CN116478880A - Acremodelling acid bacteria KC 316 and application thereof - Google Patents

Acremodelling acid bacteria KC 316 and application thereof Download PDF

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Publication number
CN116478880A
CN116478880A CN202310450899.6A CN202310450899A CN116478880A CN 116478880 A CN116478880 A CN 116478880A CN 202310450899 A CN202310450899 A CN 202310450899A CN 116478880 A CN116478880 A CN 116478880A
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amycolatopsis
fermentation
ethyl acetate
water
fermentation product
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曹艳茹
朱国兴
陈秀
孔德军
张悦
何江波
刘子超
侯秀丽
苏源
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Kunming University
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Kunming University
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P21/00—Plant growth regulators
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P5/00—Nematocides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D11/00—Solvent extraction
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
    • C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture

Abstract

The invention discloses a amycolatopsis mansoni (Amycolatopsis tucumanensis) KC 316 strain and application thereof, and the microorganism preservation registration number of the amycolatopsis mansoni strain is CGMCC No.26012. The invention provides a preparation method for preparing a amycolatopsis tussilago KC 316 fermentation product. Experiments prove that the fermentation product of the amycolatopsis tuca KC 316 can promote the growth of crops and has high activity of killing root-knot nematodes. The strain screened by the invention is reported to be used for preventing and controlling nematode diseases for the first time, and has the effect of promoting plant growth. Compared with the chemical synthesis pesticide control used at present, the pesticide has the advantages of high efficiency, low toxicity, no environmental pollution, difficult generation of drug resistance and the like because the pesticide is derived from natural environment, and has important significance for sustainable development of agriculture.

Description

Acremodelling acid bacteria KC 316 and application thereof
Technical Field
The invention relates to the technical field of microbial engineering, in particular to amycolatopsis tussilago mandshurica KC 316 and application thereof.
Background
Amycolatopsis belongs to the Pseudonocardiaceae family, and secondary products of a plurality of strains of Amycolatopsis have novel biological activities, and most of Amycolatopsis are important antibiotics with strong antibacterial ability, so that Amycolatopsis is widely applied to the industrial production of antibiotics. In 2010, it was reported by Albarreaci n et al that amycolatopsis in Tuckmann (Amycolatopsis tucumanensis) was able to synthesize a low molecular weight cysteine-rich protein in cells, which has a strong binding capacity with copper, thus having a good intracellular copper accumulation capacity, and effectively reducing the effective copper content of plants in copper contaminated areas for the treatment of environmental copper contamination.
In recent years, rapid development of single planting and intensive greenhouse vegetable planting over a long period of time has led to frequent occurrence of root knot nematodes. The annual root-knot nematode hazard in China is directly and economically lost by 500 hundred million yuan, so that the agricultural development is severely restricted. The root knot nematode damages the epidermal cells of the root through the oral needle, so that the tissue cells of the host plant are diseased to generate giant cells, and a root knot is formed. At the same time, the normal physiological metabolism of the plants is blocked, the water and inorganic salt of the plants are not absorbed enough, and the phenomena of plant wilting, leaf yellowing, delay in the waiting period, yield reduction and the like appear. In addition, the mouth needles of the root-knot nematodes bring mechanical damage to the epidermal cells of the host plants, provide an infection way for other pathogenic bacteria in the soil, cause various pathogenic bacteria to invade the host plants, form compound diseases, and cause slow plant growth, defoliation and even dead.
At present, the prevention measures of the root-knot nematodes mainly comprise agricultural prevention, physical prevention, chemical prevention and biological prevention. Agricultural control mainly controls root-knot nematodes through rotation, intercropping, soil plowing, grafting resistance stock field management and other modes, however, agricultural control is limited by land resources and the quantity of the stocks, and is difficult to carry out on a large scale. Physical control mainly eliminates the eggs of the linear worms in the soil through sunlight disinfection, soil hot water treatment and the like, but simultaneously eliminates part of beneficial bacterial groups in the soil and destroys the soil aggregate structure and the ecological system. The chemical prevention and control means are widely applied at present, and are mainly performed through chemical pesticides artificially synthesized by methyl bromide, fosthiazate, carbofuran, aldicarb and the like, but the long-term use of chemical agents is easy to cause the problems of nematode resistance, environmental pollution and the like, and residual chemical agents enter food chains to cause toxicity to people and animals, so that a plurality of chemical nematicides such as carbofuran, methyl bromide fumigant and the like are forbidden by the national plaintext. The biological control is to control the root-knot nematode by using plants, microorganisms or metabolites produced by the microorganisms, so that the problems of toxic residues, environmental harm and the like of chemical control are reduced, the control effect which is easy to degrade and difficult to generate drug resistance is achieved, and part of functional microorganisms also show promotion effect in the aspect of crop growth and development. Therefore, the screening of the novel difunctional strain with nematicidal activity and growth promoting effect has important significance for sustainable development of agriculture.
No report of the nematicidal activity and plant growth promotion function of the amycolatopsis Guan Tuku is found through searching.
Disclosure of Invention
Aiming at the technical problems, the invention aims to provide a amycolatopsis tussilago KC 316 and application thereof.
The invention provides a amycolatopsis in a gallery, which has a classification name of (Amycolatopsis tucumanensis) KC 316 and is preserved in China general microbiological culture collection center (CGMCC) of China Committee for culture Collection of microorganisms (CGMCC, address: north Chen West Lu No. 1, 3 of the Chachiensis area of Beijing, and China center for sciences of microorganisms) on 11 month 02 of 2022, and the preservation registration number is CGMCC No.26012.
The invention also provides a preparation method of the fermentation product of the amycolatopsis in the gallery, which comprises the following steps:
s1, activating a strain, namely streaking a amycolatopsis tussilago strain preserved in a glycerol pipe onto a solid medium of an improved international streptomycete No.2 medium (I SP 2), and culturing for 3-5d;
s2, preparing seed liquid, namely inoculating single bacterial colonies of the amycolatopsis in the gallery activated by S1 into a seed culture medium, performing shake culture at 25-40 ℃ and 80-200rpm, and fermenting for 3-5d to prepare the seed liquid;
s3, large-scale fermentation, namely inoculating 5mL of the seed solution prepared in the step S2 into 100mL of a fermentation medium, carrying out shake culture at 25-40 ℃ and 80-200rpm, and carrying out shaking fermentation for 5-9d to obtain a fermentation liquid;
s4, centrifuging, namely centrifuging the fermentation liquor obtained in the step S3 at 4000rpm for 10min, and collecting a fermentation liquor supernatant;
s5, extracting fermentation products, namely extracting the fermentation liquor supernatant obtained in the step S4 by adopting an organic reagent, and carrying out reduced pressure distillation on the extract liquor to remove the organic solvent, wherein the obtained products are fermentation crude extracts.
Further, the specific method in S5 is as follows: extracting with equal volume of ethyl acetate, standing until the water phase and the ethyl acetate phase are layered, and collecting the ethyl acetate phase; extracting the residual water phase with equal volume of ethyl acetate again, standing until the water phase and the ethyl acetate phase are layered, and collecting the ethyl acetate phase at the upper layer; extracting the residual water phase with equal volume of ethyl acetate for the third time, standing until the water phase and the ethyl acetate phase are layered, and collecting the ethyl acetate phase; combining ethyl acetate phases obtained by three extractions, and distilling at 40-50 ℃ under reduced pressure to remove the organic solvent, thus obtaining the substance which is the fermentation crude extract.
Further, the preparation method of the fermentation medium in the step S3 comprises the following steps: 5g glucose, 5g yeast extract, 5g malt extract powder, dissolving in water, adjusting pH to 7.5, adjusting agar to 18-20g, diluting with water to 1L, and sterilizing at 121deg.C for 15 min.
Preferably, the preparation method of the seed culture medium in the step S2 comprises the following steps: dissolving 5g yeast extract, 10g malt extract and 4g glucose in water, adjusting pH to 7.0-7.2, diluting to 1L with water, sterilizing at 121deg.C for 30 min, and collecting seed solution OD 600 nm is 1.6-2.0.
Preferably, the preparation method of the modified Streptomyces internationis No.2 medium (I SP 2) in the step S1 is as follows: 5g glucose, 5g yeast extract, 5g malt extract powder, adjust pH to 7.5, and volume to 1L with water.
Preferably, the preparation method of the solid medium in step S1 is as follows: 5g glucose, 5g yeast extract, 5g malt extract powder, dissolving in water, adjusting pH to 7.5, adjusting agar to 18-20g, diluting with water to 1L, and sterilizing at 121deg.C for 15 min.
The invention also protects the application of the fermentation product of the amycolatopsis tuca in preparing nematicide.
The invention also protects the application of the fermentation product of the amycolatopsis tussilagomannii in preparing a crop growth promoting microbial inoculum.
Preferably, the nematode is a root-knot nematode.
Compared with the prior art, the invention has the following beneficial effects:
(1) The invention provides a amycolatopsis tussilago KC 316, which is reported to be used for preventing and treating nematode diseases for the first time.
(2) The fermentation product of the amycolatopsis mandshurica KC 316 has the effect of promoting plant growth besides killing nematodes.
(3) Compared with the existing chemical synthesis pesticide control, the fermentation broth of the invention is from natural environment, and has the advantages of high efficiency, low toxicity, no environmental pollution, difficult generation of drug resistance and the like.
Drawings
FIG. 1 is a phylogenetic tree diagram;
FIG. 2 is a graph showing the nematicidal activity results of crude extracts;
FIG. 3 is a graph showing rooting promoting effect of KC 316 fermentation broth;
FIG. 4 shows the effect of KC 316 fermentation broth on stem growth promotion;
FIG. 5 is a graph showing the effect of KC 316 fermentation broth.
Detailed Description
The following detailed description of the invention is provided in connection with the accompanying drawings that are presented to illustrate the invention and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the invention in any way.
The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are carried out according to techniques or conditions described in the literature in the field or according to the product specifications. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified. DMSO is known as dimethylsulfoxide. Unless otherwise indicated, the quantitative tests in the examples below were all performed in triplicate, and the results averaged.
EXAMPLE 1 isolation, identification and preservation of strains
1.1A strain was isolated from the harvested plant rhizosphere soil of the Sedan mountain and designated strain KC 316.
1.2 identification of Strain KC 316
Bacterial colony of bacterial strain KC 316 on the 'international streptomycete No.2 culture medium' (I SP 2) improved culture medium is light yellow, dry, opaque and dry powder on the surface, the bacterial colony is tightly connected with the culture medium, the edge is irregular, and the bacterial colony is not easy to pick up.
The phylogenetic tree constructed by the 16S rRNA gene sequence of the strain KC 316 is shown in FIG. 1. Strains KC 316 and Amycolatopsis tucumanensis ABO T One branch was collected and its similarity was 99.72%. KC 316 was identified as Amycolatopsis tucumanensis in conjunction with a physiobiochemical assay.
1.3 preservation of Strain KC 316
The strain KC 316 is identified as amycolatopsis tuca (Amycolatopsis tucumanensis) and is preserved in China general microbiological culture Collection center (CGMCC) of 11/02/2022, wherein the address is 1/3 of North West Lu in the Chaoyang area of Beijing city, and the preservation registration number is 26012 of CGMCC.
Example 2 preparation of amycolatopsis Vibrio fermentation products
2.1 activation of strains: the KC 316 strain preserved in the glycerol pipe is taken, the plate is streaked onto a modified solid culture medium of Streptomyces internationis No.2 (I SP 2), and is cultured for 3-5 days, and the preparation method of the modified I SP 2 culture medium is as follows: 5g glucose, 5g yeast extract, 5g malt extract powder, adjusting pH to 7.5, adjusting agar to 18-20g, fixing volume to 1L with water, and sterilizing at 121deg.C for 15 min;
2.2 preparation of seed liquid: inoculating the single colony activated in 2.1 into a seed culture medium, and performing shaking table fermentation at 25-40 ℃ and 80-200rpm for 3-5d to prepare seed liquid; the seed liquid is OD 600 nm is 1.6-2.0;
2.3 inoculating 5mL of the seed solution in 2.2 into 100mL of fermentation medium, fermenting at 25-40 ℃ at 80-200rpm for 5-9d by a shaking table, and obtaining a product which is fermentation liquor; the fermentation medium comprises the following components: 5g glucose, 5g yeast extract, 5g malt extract, dissolving in water, adjusting pH to 7.5, diluting to 1L with water, sterilizing at 121deg.C for 15 min;
2.4 centrifuging the fermentation broth obtained in the above step at 4000rpm for 10min, collecting supernatant, extracting with equal volume of ethyl acetate, standing until the water phase and the ethyl acetate phase are layered, and collecting ethyl acetate phase; extracting the residual water phase with equal volume of ethyl acetate again, standing until the water phase and the ethyl acetate phase are layered, and collecting the ethyl acetate phase; extracting the residual water phase with equal volume of ethyl acetate for the third time, standing until the water phase and the ethyl acetate phase are layered, and collecting the ethyl acetate phase; combining ethyl acetate phases obtained by three extractions, and distilling at 40-50 ℃ under reduced pressure to remove the organic solvent, thus obtaining the product which is the crude extract.
EXAMPLE 3 lethal effect of Acremodelling bacteria KC 316 fermentation product on root knot nematode
Dissolving the crude extract prepared in example 2 in DMSO to obtain a crude extract concentration of 500. Mu.g/. Mu.L; taking 96-well plates, adding 25 mu L of Java root-knot nematode suspension (60-80), 100 mu L of crude extract solution and 75 mu L of sterile water into each well, gently stirring and mixing by using a pipette, standing at 20 ℃ for 12 hours, and then counting the nematode mortality. An equal volume of DMSO was used as a negative control instead of the crude extract solution. Three replicates were performed, 3 replicates were set for each replicate, and the results averaged. As a result, the mortality rate of nematodes in the negative control treatment was 6.5% and the mortality rate of nematodes in the crude extract treatment group was 86.3% (see FIG. 2 for details).
Example 4 action of fermentation liquor of amycolatopsis mandshurica KC 316 on promoting crop growth
Taking the fermentation broth obtained in step 2.3 of example 2, diluting the fermentation broth to 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 6 concentration gradients. Taking sterile distilled water as a negative control, respectively taking the 6 concentration gradients and 100mL of fermentation stock solutions, and detecting the effect of fermentation solutions with different concentrations on germination of cucumber seeds: soaking cucumber seeds in warm water at 40-50deg.C for 12-24 hr, taking out, placing in 9cm culture dish (5 grains/dish×5) paved with sterilized filter paper, soaking the filter paper with 3mL fermentation liquid with corresponding concentration gradient, placing in illumination incubator at 28deg.C and humidity of 70%, culturing for 12 hr in dark, and alternately treating with 12 hr illumination. 2mL of fermentation broth was supplemented every 2d, observations were made daily, 14d was recorded continuously, and the root length and shoot length were recorded. 3 replicates were performed, at least 15 replicates were set for each replicate, and the results averaged. The cucumber roots treated by the negative control treatment have a length of 10.93cm, a stem length of 2.74cm, and the cucumber roots treated by the fermentation liquor treatment group have a length of 15.73cm and a stem length of 4.17cm (see figures 3-5 for details), which shows the effect of KC 316 fermentation liquor on promoting root growth and stem growth.

Claims (10)

1. The amycolatopsis tussilaginosa is characterized in that: the classification name is (Amycolatopsis tucumanensis) KC 316, which has been preserved in China general microbiological culture Collection center (CGMCC) at 11 and 02 of 2022, and the preservation registration number is CGMCC No.26012.
2. A fermentation product of amycolatopsis tussilaginosa according to claim 1, which is prepared by a process comprising the steps of:
s1, activating a strain, namely streaking a amycolatopsis tussilago strain preserved in a glycerol pipe onto a modified Streptomyces internationis No.2 culture medium (ISP 2) solid culture medium, and culturing for 3-5d;
s2, preparing seed liquid, namely inoculating single bacterial colonies of the amycolatopsis in the gallery activated by S1 into a seed culture medium, performing shake culture at 25-40 ℃ and 80-200rpm, and fermenting for 3-5d to prepare the seed liquid;
s3, large-scale fermentation, namely inoculating 5mL of the seed solution prepared in the step S2 into 100mL of a fermentation medium, carrying out shake culture at 25-40 ℃ and 80-200rpm, and carrying out shaking fermentation for 5-9d to obtain a fermentation liquid;
s4, centrifuging, namely centrifuging the fermentation liquor obtained in the step S3 at 4000rpm for 10min, and collecting a fermentation liquor supernatant;
s5, extracting fermentation products, namely extracting the fermentation liquor supernatant obtained in the step S4 by adopting an organic reagent, and carrying out reduced pressure distillation on the extract liquor to remove the organic solvent, wherein the obtained products are fermentation crude extracts.
3. The fermentation product of amycolatopsis tussilaginosa according to claim 2, characterized in that: the specific method in the S5 is as follows: extracting with equal volume of ethyl acetate, standing until the water phase and the ethyl acetate phase are layered, and collecting the ethyl acetate phase; extracting the residual water phase with equal volume of ethyl acetate again, standing until the water phase and the ethyl acetate phase are layered, and collecting the ethyl acetate phase at the upper layer; extracting the residual water phase with equal volume of ethyl acetate for the third time, standing until the water phase and the ethyl acetate phase are layered, and collecting the ethyl acetate phase; combining ethyl acetate phases obtained by three extractions, and distilling at 40-50 ℃ under reduced pressure to remove the organic solvent, thus obtaining the substance which is the fermentation crude extract.
4. The fermentation product of amycolatopsis tussilaginosa according to claim 2, characterized in that: the preparation method of the fermentation medium in the step S3 comprises the following steps: 5g glucose, 5g yeast extract, 5g malt extract powder, dissolving in water, adjusting pH to 7.5, adjusting agar to 18-20g, diluting with water to 1L, and sterilizing at 121deg.C for 15 min.
5. The fermentation product of amycolatopsis tussilaginosa according to claim 2, characterized in that: the preparation method of the seed culture medium in the step S2 comprises the following steps: dissolving 5g yeast extract, 10g malt extract and 4g glucose in water, adjusting pH to 7.0-7.2, diluting to 1L with water, sterilizing at 121deg.C for 30 min, and collecting seed solution OD 600 nm is 1.6-2.0.
6. The fermentation product of amycolatopsis tussilaginosa according to claim 2, characterized in that: the preparation method of the modified Streptomyces internationis No.2 medium (ISP 2) in the step S1 is as follows: 5g glucose, 5g yeast extract, 5g malt extract powder, adjust pH to 7.5, and volume to 1L with water.
7. The fermentation product of amycolatopsis tussilaginosa according to claim 2, characterized in that: the preparation method of the solid culture medium in the step S1 comprises the following steps: 5g glucose, 5g yeast extract, 5g malt extract powder, dissolving in water, adjusting pH to 7.5, adjusting agar to 18-20g, diluting with water to 1L, and sterilizing at 121deg.C for 15 min.
8. Use of a fermentation product of amycolatopsis tuca according to any one of claims 2-7 for the preparation of nematicide.
9. Use of a fermentation product of amycolatopsis tucable according to any one of claims 2-7 in the preparation of a crop growth promoting inoculant.
10. The use according to claim 8, characterized in that: the nematode is root-knot nematode.
CN202310450899.6A 2023-04-25 2023-04-25 Acremodelling acid bacteria KC 316 and application thereof Pending CN116478880A (en)

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