CN112877220B - Trichoderma harsii and application thereof - Google Patents
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Abstract
The invention discloses trichoderma harsii and application thereofThe Latin chemical name of the Trichoderma virgatum is Trichoderma strigosum, the strain number TR2125 and the preservation number of CGMCC No. 21430. The fermentation liquor obtained by fermenting the trichoderma harsii is used for preparing a microbial agent, and the obtained microbial agent has a good prevention and treatment effect on tomato damping-off; the storage is not easy to agglomerate, the application is free from dust pollution, and the application is convenient; good dispersion, high viable count, spore content of 3 × 10 8 CFU/g, and its chlamydospore content is still 10 after 12 months of storage 8 CFU/g is higher than the standard.
Description
Technical Field
The invention relates to the technical field of microorganisms, and particularly relates to trichoderma harsii and application thereof.
Background
Trichoderma (including Trichoderma harzianum) is a microorganism in nature with inhibitory and killing functions against a variety of phytopathogens. The excellent characteristics of trichoderma are utilized by human beings to prepare biological pesticides and biological fertilizers with different formulations through a specific processing technology. The trichoderma has the characteristics of wide distribution, easy separation and culture, capability of generating chlamydospores with strong stress resistance, easy storage, convenient use and the like, and has the following effects: preventing and treating various plant diseases and promoting plant growth.
Tomatoes are important economic crops in China. Tomato damping-off is an important disease caused by Pythium ultimum Trow of the subdivision flagellata. Typical symptoms are sudden collapse (seed rot) before emergence of seedlings and a large amount of water stain type disease spots at the base of seedling stems in the early planting stage, so that the seedlings are withered and die, melon and fruit are damaged, wet rot is caused, and serious economic loss is caused.
The existing microbial preparation has poor control effect on tomato damping-off, or has the defects of easy caking during storage, dust pollution during application and inconvenient application.
Disclosure of Invention
The invention aims to provide trichoderma harsii and application thereof, aiming at the defects in the prior art.
In order to achieve the purpose, the invention adopts the technical scheme that:
the first aspect of the invention provides Trichoderma harsii, which has the Latin chemical name of Trichoderma strigosum, the strain number TR2125 and the preservation number of CGMCC No. 21430.
The Trichoderma harsii (Trichoderma koninaceum) strain is preserved in China general microbiological culture Collection center (CGMCC), and the preservation address is as follows: xilu No. 1 Hospital No. 3, Beijing, Chaoyang, Beicheng; the preservation date is 2021, 01 month and 13 days.
The second aspect of the present invention provides a Trichoderma reesei fermentation broth, which is prepared by the Trichoderma reesei (Trichoderma strigosum) fermentation method, and comprises the following steps:
inoculating the trichoderma harsii into a PDA culture medium, and culturing at constant temperature;
step two, transferring the strain cultured in the step one into a culture solution for shake flask culture for 24-30h to obtain a first-stage seed solution;
step three, sterilizing the culture solution in a middle-stage fermentation tank at high temperature and high pressure, then pouring the first-stage seed solution obtained in the step two into the middle-stage fermentation tank, pressurizing and introducing sterile air for fermentation for 6-7 days to obtain trichoderma harsii fermentation liquor;
in the second step and the third step, the culture solution comprises the following components in parts by weight:
further, in the second step and the third step, the culture solution comprises the following components in parts by weight:
further, in the third step, the fermentation conditions are as follows: 28-30 ℃, 0.4-0.5 kg pressure, 1500-1800L/H air flow and 120-150 rpm rotation speed.
Further, in the third step, the specific steps of the high-temperature and high-pressure sterilization are as follows: sterilizing at 121 deg.C under 15 pounds pressure for 20 min.
Further, in the third step, after the high-temperature and high-pressure sterilization, the medium-level fermentation tank keeps positive pressure and is cooled to 28-30 ℃.
Further preferably, in the third step, after the high-temperature and high-pressure sterilization, the medium-grade fermentation tank is kept at positive pressure and cooled to 30 ℃.
Further, the medium-sized fermenter means a fermenter of more than 1L and less than 1 ton size, which is common knowledge in the art.
Further, before the high-temperature and high-pressure sterilization, a defoaming agent is added into the middle-stage fermentation tank; the volume of antifoam added in a 100L format fermenter is preferably 50 ml.
The third aspect of the invention provides a microbial agent, which contains the trichoderma harsii fermentation liquor, and comprises the following components in percentage by weight:
the microbial agent is prepared by the following method: uniformly mixing the carrier, the mineral potassium humate, the turf, the wetting agent, the dispersing agent, the disintegrating agent, the binder and the humectant according to the proportion, spraying the trichoderma bristlense fermentation liquor, and uniformly stirring until the water content of the mixed material is 40-45%; and (3) after the pelleting is carried out by the pelleting machine, dehumidifying for 3 hours by a dehumidifier until the water content of the granules is 8-10%, and obtaining the finished product of the microbial agent.
Further, the humectant is one or more of glycerol, butanediol, polyethylene glycol, propylene glycol, xylitol and polypropylene glycol.
Further, the binder is one or more of methylcellulose and hydroxypropyl methylcellulose.
Further, the disintegrating agent is one or more of starch, microcrystalline cellulose and sodium alginate.
Further, the dispersing agent is one or more of NNO and Wetting agent HT-100; the wetting agent is one or more of K12, peregal, fatty alcohol-polyoxyethylene ether, sulfonate, sodium dodecyl benzene sulfonate and organic silicon.
Further, the carrier is one or more of diatomite, kaolin, bentonite and pottery clay.
In the microbial agent of the present invention, the biologically active ingredient having a biocontrol effect is a mycelium, a conidium or a chlamydospore, and preferably a chlamydospore.
By adopting the technical scheme, compared with the prior art, the invention has the following technical effects:
the microbial agent containing the trichoderma harsii fermentation liquor has a good prevention and treatment effect on tomato damping-off; the storage is not easy to agglomerate, the application is free from dust pollution, and the application is convenient; good dispersion, high viable count, spore content of 3 × 10 8 CFU/g, and its chlamydospore content is still 10 after 12 months of storage 8 CFU/g is higher than the standard.
The preparation process of the microbial agent is simple and is easy to realize continuous production.
Drawings
FIG. 1 is an electron micrograph of a conidiophores of Trichoderma harzianum according to the invention;
FIG. 2 is an electron micrograph of a conidia morphology of Trichoderma harzianum according to the present invention;
FIG. 3 is a photograph of a PDA plate morphology of Trichoderma harzianum according to the invention.
Detailed Description
The invention is further described with reference to the following drawings and specific examples, which are not intended to be limiting. It should be noted that the embodiments and features of the embodiments of the present invention may be combined with each other without conflict.
In the present invention, the pathogenic bacteria: tomato damping-off bacteria, deposited in Wanlihua Biotech, Inc., Shanghai. Tomato variety: tomato L-402 (Liaoning horticulture sprout Co.).
Example 1
The embodiment provides Trichoderma harsii which is collected and separated from panax notoginseng rhizosphere soil in Shanshan city of Yunnan province, the Latin school name of the Trichoderma harsii is Trichoderma strigosum, the strain number is TR2125, and the preservation number is CGMCC No. 21430.
The Trichoderma hirsutum (Trichoderma strain) strain is preserved in China general microbiological culture collection center (CGMCC), and the preservation address is as follows: xilu No. 1 Hospital No. 3, Beijing, Chaoyang, North; the preservation date is 2021, 01 month and 13 days.
Classification and identification of trichoderma harsii:
the growth speed of the trichoderma harsii TR2125 colony is medium and relatively high, and the diameter of the colony after three days of growth is 4-6 cm; most of the hyphae are endogenetic, and aerial hyphae with limited growth on PDA culture medium are curly hair or felt, and are white. Conidia are typically generated in a near-hemispherical spore-forming cluster, and the diameter of the spore-forming cluster can reach 4 mm; the spore-forming clusters are large in number, are usually arranged in a ring shape or are gathered at the edge of a flat plate, occasionally aggregate into a large spore-forming area (PDA), and are in a burr shape on the surface under the observation of a low power lens due to a plurality of straight and hard javelin-shaped conidiophore top sterile extensions, and particularly on an MA culture medium, burr structures are more obvious. The spore forming clusters are initially white and quickly turn to dark blue-green. The colony was colorless to white or dark yellow (old) on the reverse side as an observed exudate.
Referring to FIGS. 1 to 3, the Trichoderma harzianum TR2125 hyphae are transparent, occasionally have yellow contents, have smooth walls and diameters of 2 to 9 microns, have an intrabasal hyphae diameter of 14 microns, mostly have intercropped chlamydospores in aged cultures, frequently exist singly, occasionally are arranged in short chains, are transparent to light green, have a subspheroid shape, mostly have diameters of 5 to 12 (16) microns, have transparent walls and have wall thicknesses of 2.5 microns. Conidiophores are transparent, straight and short, have smooth walls, occasionally have obvious irregular wall thickening near the basal part, the diameter of the basal part is 6 mu m, the conidiophores gradually become thin, the diameter of the whole fertile area is 3-4 mu m, the conidiophores occasionally can be expanded on branch points, and the diameter of the conidiophores is 8 mu m; the high-degree branches are short in primary branches, 2-5 cells mostly grow out at acute angles, 2-3 cells are arranged in a vortex shape, the cells are reflected to the top end of the stem, complex secondary branches are formed, 1-2 cells are branched at the final stage, 2-3 cells are arranged in a vortex shape, the top cells are approximately cylindrical, the size of the top cells is 4.5-6.8 multiplied by 2.8-3.6 mu m, the upper portion of the stem is in a straight and hard javelin shape, and few waves are formed. The hypha section is sterile when the hyphae section is from 1 to 3 times on the fertile upper part to 50 to 100 mu m at the top, most of the single colonized conidiophores without ornaments grow into single colonized conidiophores with the length of 8 to 15 mu m and the width of 1 to 2 mu m, the diameter of the conidiophores is from 3 to 4 mu m to 1 mu m near the top, the conidiophores are not obvious, the distance between the septa is long and about 12 to 26 mu m, the phialides or the ampoules are in a shape of a bottle stalk or an ampoule, the conidiophores are from 3.8 to 7.0 multiplied by 2.1 to 3.4 mu m, the base parts are obviously contracted and gradually become colonized conidiophores, the diameter of the conidiophores is 1 mu m, the conidiophores are usually 2 to 4.8 multiplied by 1.8 to 2.5 mu m (average 4.1 multiplied by 2.1 mu m), the two ends are blunt, or the base parts are slightly sharp, green and the wall is smooth.
Identification of the molecular biological species of trichoderma harsii:
the DNA of TR2125 is amplified to a sequence segment obtained by an ITS1-5.8S-ITS2 region sequence by using primers ITS4 and ITS5, and the sequence segment of TR2125 is compared with the sequence segment of trichoderma harzianum by using data in a DNA sequence database established by NCBI Gen Bank, so that the similarity of the sequence segment of TR2125 and the trichoderma harzianum reaches 99 percent.
The ITS1-5.8S-ITS2 gene sequence of TR2125 is as follows:
TTCCTCCGGCTTATTGATATGCTTAAGTTCAGCGGGTATTCCTACCTGATCCGAGGTCAACATTTCAGAAAGTTGGGTGTTTTACGGACGTGGACGCGCCGCGCTCCCGGTGCGAGTTGTGCAAACTACTGCGCAGGAGAGGCTGCGGCGAGACCGCCACTGTATTTCGGGGCCGGGATCCCGTCTTAGGGGTTCCCGAGGTCCCCAACGCCGACCCCCCGGAGGGGTTCGAGGGTTGAAATGACGCTCGGACAGGCATGCCCGCCAGAATACTGGCGGGCGCAATGTGCGTTCAAAGATTCGATGATTCACTGAATTCTGCAATTCACATTACTTATCGCATTTCGCTGCGTTCTTCATCGATGCCAGAACCAAGAGATCCGTTGTTGAAAGTTTTGATTCATTTTGAATTTTTGCTCAGAGCTGTAAGTAAAAACGTCCGCGAGGGGACTACAGTAAAGAGTTTGGTTGGTCCCTCCGGCGGGCGCCTGGTTCCGGGGCTGCGACGCGCCCGGGGCGTGACCCCGCCGAGGCAACAGTTTGGTAACGTTCACATTGGGTTTGGGAGTTGTAAACTCGGTAATGATCCCTCCGCTGGTTCACCAACGGAGACCTTGTTACGATTTTTACTTCCAA
the sequence section obtained by amplifying the Tef1 alpha sequence of the TR2125 by using primers Ef728 and Tef1 alpha R is compared by data in a DNA sequence database established by NCBI Gen Bank, and the similarity between the Tef1 alpha sequence of the TR2125 and the Trichoderma bristle reaches 99 percent.
The Tef1 α sequence of TR2125 is as follows:
TTGCCATCCCTTTGGAGATACCAGCCTCGAACTCACCAGTACCGGCAGCGATAATCAGGATAGCGCAGTCAGCCTGGGAGGTACCAGTGATCATGTTCTTGATGAAATCACGGTGACCGGGAGCGTCTGTAGAAGGGTACGTTAGAATGATGATTTCGAGGTGTGCATGACGAAACTGAAAACATACCAATGACGGTGACATAGTACTTGGGAGTCTCGAACTTCCAGAGGGCAATGTCGATGGTGATACCACGCTCACGCTCGGCCTTGAGCTTGTCAAGAACCCACGCATACTTGAAGGAACCCTTGCCGAGTTCGGCGGCTTCCTATTGATTGAAACATGATCAGCACAATGAATTCACAAGAGATTGAGGCACAGACGATGCGATGGAATGAACAGAATGTGTTGGGGCGATGAGGGACAGTGGCGATAGCGGGGTTTGCCCAAAAAAATTGGCACCCCACTAAAAGCCAAACAAGGCAGCCAGAAAATTCTGCTGTGTCAAAGGAGGGGTAGACAACCAAAGCGGGGTGTGACGCTGGTTGAAAAAAAAAATGGCAAGACTGAGAACAGAATTGTCGGACAGTCGGGCAAGAGTGCAAGATGAGAAAAGCAGTGAATTGAGCTTACCTTCTCGACTTTCTCGATGA
the separated strain can be determined to be the trichoderma harzianum strain by molecular biological strain identification and combining morphological characteristics.
The conidiophore stage of the above-mentioned Trichoderma harzianum strain is Deuteromycotina (Deuteromycotina), Hyphomycetes (Hyphomycetes), Trichosporoles (Hyphomycetales), Blastomyces (Moniliaceae), Trichoderma (Trichoderma), and its active state is Ascomycotina (Ascomycotina), Hypocrea (Hypocrea).
Example 2
The embodiment provides a trichoderma harsii fermentation liquid, and a preparation method of the trichoderma harsii fermentation liquid comprises the following steps:
step one, inoculating the trichoderma harcalii separated in the embodiment 1 into a PDA culture medium, and culturing at constant temperature;
step two, transferring the strain cultured in the step one into a culture solution for shake flask culture for 24 hours to obtain a first-stage seed solution;
step three, sterilizing the culture solution in the middle-stage fermentation tank at the temperature of 121 ℃ under the pressure of 15 pounds for 20min, keeping positive pressure, cooling to 30 ℃, then pouring the first-stage seed solution obtained in the step two into the middle-stage fermentation tank, introducing sterile air under pressure, and fermenting for 6 days at the temperature of 28-30 ℃, the pressure of 0.4-0.5 kg, the air flow of 1500-1800L/H and the rotating speed of 120-150 rpm to obtain the trichoderma harsii fermentation liquor;
in the second step and the third step, the culture solution comprises the following components in parts by weight:
example 3
The embodiment provides a microbial agent, which contains the trichoderma harzianum fermentation liquor obtained in the embodiment 2, and comprises the following components in percentage by weight:
the microbial agent is prepared by the following method: uniformly mixing the carrier, the mineral potassium humate, the turf, the wetting agent, the dispersing agent, the disintegrating agent, the binder and the humectant according to the proportion, spraying the trichoderma bristlense fermentation liquor, and uniformly stirring until the water content of the mixed material is 40-45%; and (3) after the pelleting is carried out by the pelleting machine, dehumidifying for 3 hours by a dehumidifier until the water content of the granules is 8-10%, and obtaining the finished product of the microbial agent.
The detected spore content is 3 × 10 8 CFU/g. According to the GB20287-2006 detection standard of agricultural microbial agents, indexes such as pH, moisture, fineness, infectious microbe rate and the like all reach the national standard, and the granules are qualified.
TABLE 1 determination of the chlamydospore content of active Trichoderma harzianum in granules with different storage times
| Storage time (moon) | 1 | 2 | 3 | 4 | 5 | 6 | 12 |
| Number of spores (10) 8 A/g) | 3.7 | 3 | 2.95 | 3.12 | 2.75 | 2.57 | 2.55 |
Table 1 shows the spore content of the active trichoderma harsii microbial inoculum stored for 12 months, and the chlamydospore content of the granules is still 10 after 12 months 8 The above.
Application example
1. Antagonism test of trichoderma harsii on tomato damping-off disease
Time and place of experiment: performed in the laboratory of Wanli Biotech Ltd in the Shanghai in 8 Yuehai of 2018.
The test method comprises the following steps:
sources of damping-off germs of tested tomatoes: the tomato damping-off germ strain is collected from the stem base of a tomato diseased plant in a Shanghai greenhouse, is separated and purified by the strain group of Wanli Biotechnology Limited, is identified as Pythium ultimum Trow of the subdivision Trichophyton, and is shown as strong pathogenicity in pathogenicity determination.
Plate confrontation experiment:
culturing Trichoderma and tomato damping-off 3d in PDA culture medium at 28 deg.C, using a hole puncher
Picking trichoderma and pathogenic bacteria dishes, placing the pathogenic bacteria dishes on one side of a culture dish plate with the diameter of 9cm, inoculating the trichoderma dishes on the other side, keeping the distance between the two fungus dishes at 4cm, and taking singly cultured pathogenic bacteria as a reference; culturing at 28 deg.C for 3 days, and observing synergistic effect. Each experiment was repeated at least 3 times. When the blank control is about to grow over the whole culture dish, measuring the control growth amount (colony radius) and the treatment growth amount (growth inhibition radius after inoculation of the trichoderma harosum), and expressing the antagonism by using the bacteriostasis rate (calculation formula: bacteriostasis rate (%) (control growth amount-treatment growth amount)/control growth amount multiplied by 100).
Results (table 2): the inhibition rate of the trichoderma harsii on tomato damping-off germs reaches 90.8 percent; the transparent bacteriostatic bandwidth is 12.0 mm; the trichoderma harsii has obvious inhibition effect on tomato damping-off germs and biological control potential for preventing and treating the tomato damping-off germs.
TABLE 2 antagonistic action test results of Mucor hirsutum on tomato damping-off
| Strain name | Control growth volume (mm) | Treatment of growth (mm) | Bacteriostatic ratio (%) |
| Mucor miehei | 41.5 | 3.8 | 90.8 |
2. Comparison test of trichoderma harsii microbial agent on control effect of tomato damping-off
And (3) microbial agent: the trichoderma harsii granules prepared in example 3 were applied in 5g per pot of soil.
Chemical agents: withered water aqua (Hebei Wante Biochemical Co., Ltd.); diluted 500 times with water.
Blank control: clean water
The test method comprises the following steps: the method is carried out in artificial climate room of Shanghai Wanli Biotechnology Limited company in 2019 and 5 months. Culturing tomato seedling (tomato L-402, produced by gardening research institute of agricultural science institute of Liaoning, Liaoning gardening Miao corporation) in sunlight greenhouse, transferring to temperature and humidity control culture room when 5 pieces of multiple leaves grow out, selecting tomato seedling with consistent size, and damaging main root to form wound. Then 5g of trichoderma harsii granules are applied to each pot of soil in a hole mode. Each treatment was repeated 3 times, 50 replicates each, with a control of water and chemicals. The investigation was started after wilting occurred in the control treatment, and was performed every 5 days for a total of 3 times. And calculating the potted plant test result according to the disease rate.
The plant disease rate and the prevention and treatment effect are calculated according to the formula:
results (table 3): the in-vivo bioassay result shows that the trichoderma microbial inoculum has the advantages of remarkably inhibiting the development of tomato damping-off germs, having the prevention effect of 78.22 percent and being superior to the prevention effect of chemical contrast agents (65.32 percent). The trichoderma microbial inoculum has good control effect on the tomato damping-off.
TABLE 3 comparative test results of the effect of the Mucor hirsuta granules on controlling tomato damping-off
| Treatment of | Average disease percentage (%) | Average control effect (%) |
| Mucor bristlegrass granules | 18 | 78.22 |
| Chemical agent | 28.67 | 65.32 |
| Blank control | 82.67 | -- |
3. Evaluation of growth promoting effect of trichoderma harsii granules on tomato plants
And (3) microbial agent: the trichoderma harsii microbial inoculum prepared in example 3 was applied in 5g per pot of soil.
Fertilizer: a humic acid-containing macroelement water-soluble fertilizer (Shanghai Wanli Biotech limited); diluted 1000 times with water.
Blank control: clean water
The test method comprises the following steps: performed in artificial climate room of Wanli Biotechnology Limited, Shanghai, 8 months in 2019. Culturing tomato seedlings (tomato L-402, produced by gardening research institute of agricultural science institute of Liaoning, Liaoning horticulture Miao corporation) in sunlight greenhouse, transferring to temperature and humidity control culture room after 5 pieces of compound leaves grow out, selecting tomato seedlings with consistent size, and performing hole application with Trichoderma bristlegrass microorganism granules at 5g per pot of soil. Each treatment was repeated 3 times, 50 replicates each, with a control of clear water and chemical fertilizer. During the period, the tomato pest and disease damage management is strengthened, and the moisture is supplemented timely. After 40 days, the plant height, the fresh weight of the overground part and the fresh weight of the underground part of the tomato are measured.
Results (table 4): from the results in table 4, it can be seen that there is a significant difference between the plant height, the fresh weight of the overground part, the fresh weight of the underground part and the blank control of the tomatoes treated with the humic acid-containing macroelement water-soluble fertilizer and the trichoderma harzianum granules. By integrating the indexes of plant height, fresh weight, dry weight and the like, the trichoderma harosum has certain growth promotion effect on tomato plants.
TABLE 4 influence of Trichoderma harzianum granules on plant height and fresh weight of potted tomatoes
| Treatment of | Plant height (cm) | Fresh weight of overground part (g) | Fresh weight of underground (g) | Dry weight of aerial parts (g) | Fresh weight of underground (g) |
| Humic acid-containing water-soluble fertilizer | 48.22±6.92c | 26.99±2.08b | 5.56±2.12c | 2.70±0.58b | 0.55±0.25a |
| Mucor bristlegrass granules | 43.00±2.60b | 28.16±5.66ab | 4.64±1.25b | 2.16±1.06ab | 0.45±0.18a |
| Blank control | 39.88±2.06a | 23.70±5.92a | 3.21±0.99a | 1.59±0.41a | 0.41±0.14a |
While the invention has been described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention.
Claims (10)
1. Trichoderma reesei is characterized in that the Latin chemical name of the Trichoderma reesei is Trichoderma strigosum, the strain number TR2125 is and the preservation number is CGMCC No. 21430.
2. The trichoderma harsii fermentation liquor is characterized in that the preparation method of the trichoderma harsii fermentation liquor comprises the following steps:
step one, inoculating the trichoderma harpacum with the preservation number of CGMCC No.21430 into a PDA culture medium, and culturing at constant temperature;
step two, transferring the strain cultured in the step one into a culture solution for shake flask culture for 24-30h to obtain a first-stage seed solution;
step three, sterilizing the culture solution in a middle-stage fermentation tank at high temperature and high pressure, then pouring the first-stage seed solution obtained in the step two into the middle-stage fermentation tank, pressurizing and introducing sterile air for fermentation for 6-7 days to obtain trichoderma harsii fermentation liquor;
in the second step and the third step, the culture solution comprises the following components in parts by weight:
3. the trichoderma harsii fermentation broth of claim 2, wherein in step two and step three, the broth comprises the following components in weight ratio:
4. a microbial inoculant comprising the trichoderma reesei fermentation broth as claimed in any one of claims 2 to 3, comprising the following components in weight percent:
5. the microbial agent according to claim 4, wherein the microbial agent is prepared by the following method: uniformly mixing the carrier, the mineral potassium humate, the turf, the wetting agent, the dispersing agent, the disintegrating agent, the binder and the humectant according to the proportion, spraying the trichoderma bristlense fermentation liquor, and uniformly stirring until the water content of the mixed material is 40-45%; and (3) after the pelleting is carried out by the pelleting machine, dehumidifying for 3 hours by a dehumidifier until the water content of the granules is 8-10%, and obtaining the finished product of the microbial agent.
6. The microbial agent according to claim 4, wherein the humectant is one or more of glycerol, butanediol, polyethylene glycol, propylene glycol, xylitol and polypropylene glycol.
7. The microbial agent according to claim 4, wherein the binder is one or more of methylcellulose and hydroxypropyl methylcellulose.
8. The microbial agent according to claim 4, wherein the disintegrating agent is one or more of starch, microcrystalline cellulose and sodium alginate.
9. The microbial agent according to claim 4, wherein the dispersant is one or more of NNO, Wetting agent HT-100; the wetting agent is one or more of K12, peregal, fatty alcohol-polyoxyethylene ether, sulfonate, sodium dodecyl benzene sulfonate and organic silicon.
10. The microbial agent according to claim 4, wherein the carrier is one or more of diatomite, kaolin, bentonite and pottery clay.
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| CN103484384B (en) * | 2013-10-08 | 2015-07-01 | 天津市植物保护研究所 | Trichoderma atroviride preparation for preventing and controlling vegetable fungal diseases and preparation method of trichoderma atroviride preparation thereof |
| CN107259203A (en) * | 2016-04-07 | 2017-10-20 | 吴政宏 | Trichoderma is given into the application in the feed and/or preparation for preparing the shrimp for being used to treat or prevent vibrio infection |
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