CN114304190A - Preparation method of trichoderma wettable powder - Google Patents
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Abstract
The invention provides a preparation method of trichoderma wettable powder, which comprises the following steps: the preparation method comprises the steps of S1 conidium suspension preparation, S2 fermentation matrix preparation, S3 conidium powder preparation and S4 wettable powder preparation, wherein the composition proportion of the wettable powder is determined to be 20% of the conidium powder, AB-31% of the conidium powder, NNO 5% of the dispersant and 1% of ascorbic acid, and the wetting time, the mass suspension rate and the like of the wettable powder are determined to meet the relevant standards of the wettable powder for pesticides, so that the wettable powder has important significance for the development and industrial production of T.brev T069 products, and the composition of the wettable powder for the strains is determined, thereby laying a foundation for the production development and field application and pre-evaluation of the T.brev T069.
Description
Technical Field
The invention relates to the technical field of microbial fermentation, in particular to a preparation method of trichoderma wettable powder.
Background
In traditional agriculture of China, continuous cropping phenomena are very common, which can cause serious continuous croppingObstacles, namely reduction of soil nutrients, accumulation of a large amount of pathogenic bacteria, annual increase of morbidity and disease severity, and serious influence on the yield and quality of crops. In order to better control diseases, people often use a great amount of pesticides and fertilizers, but the effect is not obvious. And the long-term use of a large amount of chemical agents can seriously affect the ecological environment and the human health[1]Moreover, pathogenic bacteria are easy to generate drug resistance, and the difficulty of preventing and controlling diseases is increased. Biological control is an environment-friendly disease control method capable of replacing chemical pesticides, and screening high-efficiency biocontrol strains and preparing the high-efficiency biocontrol strains into high-efficiency biocontrol preparations have an important effect on promoting green development of agriculture.
Trichoderma spp is a biocontrol fungus with wider application, and can inhibit 29 plant pathogenic fungi of 18 genera such as Botrytis spp, Fusarium spp and Rhizoctonia spp to different degrees through various mechanisms such as competition, antagonism and parasitism [3-4 ]. In addition, trichoderma can also induce the resistance of plants by releasing effector factors or secreting hydrolytic enzymes, and is widely applied to the aspects of seed coating treatment, soil improvement, foliage spraying and the like [5-6 ]. At present, how to rapidly produce a large amount of propagules and prepare a stable and reliable preparation is the research focus of further popularization and application of trichoderma biocontrol.
Before the 21 st century, the application of three trichoderma propagules, namely mycelium, conidium and chlamydospore, in biocontrol agents has been studied for a long time. The mycelium has poor stress resistance and low survival rate, and chlamydospores have strong storage capacity, but harsh culture conditions and long growth period. In contrast, conidia are fast and simple to produce, and have good stress resistance to the environment, and are the main types of the current commercial trichoderma preparations.
Disclosure of Invention
Therefore, the invention provides a preparation method of trichoderma wettable powder, which solves the problems.
The technical scheme of the invention is realized as follows: a preparation method of trichoderma wettable powder comprises the following steps:
s1, preparation of conidium suspension: inoculating the strain T.brev T069 to a PDA culture medium for activated culture, culturing at the constant temperature of 20-30 ℃ for 7-11 days to generate a large amount of green spores, washing the spores with sterile water, and diluting to 0.8-1.2 multiplied by 108CFU/mL seed liquid is reserved;
s2, preparation of a fermentation substrate: collecting the cassava peels after girdling, cutting the cassava peels into small pieces, drying, crushing, sieving by using cassava peel residues of 10-100 meshes, uniformly mixing, weighing the cassava peel residues, dissolving the cassava peel residues in water, and filling the cassava peel residues into a tissue culture bottle for sterilization to obtain a fermentation substrate;
s3, preparation of conidium powder: adding the T.brev T069 spore suspension into the tissue culture bottle according to the inoculation amount of 0.3-1.2% of the volume percentage of the fermentation substrate, shaking uniformly, statically culturing in a constant-temperature incubator at 25-30 ℃ for 2-4 days, taking out and shaking uniformly to break hyphae once in 1-2 days, then drying the cassava peel residue fermented by the T.brev T069 in the air, crushing for 3-8 s by using a crusher, crushing for 1-3 times at intervals of 3-7 min, and then sieving by using a 300-400-mesh sieve to obtain T.brev T069 conidium powder;
s4, preparation of wettable powder: mixing the spore powder with a wetting agent, a dispersing agent, an ultraviolet protective agent and a carrier to prepare wettable powder, wherein the wettable powder comprises the following raw materials in percentage by mass: 12-25% of spore powder, 0.9-1.3% of wetting agent, 3-8% of dispersing agent, 0.8-1.2% of ultraviolet protective agent and the balance of carrier.
Further, preparation of the S1 conidium suspension: inoculating strain T.brev T069 to PDA culture medium, activating, culturing at 28 deg.C for 9 days to obtain a large amount of green spores, washing with sterile water, and diluting to 1 × 108CFU/mL seed solution is ready for use.
Further, the preparation of the S2 fermentation substrate: the mass-volume ratio g/mL of the cassava skin residues to the water is 5-9: 10.
further, the S3 conidium powder is prepared by adding a T.brev T069 spore suspension into a tissue culture bottle according to the inoculation amount of 1%, shaking uniformly, standing and culturing in a constant-temperature incubator at 28 ℃ for 3d, taking out and shaking uniformly to break hyphae once in the 2d, then drying cassava peel residues fermented by the T.brev T069, crushing for 5S by a crusher, crushing for 2 times at the crushing interval of 5min, and then sieving with a 325-mesh sieve to obtain the T.brev T069 conidium powder.
Further, the wettable powder of S3 comprises the following raw materials by mass percent: 20% of spore powder, 1% of wetting agent, 5% of dispersing agent, 1% of ultraviolet protective agent and the balance of carrier.
Further, the carrier is one of white carbon black, diatomite, kaolin, talcum powder and cassava peel powder, and the concentration of the carrier is 1000-5000 mug/ml.
Further, the wetting agent is one of sodium dodecyl benzene sulfonate, sodium dodecyl sulfate, nekal BX and AB-3.
Further, the dispersant is one of sodium carboxymethylcellulose, sodium lignosulfonate and sodium methylene dinaphthalene sulfonate.
Further, the ultraviolet protective agent is one of dextrin, ascorbic acid and fluorescein sodium, and the concentration of the ultraviolet protective agent is 100-1000 mu g/ml.
Compared with the prior art, the invention has the beneficial effects that:
according to the invention, a biocontrol T.brev T069 strain with strong bacteriostasis, high growth speed and high spore yield is screened at the early stage, cassava peel residue is taken as a fermentation substrate, conidium powder of the T.brev T069 is produced by adopting a solid fermentation technology, the composition proportions of the wettable powder, namely the spore powder are 20 percent, AB-31 percent, the dispersing agent NNO5 percent and the ascorbic acid 1 percent, are determined by measuring the performance indexes of different types and proportions of the auxiliary agents and the biocompatibility between the auxiliary agents and the T.brev T069, and the wetting time, the mass suspension rate and other aspects of the wettable powder are determined to be in accordance with the relevant standards of the pesticide wettable powder.
The invention provides a preparation method of trichoderma wettable powder and a novel wettable powder formula, which have important significance on the development and industrial production of T.brev T069 products, determine the composition of the strain wettable powder and lay a foundation for the production development, field application and pre-evaluation of biocontrol T.brev T069.
Drawings
Fig. 1 is a t.brev T069 fermentation process using cassava peel residue, in which a: dried cassava peel, B: crushed cassava peel, C: sterilized cassava peels, D: fermentation 2d, E: fermentation 3 d;
figure 2 is the effect of different vectors on t.brev T069 hyphal growth (36 h);
FIG. 3 is a graph showing the effect of varying concentrations of humectant on T.brev T069 hyphal growth (36 h);
figure 4 is the effect of different concentrations of dispersant on t.brev T069 hyphal growth (36 h);
figure 5 is the effect of different concentrations of uv protectant on t.brev T069 hyphal growth (24 h).
Detailed Description
In order to better understand the technical content of the invention, specific examples are provided below to further illustrate the invention.
The experimental methods used in the examples of the present invention are all conventional methods unless otherwise specified.
The materials, reagents and the like used in the examples of the present invention can be obtained commercially without specific description.
The test strain T.brev T069 is provided by the research center of Green agricultural biological preparation creation engineering (plant protection institute of Hainan university) in Hainan province.
Example 1
A preparation method of trichoderma wettable powder comprises the following steps:
s1, preparation of conidium suspension: inoculating strain T.brev T069 to PDA culture medium, activating, culturing at 20 deg.C for 7 days to obtain a large amount of green spores, washing with sterile water, and diluting to 0.8 × 108CFU/mL seed liquid is reserved;
s2, preparation of a fermentation substrate: collecting the peeled cassava skin, cutting into small pieces, drying, pulverizing, sieving with 10 mesh cassava skin residue, mixing, weighing the cassava skin residue, dissolving in water, and sterilizing in a tissue culture bottle to obtain a fermentation substrate;
s3, preparation of conidium powder: adding the T.brev T069 spore suspension into the tissue culture bottle according to the inoculation amount of 0.3% of the volume percentage of the fermentation substrate, shaking uniformly, performing static culture in a constant-temperature incubator at 25 ℃ for 2d, taking out and shaking uniformly to break hyphae once in the 1 st d, then drying cassava peel residues fermented by the T.brev T069 in the air, crushing for 3s by a crusher, crushing for 1 time at the crushing interval of 3min, and then sieving by a 300-mesh sieve to obtain T.brev T069 conidia powder;
s4, preparation of wettable powder: mixing the spore powder with a wetting agent, a dispersing agent, an ultraviolet protective agent and a carrier to prepare wettable powder, wherein the wettable powder comprises the following raw materials in percentage by mass: 12 percent of spore powder, 30.9 percent of wetting agent AB-30, 0.8 percent of dispersant NNO3 percent of ultraviolet protective agent ascorbic acid, and the balance of diatomite, wherein the concentration of the diatomite is 1000 mu g/ml, and the concentration of the ultraviolet protective agent ascorbic acid is 100 mu g/ml.
Example 2
A preparation method of trichoderma wettable powder comprises the following steps:
s1, preparation of conidium suspension: inoculating strain T.brev T069 to PDA culture medium, activating, culturing at 30 deg.C for 11 days to obtain a large amount of green spores, washing with sterile water, and diluting to 1.2 × 108CFU/mL seed liquid is reserved;
s2, preparation of a fermentation substrate: collecting the cassava peels after girdling, cutting the cassava peels into small pieces, drying, crushing, sieving by using cassava peel residues of 10-100 meshes, uniformly mixing, weighing the cassava peel residues, dissolving the cassava peel residues in water, and filling the cassava peel residues into a tissue culture bottle for sterilization to obtain a fermentation substrate;
s3, preparation of conidium powder: adding the T.brev T069 spore suspension into the tissue culture bottle according to the inoculation amount of 1.2% of the volume percentage of the fermentation substrate, shaking uniformly, performing static culture in a constant-temperature incubator at 30 ℃ for 4d, taking out and shaking uniformly to break hyphae once in the 2d, then drying cassava peel residues fermented by the T.brev T069 in the air, crushing for 8s by a crusher, crushing for 3 times at the crushing interval of 7min, and then sieving by a 400-mesh sieve to obtain T.brev T069 conidia powder;
s4, preparation of wettable powder: mixing the spore powder with a wetting agent, a dispersing agent, an ultraviolet protective agent and a carrier to prepare wettable powder, wherein the wettable powder comprises the following raw materials in percentage by mass: 25% of spore powder, 31.3% of wetting agent AB-3%, 8% of dispersant NNO, 1.2% of ultraviolet protective agent ascorbic acid, the balance of diatomite, the concentration of the diatomite is 5000 mug/ml, and the concentration of the ultraviolet protective agent ascorbic acid is 1000 mug/ml.
Example 3
A preparation method of trichoderma wettable powder comprises the following steps:
s1, preparation of conidium suspension: inoculating strain T.brev T069 to PDA culture medium, activating, culturing at 28 deg.C for 9 days to obtain a large amount of green spores, washing with sterile water, and diluting to 1.0 × 108CFU/mL seed liquid is reserved;
s2, preparation of a fermentation substrate: collecting the cassava peels after girdling, cutting the cassava peels into small pieces, drying, crushing, sieving by using cassava peel residues of 10-100 meshes, uniformly mixing, weighing the cassava peel residues, dissolving the cassava peel residues in water, and filling the cassava peel residues into a tissue culture bottle for sterilization to obtain a fermentation substrate;
s3, preparation of conidium powder: adding the T.brev T069 spore suspension into the tissue culture bottle according to the inoculation amount of 1% of the fermentation substrate by volume percent, shaking uniformly, performing static culture in a constant-temperature incubator at 28 ℃ for 3d, taking out and shaking uniformly to break hyphae once in the 2d, then drying cassava peel residues fermented by the T.brev T069 in the air, crushing for 5s by a crusher, crushing for 2 times at intervals of 5min, and then sieving by a 325-mesh sieve to obtain T.brev T069 conidium powder;
s4, preparation of wettable powder: mixing the spore powder with a wetting agent, a dispersing agent, an ultraviolet protective agent and a carrier to prepare wettable powder, wherein the wettable powder comprises the following raw materials in percentage by mass: 20% of spore powder, 31% of wetting agent AB-31%, 5% of dispersant NNO, 1% of ultraviolet protective agent ascorbic acid, and the balance of diatomite, wherein the concentration of the diatomite is 5000 mug/ml, and the concentration of the ultraviolet protective agent ascorbic acid is 100 mug/ml.
Biocompatibility determination of one, different vectors
After sieving diatomite, kaolin, talcum powder, white carbon black and cassava peel powder by a 325-mesh sieve, respectively adding the sieved powder into 100mLPDA according to the concentration of 1000, 2500 and 5000 mu g/mL, punching a 5mm bacterial cake by using a sterile puncher and inoculating the bacterial cake to the center of a PDA (personal digital assistant) plate containing a carrier, and verifying the influence of the carrier on the colony diameter of T.brev T069. Preparing spore germination liquid (20 g of sucrose, 5g of soybean peptone, 5g of potassium dihydrogen phosphate and 1000mL of water), subpackaging 45mL of the spore germination liquid into a conical flask with the capacity of 250mL, adding a plurality of carriers according to the concentration, sterilizing at 121 ℃ for 20min, and cooling for later use. 5mL of the solution was aspirated at a concentration of 1X 107Adding CFU/mL T.brev spore suspension into spore germination solution containing different carriers, culturing at 28 deg.C for 12h at 120r/min, measuring germination number (the standard of spore germination is that the visible tube length is equal to or greater than the spore diameter) with a blood counting plate under microscope, and repeating for 5 times. The effect of different concentrations of vector on spore germination and hypha growth of t.brev T069 is shown in table 1,
table 1 biocompatibility determination of different vectors with t.brev T069 (P <0.05)
Different letters indicate significant difference (P <0.05)
As can be seen from table 1 and fig. 2, in the culture medium with different concentrations of white carbon black, the spore germination rate and hypha colony diameter of t.brev T069 are significantly lower than those of the control (P < 0.05). In diatomite culture media with different concentrations, the spore germination rate of T.brev T069 is lower than that of a control, and the difference is not significant when the concentration is 2500 mug/mL; the effect on hyphal growth also differed between 24 and 36h, with the colony diameter being greatest at a concentration of 5000. mu.g/mL, significantly higher than the control. In kaolin culture media with different concentrations, the spore germination rate of T.brev T069 is lower than that of a control; at concentrations of 1000 and 5000. mu.g/mL, the colony diameters were significantly higher than the control. In culture media with different concentrations of talcum powder, the spore germination rate of T.brev T069 is lower than that of a control; the colony diameter at 5000. mu.g/mL concentration for 36h was significantly lower than the control. In cassava peel powder culture media with different concentrations, the difference between the spore germination rate and the control is not obvious; the colony diameters at 36h were all significantly lower than the control. In comprehensive comparison, the influence of the diatomite on the growth of hyphae is optimal.
Second, screening of wetting agent and determination of biocompatibility
For measuring the wetting performance, referring to GB/T5451-2001, the four wetting agents of nekal BX, Sodium Dodecyl Sulfate (SDS), Sodium Dodecyl Benzene Sulfonate (SDBS) and AB-3 are respectively added into diatomite according to the proportion of 1%, 3% and 5% (w/w) and the proportion of 20% of spore powder, and the mixture is uniformly mixed. Thereafter 5.0g of the sample was weighed and added to the surface of a 150mL beaker containing 100mL of standard water and the time required for complete wetting of the sample was recorded and repeated 3 times with the sample without wetting agent as a control and the results are given in table 2.
TABLE 2 wetting Performance and biocompatibility measurements of different wetting agents
As can be seen from Table 2, there are more significant differences in the wetting properties of the four wetting agents at three different concentrations, 1%, 3% and 5%. Among them, 5% Sodium Dodecyl Sulfate (SDS) was the most excellent wetting property, and the wetting time was 3.83. + -. 0.22s, but biocompatibility to T.brev T069 was poor (FIG. 3). In addition, when Sodium Dodecyl Benzene Sulfonate (SDBS) with different concentrations is used as a wetting agent, the wetting time is longer; spores did not germinate at different concentrations of BX as a humectant. Therefore, none of the three substances can be used as wetting agent AB-3, the wetting time is 6.52-10.08s at three concentrations, only after SDS, at the same time, the spore germination rate of T.brev T069 is more than 80% at 1% concentration, the colony diameter is 6.80cm at 36h, and 1% of AB-3 is selected as the wetting agent for comprehensive evaluation.
Thirdly, measuring the suspension characteristic and biocompatibility of the dispersing agent
The method comprises the following steps of respectively adding 1%, 3% and 5% (w/w) of dispersing agents including sodium methylenedinaphthalenesulfonate NNO, sodium lignosulfonate and sodium carboxymethylcellulose (CMC-Na) into spore powder 20%, and supplementing the spore powder with diatomite to 100%. 2.5g of sample was weighed, and the suspension characteristics of the 3 dispersed suspending agents were determined according to the method of GB/T14825-2006, which was repeated 3 times, and the results are shown in Table 3.
TABLE 3 suspension characteristics and biocompatibility measurements with different concentrations of dispersant
As can be seen from table 3, the suspension percentage was significantly improved compared to the control (suspension percentage 42.45%) without the dispersant after the addition of 3 different concentrations of the dispersant, and the suspension percentage was improved with the increase in concentration in the same dispersant. The suspension property is preferably CMC-Na, and the suspension rate can reach 82.03% at 5%, which is obviously higher than that of other concentrations of suspending agents. However, the reagent is viscous and has a dissolution time in water of more than 10 min. When sodium lignosulfonate with different concentrations is used as a suspending agent, the hypha diameter is 7.00-7.33 cm at 36h, the effect is the best, but the suspension rate is only 55.81-60.44%, and the performance is the worst. When the NNO is at the concentration of 5%, the suspension rate can reach 73.04%, which is next to the CMC-Na with the concentration of 5%; although the hypha diameter was 6.87cm at 36h, which is slightly smaller than the control, the difference was not significant. Therefore, a compromise was made to select 5% NNO as the dispersant.
Fourth, measuring the protection characteristic and biocompatibility of the ultraviolet protective agent
Adding fluorescein sodium, dextrin and ascorbic acid at concentrations of 100, 500 and 1000 μ g/mL to 106CFU/mL in t.brev spore suspension. And (3) dripping 100 mu L of the mixture on a concave glass, irradiating for 10min at a position of 40cm under an ultraviolet lamp (30W), culturing at 28 ℃ in the dark for 12h, determining the germination rate of the spores, and repeating for 5 times. The results are shown in Table 4 below, using control 1, which was prepared by mixing with sterile water and then irradiating the mixture without irradiation, and using control 2, which was prepared by mixing with sterile water and then irradiating the mixture with irradiation.
TABLE 4 determination of UV protectant Properties and biocompatibility at various concentrations
As can be seen from Table 4, the difference in spore germination rates before and after UV irradiation was significant, and the germination rate after irradiation was only 5.4%. Of the three ultraviolet protective agents, ascorbic acid with the best protective performance of 100 mu g/mL has the spore germination rate of 85.6 percent after ultraviolet irradiation for 10min, and is obviously higher than that of other ultraviolet protective agents with other concentrations and types. Meanwhile, in the determination of the spore germination of the biocompatibility, the spore germination rate of T069 is 93.4 +/-1.82%, and the ascorbic acid has no significant difference with CK; the diameter of the hypha is 5.0cm, which is obviously higher than that of a control, and the hypha also has a certain promotion effect on the growth of the hypha. Thus, 100. mu.g/mL ascorbic acid was chosen as the UV protectant.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (9)
1. A preparation method of trichoderma wettable powder is characterized by comprising the following steps: the method comprises the following steps:
s1, preparation of conidium suspension: inoculating the strain T.brev T069 to a PDA culture medium for activated culture, culturing at the constant temperature of 20-30 ℃ for 7-11 days to generate green spores, washing the spores with sterile water, and diluting to 0.8-1.2 × 108CFU/mL seed liquid is reserved;
s2, preparation of a fermentation substrate: collecting the cassava peels after girdling, cutting the cassava peels into small pieces, drying, crushing, sieving by using cassava peel residues of 10-100 meshes, uniformly mixing, weighing the cassava peel residues, dissolving the cassava peel residues in water, and filling the cassava peel residues into a tissue culture bottle for sterilization to obtain a fermentation substrate;
s3, preparation of conidium powder: adding the T.brev T069 spore suspension into the tissue culture bottle according to the inoculation amount of 0.3-1.2% by volume of a fermentation substrate, shaking uniformly, performing static culture in a constant-temperature incubator at 25-30 ℃ for 2-4 days, taking out and shaking uniformly for 1-2 days to break hyphae once, then drying cassava peel residues fermented by the T.brev T069 in the air, crushing for 3-8 s by using a crusher for 1-3 times at a crushing interval of 3-7 min, and then sieving by using a 300-400-mesh sieve to obtain T.brev T069 conidium powder;
s4, preparation of wettable powder: mixing the spore powder with a wetting agent, a dispersing agent, an ultraviolet protective agent and a carrier to prepare wettable powder, wherein the wettable powder comprises the following raw materials in percentage by mass: 12-25% of spore powder, 0.9-1.3% of wetting agent, 3-8% of dispersing agent, 0.8-1.2% of ultraviolet protective agent and the balance of carrier.
2. The method for preparing a wettable powder of trichoderma as claimed in claim 1, wherein: preparation of the S1 conidia suspension: inoculating strain T.brevt069 to PDA culture medium, activating, culturing at 28 deg.C for 9 days to obtain a large amount of green spores, washing with sterile water, and diluting to 1 × 108CFU/mL seed solution is ready for use.
3. The method for preparing a wettable powder of trichoderma as claimed in claim 1, wherein: preparation of the S2 fermentation substrate: the mass-volume ratio g/mL of the cassava skin residues to the water is 5-9: 10.
4. the method for preparing a wettable powder of trichoderma as claimed in claim 1, wherein: the preparation method of the S3 conidium powder comprises the steps of adding a T.brev T069 spore suspension into a tissue culture bottle according to the inoculation amount of 1%, shaking uniformly, carrying out standing culture in a constant-temperature incubator at 28 ℃ for 3d, taking out the spore suspension and shaking uniformly to break hyphae once in the 2 nd day, then airing cassava peel residues fermented by the T.brev T069, crushing for 5S by a crusher, crushing for 2 times at the crushing interval of 5min, and then sieving by a 325-mesh sieve to obtain the T.brev T069 conidium powder.
5. The method for preparing a wettable powder of trichoderma as claimed in claim 1, wherein: the wettable powder of S3 comprises the following raw materials in percentage by mass: 20% of spore powder, 1% of wetting agent, 5% of dispersing agent, 1% of ultraviolet protective agent and the balance of carrier.
6. A process for the preparation of wettable powders of trichoderma as claimed in claim 1 or 5, wherein: the carrier is one of white carbon black, diatomite, kaolin, talcum powder and cassava peel powder, and the concentration of the carrier is 1000-5000 mu g/ml.
7. A process for the preparation of wettable powders of trichoderma as claimed in claim 1 or 5, wherein: the wetting agent is one of sodium dodecyl benzene sulfonate, sodium dodecyl sulfate, nekal BX and AB-3.
8. A process for the preparation of wettable powders of trichoderma as claimed in claim 1 or 5, wherein: the dispersant is one of sodium carboxymethylcellulose, sodium lignosulfonate and sodium methylene dinaphthalene sulfonate.
9. A process for the preparation of wettable powders of trichoderma as claimed in claim 1 or 5, wherein: the ultraviolet protective agent is one of dextrin, ascorbic acid and fluorescein sodium, and the concentration of the ultraviolet protective agent is 100-1000 mu g/ml.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113832063A (en) * | 2021-09-29 | 2021-12-24 | 海南大学 | Preparation method and application of streptomyces coprostanus BWL-H1 microbial inoculum |
| CN113832063B (en) * | 2021-09-29 | 2023-11-24 | 海南大学 | Preparation method and application of Streptomyces faecalis BWL-H1 microbial inoculum |
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