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Paecilomyces lilacinus and its large-scale preparation method

A technology of Paecilomyces lilacinus and Paecilomyces lilacinus is applied in the directions of biochemical equipment and methods, microorganism-based methods, chemicals for biological control, etc., and can solve problems such as environmental pollution, easy occurrence of fires, etc. Achieve the effect of reducing production costs, avoiding sticky conditions, and improving labor efficiency

Active Publication Date: 2020-03-20
山东和众康源生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Corn is one of the main crops in my country. Except for a small part of corn stalks used as feed, most of them are burned, which not only causes environmental pollution, but also easily causes fire hazards and other hidden dangers.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Isolation and screening of Paecilomyces lilacinus strains:

[0029] 1. Czapek's medium (w / v):

[0030] Sucrose 3%, NaNO 3 0.3%, K 2 HPO 4 0.1%, MgSO 4 0.05%, KCl 0.05%, FeSO 4 0.001%, agar 2%, pH6.0~6.5.

[0031] 2. Soil sampling:

[0032] Soil samples were collected from wheat roots infested with cereal cyst nematode.

[0033] 3. Separation:

[0034] Put 1g of soil sample into a Erlenmeyer flask filled with 100mL sterile normal saline and glass beads and shake fully. On the Czapek's medium plate, culture at 26°C for 4-5 days, and transfer the single colony whose colony shape, color and mycelial morphology are consistent with Paecilomyces lilacinus and grow faster to the new medium Continue to cultivate until a large number of spores are produced.

[0035] 4. Primary screening:

[0036] The spores in the plate were eluted with sterile physiological saline and diluted 10 times, and 100 μL of each dilution was evenly spread on the Czapek's medium plate, and...

Embodiment 2

[0041] Molecular biology identification of Paecilomyces lilacinus:

[0042] 1. DNA extraction:

[0043] Genomic DNA of the screened strains was extracted with the Biospin Fungal Genomic DNA Extraction Kit.

[0044] 2. ITS sequence amplification:

[0045] The fungal ITS sequence universal primers were used to amplify the ITS 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.

[0046] 3. PCR product purification:

[0047] The PCR product was separated by 1.0% agarose gel electrophoresis, and the main PCR product with a size of about 700 bp on the electrophoresis gel was cut out, and the PCR product was purified using the PCR p...

Embodiment 3

[0055] Large-scale production of Paecilomyces lilacinus:

[0056] 1. Strain: The Paecilomyces lilacinus used is preserved under CGMCC No.12080.

[0057] 2. Incline medium: PDA medium.

[0058] 3. Liquid medium: glucose 2%, yeast powder 0.5%, ammonium sulfate 0.5%, diammonium hydrogen phosphate 0.5%, corn steep liquor 0.3%, corn flour 0.2%, urea 0.1%.

[0059] 4. Solid medium: 24% corn stalks, 16% rice bran, 60% corn flour, 0.1% (NH 4 ) 2 SO 4 The aqueous solution adjusts the water content to about 50%.

[0060] 5. Inoculate the slant of Paecilomyces lilacinus cultured at 26°C for 9 days into a 1L shake flask containing 400 mL of liquid medium, and culture at 26°C and 150 r / min for 7 days.

[0061] 6. Inoculate the seed liquid in 5 1L shake flasks into a 100L seed fermenter with 60L liquid medium, and cultivate for 6 days at 26°C and 150r / min.

[0062] 7. Autoclave the dry solid medium. Sterilization conditions are: 121°C, 1h. After the sterilization is completed, turn ...

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Abstract

The invention relates to Paecilomyces lilacinus and its large-scale preparation method, and belongs to the technical field of preparation of Paecilomyces lilacinum. The Paecilomyces lilacinus strain of the present invention is Paecilomyces lilacinus strain INTR-2 in the genus Paecilomyces, and the preservation number of this strain is CGMCC No. 12080. The described large-scale preparation method of Paecilomyces lilacinus adopts a liquid-solid two-phase fermentation process, through liquid medium seed fermentation and solid medium sub-disk fermentation with corn straw as the main raw material. The solid-state fermentation process uses central air conditioning. , temperature control system and high-pressure micro-mist humidifier to control temperature and humidity, and the fermentation products are dried at low temperature and then ground into powder. The Paecilomyces lilacinus of the present invention has good control effect on various nematodes and is used to produce high-standard organic agricultural products; at the same time, the present invention provides a simple and easy large-scale preparation method with high efficiency and low cost. .

Description

technical field [0001] The invention relates to Paecilomyces lilacinus and a large-scale preparation method thereof, belonging to the technical field of Paecilomyces preparation. Background technique [0002] Paecilomyces lilacinus belongs to the subphylum Deuteromyeotina, HypHomyeetes, HypHomycetales, Monilaeaeae, and Paecilomyces. A high-efficiency, broad-spectrum biocontrol fungus, which has good control effects on various nematodes, such as root-knot nematodes, cyst nematodes, golden nematodes, and heteroderma nematodes. In addition, Paecilomyces lilacinus can adapt to different climatic conditions, has a high parasitic rate on nematode eggs, can effectively control the number of soil nematodes, does not pollute the environment, has rich sources of production raw materials, is harmless to humans and animals, and has no target organisms. Easy to produce drug resistance and other advantages, it is the first choice for the production of high-standard organic agricultural p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14A01P5/00C12R1/79
CPCC12N1/14C12N1/145C12R2001/79
Inventor 周长春许正宏陆震鸣史劲松房华周凤云
Owner 山东和众康源生物科技有限公司
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