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Method for producing green trichoderma viride through liquid/solid double-phase fermentation

A technology of Trichoderma viride and liquid fermentation, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of hindering the production process of Trichoderma viride, low spore content of inoculants, and long production cycle, etc. It is suitable for mass popularization and application, with low cost and high production efficiency.

Inactive Publication Date: 2015-03-25
ENVIRONMENT & PLANT PROTECTION INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The existing cultivation methods of Trichoderma viride all have the problems of long production cycle, low spore content and high cost, which hinder the production process of Trichoderma viride

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1, liquid-solid two-phase fermentation culture Trichoderma viride

[0030] 1. Primary plate culture

[0031] Inoculate Trichoderma viride H06 on a PDA medium plate and culture at 28°C until the mycelia cover the plate and produce spores (usually 4-5 days).

[0032] PDA medium: potato 200g, peptone 10g, glucose 20g, agar 20g, dilute to 1000ml with water.

[0033] 2. Liquid seed culture

[0034] Put 400ml of liquid seed culture medium into a 1000ml Erlenmeyer flask, and then inoculate the plate bacteria obtained in step 1. The inoculum amount is 1 / 6 culture dish plate bacteria (petri dish size: 90mm) in each Erlenmeyer flask, at 28°C, Cultivate at 180rpm for 3-4 days to obtain seed liquid (the bacterial concentration in the seed liquid is 7×10 8 CFU / mL).

[0035] Liquid seed medium: take 20g of sucrose, NH 4 NO 3 5g, MgSO 4 ·7H 2 O1.5g and KH 2 PO 4 2.5g, dissolve in water and dilute to 1000mL with water; pH=6.5; autoclave at 121°C for 30 minutes, coo...

Embodiment 2

[0054] Embodiment 2, liquid-solid two-phase fermentation culture Trichoderma viride

[0055] 1. Primary plate culture

[0056] With the step 1 of embodiment 1.

[0057] 2. Liquid seed culture

[0058] Put 400ml of liquid seed culture medium into a 1000ml Erlenmeyer flask, and then inoculate the plate bacteria obtained in step 1. The inoculum amount is 1 / 6 culture dish plate bacteria (petri dish size: 90mm) in each Erlenmeyer flask, at 28°C, Cultivate at 180rpm for 3-4 days to obtain seed liquid (the bacterial concentration in the seed liquid is 6.2×10 8 CFU / mL).

[0059] Liquid seed culture medium: with the liquid seed culture medium in the step 2 of embodiment 1.

[0060] 3. Liquid fermentation production

[0061] In 100 liters of mechanically stirred fermenters, 60 liters of liquid fermentation medium are charged, and then the seed liquid obtained in inoculation step 2 is inoculated at 5%.

[0062] Culture conditions: 25°C, air flow with a ventilation rate of 10L / min, ...

Embodiment 3

[0077] Embodiment 3, liquid-solid two-phase fermentation culture Trichoderma viride

[0078] 1. Primary plate culture

[0079] With the step 1 of embodiment 1.

[0080] 2. Liquid seed culture

[0081] Put 400ml of liquid seed culture medium into a 1000ml Erlenmeyer flask, and then inoculate the plate bacteria obtained in step 1. The inoculum amount is 1 / 6 culture dish plate bacteria (petri dish size: 90mm) in each Erlenmeyer flask, at 28°C, Cultivate at 180rpm for 3-4 days to obtain seed liquid (the bacterial concentration in the seed liquid is 8.5×10 8 CFU / mL).

[0082] Liquid seed culture medium: with the liquid seed culture medium in the step 2 of embodiment 1.

[0083] 3. Liquid fermentation production

[0084] In 100 liters of mechanically stirred fermenters, 60 liters of liquid fermentation medium are charged, and then the seed liquid obtained in inoculation step 2 is inoculated at 5%.

[0085] Culture conditions: 28°C, air flow with a ventilation rate of 10L / min, a ...

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Abstract

The invention discloses a method for producing green trichoderma viride through liquid / solid double-phase fermentation. A method for cultivating trichoderma viride comprises the following steps: (1) performing liquid fermentation on the trichoderma viride to obtain a liquid fermentation product; and (2) performing solid fermentation on the liquid fermentation product. The method for cultivating trichoderma viride has the advantages of economical and practical production process, high production efficiency, small investment and small occupied area, can realize large-scale production, avoids the three-waste problem, is suitable for wide popularization and application, and can generate huge economic benefit.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation application, in particular to a method for producing Trichoderma viride by liquid-solid two-phase fermentation. Background technique [0002] Banana Fusarium wilt is a soil-borne devastating disease caused by Fusarium oxysporum, which has seriously endangered the production of bananas and the banana industry. At present, comprehensive prevention and control technology is the main method for the control of banana Fusarium wilt, and Trichoderma spp., as an important plant disease biocontrol fungus, has been widely concerned. Trichoderma viride is widely distributed in nature, can grow rapidly on a variety of nutrient substrates, has a heavy parasitic effect on a variety of plant pathogens, can effectively compete for nutrients and living space, and can also produce antibiotics and attack a variety of It is an enzyme needed by pathogens, so it is considered to be a kind of biocontrol ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12R1/885
Inventor 黄俊生梁昌聪刘磊张建华郭立佳王伟伟
Owner ENVIRONMENT & PLANT PROTECTION INST CHINESE ACADEMY OF TROPICAL AGRI SCI