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Method of producing pochonia chlamydosporia spores by virtue of liquid-solid double phase fermentation

A puconia bacterium and biphasic fermentation technology, applied in the application field of microbial fermentation technology, can solve the problems of long fermentation period, low spore content, and few fermentation studies, and achieves easy operation, promotion of bacterial growth, and simple fermentation equipment. Effect

Inactive Publication Date: 2017-05-24
HUNAN TAIGU BIOTECH
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  • Summary
  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

[0004] At present, there are no domestic patents related to the production of Chlamydospora Chlamydospora spores by liquid-solid two-phase fermentation, and there are few related fermentation studies, and the existing methods generally have problems such as low spore content and long fermentation cycle

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1 The method for producing chlamydospora Proconia spores by liquid-solid two-phase fermentation

[0036] Including the following steps:

[0037] a. Strain activation: Inoculate Chlamydospora Proconia on the slope of a PDA eggplant bottle, culture at 28°C for 7 days, rinse the slope of the activated strain repeatedly with 100 mL of sterile saline, and make Chlamydospora Nia spores are fully scattered in physiological saline to obtain activated spore suspension;

[0038] b. Shake flask seed solution preparation: 2500 mL of sucrose 62.5 g, peptone 3.75 g, NaNO 3 3.75 g, KH 2 PO 4 1.25 g, K 2 HPO 4 1.25 g, MgSO 4· 7H 2 O 2.5 g, FeSO 4 ·7H 2 0.025 g, the seed culture medium of initial pH 6, after sterilizing and cooling, the spore suspension prepared in step a is inserted in the culture medium by volume ratio 10%, carries out shaking flask culture, obtains shaking flask seed liquid, Shake flask culture conditions are: temperature 26°C, shaker speed 115 r...

Embodiment 2

[0043] Example 2 The method for producing chlamydospora Proconia spores by liquid-solid two-phase fermentation

[0044] Including the following steps:

[0045] a. Strain activation: Inoculate Chlamydospora Proconia on the slope of a PDA eggplant bottle, culture it at 28°C for 5 days, and wash the activated strain slope repeatedly with 100 mL of sterile saline, so that Chlamydospora can The spores of Niacinium spores are fully scattered in the physiological saline to obtain the activated spore suspension;

[0046] b Preparation of shake flask seed solution: Fill 2500 mL of 25 g of sucrose, 1.25 g of peptone, NaNO 3 1.25 g, KH 2 PO 4 0.5 g, K 2 HPO 4 0.5 g, MgSO 4· 7H 2 O 0.625 g, FeSO 4 ·7H 2 0.025g, the seed culture medium of initial pH 5, after sterilizing and cooling, the bacterial classification after activation among the step a is inserted in the culture medium by volume ratio 8%, carry out shake flask culture, obtain shake flask seed liquid, shake flask Culture ...

Embodiment 3

[0052] Example 3 The method for producing chlamydospora Proconia spores by liquid-solid two-phase fermentation

[0053] Including the following steps:

[0054] a Strain activation: Inoculate Chlamydospora chlamydospora on the slope of a PDA eggplant bottle, culture it at 28°C for 7 days, wash the slope of the activated strain repeatedly with 100 mL sterile saline, and make Chlamydospora Nia spores are fully scattered in physiological saline to obtain activated spore suspension;

[0055] b Preparation of shake flask seed solution: 3000 mL of sucrose 120 g, peptone 7.5 g, NaNO 3 7.5 g, KH 2 PO 4 3.6 g, K 2 HPO 4 3.6 g, MgSO 4· 7H 2 O 9g, FeSO4 7H 2 0.15 g, the seed culture medium of initial pH 8, after sterilizing and cooling, insert the bacterial classification activated in step a into the culture medium by volume ratio 10%, carry out shake flask culture, obtain shake flask seed liquid, shake flask Culture conditions: temperature 28°C, shaker speed 120 r / min, culture ti...

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Abstract

The invention provides a method of producing pochonia chlamydosporia spores by virtue of liquid-solid double phase fermentation. The method comprises the steps of: preparation of a culture activated shake flask seed liquid, liquid fermentation and solid fermentation to product spores. In the steps of preparation of the culture activated shake flask seed liquid and liquid fermentation, a liquid culture medium comprises saccharose, peptone, NaNO3, KH2PO4, K2HPO4, MgSO4.7H2O and FeSO4.7H2O, wherein the initial pH is 5-8. In the step of solid fermentation to product spores, a solid culture medium comprises the main components of bran, corn straw powder and corn powder. According to the method of massively producing pochonia chlamydosporia spores by virtue of a liquid-solid double phase fermentation technology, the highest quantity of chlamydospore can reach 1.4*10<6> / g, and the highest quantity of conidia can reach 4.3*10<9> / g. The spore-matrix mixture obtained by fermentation can be directly used for preparing antibiological inoculants after being dried and crushed.

Description

technical field [0001] The invention relates to a method for producing spores of Proconia chlamydospora by liquid-solid two-phase fermentation, which belongs to the application field of microbial fermentation technology. Background technique [0002] Root-knot nematode is a highly specialized omnivorous plant pathogenic nematode, which is one of the main diseases that cause crop yield reduction. Methods for controlling root-knot nematodes mainly include chemical control, physical control, selection of disease-resistant varieties and biological control. At present, nematode control is mainly based on chemical control, but chemical control is not only high in production cost, but also highly toxic, easy to pollute the environment, and pose a threat to human and animal health, while physical control is not effective, and the selection of disease-resistant varieties takes too long. Therefore, , safe and efficient biological control has become a hot spot in the research of plant...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N3/00C12R1/645
CPCC12N3/00
Inventor 丰来谭石勇谭武贵罗志威徐滔明郑双凤张冬雪邱尧
Owner HUNAN TAIGU BIOTECH
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