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A kind of preparation method and application of trichoderma dark green microsclerotia and its preparation

A technology of Trichoderma dark green and micro sclerotia, applied in microorganism-based methods, botanical equipment and methods, biochemical equipment and methods, etc., can solve the problems of high production cost, high labor cost, poor soil persistence, etc. To achieve the effect of shortened production cycle, good control effect and strong stress resistance

Active Publication Date: 2022-04-15
BIOTECH CENT OF SHANDONG ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the active ingredients of Trichoderma biopesticides are mainly composed of mold conidia, which are usually produced by fermentation on a wet solid fermentation medium. The fermentation process needs to last for several weeks, and then post-fermentation treatment, not only the production cost High, but also subject to various constraints, such as high labor costs, poor quality control, long fermentation time, difficulty in scaling up, short shelf life of products, etc.
Currently, liquid culture research is mainly focused on the production of Trichoderma chlamydospores, but there are problems such as low fermentation yield, lack of storage stability, or poor persistence in soil

Method used

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  • A kind of preparation method and application of trichoderma dark green microsclerotia and its preparation
  • A kind of preparation method and application of trichoderma dark green microsclerotia and its preparation
  • A kind of preparation method and application of trichoderma dark green microsclerotia and its preparation

Examples

Experimental program
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Effect test

Embodiment 1

[0028] Embodiment 1 Trichoderma dark green liquid fermentation prepares micro-sclerotia

[0029] (1) Preparation of Trichoderma dark green conidia suspension

[0030] Trichoderma dark green is HB20111 ( Trichoderma atroviride ), preserved in the China General Microorganism Culture Collection Management Center, the preservation time is December 5, 2018, the preservation address is No. 3, No. 1, Beichen West Road, Chaoyang District, Beijing, and the preservation number is CGMCC No.16963.

[0031] Trichoderma dark green conidia were obtained in large quantities by the solid fermentation method, and the process was as follows: pick the Trichoderma bacteria block containing conidia from the Trichoderma dark green storage tube preserved in the -80 ℃ ultra-low temperature refrigerator, and inoculate it on a PDA solid plate, 28 Cultivate at ℃ for 5 days; select a vigorously growing single colony on the PDA solid plate to inoculate the eggplant bottle, select PDA medium, culture at 28...

Embodiment 2

[0035] The measurement of micro-sclerotia in the liquid fermented liquid of embodiment 2

[0036]Use a pipette tip with the head cut off (to avoid mycelial clogging) to draw 1ml of liquid fermentation sample from the shake flask to determine the concentration of microsclerotia, perform two serial dilutions of 1:10 to obtain a 1:100 dilution, and 100 μL Place the diluted solution on a microscope glass slide, cover it with an extra large cover glass (24×50 mm), count all the dense microsclerotia under the cover glass with a low magnification (40×) light microscope, and count their concentration in milliliters.

[0037] Harvesting of Microsclerotia of Trichoderma viridans

[0038] According to the method of Example 1, after 7 days of liquid fermentation, a large number of microsclerotia with good shape and compact hyphae are produced in the culture medium, and then harvested. Add 2.5% (w / v) diatomaceous earth to the induction medium when harvesting, and mix thoroughly; then pou...

Embodiment 3

[0040] Example 3 Trichoderma dark green micro-sclerotia sporulation ability measurement

[0041] Dilute 10 mg of micro-sclerotia-diatomaceous earth mixture dried in Example 2 and add 100 μL of sterile water to dilute it and spread it evenly on the water agar plate, repeat 3 times, culture statically at 25 ° C, and count after continuous culture for 7 days. Add sterile water containing 0.1% SDS to the plate and wash it repeatedly to elute the conidia microsclerotia-diatomaceous earth mixture, and use a pipette to draw all the liquid on each plate, record the liquid volume, and shake evenly The concentration of conidia in the eluate was determined by microscopic counting on a hemocytometer. The concentration of conidia multiplied by the volume of liquid collected, divided by 0.01, equals the number of conidia produced per gram of microsclerotia-diatomaceous earth mixture. The results showed that Trichoderma dark viridans microsclerotia-diatomaceous earth mixture can produce 4.3...

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Abstract

The invention relates to the technical field of biological preparations, in particular to a preparation method and application of Trichoderma dark green microsclerotia and its preparation. The preparation method comprises the following steps: preparation of conidia suspension of Trichoderma dark green; submerged culture of Trichoderma dark green to induce production of micro-sclerotia; harvest of micro-sclerotia; mixing micro-sclerotia with carrier and then drying to prepare dry powder preparation . The Trichoderma microsclerotium obtained by this method has the advantages of high temperature resistance, ultraviolet resistance, and storage resistance, and can be produced by large-scale liquid fermentation. The process is simple and the cost is low, and it can be used for preparation products such as Trichoderma bio-organic fertilizers and microbial agents. It has a good control effect on wheat stem rot.

Description

technical field [0001] The invention relates to the technical field of biological preparations, in particular to a preparation method and application of Trichoderma dark green microsclerotia and its preparation. Background technique [0002] Trichoderma is an important biocontrol fungus, which has been widely used in the biological control of plant pathogenic fungi in agricultural production. Trichoderma can directly antagonize pathogens, and can also induce plant resistance to resist pathogenic fungi. At the same time, most Trichodermas also have the ability to promote The function of plant growth is a biocontrol bacterium with great application prospects. At present, the active ingredients of Trichoderma biopesticides are mainly composed of mold conidia, which are usually produced by fermentation on a wet solid fermentation medium. The fermentation process needs to last for several weeks, and then post-fermentation treatment, not only the production cost However, it is al...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14A01N63/38A01P3/00C12R1/885
CPCC12N1/14A01N63/38
Inventor 扈进冬台少华吴远征魏艳丽李纪顺
Owner BIOTECH CENT OF SHANDONG ACAD OF SCI
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