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Culture medium used for producing metarhizium anisopliae spore powder and production method of high-content spore powder of metarhizium anisopliae

A technology of Metarhizium anisopliae and culture medium, applied in the field of biological fermentation, can solve the problems of high cost of culture medium and competition for food

Active Publication Date: 2020-07-31
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] It can be seen from the above medium formula that most of the medium contains food crops, such as corn, rice and other ingredients, so that the components of the medium will compete with others for food, or use a large amount of raw materials with high cost such as traditional Chinese medicine or peptone, resulting in The cost of culture medium remains high

Method used

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  • Culture medium used for producing metarhizium anisopliae spore powder and production method of high-content spore powder of metarhizium anisopliae
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  • Culture medium used for producing metarhizium anisopliae spore powder and production method of high-content spore powder of metarhizium anisopliae

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The effect of embodiment 1pH on the mycelia growth of Metarhizium anisopliae

[0032] Use a puncher to take the above-mentioned rejuvenated Metarhizium anisopliae colony with a diameter of 5 mm at the edge, and inoculate it into PDA cultures with different pH values ​​(pH value 3, 4, 5, 6, 7, 8, 9, 10, natural (pH)). medium, 27±1°C, cultured for 5 days, then measured the diameter of the colony, and calculated the amount of mycelial growth, the results were as follows figure 1 shown. from figure 1 It can be seen that the hyphae can grow normally within the pH range of 5.0 to 9.0. Although overacidity and overalkalinity affect the germination of Metarhizium anisopliae spores, the mycelium growth is not significantly stressed. The mycelia grow best when the pH of the medium is 6. Therefore, pH 6 was selected as the optimal pH of the medium.

Embodiment 2

[0033] Embodiment 2 liquid culture condition screening

[0034] Add K to liquid potato glucose medium (potato 200g, glucose 20g, water 1000mL) 2 HPO 4 1g, MgSO 4 0.5 g is the control group (CK), and liquid seed culture medium with different concentrations is prepared, as shown in Table 1, adjusted to pH 6, and sterilized. Take 1×10 6 150 μL of the spore / mL Metarhizium anisopliae spore suspension was added to different numbers of media, and each number was replicated three times. 27±1°C, 220rpm shaker culture, after 96h, use a hemocytometer to measure the number of spores under a microscope, and take the average value.

[0035] Table 1 Effects of different proportions of cockroach powder and yam powder on the spore production of Metarhizium anisopliae in liquid culture

[0036]

[0037] As seen from Table 1, the addition of cockroach powder and yam obviously affects the sporulation of Metarhizium anisopliae, and the sporulation of the medium that adds cockroach powder...

Embodiment 3

[0038] Embodiment 3 solid medium screening

[0039] Prepare different proportions of yam herbaceous vine powder, cockroach powder, yam, glucose and water medium, as shown in Table 2, adjust the pH to 6, sterilize, spread a 70mm flat plate per 15g. Take 1 μL of 10000 spores / mL spore suspension and inoculate it in the center of the culture medium plate, repeat each concentration 3 times, and use PDA medium as the control group (CK). 27°C, humidity 80%, cultivated for 7 days, punched holes in the cultured solid plate with a puncher with an inner diameter of 5mm, punched 5 holes respectively to obtain 5 bacteria cakes, and the positions of the 5 bacteria cakes were basically the same in different plates, Disperse with 0.1% Tween 80 dispersant, shake at 180 r / min for 30 min at 28°C (with glass beads added), and count the number of sporulations with a hemocytometer. The statistical results are shown in Table 2.

[0040] Table 2 Effects of yam herbaceous vine powder, cockroach powde...

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Abstract

The invention provides a culture medium used for producing metarhizium anisopliae spore powder and a production method of the high-content spore powder of metarhizium anisopliae. The culture medium comprises a liquid seed culture medium and a solid culture medium which are used for culturing the metarhizium anisopliae, wherein the liquid seed culture medium is composed of the following componentsin percentage by weight: 6% of yam powder, 4% of cockroach powder, 3% of glucose, 0.1% of K2HPO4, 0.05% of MgSO4 and the balance water, and a pH value is 6; and the solid culture medium is composed ofthe following components in percentage by weight: 10-12% of yam herbaceous climber powder, 30-40% of cockroach powder, 20-30% of yam powder, 2-3% of glucose and the balance water, and a pH value is 6. The production method comprises the following steps: rejuvenating the metarhizium anisopliae; conducting liquid seed culture to obtain metarhizium anisopliae mother liquor; and conducting secondaryculture by means of the solid culture medium, and collecting spores. According to the provided production method, no grains are used, a spore yield is high, pollution is not liable to be caused in a fermentation process, and conidia and a substrate are easy to separate.

Description

technical field [0001] The invention belongs to the field of biological fermentation, and relates to a culture medium and a culture method for fungal fermentation to produce spore powder, in particular to a group of culture medium for preparing metarhizium anisopliae spore powder and a preparation method for metarhizium anisopliae high spore powder. Background technique [0002] Metarhizium spp. is an important class of entomopathogenic fungi. The preparations with it as the main biological active ingredient have the characteristics of no residue, easy production, long-lasting effect, strong pathogenicity, good application effect, and non-target Biosafety and other advantages, it has a parasitic and lethal effect on a variety of agricultural and forestry pests. As a class of biocontrol microorganisms, Metarhizium anisopliae has attracted more and more attention. Since the 1960s, more than 50 preparations have been developed and registered. [0003] The cultivation of high ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N3/00C12R1/645
CPCC12N1/14C12N3/00
Inventor 田平芳杨肖葛喜珍赵鹏
Owner BEIJING UNIV OF CHEM TECH
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