Pilot scale production method for gliocladium roseum chlamydospore by liquid fermentation

A technology of Glioma pink and chlamydospores, applied in the field of microorganisms, can solve the problems that do not involve the overall sporulation level and are limited to shake flask culture

Active Publication Date: 2014-01-01
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chinese patent (Patent No. ZL 2007 1 0195587.6) provides a method for preparing chlamydospores of the genus Gliocladium fungus, using glucose, bean cake powder, urea, etc. Also limited to shake flask cultures and does not address overall sporulation levels
However, there are few reports on the fermentation of Glioma pink

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Embodiment 1: 50 liters of fermentors prepare the formula of Glioma pink HLD-1 chlamydospore

[0015] components

[0016] Zinc sulfate

[0017] Add sucrose, bean cake powder, potassium dihydrogen phosphate, magnesium sulfate, and zinc sulfate to the fermenter sequentially according to the above proportions, then add tap water to 30 liters, and stir until all components are completely mixed.

Embodiment 2

[0018] Embodiment 2: 500 liters of fermentor liquids produce the formula of Glioma pink HLD-1 chlamydospore

[0019] components

[0020] Add sucrose, bean cake powder, potassium dihydrogen phosphate, magnesium sulfate, and zinc sulfate into the fermenter sequentially according to the above proportions, then add 300 liters of tap water, and stir while adding until all components are completely mixed and uniform.

Embodiment 3

[0021] Embodiment 3: the method for 500 liters of fermentor liquid production Glioma pink HLD-1 chlamydospores

[0022] components

[0023] First put 250 liters of tap water into the fermenter, then add sucrose, bean cake powder, potassium dihydrogen phosphate, magnesium sulfate, and zinc sulfate into the fermenter in sequence according to the above proportions, and then add tap water to 300 liters. Turn on the motor and stir until all the components are completely mixed, then add phosphoric acid to adjust the pH to 6. Autoclave at 121°C for 30 minutes, and when the sterilized culture solution cools down to room temperature, add 1% (volume ratio) to HLD -1 Shaker Flask Seed Solution (concentration of 10 8 spore / ml), after 72 hours of cultivation at 27°C, the fermentation broth containing chlamydospores was obtained, and the concentration of chlamydospores was 1.5×10 8 individual / ml.

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Abstract

The invention belongs to the technical field of microbes, and provides a pilot scale production method for gliocladium roseum chlamydospore by liquid fermentation, and the method comprises medium formula and fermentation parameter control. The liquid medium is composed of 40-50 g/L of cane sugar, 20-25 g/L of bean cake powder, 0.5-2 g/L of potassium dihydrogen phosphate, 0.2-1 g/L of magnesium sulfate and water, and the liquid medium is used for fermentation production of gliocladium roseum HLD-1 chlamydospore, wherein the inoculation amount is 0.2%-2% (volume ratio), the culture condition comprises that: pH is 4-6, the temperature is 26 DEG C-30 DEG C, the stirring speed is 180-250 r/min and the ventilatory capacity is 1:0.2-0.8. The culture time is calculated from inoculation, the spore is germinated when 8 hours pass by, and a large amount of chlamydospore is formed when the culture is 24-40 hours. When the time of liquid fermentation is 3-5 days, the concentration of chlamydospore is 1.5*108 per milliliter. The invention aims at providing a cheaper medium formula with abundant raw material source and an efficient easily-controllable liquid fermentation technology; and by using the method, the large-scale production of gliocladium roseum HLD-1 chlamydospore can be realized, and the method provides guarantee for large-area biocontrol application of gliocladium roseum HLD-1 chlamydospore.

Description

Technical field: [0001] The invention relates to the technical field of microbes, in particular to a method for producing chlamydospores of Gliocladium rosea HLD-1 in a liquid fermentation pilot scale, including the control of medium formula and fermentation parameters. Background technique: [0002] Gliocladium spp., as an important class of mycoparasites, can infect a variety of plant pathogenic fungi such as Sclerotinia, Rhizoctonia, Botrytis cinerea, etc., and can produce antibacterial substances, or through competition, induce plant It is considered as one of the most promising antagonistic microorganisms for biocontrol of plant diseases by controlling or weakening the degree of occurrence of diseases by means of resistance and other methods. At present, some commercial Gliomata preparations have been registered abroad, such as G.virens products in the United States. Mainly used for preventive treatment of soil in pots and nurseries; G.catenulatum granules jointly dev...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N3/00C12R1/645
Inventor 孙漫红董佩佩李世东
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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