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Green Trichderma of generating ethylene and culturing method therefor

A technology of Trichoderma viride and a cultivation method, which is applied in the field of fungi to achieve the effects of improving ecological indicators and living environment quality, eliminating pollution and reducing imports

Inactive Publication Date: 2007-08-29
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Filamentous fungi, especially Trichoderma such as Trichoderma reesei, have a strong ability to decompose cellulose and hemicellulose. At home and abroad, they have been committed to using Trichoderma and other microorganisms to convert plant fibers into ethanol. The production of ethylene by Trichoderma has not been reported at home and abroad

Method used

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  • Green Trichderma of generating ethylene and culturing method therefor
  • Green Trichderma of generating ethylene and culturing method therefor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The construction of embodiment 1 Trichoderma viride (T.viride) CGMCC No.1918

[0033] The ethylene-forming enzyme gene efe (the gene sequence is reported in Fukuda H, Ogawa T, Ishihara K, et al. Molec μlar cloning in Escherichia coli, expression, and nucleotide sequence for the gene for the ethylene-forming enzyme of Pseudomonas syringae pv. phaseolicola PK2. Biochemical and Biophysical Research Communications, 1992, 188: 826-832.).

[0034] The sequence of primers used in PCR is: upstream primer P1: 5'-cc cccgcgg atgaccaacctacagactttc-3' (the underline is the Sac II site), downstream primer P2: 5'-tat cccggg aactcatgagcctgtcgcg-3' (the underline is the Sma I site).

[0035] PCR conditions were: 94°C pre-denaturation for 5 min, 94°C denaturation for 30 s, 62°C annealing for 30 s, 72°C extension for 1 min, 30 cycles, and finally 72°C extension for 10 min. The PCR machine model used is PTC200 (manufactured by MJ Company).

[0036] Then the PCR product was inserted in...

Embodiment 2

[0040] The cultivation of embodiment 2 Trichoderma viride (T.viride) CGMCC No.1918 and the determination of ethylene production

[0041] 1. Training:

[0042] (1) Inoculate the spores of the Trichoderma viride in the PDA medium, and cultivate for 1 week at 30°C;

[0043] (2) After the spores grow into green, scrape off the spores and suspend them in sterile water, then inoculate the spore suspension in the liquid basic medium at an inoculum volume of 1% by volume, seal the test tube with a cotton plug, and keep at 30°C, 200r / min shaking culture for 48h;

[0044] (3) Collect mycelia by centrifugation (centrifuge model TGL-16H, purchased from Zhuhai Heima Medical Instrument Co., Ltd., centrifugation parameter is 10000rpm 1min), then transfer the mycelium to liquid cellulose induction medium, test tube Seal with a rubber stopper and culture at 30°C for 36 hours with shaking.

[0045] 2. Determination of ethylene production:

[0046] Take 100 μl of gas at the top of the test t...

Embodiment 3

[0047] The measurement results are as follows: the average value of the measured peak area is 145.9, and the amount of ethylene produced by Trichoderma viride (T.viride) CGMCC No.1918 transformed cellulose powder after 36 hours of shaker fermentation is 2.075 μl / h / g dry cells. The cultivation of embodiment 3 Trichoderma viride (T.viride) CGMCC No.1918 and the determination of ethylene production

[0048] One, culture: (1) inoculate the mycelia of described Trichoderma viride in PDA medium, cultivate 1 week at 30 ℃;

[0049] (2) After the spores grow into green, scrape off the spores and suspend them in sterile water, then inoculate the spore suspension in a solid fermentation medium at an inoculum size of 10% by volume, and cultivate it statically at 30°C for 48h, then inoculate The sealing cotton plug of the test tube was replaced with a rubber stopper, and the culture was continued at 30°C for 48 hours.

[0050] Two, ethylene production is measured: assay method and instrum...

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Abstract

The invention supplies a viridian CGMCC No.1918 that could produce ethylene. The DNA of the strain contains ethylene forming ferment gene efe. The invention also supplies the cultivating method for the strain. It uses biology material to produce ethylene that would prevent environment.

Description

technical field [0001] The invention relates to the field of fungi, in particular to an ethylene-producing Trichoderma viride and a cultivation method thereof. Background technique [0002] Ethylene is the most widely used organic raw material, used to produce organic chemical products such as plastics, fibers, rubber, etc. These products are widely used in various sectors of the national economy. Therefore, ethylene occupies an important position in the development of the national economy. In 2004, the world's ethylene production capacity was 110 million tons, my country's ethylene consumption was 15 million tons, while the domestic ethylene output was 6 million tons, and the self-sufficiency rate was only 43%. With the sustained and rapid development of my country's economy, my country's demand for ethylene continues to increase. It is estimated that by 2010, the demand for ethylene will reach more than 20 million tons, of which more than 60% will need to be imported. Mo...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N15/31C12P5/02C12R1/885
Inventor 陈三凤陶丽董红军
Owner CHINA AGRI UNIV
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