Stenotrophomonas acidurans T1T2 and uses thereof

Abstract

This invention relates to an acidophilic oligotrophomonas T1T2 and its application in the degradation of organophosphorus pesticides. The acidophilic oligotrophomonas is deposited at the China Center for Type Culture Collection (CCTCC), accession number CCTCC NO: M 20242351. The acidophilic oligotrophomonas T1T2 of this invention can effectively degrade trichlorfon residues on crops, achieving a degradation efficiency of over 75% on chili peppers. It is harmless to humans and animals and causes no environmental pollution, greatly improving the safety of agricultural products for consumption. Therefore, it has broad application prospects in the safe production of agricultural products.

Description

Technical Field

[0001] This invention relates to the fields of pesticide residue degradation and microbial technology, and in particular to the acidophilic oligotrophomonas T1T2 and its applications. Background Technology

[0002] Spraying pesticides is the most common and direct method of controlling pests and diseases in agriculture. Currently, approximately 2 million tons of pesticides are used annually worldwide to improve productivity and reduce losses caused by pests and diseases. Among them, organophosphorus pesticides are widely used in agricultural production due to their high efficiency and broad insecticidal spectrum. As the mainstay of the pesticide industry, organophosphorus pesticides rank first in terms of the number of varieties, output, and market share. Organophosphorus pesticides mainly include phosphate esters, pyrophosphate esters, or thioester compounds, possessing strong insecticidal and fungicidal effects. Among these, trichlorfon, as an early introduced organophosphorus pesticide product, is widely used by farmers. However, due to over-application, only 10% to 20% of the pesticide actually kills pests and diseases; the majority of the pesticide remains on the surface of crops or seeps into the soil, water, and air. Consuming crops with pesticide residues can lead to the accumulation of these substances in the body. Repeated exposure can cause a range of neurotoxic symptoms, such as sweating, tremors, mental confusion, and speech disorders. In severe cases, respiratory paralysis and even death can occur. Furthermore, over time, pesticides and their metabolites enter surface and groundwater, accumulating in livestock and humans through the food chain, further enhancing their toxicity. Therefore, pesticide residues in crops have become a significant concern for both food safety and environmental safety.

[0003] The main methods for degrading pesticide residues are natural degradation and chemical degradation. Natural degradation has drawbacks such as a long reaction process, slow onset of action, long safety isolation period for crops, and poor overall effectiveness. Therefore, chemical degrading agents are usually used to accelerate degradation. For example, existing technologies use titanium dioxide as a degrading agent, which can hydrolyze organophosphorus pesticides into water, carbon dioxide, and phosphate ions. However, the use of chemical degrading agents requires the introduction of large amounts of chemical substances that are not originally present in the farmland environment. The long-term impact of these chemicals on the ecological environment is unclear, posing certain risks. Summary of the Invention

[0004] Based on this, the purpose of the present invention is to provide a novel acidophilic oligotrophomonas T1T2 (Stenotrophomonas acidaminiphila), which can effectively degrade organophosphorus pesticide residues, especially trichlorfon residues, thereby achieving the goal of degrading pesticide residues in crops and improving crop safety.

[0005] The specific technical solution is as follows:

[0006] A type of acidophilic oligotrophomonas T1T2 has been deposited at the China Center for Type Culture Collection (CCTCC; address: China Center for Type Culture Collection, Wuhan University, Wuhan, Hubei Province, China), with accession number CCTCC NO: M20242351 and deposit date: October 28, 2024.

[0007] Compared to existing technologies, this invention provides a novel acidophilic oligotrophomonas T1T2, which can effectively degrade trichlorfon residues in crops, achieving a degradation rate of over 75% on chili peppers. When used in chili pepper cultivation, it can effectively reduce trichlorfon residues and shorten the safe isolation period for chili peppers. The acidophilic oligotrophomonas T1T2 of this invention has broad application prospects in degrading organophosphorus pesticide residues.

[0008] Furthermore, the application of the acidophilic oligotrophomonas T1T2 in pesticide degradation.

[0009] Furthermore, the pesticide is an organophosphorus pesticide.

[0010] Furthermore, the organophosphorus pesticide includes trichlorfon.

[0011] The present invention also provides a pesticide degradation agent comprising the aforementioned acidophilic oligotrophomonas T1T2.

[0012] Furthermore, the concentration of the acidophilic oligotrophomonas T1T2 in the degrading agent is 1.0 × 10⁻⁶. 4 cfu / ml ~1.0×10 5 The cfu / ml of the degrading agent is used to degrade organophosphorus pesticides.

[0013] Furthermore, the method for preparing the degradation agent includes: inoculating the acidophilic oligotrophoblast T1T2 into a culture medium, and culturing it under constant temperature shaking conditions of 30℃~37℃ and 180~220rpm to prepare a fermentation broth, and then adjusting the concentration of acidophilic oligotrophoblast T1T2 in the fermentation broth to prepare the degradation agent.

[0014] The present invention also provides a method for degrading plant residues of organophosphorus pesticides, comprising the following steps: applying the degradation agent to the plant.

[0015] Furthermore, the organophosphorus pesticide includes trichlorfon, and is applied by spraying.

[0016] Furthermore, the amount of degradation agent applied to each plant at one time is 10–30 ml. Attached Figure Description

[0017] Figure 1This is a colony morphology diagram of the acidophilic oligotrophomonas T1T2 described in this invention after culturing on TTC agar solid medium for 24 hours;

[0018] Figure 2 This is an electrophoresis image of the PCR amplification product of the 16S rDNA sequence of Oligotrophomonas acidophilus T1T2 described in this invention; the left band is the nucleic acid standard band, from top to bottom: 2000bp, 1000bp, 750bp, 500bp, 250bp, 100bp; the right band is the PCR amplification product band.

[0019] Figure 3 A phylogenetic tree constructed from the 16S rDNA sequences of the acidophilic oligotrophomonas T1T2 described in this invention and similar bacterial species;

[0020] Figure 4 The graph shows the high performance liquid chromatography (HPLC) results of the standard solution of trichlorfon acetonitrile in S403.

[0021] Figure 5 The image shows the high-performance liquid chromatography (HPLC) results of the supernatant of the mixture inoculated with activated bacterial solution in the experimental group of S403.

[0022] Figure 6 This is a high-performance liquid chromatography (HPLC) result of the supernatant of the mixture of the control group and the uninoculated activated bacterial solution in S403. Detailed Implementation

[0023] To accelerate the degradation of organophosphate pesticides without introducing substances not native to the farmland environment, this invention aims to utilize biodegradation methods to degrade organophosphate pesticides, such as trichlorfon. Bacteria are organisms with exceptional biodegradation potential. Due to their rapid reproduction rate, they can quickly adapt to their environment and may even possess specialized functions. In soil environments, if organophosphate pesticides have been used, the bacteria living in that soil are likely to possess the ability to tolerate and even degrade these pesticides. Therefore, bacteria already existing in the soil have a significant potential to degrade pesticide residues on crops. Furthermore, they may more easily colonize farmland soil environments and continuously contribute to the degradation of pesticides in the soil, demonstrating great potential for environmental protection.

[0024] Therefore, this invention aims to isolate, purify, and screen soil bacteria capable of degrading organophosphorus pesticides from soil that has been treated with such pesticides. Specifically, this invention first selects non-pathogenic bacteria that are already widely present in the soil and applies these bacteria to pesticide degradation, comparing their ability to degrade organophosphorus pesticides. Ultimately, this invention selects an *Oligotrophomonas acidophilus* strain, which exhibits excellent organophosphorus pesticide degradation ability. Furthermore, this *Oligotrophomonas acidophilus* is a common soil bacterium and is non-pathogenic. In addition, some strains of *Oligotrophomonas acidophilus* can secrete antifungal compounds, synthesize antipathogenic extracellular enzymes, and reduce plant disease incidence, which is extremely beneficial for crop cultivation.

[0025] The following, in conjunction with the accompanying drawings and specific embodiments, further details the application method of the present invention, namely, the isolation and purification method of *Oligotrophomonas acidophilus* T1T2, the liquid culture method of *Oligotrophomonas acidophilus* T1T2, the identification of *Oligotrophomonas acidophilus* T1T2, the degradation effect of *Oligotrophomonas acidophilus* T1T2 on trichlorfon in shake flasks, and the degradation effect of *Oligotrophomonas acidophilus* T1T2 on residual pesticides on plant leaves.

[0026] Isolation and purification methods of Oligotrophic acidophilus T1T2

[0027] The method for isolating and purifying the acidophilic oligotrophomonas T1T2 strain of the present invention comprises the following steps:

[0028] S101: Soil dilution: Weigh 10g of soil sample, which is from ginger-growing areas in Xuwen County, Zhanjiang City, Guangdong Province; then, in a clean bench, add 90ml of sterile water to the soil sample and shake on a shaker for 60min to evenly disperse the soil sample in the diluent to form a soil suspension; after the soil is dispersed, take 100μl of the soil suspension into 900μl of sterile water and shake to mix well to obtain a 10-fold dilution, and then dilute it 10 times in sequence to obtain a 10-fold dilution. 2 10 times dilution solution 3 10 times dilution solution 4 10 times dilution solution 5 10 times dilution and 10 6 The solution was diluted multiple times, and the entire process was carried out in a clean bench.

[0029] S102: Strain culture: Take 100 μl of a 10-fold dilution, 10 2 10 times dilution solution 3 10 times dilution solution 4 10 times dilution solution 5 10 times dilution and 10 6The diluted solution was spread onto petri dishes containing TTC agar solid medium, and then the petri dishes were placed in an incubator at 28°C for 2-3 days until single colonies grew on the culture plates.

[0030] S103: Screening and purification of strains: After the culture is completed, single colonies of different shapes, colors and sizes are picked from appropriate dilution gradient plates according to their growth. The plates are then streaked again on culture dishes containing TTC agar solid medium. The culture dishes are then placed in an incubator at 28°C for 2-3 days until single colonies grow on the culture plate. The single colonies that grow are the isolated and purified acidophilic oligotrophomonas strain T1T2.

[0031] S104: Preservation of *Oligotrophomonas acidophilus* strain T1T2: The isolated and purified *Oligotrophomonas acidophilus* strain T1T2 was picked from a petri dish containing TTC agar solid medium and inoculated into a 500ml Erlenmeyer flask containing 200ml of NA liquid medium. The flask was cultured at a constant temperature of 30℃ and with shaking at 200rpm for 36 hours to obtain a seed culture. This seed culture was then mixed with a 25% sterile glycerol aqueous solution at a volume ratio of 1:4 to obtain a glycerol seed culture containing 20% ​​glycerol. After marking the tube walls, the culture was stored at -20℃ or -80℃ for long-term preservation, thus completing the strain preservation.

[0032] The morphological image of the *Oligotrophomonas acidophilus* strain T1T2 isolated in this embodiment after 24 hours of culture on a TTC agar plate is shown below. Figure 1 As shown, the colonies are orange and round, with a soft texture and a smooth surface.

[0033] The preparation method of TTC agar solid medium is as follows: Weigh 10g of tryptone, 3g of beef extract powder, 5g of sodium chloride, 15g of agar, and 0.01g of tetrazolium red (TTC). Dissolve all components in distilled water and bring the volume to 1000ml. Then dispense the solution into Erlenmeyer flasks, seal the flasks with sterile paper, and sterilize them in an autoclave at 121℃ and 101KPa for 20min. Before solidification, pour approximately 20ml of the sterilized medium into petri dishes in a laminar flow hood. After the medium cools and solidifies, you will obtain petri dishes containing TTC agar solid medium, which should be stored at 4℃.

[0034] The preparation method for NA liquid culture medium is as follows: Weigh out 10g of tryptone, 3g of beef extract, and 5g of sodium chloride. Dissolve each component in distilled water by stirring, adjust the pH to 7.0-7.2, and bring the volume to 1000ml. Dispense the solution into Erlenmeyer flasks, seal the flasks with sterile paper, and autoclave at 121℃ and 101KPa for 20min.

[0035] Liquid culture method for Oligotrophomonas acidophilus T1T2

[0036] Oligotrophic acidophilus T1T2 can be cultured in NA liquid medium or LB liquid medium, which will be described below:

[0037] Culture method of Oligotrophic acidophilus T1T2 using NA liquid medium

[0038] S201A: Strain Activation: The glycerol seed culture stored at -20℃ in S104 was inoculated into NA liquid medium at a volume ratio of glycerol seed culture to NA liquid medium of 1:100. The medium was then incubated at a constant temperature of 30℃ with shaking at a speed between 180 rpm and 220 rpm for 36 hours to obtain activated bacterial culture. In this embodiment, the shaking speed is preferably 200 rpm.

[0039] S202A: Liquid culture: Take the activated bacterial solution and inoculate it into 500ml of NA liquid culture medium at a volume ratio of 1:100 (spark bacterial solution to NA liquid culture medium). Then, carry out isothermal shaking culture at 30℃ and a shaking speed of 180rpm-220rpm (preferably 200rpm in this embodiment) for 24h-48h (preferably 36h in this embodiment) to obtain a high-density bacterial suspension.

[0040] Culture method of Oligotrophic acidophilus T1T2 using LB liquid medium

[0041] S201B: Strain activation: Take the glycerol seed culture stored at -20℃ in S104 and inoculate it into LB liquid medium at a volume ratio of 1:100 (glycerol seed culture to LB liquid medium). Incubate at 37℃ and 200 rpm for 36 hours to obtain activated bacterial culture.

[0042] S202B: Liquid culture: Take the activated bacterial solution and inoculate it into 500ml of LB liquid medium at a volume ratio of 1:100 between the activated bacterial solution and LB medium. Then, carry out constant temperature shaking culture at 37℃ and a shaking speed of 180rpm to 220rpm (preferably 200rpm in this embodiment) for 24h to 48h (preferably 36h in this embodiment) to obtain a high-density bacterial suspension.

[0043] The preparation method of LB liquid culture medium is as follows: weigh 10g of tryptone, 5g of yeast extract and 10g of sodium chloride, stir and dissolve in distilled water, adjust the pH to 7.2-7.4 and make up to 1000ml, dispense into conical flasks and autoclave at 121℃ and 101KPa for 20min.

[0044] Species identification of Oligotrophomonas acidophilus T1T2

[0045] S301: Extracting bacterial genomic DNA

[0046] Genomic DNA was extracted using the Omega Bacterial DNA Kit (D3350-01). First, 2 ml of the seed culture described in S104 was placed in a sterile 2 ml centrifuge tube and centrifuged at 12000 rpm for 2 min. The supernatant was discarded, and the precipitate was retained. Then, 100 μl of 1×TE Buffer was added to the precipitate, vortexed, and 10 μl of lysozyme was added and mixed. The mixture was incubated at 37°C for 10 min. Next, 100 μl of BTL Buffer and 20 μl of proteinase K were added, mixed, and incubated at 55°C for 1 h, vortexing three times during incubation. Finally, 5 μl of RNase A was added, mixed, and allowed to stand at room temperature for 5 min. The mixture was then centrifuged at 10000 rpm for 2 min, and 200 μl of the supernatant was transferred to a new sterile 1.5 ml centrifuge tube. 200 μl of BTL Buffer was added... Add buffer, mix well, and incubate at 65℃ for 10 min; add 200 μl of anhydrous ethanol, vortex to mix, transfer all sample to the adsorption column, centrifuge at 10000 rpm for 2 min, discard the supernatant and adsorption column, and place the adsorption column into a new collection tube; add 500 μl of HBC buffer to the adsorption column, centrifuge at 10000 rpm for 2 min, and discard the supernatant; add 700 μl of DNA Wash Buffer to the adsorption column, centrifuge at 10000 rpm for 2 min, discard the supernatant, and repeat twice; place the empty adsorption column back into the collection tube, centrifuge at 10000 rpm for 2 min; add 30 μl to 50 μl of Elution Buffer (preheated at 65℃) to the adsorption column to dissolve the DNA precipitate, thus obtaining bacterial genomic DNA, and store at -20℃ for later use.

[0047] S302: Amplification of 16S rDNA sequence via polymerase chain reaction (PCR)

[0048] Using the genomic DNA obtained in step S301 as a template, PCR amplification was performed using Eubac27F and Eubac1492R primers. The PCR reaction system (50 μl) included: 0.25 μl TaKaRa LA Taq (5 U / μl), 2.5 μl 10×LA Taq Buffer II (Mg2+). 2+Plus), 4 μl dNTPs Mixture (2.5 mM each), 1 μl genomic DNA, 0.5 μl upstream primer Eubac27F (10 μM), 0.5 μl downstream primer Eubac1492R (10 μM), and 16.25 μl ddH2O. The sequence of the upstream primer Eubac27F is 5'-agagtttgat cctggctcag-3' (SEQ ID NO: 1); the sequence of the downstream primer Eubac1492R is 5'-ggttaccttg ttacgactt-3' (SEQ ID NO: 2).

[0049] The PCR reaction procedure was as follows: pre-denaturation at 94℃ for 5 min; denaturation at 94℃ for 30 s, annealing at 55℃ for 30 s, extension at 72℃ for 1 min, and repeated the denaturation, annealing and extension processes 30 times; extension at 72℃ for another 10 min, and storage at 4℃ to obtain the PCR product.

[0050] S303: Perform nucleic acid electrophoresis on PCR products.

[0051] Take 5 μl of the PCR product obtained in step S302, spot it onto a 1.2% agarose gel, and perform nucleic acid electrophoresis at 120V for 25 min. The electrophoresis results are as follows. Figure 2 As shown, the 16S rDNA fragment amplified using the genomic DNA of Oligotrophomonas T1T2 obtained in this invention as a template has a single and high-brightness band, and the 16S rDNA sequence length is approximately 1500 bp.

[0052] S304: 16S rDNA sequence sequencing

[0053] Take 30 μl of the PCR product obtained in step S302 and send it to Guangzhou Qingke Biotechnology Co., Ltd. for bidirectional sequencing. The sequencing results show that the 16S rDNA sequence of the Oligotrophomonas acidophilus strain described in this invention is 1404 bp in length, and the specific sequence is shown in SEQ ID NO: 3.

[0054] The 16S rDNA sequence (SEQ ID NO: 3) of the *Oligotrophomonas acidophilus* strain obtained from sequencing was input into NCBI for BLAST alignment. Sequences with high similarity to SEQ ID NO: 3 were downloaded, and a phylogenetic tree based on SEQ ID NO: 3 and the downloaded 16S rDNA segment sequence was constructed using the Neighbor-Joining Method (NJ) in MEGA11 software. The results are as follows: Figure 3As shown. According to the phylogenetic tree, the 16S rDNA sequence of the *Stenotrophomonas acidaminiphila* strain T1T2 isolated and purified in this invention has a 99% sequence similarity to that of *Stenotrophomonas acidaminiphila* strain A2 (Accession No. OQ244463.1). Based on the phylogenetic tree and sequence similarity of the 16S rDNA sequence, the strain isolated in this invention belongs to the *Stenotrophomonas acidaminiphila* species, and is named *Stenotrophomonas acidaminiphila* T1T2 in this invention.

[0055] The effect of Oligotrophozoites acidophilus T1T2 on the degradation of trichlorfon in shake flasks

[0056] S401: Strain activation: Take the glycerol seed culture stored at -20℃ in S104 and inoculate it into NA liquid culture medium at a volume ratio of glycerol seed culture to NA liquid culture medium of 1:100. Incubate at 30℃ and shaking speed of 200rpm for 36h to obtain activated bacterial culture.

[0057] S402 inoculation: Under aseptic conditions, trichlorfon was added to MSM liquid medium to a final concentration of 500 mg / L. In the experimental group, the activated bacterial solution from S401 was added to MSM liquid medium containing 500 mg / L trichlorfon solution at a ratio of 1:20 and cultured at a constant temperature of 30℃ and a shaking speed of 200 rpm for 24 h to obtain a mixture inoculated with activated bacterial solution. In the control group, NA medium was added to MSM liquid medium containing 500 mg / L trichlorfon solution at a ratio of 1:20 and cultured under the same conditions as the experimental group for 24 h to obtain a mixture without activated bacterial solution.

[0058] S403 Sampling: 2 ml of supernatant was taken from the mixture of the experimental group inoculated with activated bacterial solution and the mixture of the control group without activated bacterial solution. After filtration through a 0.22 μm filter membrane, the mixture was analyzed by high performance liquid chromatography. The standard sample was a 500 mg / L trichlorfon acetonitrile solution. Chromatographic conditions: methanol:water (70:30, V / V) was used as the mobile phase, the wavelength was 210 nm, and the flow rate was 1.0 mL / min with an equal gradient elution.

[0059] The results showed that the peak diagram of the standard sample indicated that the characteristic peak of trichlorfon appeared at approximately 11.8 min. Figure 4 The experimental group was inoculated with a mixture containing activated bacterial solution, and the characteristic peak of trichlorfon was observed. Figure 5 The characteristic peak of trichlorfon in the mixture of the control group and the uninoculated control group ( Figure 6The peak height decreased and the peak area decreased by 76.80% compared to the previous peak, indicating that the trichlorfon content decreased by approximately 76.80%.

[0060] The above results indicate that the acidophilic oligotrophomonas T1T2 has the function of degrading trichlorfon, which can be used to reduce trichlorfon residues in crops and shorten the safe isolation period of crops.

[0061] The preparation method for MSM liquid culture medium is as follows: Weigh 1g of ammonium nitrate, 0.5g of magnesium sulfate heptahydrate, 0.5g of ammonium sulfate, 0.5g of potassium dihydrogen phosphate, 0.5g of sodium chloride, and 1.5g of dipotassium hydrogen phosphate. Dissolve them in distilled water by stirring, adjust the pH to 7.0-7.2, and bring the volume to 1000ml. Dispense the solution into Erlenmeyer flasks, seal the flasks with sterile paper, and then autoclave them at 121℃ and 101KPa for 20min.

[0062] Effects of Oligotrophic acidophilus T1T2 on the degradation of pesticide residues on plant leaves

[0063] S501 strain activation: Take the glycerol seed solution stored at -20℃ as described in S104 and inoculate it into NA liquid culture medium at a volume ratio of glycerol seed solution to NA liquid culture medium of 1:100. Place it in a constant temperature shaker and culture at 30℃ for 36 hours to obtain activated bacterial solution.

[0064] Preparation of fermentation broth for S502 acidophilic oligotrophomonas T1T2: The activated broth obtained from S501 was inoculated into NA liquid medium at a volume ratio of 1:100 (activated broth to NA liquid medium). Fermentation was carried out at 30℃ and 200 rpm with constant temperature shaking until the bacterial density reached 1.0 × 10⁻⁶. 5 cfu / ml ~1.0×10 6 Stop culturing at cfu / ml and obtain fermentation broth;

[0065] S503 fermented bacterial solution was sprayed onto chili pepper leaves to degrade residual trichlorfon: In the experimental group, the fermented bacterial solution obtained from S502 was diluted 10 times to prepare a degradation agent, which was then sprayed onto the chili pepper leaves at a dosage of 20 ml per plant. The control group directly sprayed the leaves with water without fermented bacterial solution using the same method and dosage. The degradation effect test in this step was conducted at the South China Agricultural University Experimental Base in Ningxi, Zengcheng District, Guangzhou City, in an experimental greenhouse. The experiment was performed in three biological replicates, with each experimental group and control group consisting of 8 chili pepper plants with a growth period of 90 days. Forty-eight hours before spraying the degradation agent and water, a 2.0 g / L trichlorfon aqueous solution was sprayed evenly onto the leaf surface of both the experimental and control groups at a dosage of 20 ml per plant.

[0066] S504 leaf trichlorfon residue detection: 48 hours after spraying the degradation agent, 2.0 g of leaves from each chili pepper plant were extracted with 10 ml of acetonitrile solution using ultrasonic extraction at 150 W for 3 minutes. The extracts were mixed according to the groups and analyzed by high-performance liquid chromatography (HPLC). Chromatographic conditions: methanol:water (70:30, V / V) as the mobile phase, wavelength 210 nm, flow rate 1.0 mL / min, and isogradient elution. The characteristic peak area of ​​trichlorfon in the experimental group was 18.3 ± 5.3 mAU*s, and the characteristic peak area of ​​trichlorfon in the control group was 77.1 ± 8.3 mAU*s. The trichlorfon degradation rate was calculated according to the formula: Therefore, the degradation rate of trichlorfon in the experimental group was calculated to be over 75%.

[0067] The above results indicate that the degradation agent containing the fermentation broth of Oligotrophomonas acidophilus T1T2 has the function of degrading trichlorfon on plant leaves, and can be used to reduce trichlorfon residues in crops and shorten the safe isolation period of crops.

[0068] In summary, compared with the prior art, the present invention has the following advantages and effects:

[0069] 1. Oligotrophomonas acidophilus T1T2 has a good effect on degrading organophosphorus pesticides. On the leaves of crops such as chili pepper, the degradation rate of organophosphorus pesticide trichlorfon reaches more than 75%, so it can be used for the degradation of pesticide residues in plants.

[0070] 2. The application of Oligotrophomonas acidophilus T1T2 in pesticide degradation has the advantages of rapid effect and good environmental compatibility. It can effectively shorten the waiting time for pesticide natural degradation, improve production efficiency, and also ensure food safety.

[0071] 3. The degradation agent containing Oligotrophic acidophilus T1T2 has a simple formulation and a simple fermentation culture method, requiring no special equipment or harsh culture conditions, thus resulting in low production costs.

[0072] 4. Oligotrophomonas acidophilus T1T2, as a soil-derived beneficial crop bacterium, can not only degrade pesticide residues on crops, but also more easily colonize the soil after falling into the ground, continuously degrading pesticide residues in the soil and being environmentally friendly.

[0073] 5. Even after rinsing with water, some residue of biodegrading agents on crops is unavoidable. Compared to the stability of chemical biodegrading agents, biodegrading agents can be killed by cooking and are virtually harmless to humans, thus making them safer.

[0074] Experimental methods in the embodiments of this invention that do not specify specific conditions are generally performed under conventional conditions or as recommended by the manufacturer. All commonly used chemical reagents used in the embodiments are commercially available products. Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to limit the invention.

[0075] The embodiments described above are merely examples of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention.

Claims

1. An acidophilic oligotrophomonas ( Stenotrophomonas acidaminiphila T1T2, characterized in that: The Stenotrophomonas acidaminiphila T1T2 is preserved in China Center for Type Culture Collection, and the preservation number is CCTCC NO: M20242351.

2. The Stenotrophomonas acidaminiphila T1T2 of claim 1 is applied in pesticide degradation, and the pesticide is an organophosphorus pesticide, and the organophosphorus pesticide is trichlorphon.

3. A degrading agent for degrading a pesticide, characterized by comprising: The degradation agent contains the Stenotrophomonas acidaminiphila T1T2 of claim 1, and the pesticide degradable by the degradation agent is trichlorphon.

4. The degrading agent according to claim 3, characterized by: The concentration of the degrader is 1.0 x 10 4 cfu / ml ~ 1.0 x 10 5 cfu / ml.

5. The degrading agent according to claim 3, wherein: The preparation method of the degradation agent comprises the following steps: inoculating the Stenotrophomonas acidaminiphila T1T2 into a culture medium, and culturing at 30-37 DEG C and 180-220 rpm to prepare a fermentation broth, and then adjusting the concentration of the Stenotrophomonas acidaminiphila T1T2 in the fermentation broth to prepare the degradation agent.

6. A method for degrading an organophosphorus pesticide, characterized by, The organophosphorus pesticide is trichlorphon, and the degradation method comprises the following steps: applying the degradation agent containing the degradation agent of claim 3 on plants by spraying.

7. The degradation method of claim 6, wherein: The single application amount of the degradation agent for each plant is 10-30 ml.

Images