Stenotrophomonas acidophilus T1T2 and application thereof

Abstract

The invention relates to stenotrophomonas acidophilus T1T2 and application of the stenotrophomonas acidophilus T1T2 in degradation of organophosphorus pesticides. The stenotrophomonas acidophilus is preserved in the China Center for Type Culture Collection, and the preservation number of the stenotrophomonas acidophilus is CCTCC NO: M 20242351. The stenotrophomonas acidophila T1T2 can effectively degrade dipterex residues on crops, the degradation efficiency of dipterex on peppers reaches 75% or above, and the stenotrophomonas acidophila T1T2 is harmless to people and livestock and free of pollution to the environment, so that the food safety of the crops is greatly improved, and therefore the stenotrophomonas acidophila T1T2 has wide application prospects in the aspect of safe production of agricultural products.

Description

Technical Field

[0001] The invention relates to the fields of pesticide residue degradation and microbial technology, in particular to acidophilic oligotrophic monosaccharides T1T2 and applications thereof. Background Art

[0002] Spraying pesticides is the most commonly used and most direct method of preventing and controlling pests and diseases in agriculture. At present, about 2 million tons of pesticides are used for planting every year in the world to improve productivity and reduce losses caused by pests and diseases. Among them, organophosphorus pesticides have been widely used in agricultural production due to their high efficiency and wide insecticide spectrum. As the main body of the pesticide industry, organophosphorus pesticides rank first among all pesticides in terms of the number of varieties, output and market share. Organophosphorus pesticides mainly include phosphates, pyrophosphates or sulfide compounds, which have strong insecticidal and bactericidal effects. Among them, trichlorfon, as an early organophosphorus pesticide product, is widely used by farmers. However, due to excessive application of pesticides, the amount of pesticides actually used to kill pests and diseases only accounts for 10% to 20% of the amount applied, and most of the remaining pesticides remain on the surface of crops or penetrate into the soil, water and air. When humans eat crops with pesticide residues, related substances will accumulate in the body. Especially after repeated exposure, a series of neurotoxic symptoms will appear, such as sweating, tremors, mental confusion, speech disorders, and in severe cases, respiratory paralysis and even death. At the same time, as time goes by, pesticides and their metabolites enter surface water and groundwater, and accumulate in livestock and humans through the food chain, further enhancing their toxic effects on humans. As a result, the problem of pesticide residues in crops has also become a food safety and environmental safety issue that has attracted much attention.

[0003] The main methods for degrading pesticide residues are natural degradation and chemical degradation. The natural degradation method has the disadvantages of a long reaction process, slow effect, long crop safety isolation period, and poor effect. Therefore, chemical degradation agents are usually used to accelerate degradation. For example, the prior art uses titanium dioxide as a degradation agent, which can hydrolyze organophosphorus pesticides into water, carbon dioxide, and phosphate ions. However, the use of chemical degradation agents requires the introduction of a large amount of chemical substances that do not originally exist in the farmland environment. The subsequent long-term impact of these chemical substances on the ecological environment is unclear, and there are certain risks. Summary of the invention

[0004] Based on this, the purpose of the present invention is to provide a new acidophilic oligotrophic mononas T1T2 (Stenotrophomonas acidaminiphila), which can effectively degrade organophosphorus pesticide residues, especially has a good degradation effect on trichlorfon residues, so as to achieve the purpose of degrading pesticide residues in crops and improving crop safety.

[0005] The specific technical solution is:

[0006] A kind of Stenotrophomonas acidaminiphila T1T2 (Stenotrophomonas acidaminiphila T1T2), deposited in China Center for Type Culture Collection (abbreviated as: CCTCC; address: China Center for Type Culture Collection, Wuhan University, Wuhan City, Hubei Province, China), with the deposit number of CCTCC NO:M20242351 and the deposit date of October 28, 2024.

[0007] Compared with the prior art, the present invention provides a new acidophilic oligotrophic mononas T1T2, which can effectively degrade the trichlorfon residues in crops, and the degradation rate on peppers is as high as 75% or more. When used in pepper planting, it can effectively reduce the trichlorfon residues in peppers and shorten the safe isolation period of peppers. The acidophilic oligotrophic mononas T1T2 of the present invention has broad application prospects in degrading organophosphorus pesticide residues.

[0008] Furthermore, the acidophilic oligotrophic mononas T1T2 is used in the degradation of pesticides.

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

[0010] Furthermore, the organophosphorus pesticide includes trichlorfon.

[0011] The present invention also provides a degradation agent capable of degrading pesticides, comprising the acidophilic oligotrophic mononas T1T2.

[0012] Furthermore, the concentration of the acidophilic oligotrophic mononas T1T2 in the degradation agent is 1.0×10 4 cfu / ml~1.0×10 5 cfu / ml, and the pesticide that can be degraded by the degradation agent is an organophosphorus pesticide.

[0013] Furthermore, the preparation method of the degradation agent includes: inoculating the acidophilic oligotrophic mononas T1T2 into a culture medium, culturing with constant temperature shaking at 30°C to 37°C and 180 to 220 rpm to prepare a fermentation liquid, and then adjusting the acidophilic oligotrophic mononas T1T2 concentration of the fermentation liquid to prepare the degradation agent.

[0014] The present invention also provides a method for degrading organic phosphorus pesticides remaining in plants, comprising the following steps: applying the degradation agent on the plants.

[0015] Furthermore, the organophosphorus pesticide includes trichlorfon, and the application method is spraying.

[0016] Furthermore, the single application amount of the degradation agent to each plant is 10 to 30 ml. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1This is a colony morphology diagram of the acidophilic oligotrophic mononas T1T2 described in the present invention after being cultured on TTC agar solid medium for 24 hours;

[0018] Figure 2 The figure is a nucleic acid electrophoresis diagram of the PCR amplification product of the 16S rDNA sequence of the acidophilic oligotrophic mononas T1T2 of the present invention; the left band is the nucleic acid standard band, which is 2000bp, 1000bp, 750bp, 500bp, 250bp, and 100bp from top to bottom; the right band is the band of the PCR amplification product;

[0019] Figure 3 A phylogenetic tree constructed from the 16S rDNA sequences of the acidophilic oligotrophic mononas T1T2 and similar species of bacteria;

[0020] Figure 4 The figure is the detection result of the high performance liquid chromatography of the trichlorfon acetonitrile solution standard solution in S403;

[0021] Figure 5 This is a graph showing the results of high performance liquid chromatography of the supernatant of the mixed solution inoculated with the activated bacterial solution in the experimental group in S403;

[0022] Figure 6 This is a graph showing the results of high performance liquid chromatography of the supernatant of the mixed solution of the control group in S403 that was not inoculated with the activated bacterial solution. DETAILED DESCRIPTION

[0023] In order to accelerate the degradation of organophosphorus pesticides without introducing additional substances that do not belong to the farmland environment, the present invention hopes to use a biodegradation method to degrade organophosphorus pesticides represented by trichlorfon. Bacteria are a type of organism with great biodegradation potential. Due to their fast reproduction rate, they can usually adapt quickly to the environment in which they live, and may even have special functions. In the soil environment, if the soil has been used with organophosphorus pesticides, then the bacteria living in the soil there are likely to have the ability to tolerate organophosphorus pesticides or even degrade organophosphorus pesticides. Therefore, bacteria that originally live in the soil have great potential to degrade residual pesticides on crops. At the same time, it may also be easier to colonize in the farmland soil environment, and continue to play a role in degrading pesticides in the soil, which has great potential for protecting the environment.

[0024] Therefore, the present invention hopes to separate, purify and screen soil bacteria that can degrade organophosphorus pesticides from soil to which organophosphorus pesticides have been applied. Specifically, the present invention first selects strains that are already widely present in the soil and are non-pathogenic, and applies these strains to the degradation of pesticides to compare their ability to degrade organophosphorus pesticides. Finally, the present invention selects a strain of acidophilic oligotrophic mononas, which has good organophosphorus pesticide degradation ability, and the acidophilic oligotrophic mononas is a common soil bacterium without pathogenicity. In addition, some strains of acidophilic oligotrophic mononas can secrete antifungal compounds, synthesize anti-pathogenic extracellular enzymes, and reduce plant morbidity, which is extremely beneficial to crop planting.

[0025] The following, in conjunction with the accompanying drawings and specific embodiments, introduces the separation and purification method of acidophilic oligotrophic mononas T1T2, the liquid culture method of acidophilic oligotrophic mononas T1T2, the strain identification of acidophilic oligotrophic mononas T1T2, the application effect of acidophilic oligotrophic mononas T1T2 on trichlorfon in a shake flask, and the application effect of acidophilic oligotrophic mononas T1T2 on residual pesticides on plant leaves, to further explain in detail the acidophilic oligotrophic mononas T1T2 of the present invention and its application method for degrading organophosphorus pesticides represented by trichlorfon.

[0026] Method for separation and purification of acidophilic oligotrophic mononas T1T2

[0027] The method for separating and purifying the acidophilic oligotrophic mononas T1T2 strain of the present invention comprises the following steps:

[0028] S101: diluting the soil: weighing 10 g of soil sample, the soil sample is from the soil of the ginger planting area in Xuwen County, Zhanjiang City, Guangdong Province; then, in an ultra-clean workbench, adding 90 ml of sterile water to the soil sample, placing it on an oscillator and oscillating for 60 minutes, so that the soil sample is evenly dispersed in the diluent to form a soil suspension; after the soil is dispersed, 100 μl of the soil suspension is drawn into 900 μl of sterile water and shaken to obtain a 10-fold dilution, and then diluting it 10 times in sequence to obtain 10 2 Dilution, 10 3 Dilution, 10 4 Dilution, 10 5 times dilution and 10 6 The whole process was carried out in an ultra-clean workbench.

[0029] S102: strain culture: take 100 μl of 10-fold dilution, 10 2 Dilution, 10 3 Dilution, 10 4 Dilution, 10 5 times dilution and 10 6The 100-fold dilutions were spread on culture plates containing TTC agar solid culture medium, and then the culture plates were placed in a 28°C incubator for constant temperature culture for 2 to 3 days until single colonies grew on the culture plates.

[0030] S103: Screening, separation and purification of strains: After the cultivation is completed, single colonies of different shapes, colors, sizes, etc. are picked one by one from the appropriate dilution gradient plates according to the growth of single colonies, and streaked again on the culture dishes containing TTC agar solid culture medium, and then the culture dishes are placed in a 28°C incubator for constant temperature cultivation for 2 to 3 days until single colonies grow on the culture plate. The grown single colonies are the isolated and purified Acidophilus Oligotrophomonas T1T2 strains.

[0031] S104: Preservation of the strain of Acidophilus Oligotrophomonas T1T2: The isolated and purified Acidophilus Oligotrophomonas T1T2 strain was picked from a culture dish containing TTC agar solid culture medium, inoculated into a 500ml conical flask containing 200ml NA liquid culture medium, and cultured for 36 hours under the culture conditions of constant temperature of 30°C and shaking at 200rpm to obtain a seed solution. The seed solution and 25% sterile glycerol aqueous solution were pipetted and mixed in a volume ratio of 1:4 to obtain a glycerol seed solution containing 20% ​​glycerol. After marking on the tube wall, it was stored at -20°C, or stored at -80°C for a long time to complete the strain preservation.

[0032] The morphology of the Acidophilus Oligotrophomonas T1T2 strain isolated in this example after being cultured on a TTC agar solid culture plate for 24 hours is shown in the figure below. Figure 1 As shown, the colonies are orange and round in shape, with a soft texture and a smooth surface.

[0033] The preparation method of TTC agar solid culture medium is as follows: weigh 10g of tryptone, 3g of beef extract powder, 5g of sodium chloride, 15g of agar, and 0.01g of red tetrazolium (TTC), stir and dissolve each component in distilled water and then adjust the volume to 1000ml. Then dispense into conical flasks, seal the conical flasks with sterilization paper, place them in a sterilizer for 121°C, 101KPa high-pressure steam sterilization for 20min, pour the sterilized culture medium into a culture dish in a clean bench before solidification, and pour about 20ml of culture medium into each culture dish. After the culture medium is cooled and solidified, a culture dish containing TTC agar solid culture medium is obtained and stored at 4°C.

[0034] The preparation method of NA liquid medium is as follows: weigh out: 10g of tryptone, 3g of beef extract, and 5g of sodium chloride. Stir and dissolve each component in distilled water, adjust the pH to 7.0-7.2 and make up to 1000ml, dispense into conical flasks, seal the conical flasks with sterilization paper, and sterilize them with high-pressure steam at 121℃ and 101KPa for 20min.

[0035] Liquid Culture Method of Acidophilus Oligotrophomonas T1T2

[0036] Acidophilic oligotrophic monocytogenes T1T2 can be cultured in NA liquid medium or LB liquid medium, which are introduced below:

[0037] Cultivation method of acidophilic oligotrophic mononas T1T2 using NA liquid medium

[0038] S201A: Activation of bacterial strains: The glycerol seed liquid stored at -20°C in S104 is inoculated into the NA liquid culture medium at a volume ratio of 1:100 of the glycerol seed liquid to the NA liquid culture medium, and the culture is carried out at a constant temperature of 30°C and an oscillation speed between 180 rpm and 220 rpm for 36 hours to obtain an activated bacterial liquid. In this embodiment, the oscillation speed is preferably 200 rpm.

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

[0040] Cultivation method of acidophilic oligotrophic mononas T1T2 using LB liquid medium

[0041] S201B: Activation of bacterial strains: Take the glycerol seed solution stored at -20°C in S104 and inoculate it into LB liquid culture medium at a volume ratio of 1:100 between the glycerol seed solution and the LB liquid culture medium, and place it in a constant temperature of 37°C and an oscillation speed of 200 rpm for 36 hours to obtain an activated bacterial solution.

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

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

[0044] Identification of the strain of Acidophilus Stenotrophomonas T1T2

[0045] S301: Extraction of bacterial genomic DNA

[0046] Use Omega Bacterial DNA Kit (D3350-01) to extract genomic DNA. First, take 2 ml of the seed solution described in S104 and place it in a sterile 2 ml centrifuge tube, centrifuge at 12000 rpm for 2 min, discard the supernatant and retain the precipitate; then, add 100 μl 1×TE Buffer to the precipitate, vortex to mix, add 10 μl lysozyme to mix, and warm at 37°C for 10 min; add 100 μl BTL Buffer and 20 μl proteinase K, mix, warm at 55°C for 1 h, and oscillate to mix three times in between; add 5 μl RNase A enzyme, mix, stand at room temperature for 5 min, centrifuge at 10000 rpm for 2 min, take 200 μl supernatant and transfer it to a new sterile 1.5 ml centrifuge tube; add 200 μl BTL Buffer, mix well, and place in a 65℃ warm bath for 10min; add 200μl anhydrous ethanol, vortex mix well, transfer all the samples to the adsorption column, centrifuge at 10000rpm for 2min, discard the supernatant and the adsorption column, and place the adsorption column in a new collection tube; add 500μl HBC Buffer to the adsorption column, centrifuge at 10000rpm for 2min, and discard the supernatant; add 700μl DNA Wash Buffer to the adsorption column, centrifuge at 10000rpm for 2min, discard the supernatant, and repeat twice; put the empty adsorption column back into the collection tube, centrifuge at 10000rpm for 2min; add 30μl~50μl Elution Buffer (preheated at 65℃) to the adsorption column to dissolve the DNA precipitate, and obtain bacterial genomic DNA, which is stored at -20℃ for later use.

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

[0048] The genomic DNA obtained in step S301 was used as a template, and Eubac27F and Eubac1492R were used as primers for PCR amplification. The PCR reaction system (50 μl) included: 0.25 μl TaKaRa LA Taq (5 U / μl), 2.5 μl 10× LA Taq Buffer II (Mg 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), 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 program was as follows: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s, annealing at 55°C for 30 s, and extension at 72°C for 1 min. The process of denaturation, annealing, and extension was repeated 30 times; extension at 72°C for another 10 min, and storage at 4°C to obtain the PCR product.

[0050] S303: Electrophoresis of PCR products

[0051] 5 μl of the PCR product obtained in step S302 was spotted on a 1.2% agarose gel and subjected to nucleic acid electrophoresis at 120 V for 25 min. Figure 2 As shown, the 16S rDNA fragment band amplified using the genomic DNA of the acidophilic oligotrophic mononas T1T2 obtained in the present invention as a template is single and has high brightness, and the length of the 16S rDNA sequence is about 1500 bp.

[0052] S304: 16S rDNA sequencing

[0053] 30 μl of the PCR product obtained in step S302 was sent to Guangzhou Qingke Biotechnology Co., Ltd. for bidirectional sequencing. The sequencing results showed that the 16S rDNA sequence of the acidophilic oligotrophic mononas strain of the present invention was 1404 bp in length, and the specific sequence was shown in SEQ ID NO: 3.

[0054] The 16S rDNA sequence (SEQ ID NO: 3) of the acidophilic oligotrophic monocytic strain obtained by sequencing was input into NCBI, and BLAST comparison was performed. The sequences with high similarity to SEQ ID NO: 3 were downloaded, and the neighbor-joining method (NJ method) was used in the software MEGA11 to construct a phylogenetic tree based on SEQ ID NO: 3 and the downloaded 16S rDNA segment sequence. The results are shown in the figure. Figure 3As shown. According to the phylogenetic tree, the acidophilic oligotrophomonas T1T2 strain isolated and purified by the present invention has a sequence similarity of 99% with that of Stenotrophomonas acidaminiphila strain A2 (Accession No OQ244463.1) in its 16S rDNA sequence. According to the phylogenetic tree and sequence similarity of the 16S rDNA sequence, the strain isolated by the present invention belongs to the species Stenotrophomonas acidaminiphila, and the present invention names it as acidophilic oligotrophomonas T1T2 (Stenotrophomonas acidaminiphila T1T2).

[0055] Degradation effect of trichlorfon by acidophilic oligotrophic monosaccharides T1T2 in shake flasks

[0056] S401: Activation of bacterial strains: Take the glycerol seed liquid stored at -20°C in S104 and inoculate it into the NA liquid culture medium at a volume ratio of 1:100 of the glycerol seed liquid to the NA liquid culture medium, and perform shaking culture at a constant temperature of 30°C and a shaking speed of 200 rpm for 36 hours to obtain an activated bacterial liquid;

[0057] S402 inoculation: under a sterile environment, trichlorfon was added to the MSM liquid culture medium to a final concentration of 500 mg / l. The experimental group took the activated bacterial solution in S401 and added it to the MSM liquid culture medium containing 500 mg / l trichlorfon solution at a ratio of 1:20, and cultured at a constant temperature of 30°C and a shaking speed of 200 rpm for 24 hours to obtain a mixed solution inoculated with the activated bacterial solution. The control group added NA culture medium to the MSM liquid culture medium containing 500 mg / l trichlorfon solution at a ratio of 1:20 and cultured for 24 hours under the same conditions as the experimental group to obtain a mixed solution not inoculated with the activated bacterial solution.

[0058] S403 sampling: 2 ml of supernatant was taken from the mixed solution inoculated with activated bacterial solution in the experimental group and the mixed solution inoculated with no activated bacterial solution in the control group, and then filtered through a 0.22 μm filter membrane for high performance liquid chromatography detection. The standard sample was a 500 mg / l solution of trichlorfon in acetonitrile. The chromatographic conditions were: methanol: water (70:30, V / V) as the mobile phase, a wavelength of 210 nm, and an isocratic elution at a flow rate of 1.0 mL / min.

[0059] The results showed that the peak graph of the standard sample showed that the characteristic peak of trichlorfon appeared at around 11.8min ( Figure 4 ), the characteristic peak of trichlorfon in the experimental group inoculated with the mixed solution of activated bacterial solution ( Figure 5 ) and the mixed solution of the control group without inoculation of activated bacteria solution ( Figure 6) peak height decreased and the peak area decreased by 76.80%, indicating that the trichlorfon content decreased by about 76.80%.

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

[0061] Among them, the preparation method of MSM liquid culture medium is: 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 potassium dihydrogen phosphate, stir and dissolve in distilled water, adjust the pH to 7.0-7.2 and make up to 1000ml. Dispense into conical flasks, then seal the conical flasks with sterilization paper and sterilize them with high-pressure steam at 121°C and 101KPa for 20min.

[0062] Effect of Acidophilic Oligotrophomonas T1T2 on Degradation of Pesticide Residues on Plant Leaves

[0063] S501 strain activation: take the glycerol seed liquid stored at -20°C described in S104 and inoculate it into the NA liquid culture medium at a volume ratio of 1:100, place it in a constant temperature shaker at 30°C for 36 hours to obtain an activated bacterial liquid;

[0064] Preparation of fermentation broth of S502 Acidophilus Oligotrophomonas T1T2: The activated bacterial broth obtained in S501 was inoculated into NA liquid medium at a volume ratio of 1:100, and the culture was cultured at 30°C and 200 rpm under constant temperature shaking until the bacterial density reached 1.0×10 5 cfu / ml~1.0×10 6 cfu / ml to stop the culture and obtain the fermentation liquid;

[0065] S503 fermented bacteria liquid was sprayed on pepper leaves to degrade residual trichlorfon: the experimental group diluted the fermented bacteria liquid obtained by S502 by 10 times to make a degradation agent, and sprayed it on pepper leaves with a dosage of 20ml per plant. The control group directly used clean water without fermented bacteria liquid to spray the leaves in the same method and the same dosage. The degradation effect test of this step was carried out at the experimental base of South China Agricultural University in Ningxi, Zengcheng District, Guangzhou. The experimental land was an experimental greenhouse. The experiment was repeated three times biologically, and each experimental group and control group were 8 chili peppers with a growth period of 90 days. 48 hours before spraying the degradation agent and clean water, a trichlorfon aqueous solution with a concentration of 2.0g / l was evenly sprayed on the surface of the plant leaves of the experimental group and the control group with a dosage of 20ml per plant.

[0066] S504 leaf trichlorfon residue detection: 48 hours after spraying the degradation agent, take 2.0g of leaves from each chili pepper plant, add 10ml acetonitrile solution and ultrasonically extract for 3 minutes at a power of 150W. The extracts are mixed by group and tested by high performance liquid chromatography. The chromatographic conditions are: methanol: water (70:30, V / V) as the mobile phase, the wavelength is 210nm, the flow rate is 1.0mL / min and the gradient elution is carried out. The characteristic peak area of ​​trichlorfon in the experimental group was measured to be 18.3±5.3mAU*s, and the characteristic peak area of ​​trichlorfon in the control group was 77.1±8.3mAU*s. The degradation rate of trichlorfon was calculated according to the formula: Therefore, it was calculated that the degradation rate of trichlorfon in the experimental group was more than 75%.

[0067] The above results indicate that the degradation agent containing the fermentation liquid of Acidophilus Oligotrophomonas 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. Acidophilic oligotrophic monosaccharides T1T2 has a good effect on the degradation of organophosphorus pesticides. The degradation rate of organophosphorus pesticide trichlorfon on the leaves of crops represented by chili peppers is more than 75%, so it can be used for the degradation of plant residual pesticides.

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

[0071] 3. The degrading agent containing acidophilic oligotrophic mononas T1T2 has a simple formula, and the fermentation culture liquid is simple to culture, without the need for special equipment or harsh culture conditions, so the production cost is low.

[0072] 4. Acidophilic Oligotrophomonas T1T2 is a soil-derived beneficial bacterium for crops. In addition to being able to degrade residual pesticides on crops, it can also more easily colonize in the soil after falling into the ground, continuously degrading residual pesticides in the soil and being continuously friendly to the environment.

[0073] 5. Even after washing with clean water, the residues of degrading agents on crops will inevitably remain. Compared with the stability of chemical degrading agents, biodegrading agents can be killed by cooking and are almost harmless to the human body, so they are safer.

[0074] The experimental methods in the embodiments of the present invention that do not specify specific conditions are usually carried out under conventional conditions or according to the conditions recommended by the manufacturer. The various common chemical reagents used in the embodiments are all commercially available products. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those generally understood by those skilled in the art of the present invention. The terms used in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0075] The above-mentioned embodiments only express several implementation methods of the present invention, and the description is relatively specific and detailed, but it cannot be understood as limiting the scope of the invention patent. It should be pointed out that for ordinary technicians in this field, several modifications and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention.

Claims

1. An acidophilic oligotrophic mononas T1T2, characterized in that: The acidophilic oligotrophic mononas T1T2 is deposited in China Center for Type Culture Collection with a deposit number of CCTCC NO: M 20242351.

2. Use of the acidophilic oligotrophic mononas T1T2 according to claim 1 in pesticide degradation.

3. The use according to claim 2, characterized in that: The pesticide is an organophosphorus pesticide.

4. The use according to claim 3, characterized in that: The organophosphorus pesticides include trichlorfon.

5. A degradation agent for degradable pesticides, characterized in that: It comprises the acidophilic oligotrophic mononas T1T2 as described in claim 1.

6. The degradation agent according to claim 5, characterized in that: The concentration of the degradation agent, Acidophilus oligotrophic mononas T1T2, is 1.0×10 4 cfu / ml~1.0×10 5 cfu / ml, and the pesticide that can be degraded by the degradation agent is an organophosphorus pesticide.

7. The degradation agent according to claim 6, characterized in that: The preparation method of the degradation agent comprises: inoculating the acidophilic oligotrophic mononas T1T2 into a culture medium, culturing the culture medium under constant temperature shaking conditions of 30° C. to 37° C. and 180 to 220 rpm to prepare a fermentation liquid, and then adjusting the acidophilic oligotrophic mononas T1T2 concentration of the fermentation liquid to prepare the degradation agent.

8. A method for degrading organophosphorus pesticides, characterized in that: The following steps are involved: The degrading agent according to claim 5 is applied to plants.

9. The degradation method according to claim 8, characterized in that: The organophosphorus pesticide includes trichlorfon, and the application method is spraying.

10. The degradation method according to claim 8, characterized in that: The single application amount of the degrading agent to each plant is 10 to 30 ml.

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