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Method for nondestructively detecting erwinia chrysanthemi in situ by utilizing Raman spectrum

A Raman spectrum detection and Raman spectrum technology is applied in the field of in-situ nondestructive detection of chrysanthemum rot fungus, and can solve the problems that there is no invention report on the in-situ detection of chrysanthemum rot fungus by Raman spectrum, so as to meet the requirements of rapid detection and identification, The effect of high sensitivity and fast detection speed

Inactive Publication Date: 2016-11-16
HAINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is no report on the invention of Raman spectroscopy in situ detection technology of the chrysanthemum rot

Method used

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  • Method for nondestructively detecting erwinia chrysanthemi in situ by utilizing Raman spectrum
  • Method for nondestructively detecting erwinia chrysanthemi in situ by utilizing Raman spectrum
  • Method for nondestructively detecting erwinia chrysanthemi in situ by utilizing Raman spectrum

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Example 1: Raman Spectroscopy In-Situ Detection of Chrysanthemum rot pathogen suspension

[0025] 1) Preparation of bacteria samples to be tested: culture bacteria in NB liquid medium at 28°C with shaking at 150 rpm. The bacteria cells in the stable growth phase were collected and washed three times with sterile water. Suspend the cells in sterile water to a concentration of 10 5 CFU mL -1 . Take 10 μL of bacterial suspension and drop it on a glass slide, place it on an ultra-clean bench to dry, and use it as the bacterial sample to be tested.

[0026] 2) Raman spectrum detection of germs: Raman spectra of germs are collected using a Raman spectrum microscope. Focusing is carried out with a 50 times long focusing objective lens, the excitation wavelength is 532nm, and the exposure time is 10s. figure 2 Raman spectra were detected for Chrysanthemum rot fungi and other 6 kinds of plant pathogenic bacteria samples, among which 1 was Chrysanthemum rot fungus, and 2-7 ...

Embodiment 2

[0028] Example 2: Raman spectrum in-situ detection of Chrysanthemum rot in onion tissue

[0029] 1) Pretreatment of samples for in-situ detection: Disinfect the surface of fresh and healthy onion bulbs with 75% ethanol, take the Chrysanthemum rot bacteria cultured at 28°C for 24 hours, and dilute them with sterile water to 10 8 CFU mL -1 bacterial suspension. Use a sterile puncher to punch a hole in the center of the onion bulb, then inoculate the bacterial suspension into the center hole with a syringe, and place the onion bulb in a sterile petri dish in a 28°C incubator for 48 hours of moisture cultivation as the sample to be tested in situ .

[0030] 2) Sample preparation for in-situ detection: 24 hours after inoculation with Chrysanthemum rot bacteria, cut onion bulb rot tissue with a sterile blade, place in 2ml sterile water, chop onion tissue, and allow the bacteria to overflow into the water. After standing for a while, filter with sterile gauze, first use 3 layers o...

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Abstract

The invention discloses a method for nondestructively detecting erwinia chrysanthemi in situ by utilizing Raman spectrum. The method comprises the following concrete steps: 1) preparing a to-be-detected pathogenic bacterium sample; 2) detecting pathogenic bacterium Raman spectrum; 3) analyzing the pathogenic bacterium Raman spectrum; 4) carrying out in situ detection sample pretreatment; 5) preparing an in situ detection sample; and 6) carrying out pathogenic bacterium Raman spectrum in situ detection. The method established in the invention is used for nondestructively detecting the erwinia chrysanthemi in situ by utilizing the Raman spectrum. The method disclosed by the invention has the advantages that in situ sampling is carried out, in situ detection is carried out, a single pathogenic bacterium is detected, sensitivity is high, and the sample is not destructed; a step of separating and purifying the pathogenic bacterium is not needed, enrichment culture of the pathogenic bacterium is not needed, detection speed is high, and detection can be completed within 40 minutes; and requirement on rapid detection and identification can be met.

Description

technical field [0001] The invention relates to a method for in-situ non-destructive detection of chrysanthemum rot pathogens by using Raman spectroscopy. The invention belongs to the technical field of detection of quarantine plant pathogenic microorganisms. Background technique [0002] Chrysanthemum rot (Erwinia chrysanthemi) is an imported quarantine pest in my country, and it is also a bacterial disease that the inspection and quarantine departments focus on on imported seedlings and other propagation materials, especially on imported flower bulbs. Chrysanthemum rot belongs to Gracilicutes, Enterobacteriaceae, Erwinia, and has the following main characteristics: (1) Wide range of hosts: it can infect general vegetable crops such as potatoes, onions, celery , onion, garlic, burdock, etc., can also infect horticultural crops such as chrysanthemum, evergreen, hyacinth, philodendron, African Viola, etc., and can also infect orchid plants Phalaenopsis spp., Oncidium ( Onci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65
CPCG01N21/65
Inventor 路延笃甘琴华
Owner HAINAN UNIVERSITY