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A kind of high-efficiency preparation method of biocontrol Trichoderma chlamydospore and bacterial agent

A technology for chlamydospores and bio-control of Trichoderma, which is applied in the directions of using spores, microorganism-based methods, biochemical equipment and methods, etc., and can solve problems such as difficulty in popularization.

Active Publication Date: 2021-05-11
山东济清科技服务有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chlamydospores are only produced in large quantities under specific conditions. At present, most methods of inducing chlamydospores are aimed at specific Trichoderma strains, adjusting parameters such as nutrients and culture conditions to obtain chlamydospores. A large number of adjustments are required for different strain parameters, which is difficult to promote larger

Method used

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  • A kind of high-efficiency preparation method of biocontrol Trichoderma chlamydospore and bacterial agent
  • A kind of high-efficiency preparation method of biocontrol Trichoderma chlamydospore and bacterial agent
  • A kind of high-efficiency preparation method of biocontrol Trichoderma chlamydospore and bacterial agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A kind of efficient preparation method of biocontrol Trichoderma chlamydospore, comprises the following steps:

[0029] (1) Prepare oxalic acid mother liquor, the concentration of oxalic acid mother liquor is 0.1~0.5mol / L, store at room temperature after being filtered by 0.22 μm water phase filter membrane under aseptic condition;

[0030] (2) Prepare medium I, peel 200 g of potatoes, chop and boil in water for 30 minutes, filter the filtrate through gauze, add 20 g of glucose to the filtrate, make up 1 L of volume with water, and sterilize at 115° C. for 30 minutes to obtain medium I;

[0031] (3) Prepare medium II, glucose 5-10g, yeast powder 5-10g, peptone 10-20g, magnesium sulfate heptahydrate 1.3-1.5g, ammonium nitrate 1.5-1.9g, potassium dihydrogen phosphate 1-2g, water supplement 1L volume, autoclave at 115°C for 30 minutes to obtain Medium II;

[0032] (4) Under sterile conditions, add oxalic acid mother liquor to medium II, and adjust the pH of the medium to ...

Embodiment 2

[0039] The three Trichoderma strains to be tested are numbered Trichoderma viride ACCC31911, Trichoderma viride ACCC32489 and Trichoderma aculeatus ACCC30536 respectively in the China Agricultural Microorganism Culture Collection Center. Among them, the optimum growth temperature of Trichoderma viride is 25°C, the optimum growth temperature of Trichoderma viride is 28°C, and the optimum growth temperature of Trichoderma aculeatus is 30°C.

[0040] step one

[0041] Trichoderma strains were inoculated on PDA (200 g of potatoes, 20 g of glucose, 10 g of agar, 1 L of water) plates, and cultured in half light and half dark at a suitable temperature for each strain for 7 days to activate the strains.

[0042] step two

[0043] Take the activated Trichoderma plate, rinse the conidia into a sterile centrifuge tube with sterile water, suspend the conidia evenly with sterile water containing 0.05% Tween 80, and count to make the concentration 10 7 Conidia / mL of suspension.

[0044] ...

Embodiment 3

[0052]Trichoderma chlamydospore prepared by the present invention is prepared into wettable powder (or other conventional methods can be prepared into bacterial preparations), the steps are as follows:

[0053] Step 1: Mix the chlamydospore fermentation broth prepared in Example 1 with diatomaceous earth in a ratio of 6:1, dry at 50°C, thoroughly pulverize, and pass through a 100-mesh sieve to obtain chlamydospore powder;

[0054] Step 2: 20 parts by weight of chlamydospore powder, 5 parts by weight of sodium lauryl sulfate, 4 parts by weight of carboxymethylcellulose, 1 part by weight of dextrin, and 70 parts by weight of diatomaceous earth. Stir evenly to get the chlamydospore wettable powder.

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Abstract

The invention discloses a high-efficiency preparation method of biocontrol Trichoderma chlamydospores and bacterial agents. The preparation of biocontrol Trichoderma chlamydospores comprises the following steps: (1) preparing oxalic acid mother liquor; (2) preparing medium I, the prepared The medium I mentioned above includes potatoes, glucose and water; (3) prepare the medium II, the medium II includes glucose, yeast powder, peptone, magnesium sulfate heptahydrate, ammonium nitrate, potassium dihydrogen phosphate and water; ( 4) Add oxalic acid to medium II; (5) activate Trichoderma strain; (6) prepare conidia suspension from activated Trichoderma, and cultivate seed liquid in medium I; (7) The seed solution was inoculated into medium II supplemented with oxalic acid and cultivated to obtain a fermentation solution. The invention induces the production of Trichoderma chlamydospores by adjusting the pH, utilizes the inducing effect of oxalic acid on the biocontrol mechanism of Trichoderma, and the induced chlamydospores have good control especially for botrytis cinerea and sclerotinia and other oxalic acid secreting diseases Effect.

Description

technical field [0001] The invention relates to the technical field of preparation of biocontrol fungi, in particular to a high-efficiency preparation method of biocontrol Trichoderma chlamydospores and fungus. Background technique [0002] As a biocontrol factor, Trichoderma spp. has the characteristics of wide adaptability, broad spectrum and multi-mechanism, and has been widely used as an antagonistic fungal agent to prevent and control plant diseases and promote plant growth. In the world, more than 50 different types of Trichoderma have been registered as biopesticides or biofertilizers, accounting for 50% of the market share of fungal biocontrol agents. Currently commercial Trichoderma live preparations, the active ingredient is mainly Trichoderma conidia, because the conidia have natural defects such as poor stress resistance and poor storage resistance, the shelf life of Trichoderma live preparations is generally only 3 months. In addition to prolonging the effectiv...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N3/00C12N1/14A01P3/00C12R1/885
CPCC12N1/14C12N3/00
Inventor 吴晓青张新建张广志周方园赵晓燕周红姿谢雪迎
Owner 山东济清科技服务有限公司
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