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Extraction method for endophytic fungi in gramineous plant stem pith

A technology of gramineous plants and endophytic fungi, applied in the direction of separation of microorganisms, to achieve the effects of high fungi extraction efficiency, simple operation and low content of miscellaneous bacteria

Inactive Publication Date: 2018-12-04
邵万均
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, there is a lack of methods for extracting endophytic fungi from the stem pith of grasses

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] The extraction method of the endophytic fungus in the grass plant stem pith of the present embodiment comprises the following steps,

[0019] (1) Retrieve the Gramineae plant as a whole into the operating room, cut the plant into 5 cm segments, remove the outer leaves of the stalk, and take the stalk part;

[0020] (2) Put the stem segment obtained in step (1) into a 75% alcohol solution to clean for 60 seconds, then put it into a 1% sodium hypochlorite solution and clean it for 30 seconds, wash it twice with clear water and put it in a high-temperature and high-pressure sterilized container Dry on sterile paper for use;

[0021] (3) longitudinally cutting the stem segment obtained in step (2) with a pre-high temperature sterilized scalpel to expose the stem pith;

[0022] (4) Utilize pre-high temperature sterilized tweezers to take out the stem pith and place it on the culture medium of a glass dish, select 3 positions for each glass dish to place the stem pith;

[0...

Embodiment 2

[0026] The extraction method of the endophytic fungus in the grass plant stem pith of the present embodiment comprises the following steps,

[0027] (1) Retrieve the Gramineae plant as a whole into the operating room, cut the plant into 8 cm segments, remove the outer leaves of the stalk, and take the stalk part;

[0028] (2) put the stem segment obtained in step (1) into a 75% alcohol solution and clean it for 100 seconds, then put it into a 1% sodium hypochlorite solution and clean it for 40 seconds, wash it with clear water for 3 times and put it in a pre-high temperature and high pressure sterilized container Dry on sterile paper for use;

[0029] (3) longitudinally cutting the stem segment obtained in step (2) with a pre-high temperature sterilized scalpel to expose the stem pith;

[0030] (4) Utilize pre-high temperature sterilized tweezers to take out the stem pith and place it on the culture medium of a glass dish, select 4 positions for each glass dish to place the s...

Embodiment 3

[0034] The extraction method of the endophytic fungus in the grass plant stem pith of the present embodiment comprises the following steps,

[0035] (1) Retrieve the Gramineae plant as a whole into the operating room, cut the plant into 6 cm segments, remove the outer leaves of the stalk, and take the stalk part;

[0036] (2) put the stem segment obtained in step (1) into a 75% alcohol solution to clean for 80 seconds, then put it into a 1% sodium hypochlorite solution and clean it for 35 seconds, wash it with clear water for 3 times and put it in a high-temperature and high-pressure sterilized container Dry on sterile paper for use;

[0037] (3) longitudinally cutting the stem segment obtained in step (2) with a pre-high temperature sterilized scalpel to expose the stem pith;

[0038] (4) Utilize pre-high temperature sterilized tweezers to take out the stem pith and place it on the culture medium of a glass dish, select 3 positions for each glass dish to place the stem pith;...

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PUM

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Abstract

The invention discloses an extraction method for endophytic fungi in gramineous plant stem pith. The method includes putting stem segments into an alcohol solution with 75% to clean for 60-100 sec; then putting the stem segments into a sodium hypochlorite solution with 1% to clean for 30-40 sec; putting the stem segments on an aseptic paper to dry for later use after the stem segments is cleaned for 2-3 times by using clear water; vertically cutting the stem segments by using a scalpel to bare stem pith; taking the stem pith out by using a pair of tweezers, and placing the stem pith on culturemedia of glass dishes; selecting 3-4 positions of each glass dish to place the stem pith; sealing the glass dishes, and placing the glass dishes in a darkroom to perform culturing for 5-6 days underan environment of 25 DEG C; rejecting other microbial colonies, and performing continuous culturing for 10-15 days; and picking out the endophytic fungal colonies by using an inoculating needle to perform individual culturing so that endophytic fungus strains can be obtained. Compared with the prior art, the method has characteristics of high fungal extraction efficiency, low competitor contents and simple operation.

Description

technical field [0001] The invention relates to the field of fungi extraction, in particular to a method for extracting endophytic fungi from stem pith of gramineous plants. Background technique [0002] Poaceae (scientific name: Gramineae), also known as Poaaceae, is divided into more than 620 genera and more than 10,000 species. There are more than 190 genera and about 1,200 species in China. About 20% of the earth's land is covered with grass, and the Grass family includes a variety of plants commonly known as "so-and-so grass". However, it must be pointed out that not all grasses are grasses; similarly, not all grasses are low "grasses", such as bamboo, which can also be as high as tens of meters and form a continuous forest. The Poaceae family contains many important food crops. [0003] Endophytic fungi can infect many grasses and produce alkaloids. At present, there is still a lack of methods for extracting endophytic fungi from the stem pith of grass plants. Con...

Claims

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Application Information

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IPC IPC(8): C12N1/02
CPCC12N1/02
Inventor 邵万均
Owner 邵万均