocal microscope (ZEIS S), DK-S28 electric heating constant temperature water bath (Shanghai Jinghong Experimental Equipment Co., Ltd.) , MLR-350HT biochemical incubator (SANYO).

[0046] 2. Experimental method

[0047] Eight strains of North American soybean sudden death syndrome pathogens were used as experimental materials. The freshly prepared spore suspension was treated for 5 times at 5 water bath temperatures of 42°C, 44°C, 46°C, 48°C, and 50°C, namely 1min, 2min, 3min, 4min, and 5min, and all samples were divided into Two groups, one group was stained with propidium iodide 40-fold dilution (staining final concen...

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Abstract

The present invention discloses an activity detection method for fusarium virguliforme. According to the method, a propidium iodide staining method is adopted, a laser scanning confocal microscopy is adopted to detect, and the dead spores and the living spores of the fusarium virguliforme can be rapidly, accurately and sensitively distinguished. The method provided by the present invention can replace the traditional spore germination method, and is applicable for the departments of port inspection and quarantine, agricultural production, plant protection and the like.

Description

technical field [0001] The invention relates to a method for detecting microbial activity, in particular to a method for detecting the activity of North American soybean sudden death syndrome bacteria (Fusarium virguliforme O'Donnell et T.Aoki). Background technique [0002] North American soybean sudden death syndrome (Fusarium virguliforme O'Donnell et T.Aoki) is one of the pathogenic bacteria that cause sudden soybean death syndrome (Sudden Death Syndrome, referred to as SDS). It is mainly distributed in the United States, Canada, Argentina and other countries. There are reports of the pathogen causing harm (Wu Pinshan et al., 2003). The disease is widely distributed and harmful. From 2003 to 2005, the cumulative loss of soybean yield caused by the disease reached 1.89 million tons (Wrather et al., 2006). [0003] At present, the detection of SDS pathogens in North America is mainly based on molecular biology methods, such as PCR, etc. These methods can accurately, sensi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/30G01N21/64
Inventor 章桂明代京莎王颖程颖慧向才玉
Owner SHENZHEN AUDAQUE DATA TECH
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