CN113999778B - Trichoderma atroviride microsclerotia and preparation method and application of preparation thereof - Google Patents
Trichoderma atroviride microsclerotia and preparation method and application of preparation thereof Download PDFInfo
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Abstract
The invention relates to the technical field of biological preparations, in particular to trichoderma atroviride microsclerotia and a preparation method and application of a preparation thereof. The preparation method comprises the following steps: preparing trichoderma atroviride conidium suspension; deep culture and induction of trichoderma atroviride liquid to produce microsclerotia; harvesting the microsclerotia; mixing the microsclerotia and the carrier, drying, and preparing the dry powder preparation. The trichoderma microsclerotia obtained by the method has the advantages of high temperature resistance, ultraviolet resistance, storage resistance and the like, can be produced by large-scale liquid fermentation, has simple process and low cost, can be used for preparing preparation products containing trichoderma bio-organic fertilizer, microbial inoculum and the like, and has better control effect on wheat stem base rot.
Description
Technical Field
The invention relates to the technical field of biological preparations, in particular to trichoderma atroviride microsclerotia and a preparation method and application of a preparation thereof.
Background
Trichoderma is an important biocontrol fungus, and has been widely applied to biological control of plant pathogenic fungi in agricultural production, the Trichoderma can directly antagonize pathogenic bacteria and induce plant resistance to resist pathogenic bacteria attack, and meanwhile, most of the Trichoderma also has the function of promoting plant growth, and is a biocontrol fungus with great application prospect. Currently, trichoderma biopesticide active ingredients mainly consist of conidia of mildew, the conidia are usually generated by fermentation on a wet solid fermentation culture medium, the fermentation process needs to last for several weeks, and then fermentation post-treatment is performed, so that the production cost is high, and the production is limited by various limiting factors, such as high labor cost, poor quality control, long fermentation time, difficult scale, short shelf life of products and the like. Currently, liquid culture research is mainly focused on the production of trichoderma chlamydospore, but the problems of low fermentation yield, lack of storage stability, poor durability in soil and the like exist.
Disclosure of Invention
The invention aims to provide a method for producing microsclerotia and a preparation thereof by liquid fermentation for large-scale production of trichoderma atroviride.
In one aspect, the invention provides a preparation method of trichoderma atroviride microsclerotia, which comprises the following steps:
(1) preparing trichoderma atroviride conidium suspension;
(2) deep culture and induction of trichoderma atroviride liquid to generate microsclerotia: inoculating the conidium suspension prepared in the step (1) into an induction culture medium, and performing shake culture at 25-30 ℃ and 200rpm for 7-9 days to obtain a trichoderma atroviride micro-bacterial nuclear fermentation solution;
(3) and (3) adding a filter aid into the microsclerotia fermentation liquid after the completion of the shake culture, uniformly mixing, carrying out suction filtration, removing a supernatant, obtaining the trichoderma atroviride microsclerotia, and temporarily storing.
Further, Trichoderma atroviride is HB20111 (C: (C.))Trichoderma atroviride) The microbial inoculum is preserved in the China general microbiological culture collection management center for 12 and 5 days in 2018, the preservation address is No. 3 of Xilu No.1 of Beijing north Shangyang district, the preservation number is CGMCC No.16963, and the patent with the publication number of CN110915822A is disclosed.
Further, in the step (2), the conidium inoculation amount is 10 inoculations per 100mL induction medium6And (4) spores.
Further, in the step (2), the induction medium comprises the following components: KH (Perkin Elmer)2PO4 3.0~5.0g/L、CaCl2·2H2O 0.5~1.0g/L、MgSO4·7H2O 0.4~0.8g/L、CoCl2·6H2O34~40mg/L、MnSO4·H2O 14~20mg/L、ZnSO4·7H2O 12~16mg/L；FeSO4·7H20.1-0.4 g/L of O; 15-30 g/L of glucose; casein hydrolysate 2.0-3.0 g/L.
Further, in the step (3), the filter aid is diatomite and is added in an amount of 1-5% (w/v).
In another aspect, the present invention provides a method for preparing a trichoderma atroviride microsclerotia preparation, comprising the steps of:
mixing the microsclerotia obtained after suction filtration with a carrier, drying at low temperature to obtain a dry microsclerotia-carrier mixture, and then adding an auxiliary agent to uniformly mix to obtain the trichoderma atroviride microsclerotia dry powder preparation.
Further, the carrier is one or a combination of more of diatomite, medical stone, kaolin, starch and corn bran powder; the mass percentage of the microsclerotia after suction filtration to the carrier is 1: (10-20).
Further, the auxiliary agent is one or a combination of several of sodium carboxymethylcellulose, xanthan gum and sodium alginate, and the mass percentage of the dried microsclerotia-carrier mixture to the auxiliary agent is 1000: 1-200: 1.
Further, the low-temperature drying adopts a fluidized bed drying at 35-40 ℃ until the water content is below 5%, and then the bagged product is subpackaged.
On the other hand, the invention also provides application of the trichoderma atroviride microsclerotia preparation in preventing and treating plant fungal diseases.
The invention has the beneficial effects that:
1. the trichoderma atroviride HB20111 with the biological preservation number of CGMCC No.16963 is selected, has the function of antagonizing various plant pathogenic fungi, and the microsclerotia generated by the fermentation of the trichoderma atroviride is selected as an active ingredient for preventing and treating the wheat stem basal rot, so that the trichoderma atroviride has a good prevention and treatment effect.
2. The trichoderma atroviride microsclerotia preparation prepared by the invention has strong stress resistance, can adapt to unfavorable storage conditions such as long-term (30 days) high temperature (30-40 ℃), ultraviolet rays and the like, and has a shelf life of more than 12 months.
3. After the trichoderma atroviride microsclerotia preparation prepared by the invention is rehydrated, hyphae can quickly recover and generate conidia, so that mass propagation in soil and plant rhizosphere is realized, ecological niches are strived for with plant pathogenic bacteria, and meanwhile, antibacterial substances are secreted to inhibit the growth of the pathogenic bacteria.
4. The trichoderma atroviride micro sclerotium preparation prepared by the invention adopts a liquid fermentation process, the fermentation raw materials are all common reagents, the cost is low, the production period is shortened by 5-7 days, the large-scale production of trichoderma is easy to realize, the trichoderma atroviride micro sclerotium preparation can replace trichoderma conidium to be used as an effective component of a microbial agent, and the trichoderma atroviride micro sclerotium preparation is used for the industrial production of novel trichoderma fungicides.
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In order to more clearly illustrate the embodiments or technical solutions in the prior art of the present invention, the drawings used in the description of the embodiments or prior art will be briefly described below, and it is obvious for those skilled in the art that other drawings can be obtained based on these drawings without creative efforts.
FIG. 1 is a microscopic image of fermentation-induced culture of Trichoderma atroviride liquid for 48 h in example 1.
FIG. 2 is a microscopic image of Trichoderma atroviride liquid fermentation-induced culture for 120 h in example 1.
FIG. 3 is a microscopic image of Trichoderma atroviride liquid fermentation induction culture 168 h in example 1.
Detailed Description
In order to make those skilled in the art better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
EXAMPLE 1 preparation of Microsclerotium by liquid fermentation of Trichoderma atroviride
(1) Preparation of Trichoderma atroviride conidia suspension
Trichoderma atroviride HB20111 (Trichoderma atroviride) The microbial inoculum is preserved in the China general microbiological culture collection management center for 12 and 5 days in 2018, the preservation address is No. 3 of Xilu No.1 of Beijing, Chaoyang, and the preservation number is CGMCC No. 16963.
The trichoderma atroviride conidia are obtained in a large quantity by a solid fermentation method, and the process flow is as follows: picking trichoderma atroviride blocks containing conidia from a trichoderma atroviride storage tube stored in an ultralow temperature refrigerator at minus 80 ℃, inoculating the trichoderma atroviride blocks to a PDA solid plate, and culturing for 5 days at 28 ℃; selecting single colony growing vigorously on PDA solid plate, inoculating to eggplant bottle, selecting PDA culture medium, culturing at 28 deg.C for 5-10 days until hypha and conidium are full, and harvesting. Washing eggplant bottle surface with 0.9% physiological saline sterilized at 121 deg.C for 20min, washing fresh spore, and diluting to 1 × 108spore/mL for use.
(2) Induced culture of trichoderma atroviride microsclerotia
The formula of the induction culture medium is as follows: KH (Perkin Elmer)2PO4 3.0~5.0g/L、CaCl2·2H2O 0.5~1.0g/L、MgSO4·7H2O 0.4~0.8g/L、CoCl2·6H2O34~40mg/L、MnSO4·H2O 14~20mg/L、ZnSO4·7H2O 12~16mg/L；FeSO4·7H20.1-0.4 g/L of O; 15-30 g/L of glucose; 2.0-3.0 g/L casein hydrolysate and deionized water; the liquid culture medium is respectively dispensed into 500mL triangular flasks in an amount of 200 mL, the pH value is adjusted to 7.5, high-temperature and high-pressure sterilization is carried out at 121 ℃ for 30 min, and inoculation is carried out after cooling.
Liquid fermentation culture conditions: inoculating the prepared trichoderma atroviride conidium suspension into an induction culture medium according to the inoculation amount of 1%, performing shake culture at the temperature of 24-28 ℃ and under the condition of 150-180 rpm, and observing the microsclerotia generation condition in the induction culture medium every day after inoculation. After inoculation and shaking culture for 16 h, trichoderma atroviride conidia germinate to form hyphae, and a large number of micro hypha balls are formed after 24-48 h, wherein the color of the bacterial liquid is not changed, as shown in figure 1; after 72 h of culture, the hypha amount in the bacterial suspension is continuously increased, the hypha balls are continuously increased and become compact, and no microsclerotia is generated under the observation of an optical microscope; after 120 h of culture, the mycelium pellet becomes more compact, the edge of part of the mycelium pellet is brown, and a part of microsclerotia dormancy structures can be seen when a sample is observed under a microscope. The microsclerotia is grayish brown, smooth in edge and compact in middle, as shown in fig. 2; after 144h of culture, the number of the microsclerotia is greatly increased, hyphae begin to germinate around a small number of the microsclerotia, the number of the microsclerotia is not obviously increased after 168 h, and slender hyphae are formed around part of the microsclerotia, as shown in figure 3.
EXAMPLE 2 measurement of Microsclerotium in liquid fermentation broths
The microsclerotia concentration was determined by sucking 1ml of liquid fermentation sample from the flask using a tip with a head cut off (to avoid hyphal clogging), performing two 1:10 serial dilutions to give a 1:100 dilution, placing 100. mu.L of the dilution on a microscope glass slide, covering with an extra large cover glass (24X 50 mm), counting all dense microsclerotia under the cover with a low power (40X) light microscope, and calculating their concentration per ml.
Harvesting of Trichoderma atroviride microsclerotia
After 7 days of liquid fermentation according to the method of example 1, a large number of microsclerotia of compact hyphae with good morphology were produced in the medium, and the mycelia were harvested. When in harvest, 2.5 percent (w/v) of diatomite is added into the induction culture medium and thoroughly mixed; then pouring the mixture on 80 or 120 mesh screen, washing with running water, and removing fermentation liquor and hypha components until the microsclerotia becomes the main residue on the screen. Then, Whatman qualitative filter paper No.1 was placed in the Buchner funnel, the filter was wetted with deionized water, and then vacuum was applied to pour the microsclerotia-diatomaceous earth mixture into the Buchner funnel, removing the residual medium.
After the medium is removed, the filter cake (water content about 75%) is removed, triturated manually or with a food processor, dried overnight at room temperature in a fume hood or drying chamber, packaged and stored at 4 ℃ when the moisture of the microsclerotia-diatomaceous earth mixture is less than 5%.
Example 3 measurement of Nuclear spore-Forming ability of Trichoderma atroviride Microbacterium
10mg of the microsclerotia-diatomaceous earth mixture dried in example 2 was diluted with 100. mu.L of sterile water, applied evenly to a water agar plate, and incubated at 25 ℃ for 3 times, and after incubation for 7 days, the plate was washed repeatedly with sterile water containing 0.1% SDS to elute the microspore microsclerotia-diatomaceous earth mixture, and all the liquid on each plate was pipetted, the volume of the liquid was recorded, and after shaking evenly, the plate was counted microscopically by a blood-ball counter to determine the concentration of the microspores in the eluate. Conidia concentration multiplied by the volume of liquid collected divided by 0.01 equals the amount of conidia produced per gram of microsclerotia-diatomaceous earth mixture. The results showed that the Trichoderma atroviride microsclerotia-diatomaceous earth mixture produced 4.3X 10 per mg5And (4) spores.
Example 4 Trichoderma atroviride Microsclerotium heat resistance assay
12 parts of the microsclerotia-diatomaceous earth mixture dried in example 2, 0.1 g each, were weighed into test tubes containing 50 mL of sterile water, 12 tubes in total, and divided into four groups of 3 tubes, each group was placed in a 40 ℃, 45 ℃, 50 ℃, 55 ℃ water bath, 500. mu.L of microsclerotia diluted solution was taken out of the test tubes every 10min, evenly spread on a water agar plate, taken 4 times in total, and the conidia of Trichoderma atroviride were used as a control. Incubation was carried out at 25 ℃ and 100 microsclerotia were counted under a 24 h low power (40 ×) optical microscope to determine the germination rate of each treated microsclerotia. Control conidia 100 conidia were counted in a high power (400 ×) optical microscope and the spore germination rate was recorded and calculated.
As a result, the germination rates of the microsclerotia and the conidia treated at 40 ℃ have no significant difference with the germination rate without heat treatment, and are both more than 85%. The germination rate of the microsclerotia after 20min of treatment at 55 ℃ is obviously reduced to 72.59%, the germination rate of the microsclerotia after 40min of treatment is 45.56%, while the germination rate of the conidia after 10min of treatment is obviously reduced to 62.54%, and the germination rate of the conidia after 40min of treatment is only 16.99%. Indicating that the microsclerotia have better heat resistance than the microspores.
Example 5 measurement of ultraviolet ray resistance of Microsclerotium of Trichoderma atroviride
1 g of the microsclerotia-diatomaceous earth mixture dried in example 2 was weighed into a test tube containing 100mL of sterile water, 500. mu.L of the microsclerotia diluted solution was uniformly applied to a water agar plate, and the plate was irradiated under a UV-B ultraviolet lamp for 0, 1, 2, and 3 hours at a distance of 50 cm. The plate after UV irradiation was placed in an incubator at 25 ℃ and incubated for 24 hours in the dark, and 100 microsclerotia were counted under a low power (40 ×) optical microscope to determine the germination rate of each treated microsclerotia. The experiment was repeated 3 times with Trichoderma atroviride conidia as a control. The result shows that the germination rate is gradually reduced along with the increase of the ultraviolet irradiation time, and after 1 hour of ultraviolet irradiation, the germination rate of conidia is reduced to 73.66 percent, and the germination rate of microsclerotia is 81.37 percent; after 3h irradiation, the germination rate of conidia is sharply reduced to 22.98%, and the germination rate of microsclerotia is 58.27%, which indicates that the ultraviolet resistance of microsclerotia is superior to that of conidia.
Example 6 measurement of the Room temperature storage Capacity of Microsclerotium of Trichoderma atroviride
The microsclerotia cultured by the microsclerotia-diatomite mixture dried in the example 2 is sealed and placed at room temperature, and a part of the microsclerotia-diatomite mixture is taken out every three months to measure the germination rate, which comprises the following specific steps: diluting 10mg of dried mixture of Trichoderma microsclerotia and diatomaceous earth with 100 μ L of sterile water, uniformly spreading on water agar plates, culturing 3 water agar plates at 25 deg.C for 48 hr, and observing the recovery condition of mycelia of about 100 mixture of Trichoderma microsclerotia and diatomaceous earth on the plates under low power (40 ×) optical microscope. The result shows that the germination rate of the microsclerotia in 6 months is 90.46 percent, while the germination rate of the microsclerotia placed for 1 year is 83.53 percent in 48 hours, and thus the trichoderma atroviride microsclerotia has a better storage effect.
Example 7 preparation of Trichoderma atroviride Microsclerotium preparation
Putting the trichoderma atroviride micro-sclerotium fermentation solution harvested in the example 2 into a 50L plastic barrel, adding 2.5% (w/v) of diatomite, stirring and uniformly mixing, filtering to remove the fermentation solution, adding 2-3 times of medical stone into the solid part obtained by filtering, uniformly mixing, adding into a fluidized bed, drying by hot air at 35 ℃ until the water content is less than or equal to 5%, then adding 5-8 parts of carrier, adding 0.1-0.5% of xanthan gum to obtain the dried trichoderma atroviride micro-sclerotium preparation, and finally subpackaging into bagged products.
EXAMPLE 8 Trichoderma atroviride Microsclerotium preparation storage and spore production ability
Weighing 10 g of the trichoderma atroviride microsclerotia preparation, placing the trichoderma atroviride microsclerotia preparation into a 500mL triangular flask (containing 100mL of sterile water and sterilized glass beads), oscillating for 5 min in a shaking table, placing 100 mu L of diluent on a microscope glass slide, covering the microscope glass slide with an extra-large 24X 50 mm cover glass, counting all compact microsclerotia under the cover glass by using a low-power (40X) light microscope, and calculating the number of the microsclerotia in the microsclerotia preparation to serve as an initial value. Then, the number of microsclerotia in the microsclerotia preparation stored at room temperature, the germination rate (method described in reference example 4) and the spore productivity were measured every month, and the results showed that the germination rate was more than 80% at room temperature when the microsclerotia preparation of Trichoderma atroviride was stored for 1 year at 48 hours, and that the germination rate was 7.2X 10% per gram of the preparation under the rehydration condition (method described in reference example 3)5And (4) spores.
Example 9 spore-forming ability of Trichoderma atroviride Microsclerotium preparation in soil
The water agar plate is adopted to determine the viability and the conidium yield of the trichoderma atroviride microsclerotia preparation in natural condition soil and sterile processing soil. The microsclerotia preparation and soil are mixed according to the mass percentage of 100:1, the sporulation capacity is measured according to the method in the example 3, a soil sample containing the microsclerotia preparation is cultured on a water agar culture medium for 7 days, and the hypha protrusion or hypha germination condition is observed every day. For sterile and non-sterile soils, hyphae began to grow 3 and 5 days after culture and conidia began to form by day 7, respectively. Wherein the microsclerotia preparation inoculated into the sterile soil sample can produce 8.9 x 10 per gram of soil5Conidium, while the microsclerotia preparation inoculated on non-sterile (natural) soil can produce 9.2X 10 per gram of soil4And (4) forming conidia.
Example 10 test for controlling wheat shoot basal rot with Trichoderma atroviride Microsclerotium preparation
The test site is located in the city area of Texas city, the previous stubble of the test site is corn, and the stem base rot is serious in recent years. The trichoderma atroviride microsclerotia preparation prepared in the embodiment 7 of the invention is used before sowing, the dosage of the microbial inoculum is used according to 2kg per mu of land, the microbial inoculum is mixed with soil to 10kg and is uniformly scattered in a field, a blank control is set, and in order to eliminate the influence of other factors, a treatment group and the blank control group do not use a seed dressing agent for treatment and a chemical agent in the planting process. The test cell is 8m long, 1.5m wide and 12m in area2Each group treated 3 replicates. The wheat variety is selected as Jimai 22, which is a main cultivated variety in Shandong province.
The wheat stem base rot investigation method refers to disease classification standards such as Moya-Elizondo, E, A and the like. Grading is carried out 180 days after sowing according to the browning degree of the first internode of the wheat, wherein 0-level browning is not carried out, and the ratio of 1-level internode browning is 0% -25%, the ratio of 2-level internode browning is 25% -50%, the ratio of 3-level internode browning is 50% -75% and the ratio of 4-level internode browning is 75% -100%. Disease severity (disease index) = Σ (number of diseased plants at each stage × number of representative plants at the disease stage)/(number of representative plants at the highest disease stage × total number of investigated plants) × 100%.
The number of white spikes in each cell was investigated 220 days after sowing, and the control effects of treatment and control of the microsclerotia formulations were counted according to the white spike rate, with specific results shown in table 1. The control effect calculation method comprises the following steps: control effect (%) [ disease index of blank control area (spikelet rate) -disease index of treatment area (spikelet rate) ]/disease index of blank control area (spikelet rate) × 100%.
TABLE 1 control Effect of Trichoderma atroviride Microsclerotium preparations on wheat shoot basal rot
Through the above examples, it can be found that the trichoderma atroviride microsclerotia of the invention can be produced by using liquid submerged fermentation, and is convenient for large-scale production; meanwhile, the obtained microsclerotia has good storage resistance and can be stored for more than 1 year at room temperature; the germination rate of the dry microbial sclerotium powder after rehydration is high, a large amount of conidia can be produced, and the microbial sclerotium preparation prepared on the basis can prevent and treat wheat stem basal rot, so that the microbial sclerotium preparation has good disease prevention capability and can be used for production and application of biological organic fertilizers and microbial agents.
Although the present invention has been described in detail by referring to the preferred embodiments, the present invention is not limited thereto. Various equivalent modifications or substitutions can be made on the embodiments of the present invention by those skilled in the art without departing from the spirit and scope of the present invention, and these modifications or substitutions are within the scope of the present invention/any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (6)
1. The application of the trichoderma aureoviride microsclerotia preparation in preventing and treating the wheat stem basal rot is characterized in that the preparation method of the trichoderma aureoviride microsclerotia preparation comprises the following steps: mixing trichoderma atroviride microsclerotia with a carrier, drying at low temperature to obtain a dried microsclerotia-carrier mixture, adding an auxiliary agent, and uniformly mixing to obtain a trichoderma atroviride microsclerotia dry powder preparation;
the preparation method of the trichoderma atroviride microsclerotia comprises the following steps:
(1) preparing trichoderma atroviride conidium suspension;
(2) deep culture and induction of trichoderma atroviride liquid to generate microsclerotia: inoculating the conidium suspension prepared in the step (1) into an induction culture medium, and performing shake culture at 25-30 ℃ and 200rpm for 7-9 days to obtain a trichoderma atroviride micro-bacterial nuclear fermentation solution;
(3) adding a filter aid into the microsclerotia fermentation solution after the completion of the shake culture, uniformly mixing, carrying out suction filtration, removing a supernatant, obtaining the microsclerotia of trichoderma atroviride, and temporarily storing;
in the step (2), the induction medium comprises the following components: KH (Perkin Elmer)2PO4 3.0~5.0g/L、CaCl2·2H2O 0.5~1.0g/L、MgSO4·7H2O 0.4~0.8g/L、CoCl2·6H2O 34~40mg/L、MnSO4·H2O 14~20mg/L、ZnSO4·7H2O 12~16mg/L；FeSO4·7H20.1-0.4 g/L of O; 15-30 g/L of glucose; 2.0-3.0 g/L of casein hydrolysate;
trichoderma atroviride HB20111 (Trichoderma atroviride) The microbial inoculum is preserved in the China general microbiological culture collection management center for 12 and 5 days in 2018, the preservation address is No. 3 of Xilu No.1 of Beijing, Chaoyang, and the preservation number is CGMCC No. 16963.
2. The use of the trichoderma atroviride microsclerotia preparation as claimed in claim 1 for controlling wheat stem basal rot, wherein in the step (2), the conidium inoculation amount is 10 per 100mL of induction medium6And (4) spores.
3. The application of the trichoderma atroviride microsclerotia preparation in the control of wheat stem basal rot is described in claim 1, wherein in the step (3), the filter aid is diatomite and is added in an amount of 1% -5% (w/v).
4. The use of the trichoderma atroviride microsclerotia preparation as claimed in claim 1 for preventing and treating wheat stem basal rot, wherein the carrier is one or a combination of more of diatomite, medical stone, kaolin, starch and corn husk powder; the mass percentage of the microsclerotia after suction filtration to the carrier is 1: (10-20).
5. The application of the trichoderma atroviride microsclerotia preparation in preventing and treating wheat stem basal rot is disclosed in claim 1, and is characterized in that the auxiliary agent is one or a combination of more of sodium carboxymethylcellulose, xanthan gum and sodium alginate, and the mass percentage of the dried microsclerotia-carrier mixture to the auxiliary agent is 1000: 1-200: 1.
6. The application of the trichoderma atroviride microsclerotium preparation in the control of wheat stem basal rot is disclosed in claim 1, wherein the low-temperature drying adopts a fluidized bed for fluidized drying, the drying temperature is 35-40 ℃, the drying is carried out until the moisture content is below 5%, and then the drying is carried out to obtain bagged products.
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| CN104357338A (en) * | 2014-11-21 | 2015-02-18 | 重庆大学 | Fermentation method and applications of paecilomyce lilacinus microsclerotia |
| CN107109343A (en) * | 2014-09-18 | 2017-08-29 | (由农业部部长代表的)美利坚合众国 | Trichoderma composition and application method |
| CN110915822A (en) * | 2019-11-25 | 2020-03-27 | 山东省科学院生态研究所(山东省科学院中日友好生物技术研究中心) | Application of trichoderma atroviride HB20111 in prevention and treatment of wheat stem basal rot and sheath blight |
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Patent Citations (3)
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| CN107109343A (en) * | 2014-09-18 | 2017-08-29 | (由农业部部长代表的)美利坚合众国 | Trichoderma composition and application method |
| CN104357338A (en) * | 2014-11-21 | 2015-02-18 | 重庆大学 | Fermentation method and applications of paecilomyce lilacinus microsclerotia |
| CN110915822A (en) * | 2019-11-25 | 2020-03-27 | 山东省科学院生态研究所(山东省科学院中日友好生物技术研究中心) | Application of trichoderma atroviride HB20111 in prevention and treatment of wheat stem basal rot and sheath blight |
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| Liquid Culture Production of Fungal Microsclerotia;Mark A Jackson等;《Methods Mol Biol》;20161231;第1477卷;全文 * |
| Liquid culture production of microsclerotia and submerged conidia by Trichoderma harzianum active against damping-off disease caused by Rhizoctonia solani;Nilce N. KOBORI等;《Fungal Biol》;20141219;第119卷(第4期);全文 * |
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