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Rapid identification method for powdery mildew conidia

A technology of conidia and powdery mildew, which is applied in the field of agricultural biology, can solve the problems of difficult enrichment of samples, similar morphology, inability to pure separation and culture, and similar conidia, etc., and achieves the effect of simple method, less time, and less interference.

Active Publication Date: 2022-04-12
HAINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, plant pathogenic fungi include obligate parasitic fungi and non-obligate parasitic fungi, because obligate parasitic fungi cannot be cultured on synthetic media, and their identification faces very similar pathogenic symptoms and conidia at the species level , cannot be purely isolated and cultured, samples are difficult to enrich, and similar shapes are difficult to distinguish. Therefore, the existing research focuses on the application of Raman technology in non-obligate parasitic fungi. At present, there is no Raman technology applied to obligate parasitic fungi. research report

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  • Rapid identification method for powdery mildew conidia
  • Rapid identification method for powdery mildew conidia
  • Rapid identification method for powdery mildew conidia

Examples

Experimental program
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Embodiment 1

[0033] Example 1 Identification of conidia of plant pathogenic fungi of different genera

[0034] S1. Collection of conidia of plant pathogenic fungi: collection of conidia of Fusarium graminearum, Pestalotiopsismicrospora, Erysiphe quercicola and Colletotrichum siamense;

[0035] S2. Raman spectrum measurement of conidia: use a single crystal silicon wafer at 520cm before measurement -1 Calibrate the instrument at the peak at the peak, and then perform Raman spectrum measurement on the sample collected in step S1. The excitation light source for Raman spectrum measurement is 514nm, the laser power is 5mW, the spot diameter is 1μm, the collection time is 10s, and the spectrum measurement range is 600cm -1 ~2000cm -1 , with a resolution of 1cm -1 , the Raman spectrum of each conidia was collected with a 50X magnification objective lens; the measurement of each conidia included three biological replicates, and each biological replicate contained the Raman spectra of 10 molecul...

Embodiment 2

[0042] Example 2 Identification on the level of powdery mildew conidia

[0043] S1. Collection of plant pathogenic fungus conidia: select plants suffering from powdery mildew, and collect respectively the conidia of pathogenic bacteria on the surface of rubber leaves, PAD4 mutant Arabidopsis thaliana and cowpea leaves that have fallen ill;

[0044] S2. Raman spectrum measurement of conidia: use a single crystal silicon wafer at 520cm before measurement -1 Calibrate the instrument at the peak at the peak, and then perform Raman spectrum measurement on the sample collected in step S1. The excitation light source for Raman spectrum measurement is 514nm, the laser power is 5mW, the spot diameter is 1μm, the collection time is 10s, and the spectrum measurement range is 600cm -1 ~2000cm -1 , with a resolution of 1cm -1 , the Raman spectrum of each conidia was collected with a 50× magnification objective lens; the measurement of each conidia included three biological replicates, an...

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Abstract

The invention provides a powdery mildew conidium rapid identification method, which comprises the following five steps: conidium collection, Raman spectrum measurement, data processing, cluster analysis and identification, the operation is simple and convenient, the required time is short, the substrate signal generated by excitation wavelength during measurement is smaller, the background signal interference is small, and the identification accuracy is high. The method has great advantages for specific parasitic bacteria such as powdery mildew which is difficult to culture, rapid identification of powdery mildew is realized on the single spore level, and the identification method has extremely small demand for samples, is not afraid of existence of sample pollution and interference of other signals, and provides powerful support for prevention and control of crop powdery mildew diseases.

Description

technical field [0001] The invention relates to the technical field of agricultural biology, in particular to a rapid identification method for powdery mildew conidia. Background technique [0002] With the rapid development of microbiology and molecular biology, great progress has been made in the field of identification of plant pathogenic fungi, mainly including the rapid development of traditional diagnostic and molecular identification methods. The identification of plant pathogenic bacteria (fungi) is usually based on the symptoms of infection and the morphology of the pathogen such as sclerotia or mycelium, spore morphology and sporulation structure. The process is relatively complicated and requires profound knowledge of microorganisms or etiology. In order to improve the identification efficiency of microorganisms or pathogens, researchers have reported using molecular features and fatty acids to identify pathogens, but these methods are limited by extraction method...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65G06K9/62
Inventor 缪卫国徐鑫泽施泽坤刘文波李潇林春花
Owner HAINAN UNIVERSITY