CN111411047A - Preparation method of dark-color endophytic fungus dry powder microbial inoculum - Google Patents
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Abstract
The invention discloses a preparation method of a dark-color septate endophytic fungus dry powder microbial inoculum. The invention discloses a preparation method of a dark color endophytic fungus dry powder microbial inoculum, which comprises the following steps: drying the dark color endophytic fungi-isolating needle A2-1 at 60 deg.C to obtain the dried dark color endophytic fungi-isolating agent. The method for preparing the microbial inoculum is simple and quick and can be finished within 42 hours; no obvious influence on the activity of bacteria; the obtained microbial inoculum can be stored for a long time, still has good activity after being stored for 20 days at normal temperature, and is easy to store; compared with the traditional liquid microbial inoculum, the obtained microbial inoculum also effectively overcomes the problems of inconvenient transportation, easy pollution and the like; the utilization efficiency of the microbial inoculum obtained by the method can be improved. The method disclosed by the invention can be used for producing the A2-1 microbial inoculum in batches, plays an important role in further popularization and application of the microbial inoculum, and has a good application prospect.
Description
Technical Field
The invention relates to a preparation method of a dark-color endophytic fungus dry powder microbial inoculum, belonging to the field of microorganisms.
Background
Dark septate endophytic fungi (DSE) generally refer to a group of small soil endophytic fungi that colonize plant roots and can form typical structures such as septate hyphae and microsclerotia, but are not pathogenic to hosts.
Research shows that the deep color epiphyte with septa can promote plant growth, raise plant stress resistance and improve soil physical and chemical properties. Chinese patent application 201910505641.5 discloses that deep color intraseptal fungus needle A2-1 (the bacterial strain with the China Committee for culture Collection of microorganisms and having the preservation number of CGMCC No. 17465) can promote the growth of roots. With the gradually revealed ecological action of the dark-colored septate endophytic fungi, the research and development of the dark-colored septate endophytic fungi microbial inoculum also gradually become a hotspot and difficulty of research.
At present, the dark fungus bacterium agent with internal fungus is activated for 2 times, each time lasts for 7-10 days, the preparation process is slow, and the transfer process is easy to pollute and contaminate mixed bacteria. In addition, as the application of dark-color entomogenous fungi gradually moves from indoor to outdoor, and the liquid microbial inoculum in a laboratory scale has the problems of unstable performance, inconvenient transportation and the like, so that a method for preparing the solid microbial inoculum which is easy to transport, store and produce in a quantitative mode is urgently needed to be explored.
Disclosure of Invention
The invention aims to provide a preparation method of a dark color septate endophytic fungus dry powder microbial inoculum, and particularly relates to a preparation method of a needle A2-1 dry powder microbial inoculum.
The invention provides a preparation method of a needle A2-1 dry powder microbial inoculum, which comprises the following steps: drying the needle A2-1 at 60 deg.C to obtain dried fungus A2-1; the needle A2-1 is a bacterial strain with the preservation number of CGMCC No.17465 in the China Committee for culture Collection of microorganisms.
The drying time may be 42 hours or less.
In the above method, the moisture content of the dried fungus may be 2% to 3%, such as 3%.
In the above method, the drying of needle a2-1 at 60 ℃ may comprise: drying the mycelium of needle A2-1 at 60 deg.C.
The method also comprises pulverizing dried fungus to obtain needle A2-1.
The method also comprises sieving the pulverized fungus with a sieve with a diameter of 0.5mm or less to obtain a powdery substance with a diameter of 0.5mm or less, namely the A2-1 microbial inoculum.
The application of the method in preparing products for promoting plant growth or improving plant stress resistance or improving soil physicochemical property also belongs to the protection scope of the invention.
According to the preparation method of the needle A2-1 dry powder microbial inoculum, disclosed by the invention, the needle A2-1 dry powder microbial inoculum is obtained by drying the needle A2-1 hyphae at 60 ℃ until the water content is 3%, and the method has the following advantages: the preparation of the microbial inoculum is simple and rapid and can be finished within 42 hours; the activity of the bacteria is not obviously influenced, and the activity of the bacteria is not obviously different from that before drying; the obtained microbial inoculum can be stored for a long time, still has good activity after being stored for 20 days at normal temperature, and is easy to store; compared with the traditional liquid microbial inoculum, the obtained microbial inoculum also effectively overcomes the problems of inconvenient transportation, easy pollution and the like; the grain size of the microbial inoculum obtained by the method is less than or equal to 0.5mm, and the utilization efficiency of the microbial inoculum is further improved. The method disclosed by the invention can be used for producing the A2-1 microbial inoculum in batches, plays an important role in further popularization and use of the microbial inoculum, lays an important foundation for sustainable development of land, and has a good application prospect.
Biological material preservation instructions
Classification nomenclature of biological materials: darksidea sp.
Strain number of biological material: needle A2-1
Deposit name of biological material: china general microbiological culture Collection center
The preservation unit of the biological material is abbreviated as: CGMCC (China general microbiological culture Collection center)
Deposit unit address of biological material: west road No.1, north west of the township, beijing, ministry of sciences, china, institute of microbiology, zip code: 100101
Preservation date of biological material: 4 and 8 months in 2019
Accession number to the collection of biological materials: CGMCC No.17465
Biological material preservation instructions
Classification nomenclature of biological materials: darksidea sp.
Strain number of biological material: needle A2-7
Deposit name of biological material: china general microbiological culture Collection center
The preservation unit of the biological material is abbreviated as: CGMCC (China general microbiological culture Collection center)
Deposit unit address of biological material: west road No.1, north west of the township, beijing, ministry of sciences, china, institute of microbiology, zip code: 100101
Preservation date of biological material: 11/19/2019
Accession number to the collection of biological materials: CGMCC No.18811
Biological material preservation instructions
Classification nomenclature of biological materials: geospora sp (Pleosporales sp.)
Strain number of biological material: needle A2-8
Deposit name of biological material: china general microbiological culture Collection center
The preservation unit of the biological material is abbreviated as: CGMCC (China general microbiological culture Collection center)
Deposit unit address of biological material: west road No.1, north west of the township, beijing, ministry of sciences, china, institute of microbiology, zip code: 100101
Preservation date of biological material: 11/19/2019
Accession number to the collection of biological materials: CGMCC No.18812
Drawings
FIG. 1 is a photograph showing the configuration of a needle A2-1.
FIG. 2 is a photograph showing the configuration of the needle A2-7.
FIG. 3 is a photograph showing the configuration of the needle A2-8.
FIG. 4 shows the effect of the size of the dry powder on the growth of the microbial inoculum.
FIG. 5 shows the effect of drying temperature on the growth of microbial inoculum. Under the same culture time, the dry powder microbial inoculum prepared at the temperature of 30, 40, 50, 60, 70 and 80 ℃ is sequentially used from left to right as a wet microbial inoculum control.
Detailed Description
The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention. The experimental procedures in the following examples are conventional unless otherwise specified. Materials, reagents, instruments and the like used in the following examples are commercially available unless otherwise specified. The quantitative tests in the following examples, all set up three replicates and the results averaged. In the following examples, unless otherwise specified, the 1 st position of each nucleotide sequence in the sequence listing is the 5 'terminal nucleotide of the corresponding DNA/RNA, and the last position is the 3' terminal nucleotide of the corresponding DNA/RNA.
In the examples, the operations were all performed in a sterile operating table, and before the test operation, the operating table was irradiated with ultraviolet light for 20 min. All reagents and devices in the examples are sterile reagents and devices.
Liquid MMN medium (ph 5.5): adding CaCl20.05g、MgSO40.15g、NaCl 0.025g、FeCl30.01g、KH2PO40.5g, 0.0001g of Vitamin B1 (thiamine), (NH)4)2HPO4Mixing 0.25g, glucose 10g, citric acid 0.2g and Malt extract 10g to obtain mixture, adding distilled water to a constant volume of 1L, packaging, and autoclaving at 121 deg.C for 15 min.
Solid PDA culture medium is prepared by collecting peeled potato 200g, cutting into small pieces, adding 1L distilled water, boiling for 30min, filtering with gauze, collecting filtrate, adding glucose 20.0g and KH into the filtrate2PO43.0g、MgSO47H2O1.5 g, vitamin B110 μ g and agar 15.0g, adjusting pH to 6.0, diluting with distilled water to 1.0L, and sterilizing at 115 deg.C for 15 min.
Example 1 preparation of bacterial solution
The strains are needle A2-1, needle A2-7 and needle A2-8, which are separated and purified from the root system of needle cogongrass in the North electric winning area of the union of Silian Haote, inner Mongolia autonomous region.
The ITS sequence of the detection needle A2-1 is the sequence 1 in the sequence table, and is identified as deep color septate endophytic fungus (DSE) (Darksidea sp.), the strain is preserved in China general microbiological culture Collection center (CGMCC for short, with the address of No. 3 Xilu 1. Beicheng Kogyang area in Beijing) in 2019 and 8 months, and the preservation number is CGMCC No. 17465. The dark color has morphological characteristics of endophytic fungi: black or dark brown, the hyphae are dark in the root system and have obvious transverse septa. A photograph of the morphology of needle A2-1 in the root system is shown in FIG. 1.
The ITS sequence of probe A2-7, shown as sequence 2 in the sequence listing, was identified as a Dark Septate Endophyte (DSE) (Darksidea sp.). The needle A2-7 has been preserved in China general microbiological culture Collection center (CGMCC) No.18811 in 2019 at 19.11. The dark color has morphological characteristics of endophytic fungi: black or dark brown, the hyphae are dark in the root system and have obvious transverse septa. A photograph of the morphology of needle A2-7 in the root system is shown in FIG. 2.
The ITS sequence of the detection needle A2-8 is identified as the dark-color endophytic fungus (DSE) Geobacillus (Pleosporales sp.) as shown in the sequence 3 in the sequence table. The needle A2-8 has been preserved in China general microbiological culture Collection center (CGMCC) No.18812 in 2019, 11 and 19 months. Morphological characteristics of G.grisea (Pleosporalessp.): black or dark brown, the hyphae are dark in the root system and have obvious transverse septa. A photograph of the morphology of needle A2-8 within the root system is shown in FIG. 3.
Each strain is prepared into a bacterial liquid according to the following steps:
1. cultivation of the Strain
Activating the strain, namely picking up hyphae at the edge of the strain by using a sterile inoculating needle in an operation table, and inoculating the hyphae to a new solid PDA culture medium for culture. After new bacteria grow out, picking out edge hyphae again according to the characteristics of the color, the shape and the like of the bacterial colony, and inoculating the hyphae to a new solid PDA culture medium for culture. The steps are repeated until the colonies in the culture medium are uniform and consistent.
2. Preparation of bacterial liquid
After completion of step 1, 5mm × 5mm cake was punched out with a sterile punch, and 2 cakes were placed in a 250m L conical flask containing 150m L liquid MMN medium and cultured under shaking at 27 ℃ and 170r/min in the dark for 24 flasks co-culture.
And (3) respectively obtaining a needle A2-1 bacterial liquid, a needle A2-7 bacterial liquid and a needle A2-8 bacterial liquid after the step 2 is finished.
Example 2 influence of Water content on microbial inoculum
The bacterial liquid used was the needle a2-1 bacterial liquid of example 1, and the bacterial liquids were prepared by the following procedure:
filtering the bacterial liquid under a single-layer gauze, collecting fresh hyphae and mixing uniformly. 15g of fresh hyphae are dried at 120 ℃ for 12 hours, and the water content is calculated. And (3) randomly dividing the rest fresh hyphae into 6 parts with equal amount, placing the fresh hyphae in a clean oven, and drying the hyphae at the temperature of 30 ℃ until the water content of the hyphae is respectively 4-6% (excluding 4%, including 6%), 3-4% (excluding 3%, including 4%), 2-3% (excluding 2%, including 3%), 1-2% (excluding 1%, including 2%), 0-1% (excluding 0, including 1%) and 0, so as to obtain the solid hyphae with different water content.
Solid hyphae with different water contents are respectively inoculated in a PDA solid culture medium (the size of a culture dish is 9mm × 9mm) in an ultraclean workbench, dark culture is carried out in a constant-temperature incubator at the temperature of 28 ℃, each culture dish contains 1 bacterial piece with the same size, the water content in each range is repeated for 5 times, the day of inoculation is recorded as the day of inoculation 0, the diameter of bacterial colonies is measured in the days of inoculation 2, 5 and 9 by adopting a cross method, and a control group is replaced by fresh bacterial cakes with the same size and is repeated for 5 times.
The results (Table 1) show that the colony diameter of needle A2-1 tended to decrease after the change in water content for the same number of days of culture, and the critical water content value was 2-3%, and therefore, the water content was not less than 2% when the solid dry microbial inoculum was prepared.
TABLE 1 colony diameters (cm) of different bacterial agents
Note: in table 1, CK represents wet bacteria control; 4-6%, excluding 4%, including 6%; 3-4%, excluding 3%, including 4%; 2-3%, excluding 2%, including 3%; 1-2%, excluding 1%, including 2%; 0-1%, excluding 0, including 1%. In the same column, the data marked with the same letter has no significant difference, and the data marked with different letters has significant difference.
Example 3 Effect of powder particle size on Dry powder microbial inoculum
Solid hyphae having a water content of 3% at 30 ℃ of needle A2-1 were prepared according to the method of example 2, and the obtained solid hyphae were ground and sieved through sieves having a pore size of 5mm, 1mm and 0.5mm, respectively, to obtain dry microbial inoculum having a particle size of more than 5mm, 0.5-1mm (excluding 1mm, including 0.5mm) and less than 0.5 mm.
Inoculating dry powder microbial inoculum with different particle sizes into a PDA culture medium, culturing in a constant-temperature incubator at 28 ℃ in the dark, wherein each culture dish contains 1 particle size microbial inoculum, each microbial inoculum is repeated for 5 times, and the day of inoculation is recorded as the day of inoculation 0. The colony diameters were measured on days 2, 5 and 9 of inoculation using the cross method. The control group was replaced with fresh cakes of the same size, repeated 5 times.
The number of days of culture was plotted on the abscissa and the diameter of the colony was plotted on the ordinate as a bar graph, as shown in FIG. 4. When the bacterial is cultured on the 2 nd day, the diameter of the bacterial colony is gradually increased along with the gradual increase of the size of the bacterial powder; the microbial inoculum powder size had no significant effect on colony diameter as culture time was gradually increased to day 9. Therefore, the size of the microbial inoculum powder has no obvious influence on the growth of the microbial inoculum in a certain range. The smaller the microbial inoculum powder is, the more the microbial inoculum powder quantity under the same quality is, and the higher the microbial inoculum utilization efficiency is, so that the particle size of the prepared dry powder microbial inoculum can be less than 0.5 mm.
Example 4 Effect of drying temperature on growth of microbial inoculum
The bacterial liquid used was the needle a2-1 bacterial liquid of example 1, and the bacterial agent was prepared according to the following steps:
filtering a bacterium solution of needle A2-1 under a single-layer gauze, collecting fresh hyphae, uniformly mixing, randomly dividing the fresh hyphae into 7 parts with equal quantity, each part is 30g, drying 6 parts of hyphae at 30 ℃, 40 ℃, 50 ℃, 60 ℃, 70 ℃ and 80 ℃ respectively until the water content is 3% to obtain solid hyphae, recording the drying time, fully grinding the dried solid hyphae under different conditions respectively, sieving (0.5mm) to obtain 6 dry powder bacterial agents with the particle size of less than 0.5mm, inoculating all the obtained dry powder bacterial agents in a PDA solid culture medium (the size of a culture dish is 9mm × 9mm) in an ultraclean workbench in a constant temperature incubator at 28 ℃ for dark culture, each culture dish contains 1 bacterial agent, each treatment is repeated for 5 times, the day of inoculation is recorded as day 0 of inoculation, measuring the diameters of bacterial colonies on the 5 th and 9 th day of inoculation by adopting a cross method, and utilizing the other 1 part of fresh hyphae which is not dried as a wet bacterial agent control for inoculation, and repeating for 5 times.
And counting the drying time required at different drying temperatures, wherein the drying time required under the conditions of the drying temperature of 30 ℃, 40 ℃, 50 ℃, 60 ℃, 70 ℃ and 80 ℃ is 96, 84, 66, 42, 24 and 18 hours respectively.
The diameters of the colonies inoculated with the respective inoculants at different times are shown in Table 2, and the diameters of the colonies tend to be unchanged and then become smaller with the increase of the drying temperature on 5 th and 9 th days, wherein the critical temperature is 60 ℃, and the graph is shown in FIG. 5. Therefore, the optimal drying temperature for preparing the needle A2-1 dry powder microbial inoculum is 60 ℃, and the microbial inoculum prepared at the temperature not only ensures the activity of the dry powder microbial inoculum, but also shortens the drying time and greatly improves the preparation efficiency.
The needle A2-1 bacterial liquid in the above steps was replaced with the needle A2-7 bacterial liquid and the needle A2-8 bacterial liquid of example 1, respectively, and the drying temperature was set at 60 ℃ to obtain needle A2-7 dry powder bacterial agents and needle A2-8 dry powder bacterial agents, respectively, and the undried fresh hyphae of needle A2-7 and needle A2-8 were used as wet bacterial agent controls, respectively. The obtained microbial inoculum was cultured according to the method for culturing the microbial inoculum of the needle A2-1, and the colony diameters of the microbial inocula inoculated at different times are shown in Table 2.
When the drying temperature is 60 ℃, the diameters of bacterial colonies on the 5 th day and the 9 th day of the obtained needle A2-1 dry powder bacterial agent inoculation are 77.5 percent and 83.7 percent of the wet bacterial agent control respectively, the diameters of the bacterial colonies on the 5 th day and the 9 th day of the obtained needle A2-7 dry powder bacterial agent inoculation are 39.8 percent and 58.2 percent of the wet bacterial agent control respectively, and the diameters of the bacterial colonies on the 5 th day and the 9 th day of the obtained needle A2-8 dry powder bacterial agent inoculation are 35.1 percent and 50.5 percent of the wet bacterial agent control respectively. It is indicated that needles A2-7 and needle A2-8 are not suitable for the preparation of dry powdered inocula at a drying temperature of 60 ℃.
TABLE 2 colony diameters (cm) of different bacterial agents
Note: in Table 2, CK represents the control of wet bacteria, and data of the same species at the same incubation time are marked with different letters to indicate that the difference is significant, and marked with the same letters to indicate that the difference is not significant.
Example 5 preservation of Dry powder microbial inoculum
A dry powder microbial inoculum of the needle A2-1 strain at the drying temperature of 60 ℃ is prepared according to the method of the embodiment 4, the prepared microbial inoculum is placed in a clean self-sealing bag and is respectively stored for 10 days and 20 days at the room temperature (20 ℃), then the microbial inoculum is inoculated in a PDA culture medium and is cultured in a dark environment in a constant temperature incubator at the temperature of 28 ℃, and whether the microbial inoculum still has activity is observed.
As a result, the needle A2-1 dry powder microbial inoculum can still grow in the culture medium, which indicates that the needle A2-1 dry powder microbial inoculum can be stored for at least 20 days at room temperature.
<110> university of mineral industry (Beijing), Beijing Synbiotic ecological environmental engineering technology Limited
<120> preparation method of dark-color compartmental endophytic fungus dry powder microbial inoculum
<160>3
<170>PatentIn version 3.5
<210>1
<211>589
<212>DNA
<213>Darksidea sp.
<400>1
cttccgtagg tgaacctgcg gaaggatcat tacctggcct tgggccgctc gcgggagcta 60
gtcgcttgcg acgacgctac cgagggcgct tagcccttga ctatcacctt gactacgtgc 120
accttttgtt gtttcctcgg caggtcacct gccgccagga accccctaaa ccttttgtaa 180
tagcatccaa acttctgaaa acaaaccaaa ttatttacaa cttttaacaa tggatctctt 240
ggttctggca tcgatgaaga acgcagcgaa atgcgataag tagtgtgaat tgcagaattc 300
agtgaatcat cgaatctttg aacgcacatt gcgccccatg gtattccgtg gggcatgcct 360
gttcgagcgt catttacccc ctcaagctcc gcttggtgtt gggcgtctgt cccgcttcgc 420
gcgcggactc gccccaaagg tattggcagc ggtcgtgcca gcttctcgcg cagcacattg 480
cgcttctcga ggcaccggcg ggcccgcgtc catcaagctc accccccagt ttgacctcgg 540
atcaggtagg gatacccgct gaacttaagc atatcaataa gcggaggaa 589
<210>2
<211>589
<212>DNA
<213>Darksidea sp.
<400>2
cttccgtagg tgaacctgcg gaaggatcat tacctggcct tgggccgctc gcgggagcta 60
gtcgcttgcg acgacgctac cgagggcgct tagcccttga ctatcacctt gactacgtgc 120
accttttgtt gtttcctcgg caggtcacct gccgccagga accccctaaa ccttttgtaa 180
tagcatccaa acttctgaaa acaaaccaaa ttatttacaa cttttaacaa tggatctctt 240
ggttctggca tcgatgaaga acgcagcgaa atgcgataag tagtgtgaat tgcagaattc 300
agtgaatcat cgaatctttg aacgcacatt gcgccccatg gtattccgtg gggcatgcct 360
gttcgagcgt catttacccc ctcaagctcc gcttggtgtt gggcgtctgt cccgcttcgc 420
gcgcggactc gccccaaagg tattggcagc ggtcgtgcca gcttctcgcg cagcacattg 480
cgcttctcga ggcaccggcg ggcccgcgtc catcaagctc accccccagt ttgacctcgg 540
atcaggtagg gatacccgct gaacttaagc atatcaataa gcggaggaa 589
<210>3
<211>593
<212>DNA
<213> Geospora sp (Pleosporales sp.)
<400>3
cttccgtagg tgaacctgcg gaaggatcat tacctggcct tgggccgctc gggggagcca 60
gtcgctcgcg acggcgctgc cttgggcgct tagcccttga ctatcacctt gactacgtgc 120
accttttgtt gtttcctcgg caggtcctct gccgccagga accccctaaa cctttttgca 180
atagcatcca aacttctgaa aacaaaccaa atcatttaca acttttaaca atggatctct 240
tggttctggc atcgatgaag aacgcagcga aatgcgatat gtagtgtgaa ttgcagaatt 300
cagtgaatca tcgaatcttt gaacgcacat tgcgccccat ggtattccgt ggggcatgcc 360
tgttcgagcg tcatttaccc cctcaagctc cgcttggtgt tgggcgtctg tcccgcttcg 420
cgcgcggact cgccccaaag gtattggcag cggtcatgcc agcttctcgc gcagcacatt 480
gcgcttctcg aggcaccggc gggcccgcgt ccatcaagct ctcacccccc cagtttgacc 540
tcggatcagg tagggatacc cgctgaactt aagcatatca ataagcggag gaa 593
Claims (6)
1. The preparation method of the needle A2-1 microbial inoculum comprises the following steps: drying the needle A2-1 at 60 deg.C to obtain dried fungus A2-1; the needle A2-1 is a bacterial strain with the preservation number of CGMCCNo.17465 in the China general microbiological culture Collection center.
2. The method of claim 1, wherein: the moisture content of the dried fungus is 2% -3%.
3. The method according to claim 1 or 2, characterized in that: the drying of needle a2-1 at 60 ℃ comprises: drying the mycelium of needle A2-1 at 60 deg.C.
4. A method according to any one of claims 1-3, characterized in that: the method also comprises the step of crushing the dried fungi, wherein the obtained crushed fungi is the A2-1 fungicide.
5. The method of claim 4, wherein: the method also comprises the step of sieving the crushed fungi by a sieve with the diameter of less than or equal to 0.5mm to obtain powdery substances with the diameter of less than or equal to 0.5mm, namely the A2-1 microbial inoculum.
6. Use of the method according to any one of claims 1 to 5 for the preparation of a product for promoting plant growth or enhancing stress resistance of plants or improving the physicochemical properties of soil.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030138936A1 (en) * | 2000-05-02 | 2003-07-24 | Taiji Mizuguchi | Spray-dried microbial cells |
| US20140134153A1 (en) * | 2011-07-15 | 2014-05-15 | Kabushiki Kaisha Ouju Seiyaku | Antimicrobial agent and method for producing the same |
| CN110250210A (en) * | 2019-06-12 | 2019-09-20 | 中国矿业大学(北京) | A kind of optimal DSE strain for promoting maize seed soaking and taking root |
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2020
- 2020-04-17 CN CN202010304640.7A patent/CN111411047A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030138936A1 (en) * | 2000-05-02 | 2003-07-24 | Taiji Mizuguchi | Spray-dried microbial cells |
| US20140134153A1 (en) * | 2011-07-15 | 2014-05-15 | Kabushiki Kaisha Ouju Seiyaku | Antimicrobial agent and method for producing the same |
| CN110250210A (en) * | 2019-06-12 | 2019-09-20 | 中国矿业大学(北京) | A kind of optimal DSE strain for promoting maize seed soaking and taking root |
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| 张峰峰等: "解淀粉芽孢杆菌HN固体菌剂发酵制备技术研究", 《中国饲料》 * |
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