Quick separation and detection method of fusarium in sample and culture medium used by method

A technology for separating a culture medium and a detection method, which is applied to the rapid separation and detection method of Fusarium in a sample and the field of the culture medium used, can solve the problems of high contamination rate of other fungi, consuming a lot of manpower and material resources, and large separation workload, etc. The effect of fungal contamination, reduced detection cost, and high detection sensitivity

Active Publication Date: 2016-10-12
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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  • Quick separation and detection method of fusarium in sample and culture medium used by method
  • Quick separation and detection method of fusarium in sample and culture medium used by method
  • Quick separation and detection method of fusarium in sample and culture medium used by method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Optimal experiment for Fusarium isolation and detection

[0058] Several kinds of solid culture media were prepared, which were sterilized by high pressure and high temperature for later use. The contents of several medium components are:

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

[0060] ②PDA: 200g peeled potatoes, boiled water and filtered juice about 800ml, add 20g glucose, 20 agar powder, natural pH value, add pure water to make up to 1L; add streptomycin and cephalosporin to the final concentration of 100mg each / 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 7.0; Streptomycin and cephalosporin were added...

Embodiment 2

[0081] Example 2 Separation and detection of Fusarium in wild rice

[0082] The difference between this example and Example 1 is that: the wild rice water purchased from the vegetable market is used as the material; only two kinds of medium, solid and liquid, of the separation medium are prepared, and the component contents are: 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 to be Fusarium by microscope observation. The DNA was further expanded and cultured in the separated liquid medium, and then amplified and sequenced with fungal ITS1 and ITS4 primers; the sequenced sequences were compared with Fusarium chlamydosporum strain NZD-mf112 by Blastn on NCBI. So far, there is no relevant report on the separation or detection of Fusarium in wild water. This example shows that the present invention can be widely applied...

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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 a culture medium used therefor. Background technique [0002] Fusarium fungi can cause serious crop diseases such as wheat scab, Fusarium root rot and corn ear rot. Fusarium toxins produced by fungi of this genus are a class of 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; and Fusarium toxin detection, which is closely related to Fusarium, is an important detection item in food and food safety. [0003] The detection technology for Fusarium toxin content can only reflect the toxin concentration in the sample, and cannot detect the presence of live Fusarium bacteria in the sample; this method cannot pred...

Claims

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Application Information

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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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