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Trichoderma viride and large-scale preparation method thereof

A technology of Trichoderma viride and Trichoderma viride, applied in the direction of biochemical equipment and methods, botany equipment and methods, and microbial-based methods, can solve problems that hinder the large-scale production process of Trichoderma viride, long production cycle, sporulation Low volume and other problems, to solve the dead angle problem of cleaning and sterilization, improve labor efficiency, increase the effect of surface area

Inactive Publication Date: 2017-05-17
YIYUAN KANGYUAN BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the production process of Trichoderma viride is mainly solid-state fermentation, which generally has problems such as long production cycle, low spore production, and high cost, which hinder the large-scale production process of Trichoderma viride

Method used

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  • Trichoderma viride and large-scale preparation method thereof
  • Trichoderma viride and large-scale preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Isolation and screening of Trichoderma viride strains:

[0033] 1. Medium:

[0034] Carboxymethylcellulose sodium (CMC-Na) medium, cellulose Congo red differential medium and potato dextrose (PDB) medium.

[0035] 2. Soil sampling:

[0036] Soil samples were collected from the roots of dead trees covered by fallen leaves.

[0037] 3. Separation:

[0038] Put 1g of soil sample into a Erlenmeyer flask filled with 100mL sterile normal saline and glass beads and shake fully. On the sodium carboxymethylcellulose medium plate, culture statically at 28°C for 4-5 days, transfer the single colony with obvious transparent hydrolysis circle around it to a new medium and continue to cultivate until a large number of spores are produced.

[0039] 4. Primary screening:

[0040] The spores in the plate were eluted with sterile physiological saline and serially diluted 10 times, and 100 μL of each diluted concentration was evenly spread on the sodium carboxymethylcellulose medium ...

Embodiment 2

[0045] Molecular Biological Species Identification of Trichoderma viride:

[0046] 1. DNA extraction:

[0047] Genomic DNA of the screened strains was extracted with the Biospin Fungal Genomic DNA Extraction Kit.

[0048] 2.18S sequence amplification:

[0049] The fungal 18S sequence universal primers were used to amplify the 18S sequence of the screened strains. The PCR reaction system was: 5.0 μL of 10×PCR buffer, 4.0 μL of dNTPs (2.5 mM), 2.0 μL of primers ITS1 and ITS4, 2.0 μL of ExTaq enzyme (5U / μL), 2.0 μL of DNA template, ddH 2 O 34 μL; PCR operating program: 94°C pre-denaturation for 5 min, 95°C denaturation for 60 s, 55°C annealing for 60 s, 72°C extension for 90 s, 34 cycles, and finally 72°C final extension for 10 min.

[0050] 3. PCR product purification:

[0051] The PCR product was separated by 1.0% agarose gel electrophoresis, and the main PCR product with a size of about 1500 bp on the electrophoresis gel was cut off, and the PCR product was purified using ...

Embodiment 3

[0060] Large-scale production of Trichoderma viride:

[0061] 1. Strain: the preservation number of the Trichoderma viride strain used is CGMCC No.12079.

[0062] 2. Incline medium: PDA medium.

[0063] 3. Liquid medium: glucose 2%, yeast powder 0.5%, peptone 0.5%, dipotassium hydrogen phosphate 0.2%, magnesium sulfate 0.1%, zinc chloride 0.05%, ferrous sulfate 0.01%.

[0064] 4. Solid medium: main ingredients: flour 37.9%, bran 37.9%; auxiliary materials: corn flour 12%, soybean meal 6%, peanut shell 6%; inorganic salt solution (w / v): potassium dihydrogen phosphate 0.06% , citric acid 0.05%, ferrous sulfate 0.04%, magnesium sulfate 0.05%; the water content of the solid medium is 50%. When preparing, first dissolve the inorganic salt in water, and then stir and mix with the main material and auxiliary material.

[0065] 5. Inoculate the slant of Trichoderma viride cultured at 28°C for 9 days into a 1L shake flask containing 400 mL of liquid medium, and culture at 28°C and 1...

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Abstract

The invention relates to trichoderma viride and a large-scale preparation method thereof and belongs to the technical field of preparation of trichoderma. According to the trichoderma viride provided by the invention, a strain is trichoderma viride HS-F9 in the trichoderma, and the collection number of the strain is CGMCC (China General Microbiological Culture Collection Center) No.12079. The large-scale preparation method of the trichoderma viride, provided by the invention, adopts a liquid-solid double-phase fermentation process; seed fermentation of a liquid culture medium and disc-by-disc fermentation of a solid culture medium are adopted; in a solid fermentation process, a central air conditioner, a temperature control system and a high-pressure micro-mist atomizer are used for carrying out temperature control and humidity control; and a fermented product is dried at a low temperature and is ground to prepare powder. The trichoderma viride provided by the invention has a high antibacterial broad-spectrum property and also has a certain crop growth promotion effect; and meanwhile, the large-scale preparation method provided by the invention is high in efficiency and low in cost.

Description

technical field [0001] The invention relates to a Trichoderma viride and a large-scale preparation method thereof, belonging to the technical field of Trichoderma preparation. Background technique [0002] Plant soil-borne diseases are a very common type of plant diseases, including root rot, blight, damping-off, wilt, etc. Pathogens usually infect the roots of plants, causing diseases of the roots and even the whole plant, causing major economic loss. For the control of plant soil-borne diseases, conventional methods include chemical control and biological control. Among them, chemical control has little effect on most pests and diseases, and chemical pesticides will cause a large amount of pesticide residues, causing environmental pollution and endangering human health; at the same time, long-term use of chemical pesticides will cause pathogenic bacteria to develop resistance to pesticides, which will lead to chemical control effects. decline or even fail. Because biolo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14A01N63/04A01P5/00A01P3/00C12R1/885
CPCA01N63/30C12N1/14C12N1/145C12R2001/885
Inventor 周长春许正宏陆震鸣史劲松房华周凤云
Owner YIYUAN KANGYUAN BIO TECH
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