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A special medium for isolating endophytic Trichoderma fungus and its preparation method and application

A culture medium and a technology for growing Trichoderma, applied in the field of microorganisms, can solve the problems of difficulty in counting and picking colonies, dispersion of Trichoderma hyphae, and low separation frequency, and achieve the effects of obvious colony characteristics, easy separation, and high separation frequency.

Active Publication Date: 2021-11-19
INST OF PLANT PROTECTION NINGXIA ACAD OF AGRI & FORESTRY SCI KEY LAB OF NINGXIA PLANT DISEASE & INSECT PESTS CONTROL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing separation methods for endophytic fungi tend to disperse the Trichoderma hyphae, the separation frequency is low, there are many miscellaneous bacteria, it is difficult to count and pick single colonies, the separation types are less, and the specificity is also poor.

Method used

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  • A special medium for isolating endophytic Trichoderma fungus and its preparation method and application
  • A special medium for isolating endophytic Trichoderma fungus and its preparation method and application
  • A special medium for isolating endophytic Trichoderma fungus and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] This example provides a special medium for isolating endophytic Trichoderma fungus, and its components include potato, glucose, agar powder, chloramphenicol, streptomycin sulfate, Bengal red, pentachloronitrobenzene and water. The preparation method of this special medium is briefly described as follows:

[0025] 1) Prepare PDA medium

[0026] PDA medium: potato (peeled) 200g, glucose 20g, agar 12g, add sterilized water (or distilled water) to 1000mL, natural pH value.

[0027] The specific preparation process of PDA medium: Peel 200g of potatoes and cut them into small pieces, put them into 1000mL of water and boil until the potatoes become soft and not rotten, then filter with 8 layers of gauze, and pour the filtrate back into the pot Add agar and glucose, stir evenly and replenish water to 1000mL, and finally divide into 150mL Erlenmeyer flasks for later use.

[0028] 2) Add bacteria inhibitors

[0029] Add an appropriate amount of miscellaneous bacteria inhibitor...

Embodiment 2

[0032] This example provides a conventional method for isolating and cultivating endophytic Trichoderma using the special culture medium for isolating endophytic Trichoderma described in Example 1.

[0033] Firstly, the tissue samples such as plants, leaves and rhizomes of different crops (vegetation) collected were rinsed with tap water, then soaked in 75% ethanol for 5 minutes, then transferred to 2% sodium hypochlorite for 5 minutes, and finally rinsed with sterile water 3 to 5 times, thoroughly disinfect the surface. Afterwards, put an appropriate amount of sterilized plant or tissue samples into a high-speed crusher to crush them into pulp, use the flat plate smear method, take the juice (or add 30mL sterilized water after appropriate dilution) and evenly apply it on the special-purpose powder described in this embodiment. On the medium plate, use 0.2mL of medium per plate of 25mL, repeat 3 times for each treatment, and then place it in a mold incubator at 25°C for 3-5 da...

Embodiment 3

[0035]This example provides a comparison of the effects of using different culture media to isolate endophytic Trichoderma. In this example, three commonly used culture media were selected as controls (medium number 1-3), see Table 1 for details. According to relevant documents and preliminary experiments, utilize the conventional medium No. 1 of separating endophytic fungus, Martin's medium No. 2 and the selective medium 3 of Trichoderma in the separation soil and the medium 4 of the present invention's development, adopt tissue disinfection and smear method for the isolation of endophytic Trichoderma.

[0036] Table 1 The results of the determination of the effect of different culture media on the separation of endophytic Trichoderma fungus

[0037]

[0038] The results (table 1) of the effect measurement of different culture media for separating endophytic Trichoderma fungus show that medium 4 has a significant inhibitory effect on two kinds of miscellaneous bacteria, b...

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Abstract

The invention belongs to the technical field of microbes, and specifically discloses a special culture medium for isolating endophytic Trichoderma fungus, a preparation method and application thereof. The components of this special medium include potato, glucose, agar powder, chloramphenicol, streptomycin sulfate, Bengal red, pentachloronitrobenzene and water; the potato, glucose, agar powder and water are prepared into PDA culture base; in each liter of PDA medium, the content of chloramphenicol is 50-150 mg, the content of streptomycin sulfate is 4-8 mg, the content of Bengal red is 25-100 mg, and the content of pentachloronitrobenzene is 6-8 mg. 10 mg. The present invention can efficiently, rapidly and accurately isolate endophytic Trichoderma fungus, has a low inhibitory rate to Trichoderma fungus and has a good inhibitory effect on other miscellaneous bacteria, can form relatively single Trichoderma fungus with better colony characteristics, and isolate species More, easy to identify and purify, strong specificity.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and relates to a special medium for isolating endophytic fungi, in particular to a special medium for isolating and cultivating endophytic Trichoderma fungi. The invention also provides the preparation method of the special culture medium and its application in isolating and cultivating endophytic Trichoderma fungus. Background technique [0002] Fungi of the genus Trichoderma (Trichoderma Pers.) are widely distributed around the world. They are not only one of the important communities of soil microorganisms, but also a kind of plant endophytic fungi. Plant residues such as litter, fallen leaves and decayed wood, as well as fruiting bodies of other fungi, can be isolated. Existing research data show that Trichoderma has a wide range of application values ​​in agriculture, industry, environmental protection, etc., and its application performance in the agricultural field is particularly p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12N1/02C12R1/885
CPCC12N1/02C12N1/14
Inventor 康萍芝张丽荣白小军杜玉宁张治科马建华
Owner INST OF PLANT PROTECTION NINGXIA ACAD OF AGRI & FORESTRY SCI KEY LAB OF NINGXIA PLANT DISEASE & INSECT PESTS CONTROL
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