A rapid separation and detection method for fusarium in a sample and the culture medium used

A technology for separating culture medium and detection method, which is applied to the rapid separation and detection method of Fusarium in samples and the field of medium used, which can solve the problems of high contamination rate of other fungi, great manpower and material resources, and heavy separation workload, etc., and achieve reduction Effects of fungal contamination, reduced detection cost, and high detection sensitivity

Active Publication Date: 2019-06-07
INST OF BAST FIBER CROPS CHINESE ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0017] In summary, the detection method based on the isolation of Fusarium has the following disadvantages: a. The medium used in the current isolation method, such as PDA or PPA, etc., leads to a high contamination rate of other fungi, and multiple cycles of isolation are required to obtain pure strains; Difficult to separate
c. Due to the separation characteristics of a, the separation workload is large and requires a large amount of manpower and material resources

Method used

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  • A rapid separation and detection method for fusarium in a sample and the culture medium used
  • A rapid separation and detection method for fusarium in a sample and the culture medium used
  • A rapid separation and detection method for fusarium in a sample and the culture medium used

Examples

Experimental program
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Effect test

Embodiment 1

[0057] Example 1 Fusarium isolation and detection optimization experiment

[0058] Prepare several kinds of solid media, and prepare them for use after autoclaving and high temperature sterilization. The contents of several medium components are as follows:

[0059] ①PPA: D-galactose 1%, peptone 0.5%, KH 2 PO 4 0.1%, MgSO 4 0.05%, 2% agar powder, pH value 6.8; add pentachloronitrobenzene (PCNB) to make the final concentration of 150 μg / ml when pouring the plate, add streptomycin and cephalosporin to each final concentration of 100 mg / ml .

[0060] ②PDA: 200g peeled potato boiled water and filtered juice about 800ml, add 20g glucose, 20 agar powder, natural pH value, add pure water to 1L; add streptomycin and cephalosporin each to a final concentration of 100mg when pouring into a plate / ml.

[0061] 3. separation medium of the present invention: 1% sucrose, 1.7% agar powder, 0.2% K 2 HPO 4 , 0.25% MgSO 4 , 0.25% CaCl 2 , 0.4% CaCO 3 , pH value 7.0; Streptomycin an...

Embodiment 2

[0081] Example 2 Separation and detection of Fusarium in Zizania

[0082] The difference between this embodiment and the first embodiment is that: the wild rice stem purchased from the vegetable market is used as a material; only solid and liquid culture media for separating the culture medium are prepared, and the composition content is: 0.5% mannitol, 2% agar powder ( liquid medium without agar powder), 0.3% K 2 HPO 4 , 0.1% MgSO 4 , 0.1% CaCl 2 , 0.5% CaCO 3 , pH 6.5.

[0083] The cultured white translucent hyphae were preliminarily judged as Fusarium by microscopic observation. The isolated liquid medium was further expanded and cultured to extract DNA, and then the fungal ITS1 and ITS4 primers were used to amplify and sequence the sequence; the sequenced sequence was compared with Blastn on NCBI, and the result was 100% similar to Fusarium chlamydosporum strain NZD-mf112. So far, there is no report on the isolation or detection of Fusarium in Zizania zizania. This e...

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Abstract

The invention relates to a quick separation and detection method of fusarium in a sample and a culture medium used by the method. The method comprises the following steps: coating a sample on an isolation culture medium, wherein the isolation culture medium is free of any nitrogen source and contains 0.4-1 wt% of carbon source, 1.5-2 wt% of agar powder, 0.15-0.3 wt% of K2HPO4, 0.1-0.3 wt% of MgSO4, 0.1-0.3 wt% of CaCl2 and 0.3-0.5 wt% of CaCO3, and the pH value is 6.5-7; the culture medium also contains antibiotics which can control bacteria but not inhibit fungi, and the carbon source is composed of one or more of sucrose, glucose, D-galactose, arabinose and mannitol; and after finishing culture, selecting white semitransparent aerial hyphae from the single bacterial colony, and judging whether the white semitransparent aerial hyphae are fusarium by microscopic observation according to cellular morphological characteristics. The method can implement detection on fusarium in various samples, avoids pollution of other fungi, and thus, has the advantages of higher detection sensitivity and shorter period.

Description

technical field [0001] The invention relates to a strain separation technology, in particular to a method for rapid separation and detection of Fusarium in a sample and the culture medium used. Background technique [0002] Fusarium fungi can cause serious crop diseases, such as wheat head blight, Fusarium root rot, and corn ear rot. Fusarium toxins produced by fungi of this genus are secondary metabolites that can cause serious health problems such as vomiting, diarrhea, Kashin-Beck disease and even cancer. Therefore, fusarium detection technology plays an important role in customs quarantine and field disease prevention and control detection; and the detection of fusarium toxin, which is closely related to fusarium, is an important detection item in grain and food safety. [0003] The detection technology for Fusarium toxin content can only reflect the toxin concentration in the sample, but cannot detect whether there are live Fusarium bacteria in the sample; this method ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04
CPCC12Q1/04C12Q1/045
Inventor 李智敏严准严理高春生余永廷曾粮斌陈佳程毅孙向平薛召东
Owner INST OF BAST FIBER CROPS CHINESE ACADEMY OF AGRI SCI
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