Non-toxic coloring agent used for detecting grass endophytic fungi

An endophytic fungus and dye technology, applied in the preparation of test samples, measuring devices, sampling, etc., can solve the problems of serious environmental hazards, environmental hazards, high toxicity, etc., and achieve good dyeing effect and improve dyeing effect. Effect

Inactive Publication Date: 2017-01-25
LANZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] 2) Environmental hazards: serious hazards to the environment, causing pollution to both water bodies and the atmosphere
[0013] 3) Explosion hazard: the product is flammable, highly toxic and highly corrosive
[0014] The No. 2 solution in this detec...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] A non-toxic staining agent for the detection of grass endophytic fungi, comprising the following components: 0.325g aniline blue, 100ml water, 50ml 85% lactic acid;

[0031] The described non-toxic staining agent for the detection of grass endophytic fungi is used for the detection method of seed endophytic fungi, comprising the following steps:

[0032] Put 100 seeds into a 20ml beaker, pour 5% sodium hydroxide solution and soak overnight, rinse off the lye with tap water, add dyeing agent, heat to boiling and then cool to room temperature, take out one seed and put it in a place dripping with aniline blue On the glass slide of toxic staining agent, cover with a cover glass, press the seeds gently, and observe the hyphae under the objective lens 10 times and 40 times.

Embodiment 2

[0034] A nontoxic staining agent for detecting endophytic fungi in grasses, comprising the following components: 1 part of lactic acid, 2 parts of glycerol, 1 part of distilled water, and aniline blue with a mass fraction of 0.05%.

[0035] The non-toxic staining agent used for the detection of endophytic fungi in grasses is used for the detection of endophytic fungi in leaf sheaths, comprising the following steps: sowing grass seeds with endophytic fungi in nutrient soil, taking 6-week-old The seedlings were tested for the bacteria carrier rate. Use tweezers and a scalpel to tear off the inner epidermis of the leaf sheath, put it on a glass slide and add a stain for staining, cover it with a cover glass, and heat it on the flame of an alcohol lamp for 7 seconds. Observe endophytic fungal hyphae under a microscope.

Embodiment 3

[0037] A nontoxic staining agent for detecting endophytic fungi in grasses, comprising the following components: 1 part of lactic acid, 2 parts of glycerol, 1 part of distilled water, and aniline blue with a mass fraction of 0.05%.

[0038] The non-toxic staining agent for the detection of grass endophytic fungi is used for the detection method of stem endophytic fungi, comprising the following steps: using a scalpel to cut the grass stems longitudinally, scraping the stems with the tip of the knife The medulla was placed on a glass slide, stained with a staining agent, covered with a cover glass, and heated on the flame of an alcohol lamp for 6 seconds. After standing still for a while, the hyphae of the endophytic fungus were observed under a microscope.

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PUM

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Abstract

The invention provides a non-toxic coloring agent used for detecting grass endophytic fungi. The non-toxic coloring agent comprises aniline blue, lactic acid and solvent. The non-toxic coloring agent is free from phenol harmful to human beings and environment, coloring effect is improved by simply heating slides on alcohol lamp flame during coloring, and distinguish between endogenous fungal hyphae and cell walls is facilitated.

Description

technical field [0001] The invention relates to the technical field of fungal detection, in particular to a non-toxic staining agent for detecting endophytic fungi in grasses. Background technique [0002] Grass endophytes can live in grasses most of the time or parasitize them all the time, but they will not cause any external disease symptoms of grasses (Siegel et al., 1987). Extensively include the genus Epichloe (Epichloe) of the genus Ascomycota, the genus Balansia (Balansia) VA mycorrhizal fungi, and some genera (Clay, 1991) of the fungus half-known fungi (Clay, 1991). The herbaceous family forms a symbiotic endophytic fungus of the genus Cinnamomum and its anamorphic Neotyphodium (Huang Longyi and Huang Xiaolong, 2008) [0003] Endophytic fungi are host-specific, and endophytic fungi isolated from different hosts have different characteristics, which are related to the process of host infection by fungi (Toti et al., 1992). There are many methods for detecting endop...

Claims

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Application Information

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IPC IPC(8): G01N1/30
CPCG01N1/30G01N2001/302
Inventor 古丽君南志标李春杰
Owner LANZHOU UNIVERSITY
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