CN118086102A - Application of stenotrophomonas gondii in inhibiting rice blast bacteria - Google Patents
Application of stenotrophomonas gondii in inhibiting rice blast bacteria Download PDFInfo
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- CN118086102A CN118086102A CN202410064816.4A CN202410064816A CN118086102A CN 118086102 A CN118086102 A CN 118086102A CN 202410064816 A CN202410064816 A CN 202410064816A CN 118086102 A CN118086102 A CN 118086102A
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- 241000122971 Stenotrophomonas Species 0.000 title claims abstract description 40
- 230000002401 inhibitory effect Effects 0.000 title claims abstract description 11
- 235000007164 Oryza sativa Nutrition 0.000 title claims description 25
- 235000009566 rice Nutrition 0.000 title claims description 25
- 241000894006 Bacteria Species 0.000 title claims description 8
- 240000007594 Oryza sativa Species 0.000 title 1
- 238000000855 fermentation Methods 0.000 claims abstract description 50
- 230000004151 fermentation Effects 0.000 claims abstract description 50
- 230000001580 bacterial effect Effects 0.000 claims abstract description 31
- 239000000725 suspension Substances 0.000 claims abstract description 29
- 238000004519 manufacturing process Methods 0.000 claims abstract description 15
- 230000015572 biosynthetic process Effects 0.000 claims abstract description 12
- 210000002199 attachment cell Anatomy 0.000 claims abstract description 10
- 230000007918 pathogenicity Effects 0.000 claims abstract description 9
- 238000009629 microbiological culture Methods 0.000 claims abstract description 4
- 241000209094 Oryza Species 0.000 claims description 24
- 239000007788 liquid Substances 0.000 claims description 20
- 238000012258 culturing Methods 0.000 claims description 12
- 238000004321 preservation Methods 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 6
- 239000012880 LB liquid culture medium Substances 0.000 claims description 4
- 230000003213 activating effect Effects 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- 238000011218 seed culture Methods 0.000 claims description 4
- 239000012528 membrane Substances 0.000 claims description 3
- 238000000034 method Methods 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 241001052560 Thallis Species 0.000 claims description 2
- 238000012136 culture method Methods 0.000 claims description 2
- 239000000706 filtrate Substances 0.000 claims description 2
- 239000001963 growth medium Substances 0.000 claims description 2
- 239000002054 inoculum Substances 0.000 claims description 2
- 238000000967 suction filtration Methods 0.000 claims description 2
- 210000003127 knee Anatomy 0.000 claims 1
- 241001330975 Magnaporthe oryzae Species 0.000 abstract description 10
- 231100000956 nontoxicity Toxicity 0.000 abstract description 3
- 230000007613 environmental effect Effects 0.000 abstract description 2
- 235000010633 broth Nutrition 0.000 description 14
- 230000002265 prevention Effects 0.000 description 6
- 238000011282 treatment Methods 0.000 description 6
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 239000000126 substance Substances 0.000 description 4
- TWFZGCMQGLPBSX-UHFFFAOYSA-N Carbendazim Natural products C1=CC=C2NC(NC(=O)OC)=NC2=C1 TWFZGCMQGLPBSX-UHFFFAOYSA-N 0.000 description 3
- 239000006013 carbendazim Substances 0.000 description 3
- JNPZQRQPIHJYNM-UHFFFAOYSA-N carbendazim Chemical compound C1=C[CH]C2=NC(NC(=O)OC)=NC2=C1 JNPZQRQPIHJYNM-UHFFFAOYSA-N 0.000 description 3
- 206010059866 Drug resistance Diseases 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 241000122973 Stenotrophomonas maltophilia Species 0.000 description 2
- 241000607479 Yersinia pestis Species 0.000 description 2
- 230000003042 antagnostic effect Effects 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 239000000575 pesticide Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 231100000674 Phytotoxicity Toxicity 0.000 description 1
- 241001148470 aerobic bacillus Species 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 239000003899 bactericide agent Substances 0.000 description 1
- 238000009412 basement excavation Methods 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- -1 small molecule compounds Chemical class 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 230000005074 turgor pressure Effects 0.000 description 1
Classifications
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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
- A01P3/00—Fungicides
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- 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
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- 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
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- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
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- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
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- Virology (AREA)
- Pest Control & Pesticides (AREA)
- Biomedical Technology (AREA)
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- Plant Pathology (AREA)
- General Engineering & Computer Science (AREA)
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- Tropical Medicine & Parasitology (AREA)
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Abstract
The invention relates to an application of stenotrophomonas gondii in inhibiting rice blast fungus attachment cell formation and pathogenicity on a host, wherein the stenotrophomonas gondii is stenotrophomonas gondii WXY53 (Stenotrophomonas geniculata WXY) which is preserved in China general microbiological culture collection center (CGMCC) No. 26681, and the bacterial suspension, fermentation broth and sterile fermentation broth have the advantages of low production cost, NO toxicity, environmental protection and capability of remarkably forming the rice blast fungus attachment cell and pathogenicity on the host.
Description
Technical Field
The invention belongs to the technical field of biological pesticides, and particularly relates to a bacterial suspension, sterile fermentation liquor and fermentation liquor of stenotrophomonas gondii, and a corresponding preparation process and application thereof in agricultural pest control.
Background
Rice blast is one of three diseases of rice, and the disease is widely distributed. In the epidemic disease area, the yield is generally reduced by 10% -20%, 40% -50% can be achieved when the disease is serious, and especially serious fields and even particles are not harvested. For the prevention and control of rice blast, chemical prevention and control and resistance breeding are mainly used at present, but the prevention and control effect is poor due to the enhancement of drug resistance and the reduction of variety resistance, and the prevention and control of rice blast is still an important difficulty in production at present. The rice blast bacteria successfully infects the rice blast bacteria mainly by forming attachment cells with huge turgor pressure to penetrate the host epidermis, so that the excavation of products and techniques for efficiently inhibiting the formation of the attachment cells is an effective way for guaranteeing the efficient prevention and control of the rice blast.
Biological control is more and more paid attention to plant diseases and insect pests prevention and treatment because of the advantages of no toxicity, no harm, no pollution, difficult generation of drug resistance, high efficiency and the like. Stenotrophomonas (Stenotrophomonas) is a class of gram-negative aerobic bacteria, the species of which reaches more than 20 species, and the distribution is relatively wide. The stenotrophomonas maltophilia (Stenotrophomonas maltophilia) is the only reported pathogenic bacteria at present, is mostly a clinical source, and is derived from soil, plant endophytic sources and other oligotrophic monad strains, which do not show pathogenic capability and have good application prospects in the industrial and agricultural fields. Studies have shown that a portion of the oligotrophic monad strain is capable of degrading organic pesticides, secreting plant growth hormones, extracellular enzymes and small molecule compounds, and is highly effective in antagonizing a portion of the fungus, protecting and promoting crop growth. However, no research on antagonizing Pyricularia oryzae has been reported.
Disclosure of Invention
The invention aims to solve the technical problems of overcoming the defects of the prior art and providing a bacterial suspension, a sterile fermentation liquid and a fermentation liquid which are prepared by utilizing a stenotrophomonas gondii strain and have low production cost and application in inhibiting the formation of rice blast bacterial attachment cells and pathogenicity on rice.
In order to solve the technical problems, the invention provides a stenotrophomonas gondii strain which is stenotrophomonas gondii WXY53 (Stenotrophomonas geniculata WXY) and is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC NO: 26681, the preservation unit address is in the China academy of sciences microbiological study, and the preservation date is 2023, 2 months and 27 days, and the strain is named stenotrophomonas gondii WXY53 (Stenotrophomonas geniculata WXY).
As a general technical concept, the invention provides a bacterial suspension prepared from a stenotrophomonas gondii strain, a sterile fermentation liquor and an application of the fermentation liquor in inhibiting rice blast fungus attachment cell formation and pathogenicity on rice.
Compared with the prior art, the invention has the advantages that:
1. The stenotrophomonas gondii WXY53 has the advantages of simple production process, short culture time and low cost, and the strain can be prepared into bacterial suspension, fermentation liquor and sterile fermentation liquor required for bacteriostasis by a simple, quick and low-cost operation method;
2. the bacterial suspension, the fermentation liquor and the sterile fermentation liquor prepared by the stenotrophomonas gondii WXY53 have the characteristics of environmental friendliness, no toxicity to human and livestock, no phytotoxicity to crops, good antibacterial effect, simplicity and convenience in application and the like;
Drawings
FIG. 1 is a comparative graph of bacterial suspensions, sterile broths and broths inhibiting the formation of rice blast bacterial attachment cells in example 2 of the present invention, with 3 treatments being bacterial suspensions, sterile broths and broths, respectively, with the double star representing a significant difference (P < 0.01).
FIG. 2 is a graph showing a comparison of the pathogenicity of the bacterial suspension, sterile fermentation broth, and chemical sterilant carbendazim of example 3 of the present invention against Pyricularia oryzae on rice, with 4 treatments being bacterial suspension, sterile fermentation broth, and chemical sterilant carbendazim, respectively.
Detailed Description
The invention is further described below in connection with the drawings and the specific preferred embodiments, but the scope of protection of the invention is not limited thereby.
Example 1: preparation of stenotrophomonas gondii WXY53 bacterial suspension, sterile fermentation liquid and fermentation liquid
A strain of stenotrophomonas gondii, called stenotrophomonas gondii WXY53 (Stenotrophomonas geniculata WXY) is preserved in China general microbiological culture Collection center (CGMCC) with a preservation number of 26681 and a preservation unit address of 2023, 2 and 27 days.
In the embodiment, the suspension, the fermentation liquor and the sterile fermentation liquor of the stenotrophomonas gondii WXY53 are prepared by activating, seed culturing, production culturing and the like of the stenotrophomonas gondii WXY 53. The preparation method comprises the following steps:
(1) Activating: culturing the preserved seeds of the knee-like narrow oxygen single-cell strain according to a plate streak culture method until yellow micro-transparent single colonies appear;
(2) Seed culture: inoculating the yellow micro-transparent single colony obtained in the step (1) into an LB liquid culture medium triangular flask, culturing by shaking under the conditions that the temperature of a shaking table is 37℃, pH =7.4 and the rotating speed is 200rpm, and culturing to a logarithmic phase under the dark environment condition;
(3) And (3) production and culture: inoculating the bacterial liquid obtained after the seed culture in the step (2) into a production bottle, carrying out production culture by using an LB liquid culture medium, wherein the inoculum size of the bacterial liquid is 5% of the total volume of the production culture medium of the stenotrophomonas gondii, carrying out shaking culture at a shaking table temperature of 37℃, pH =7.4 and a rotating speed of 200rpm, and culturing to a logarithmic phase under dark environment conditions; this is the stenotrophomonas gondii fermentation broth;
(4) Preparing a bacterial suspension: centrifuging the fermentation broth obtained in the step (3) at 10000 rpm, 4 ℃ and 15: 15 mins, removing the supernatant, and suspending the thallus with clear water.
(5) Preparing a sterile fermentation liquid: and (3) centrifuging the fermentation broth obtained by the production culture in the step (3) at 10000 rpm, 4 ℃ and 15 mins, collecting the supernatant, performing suction filtration with a 0.22 mu m filter membrane to remove residual thalli in the supernatant, and collecting the filtrate to obtain the sterile fermentation broth.
Example 2: stenotrophomonas gondii WXY53 bacterial suspension, sterile fermentation liquid and fermentation liquid for inhibiting rice blast fungus attachment cell formation
Conidium of Pyricularia oryzae is collected, washed 2 times with ddH 2 O, and then the spore liquid is suspended to a certain concentration (1 x 10 5 spores/ml) by using the bacterial suspension, the sterile fermentation liquid and the fermentation liquid prepared in example 1, 30 μl is dripped on the hydrophobic membrane, and after 8 and 24 hours, the formation of attachment cells and the statistical formation rate are observed under a microscope. The formation of conidia of Pyricularia oryzae after treatment of the suspension, sterile broth and broth of stenotrophomonas gondii WXY53 was significantly inhibited (FIG. 1A), and the formation of conidia of Pyricularia oryzae after 8 hours of treatment was found to be 17.0%, 1.3% and 0.33% respectively, and found to be 29.33%, 3.0% and 2.0% respectively, which were significantly lower than that of the fresh water control (93% and 94.67%) by statistical analysis, indicating that the suspension, sterile broth and broth of WXY53 inhibit the formation of conidia of Pyricularia oryzae.
Example 3: the stenotrophomonas gondii WXY53 bacterial suspension, the sterile fermentation liquid and the fermentation liquid inhibit pathogenicity of rice blast bacteria on rice.
Healthy rice seedlings (the variety is Co-39 sensitive to rice blast bacteria) growing for about 2 weeks are selected, spores are collected from a spore-producing flat plate and added into a stenotrophomonas gonelliptica WXY53 bacterial suspension, a sterile fermentation liquid and a fermentation liquid containing 2 permillage gelatin to prepare a suspension with the final concentration of 5.0X10 4/mL, then the spore suspension is uniformly sprayed on rice leaves by using a throat sprayer, and the spore liquid suspended by clear water is used as a control. Inoculated rice was transferred to a 28 ℃ dark, high humidity (RH > 95%) vessel for cultivation 24h, and then placed in a light-dark alternating (12 h each) and high humidity vessel for further cultivation, and after 7 d the results were observed and photographed. Experimental results show that after the treatment of the stenotrophomonas gondii WXY53 bacterial suspension, the sterile fermentation liquor and the fermentation liquor, the pathogenicity of the stenotrophomonas gondii WXY53 bacterial suspension, the sterile fermentation liquor and the fermentation liquor is equivalent to that of the chemical bactericide carbendazim, and is obviously lower than that of a control (figure 2), and the WXY53 bacterial suspension, the sterile fermentation liquor and the fermentation liquor are used for inhibiting the pathogenicity of rice blast bacteria on rice.
Claims (5)
1. The stenotrophomonas gondii is stenotrophomonas gondii WXY53 (Stenotrophomonas geniculata WXY) and is preserved in China general microbiological culture Collection center with the preservation number of CGMCC NO: 26681.
2. A bacterial suspension, a sterile fermentation broth, and a fermentation broth prepared from the stenotrophomonas knee of claim 1, wherein: is mainly obtained by activating, seed culturing, producing and culturing the stenotrophomonas gondii of claim 1.
3. A process for the preparation of a bacterial suspension, a sterile fermentation broth and a fermentation broth according to claim 2 comprising the steps of:
(1) Activating: culturing the preserved strain of the stenotrophomonas gondii strain of claim 1 by a plate streak culture method until yellow micro-transparent single colony appears;
(2) Seed culture: inoculating the yellow micro-transparent single colony obtained in the step (1) into an LB liquid culture medium triangular flask, culturing by shaking under the conditions that the temperature of a shaking table is 37℃, pH =7.4 and the rotating speed is 200rpm, and culturing to a logarithmic phase under the dark environment condition;
(3) And (3) production and culture: inoculating the bacterial liquid obtained after the seed culture in the step (2) into a production bottle, carrying out production culture by using an LB liquid culture medium, wherein the inoculum size of the bacterial liquid is 5% of the total volume of the production culture medium of the stenotrophomonas gondii, carrying out shaking culture at a shaking table temperature of 37℃, pH =7.4 and a rotating speed of 200rpm, and culturing to a logarithmic phase under dark environment conditions; this is the stenotrophomonas gondii fermentation broth;
(4) Preparing a bacterial suspension: centrifuging the fermentation broth obtained in the step (3) at 10000 rpm, 4 ℃ and 15 mins, removing the supernatant, and suspending the thallus with clear water;
(5) Preparing a sterile fermentation liquid: and (3) centrifuging the fermentation broth obtained by the production culture in the step (3) at 10000 rpm, 4 ℃ and 15 mins, collecting the supernatant, performing suction filtration with a 0.22 mu m filter membrane to remove residual thalli in the supernatant, and collecting the filtrate to obtain the sterile fermentation broth.
4. A bacterial suspension, sterile fermentation broth and use of a fermentation broth according to claim 3 for inhibiting the formation of rice blast bacterial attachment cells.
5. A bacterial suspension, sterile fermentation broth and use of a fermentation broth according to claim 3 for inhibiting the pathogenicity of rice blast bacteria on rice.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410064816.4A CN118086102A (en) | 2024-01-17 | 2024-01-17 | Application of stenotrophomonas gondii in inhibiting rice blast bacteria |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410064816.4A CN118086102A (en) | 2024-01-17 | 2024-01-17 | Application of stenotrophomonas gondii in inhibiting rice blast bacteria |
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| Publication Number | Publication Date |
|---|---|
| CN118086102A true CN118086102A (en) | 2024-05-28 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202410064816.4A Pending CN118086102A (en) | 2024-01-17 | 2024-01-17 | Application of stenotrophomonas gondii in inhibiting rice blast bacteria |
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| Country | Link |
|---|---|
| CN (1) | CN118086102A (en) |
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- 2024-01-17 CN CN202410064816.4A patent/CN118086102A/en active Pending
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