CN116831130A - Application of 1-methyl-5-mercapto-1H-tetrazole in preventing and treating citrus canker - Google Patents

Application of 1-methyl-5-mercapto-1H-tetrazole in preventing and treating citrus canker Download PDF

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CN116831130A
CN116831130A CN202310746981.3A CN202310746981A CN116831130A CN 116831130 A CN116831130 A CN 116831130A CN 202310746981 A CN202310746981 A CN 202310746981A CN 116831130 A CN116831130 A CN 116831130A
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compound
tetrazole
mercapto
methyl
pathogenic bacteria
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崔紫宁
王鑫
张钰晴
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South China Agricultural University
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION 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
    • A01N43/00Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/713Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with four or more nitrogen atoms as the only ring hetero atoms
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01PBIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P1/00Disinfectants; Antimicrobial compounds or mixtures thereof

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  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Plant Pathology (AREA)
  • Environmental Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Pest Control & Pesticides (AREA)
  • General Health & Medical Sciences (AREA)
  • Dentistry (AREA)
  • Agronomy & Crop Science (AREA)
  • Health & Medical Sciences (AREA)
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  • General Chemical & Material Sciences (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

The invention belongs to the technical field of plant disease control, and particularly relates to application of 1-methyl-5-mercapto-1H-tetrazole in controlling citrus canker. The compound has strong inhibition effect on hpa1 gene promoter in a pathogenic bacteria III type secretion system, does not influence the normal growth of pathogenic bacteria, can strongly inhibit virulence factors of plant pathogenic bacteria while not influencing the growth of plant pathogenic bacteria, remarkably reduces the pathogenicity of plant pathogenic bacteria, and achieves the effect of preventing and treating citrus canker; the composition also has the effect of reducing the generation of drug resistance of pathogenic bacteria to the compound, and has longer effective service life in the aspect of controlling plant diseases, and wide application prospect.

Description

Application of 1-methyl-5-mercapto-1H-tetrazole in preventing and treating citrus canker
Technical Field
The invention belongs to the technical field of plant disease control. More particularly relates to the application of 1-methyl-5-mercapto-1H-tetrazole in preventing and treating citrus canker.
Background
Citrus is the first major fruit in the world, and is also the most important fruit tree with the widest cultivation area and the most economic status in the south of China. Citrus canker is one of the most serious bacterial diseases on citrus, caused by the gram-negative bacterium citrus canker Xanthomonas citri subsp. The citrus canker is easy to prevent and treat, and once the citrus canker is infected, the citrus canker is toxic for life. In China, the average economic loss caused by citrus canker is as high as 5 hundred million yuan each year, and the healthy development of the citrus industry is greatly threatened. Therefore, the development of new effective disease control drugs is one of the key problems that need to be solved urgently.
Traditional antibiotics are usually targeted by the key factors of bacterial survival, but the long-term irregular use of antibiotics has made bacterial resistance more and more common and serious. In order to solve the problem of drug resistance, chinese patent application CN111713514A discloses a medicament for citrus canker, which is prepared by combining an organic copper bactericide, nano silver sol and plant volatile oil, and improves the sterilizing capability and delays the generation of drug resistance of pathogenic bacteria through the synergistic effect of the organic copper bactericide, the nano silver sol and the plant volatile oil. Although the method delays the generation of drug resistance, with long-term use, citrus canker bacteria gradually cause the generation of drug resistance of pathogenic bacteria according to the rule of being out of priority in the nature, and the long-term use is not facilitated.
Therefore, there is an urgent need to provide a method for controlling citrus canker, which has a good anti-citrus canker effect and does not cause the citrus canker to have drug resistance even after long-term use.
Disclosure of Invention
The invention aims to overcome the defect and the defect that the traditional method for preventing and treating the citrus canker is easy to cause the drug resistance of pathogenic bacteria, and provides the application of the 1-methyl-5-mercapto-1H-tetrazole in preventing and treating the citrus canker, so that the effects of good anti-citrus canker and no drug resistance of the citrus canker can be achieved after long-term use.
The invention aims to provide an application of 1-methyl-5-mercapto-1H-tetrazole as a plant pathogenic bacteria type III secretion system inhibitor.
Another object of the present invention is to provide a composition for controlling plant diseases.
The above object of the present invention is achieved by the following technical scheme:
the natural product is a treasury which is presented to human beings by nature, people can obtain prevention and treatment medicines, medical and agricultural antibiotics for curing diseases seriously endangering human health, develop efficient low-toxicity pesticides, plant growth hormone and other substances with economic value, and the natural product is environment-friendly. The inventor finds that the natural active product 1-methyl-5-mercapto-1H-tetrazole has remarkable control effect on citrus canker by inhibiting the expression of the hpa1 gene promoter in a plant pathogenic bacteria type III secretion system.
Therefore, the invention claims the application of 1-methyl-5-mercapto-1H-tetrazole in preventing and treating citrus canker, wherein the 1-methyl-5-mercapto-1H-tetrazole is called compound Z-10 for short, and has the following structure:
the prevention and treatment is prevention and/or treatment, namely prevention and/or treatment of citrus canker, and can prevent plants from being affected by the citrus canker.
Further, the pathogenic bacteria of the citrus canker is citrus canker Xanthomonas citri subsp.
Still further, the compound Z-10 inhibits the plant pathogenic bacteria type III secretion system. The type III secretion system (T3 SS) is a key virulence factor in gram-negative pathogenic bacteria, and the expression of the main regulatory factors, hrpG and HrpX, the assembly of needle structures and the expression of effector proteins are the main causes of pathogenicity of such bacteria. The compound Z-10 acts on virulence factors of bacteria, takes the virulence factors as targets, reduces pathogenicity of the bacteria, does not influence normal growth of the bacteria, can achieve the effect of reducing drug resistance of the bacteria, and realizes long-term and effective prevention and control of citrus canker.
Further, the compound Z-10 inhibits the expression of the hpa1 gene promoter in a plant pathogenic bacteria type III secretion system.
Furthermore, the 1-methyl-5-mercapto-1H-tetrazole can be pharmaceutically acceptable salt or hydrate thereof, and the pharmaceutically acceptable salt or hydrate of the 1-methyl-5-mercapto-1H-tetrazole can be used as a precursor to achieve an activity similar to that of the 1-methyl-5-mercapto-1H-tetrazole.
Based on this, the invention also claims the use of 1-methyl-5-mercapto-1H-tetrazole, said 1-methyl-5-mercapto-1H-tetrazole being abbreviated as compound Z-10, as an inhibitor of the type III secretion system of phytopathogens, having the following structure:
further, the 1-methyl-5-mercapto-1H-tetrazole can be pharmaceutically acceptable salt or hydrate thereof, and the pharmaceutically acceptable salt or hydrate of the 1-methyl-5-mercapto-1H-tetrazole can be used as a precursor to achieve an activity effect similar to that of the 1-methyl-5-mercapto-1H-tetrazole.
Further, the inhibitor is in a solid preparation or a liquid preparation. Preferably, the solid formulation comprises a direct use solid formulation, a dispersible solid formulation, a soluble solid formulation; the liquid preparation comprises a liquid preparation, a dispersion liquid preparation, an emulsion preparation, a suspension preparation and a multiphase preparation.
In addition, the invention also provides a composition for preventing and treating plant diseases, which contains 1-methyl-5-mercapto-1H-tetrazole or pharmaceutically acceptable salt or hydrate thereof and one or more of bactericides, biocontrol bactericides, plant growth regulators and pesticides.
Further, the plant disease is citrus canker. The pathogenic bacteria of the citrus canker is citrus canker Xanthomonas citri subsp.
Preferably, the bactericide comprises: thiabendazole, oxine-copper, zinc thiazole, spring terramycin, mancozeb, copper hydroxide, bordeaux mixture and the like.
Preferably, the biocontrol microbial agents include bacillus subtilis, bacillus amyloliquefaciens, bacillus methylotrophicus and the like.
Preferably, the plant growth regulator comprises: gibberellin, benzylaminopurine, sodium nitrophenolate, brassin, and the like.
Preferably, the insecticide comprises: etoxazole, spirodiclofen, avermectin, etoxazole and the like.
The invention has the following beneficial effects:
the invention provides an application of 1-methyl-5-mercapto-1H-tetrazole in preventing and treating citrus canker, which has a strong inhibition effect on hpa1 gene promoters in pathogenic bacteria type III secretion systems, does not influence normal growth of pathogenic bacteria, can strongly inhibit virulence factors of plant pathogenic bacteria while not influencing growth of the plant pathogenic bacteria, remarkably reduces pathogenicity of the plant pathogenic bacteria, and achieves the effect of preventing and treating citrus canker; the composition also has the effect of reducing the generation of drug resistance of pathogenic bacteria to the compound, and has longer effective service life in the aspect of controlling plant diseases, and wide application prospect.
Drawings
FIG. 1 is a graph showing the growth of Xcc jx-6 in NB medium or XVM medium after addition of Compound Z-10 in example 2 of this invention.
FIG. 2 is a graph showing the effect of compound Z-10 of example 4 of the present invention on tobacco HR after treatment with Xcc jx-6.
FIG. 3 is a graph showing the control of citrus canker by different concentrations of Compound Z-10 in example 5 of the present invention.
FIG. 4 is a graph showing the results of the coating of the number of Xcc jx-6 colonies in citrus leaves after inoculation with different concentrations of Compound Z-10 in example 5 of the present invention.
FIG. 5 is a graph showing the statistical results of the number of Xcc jx-6 colonies in citrus leaves after inoculation with different concentrations of compound Z-10 in example 5 of the present invention.
FIG. 6 is a graph showing the results of the coating of the number of Xcc jx-6 colonies in citrus leaves after the compound Z-10 of example 6 of the present invention was inoculated with biocontrol strain HN-8.
FIG. 7 is a graph showing the statistical result of the number of Xcc jx-6 colonies in citrus leaves after compound Z-10 and biocontrol strain HN-8 in example 6 of the present invention are inoculated.
Detailed Description
The invention is further illustrated in the following drawings and specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
The citrus canker caused by citrus canker pathogen (Xanthomonas citri subsp. Citri, xcc) is an important bacterial disease on citrus, and the development of novel and effective disease control novel medicines is one of the key problems which need to be solved urgently. The type III secretion system (T3 SS) is a key virulence factor of Xcc, is highly homologous and conserved in Xanthomonas, can reduce pathogenicity of the Xanthomonas when taking the T3SS as a target, reduces occurrence of diseases without affecting growth of the Xcc, can delay drug resistance of pathogenic bacteria to drugs, and is beneficial to long-term effective use of the drugs.
The following examples are examples of test compounds for inhibition of T3SS by citrus canker bacteria, test pathogenic bacteria: xcc wild-type strain Xcc jx-6, and strain Xcc jx-6 with a reporter plasmid (lux reporter system containing hpa1 promoter sequence, regulated by T33SS regulatory factor), mainly cause citrus canker. Compound Z-10 was dissolved in dimethyl sulfoxide (DMSO) to prepare a test solution at a concentration of 200 mM.
Reagents and materials used in the following examples are commercially available unless otherwise specified.
EXAMPLE 1 inhibition of the hpa1 Gene promoter in the T3SS System by Compound Z-10
1. The experimental method comprises the following steps:
xcc jx-6 of the reporter plasmid was streaked on NA plates, single colonies were picked up 2-3 days later and placed in NB medium, kanamycin (final concentration 50. Mu.g/mL) was added, and cultured at 28℃and 200rpm until OD was reached 600 About 1.0, cells were collected by centrifugation, and XVM culture medium (20 mM NaCl,10mM (NH 4)) 2 SO 4 ,5mM MgSO 4 ,1mM CaCl 2 ,0.16mM KH 2 PO 4 ,0.32mM K 2 HPO 4 ,0.01mM FeSO 4 10mM fructose, 10mM sucrose, 0.03% Casamino Acids, pH 6.7), and then re-suspended in XVM2 medium, kanamycin was added to adjust the OD 600 Adding compound Z-10 to a final concentration of 200 μm to 0.1, mixing thoroughly, and taking the same volume of DMSO as solvent as reference to obtain bacterial suspension containing the compoundSub-packaging into black 96-well plates with transparent bottoms, culturing XVM2 with the same volume as a background at 150 mu L of each well for subsequent calculation of inhibition rate, and setting 3 parallel per treatment; 96-well plates were treated at 28℃and 200rpm for 16h. Measuring self-luminescence value (LUX) and OD respectively with multifunctional enzyme-labeled instrument 600 After subtraction of the background value of XVM medium, inhibition of the compound was calculated using the formula. The calculation formula is as follows:
2. experimental results:
the results are shown in Table 1.
TABLE 1 inhibition of the hpa1 Gene promoter in T3SS System by Compounds Z-10
Compounds of formula (I) LUX/OD 600 Inhibition rate%
DMSO 1811.70±94.29 -
Z-10 203.61±28.13* 88.76
Note that: ", represents a significant difference compared to DMSO-treated group, indicating that the compound has a strong inhibitory effect on hpa1 promoter.
As can be seen from the table, the compound Z-10 has a strong inhibitory effect on the hpa1 promoter.
EXAMPLE 2 Effect of Compound Z-10 on growth of wild-type Strain Xcc jx-6
1. The experimental method comprises the following steps:
the growth curves for Xcc jx-6 in the presence of compound Z-10 in enriched NB and T3SS induction medium XVM2 were determined, respectively. Streaking and activating Xcc jx-6 wild strain on NA plate, picking single colony to NB culture medium after 2-3 days, growing to OD 600 About 1.0, cells were collected by centrifugation, resuspended in NB medium or XVM medium, and OD adjusted 600 To 0.1, compound Z-10 was added separately, mixed well to a final concentration of 200. Mu.M, placed in 96 well plates, and 3 replicates were set per treatment with equal volumes of DMSO as solvent control. Culturing at 28deg.C under 200rpm, measuring data (band plate cover) with multifunctional enzyme-labeled instrument every 4 hr, and measuring to 48 hr.
2. Experimental results:
the experimental results are shown in FIG. 1, wherein the left graph shows the growth curve of Xcc jx-6 in NB medium supplemented with Compound Z-10, and the right graph shows the growth curve of Xcc jx-6 in XVM medium supplemented with Compound Z-10. The results showed that compound Z-10 did not show significant inhibition of Xcc jx-6 growth in either NB medium or XVM2 medium during the slow, log and stationary phases of Xcc jx-6 growth as compared to control DMSO.
EXAMPLE 3 determination of Minimum Inhibitory Concentration (MIC) of Compound Z-10 for Xcc jx-6
1. The experimental method comprises the following steps:
xcc jx-6 Strain was cultured in NB Medium to OD 600 About 0.6 to 0.8, collecting the cells by centrifugation, re-suspending the cells with NB medium, and adjusting the OD 600 To 0.2, the concentration of the bacterial liquid is approximately 2X 10 8 CFU/mL, the bacterial suspension was diluted to 4X 10 in a ratio of 1:500 5 Concentration of CFU/mL; adding a certain volume of Z-10 mother liquor into diluted bacterial suspension, and fully and uniformly mixing to ensure that the initial compound concentration is 12800 mu M; spring day with initial concentration of 12800 mug/mL is used as positive reagent control; equal volume of DMSO plusInto the bacterial suspension as a solvent control; the same volume of NB medium without bacteria was used as a blank, and the compound concentration was diluted to 6.25. Mu.M or 6.25. Mu.g/mL (total 12 gradients) in 96-well plates by a 2-fold gradient dilution method, and left at 28℃for 1-2 days. And taking the turbidity of the solvent contrast as a reference standard, wherein in the compound Z-10 treatment group, the compound concentration corresponding to the clear bacterial liquid in the micropores is the Minimum Inhibitory Concentration (MIC).
2. Experimental results:
the results are shown in Table 2.
TABLE 2 determination of Minimum Inhibitory Concentration (MIC) of Compound Z-10 for Xcc jx-6
As can be seen from the table, the minimum inhibitory concentration of the compound Z-10 of the present invention on Xcc jx-6 was 743. Mu.g/mL.
EXAMPLE 4 Effect of Compound Z-10 on the allergic response (HR) of Xcc jx-6 on tobacco
1. The experimental method comprises the following steps:
xcc jx-6 strain was cultured in NB medium to OD 600 About 0.6 to 0.8, centrifugally collecting the thalli, re-suspending the thalli with sterile water, and adjusting the OD 600 To 0.5, compound Z-10 was added and mixed well to a final concentration of 200. Mu.M, treated with an equal volume of DMSO as a solvent control, sterile water as a blank (CK), and treated at 28℃for 2h. And then inoculating the treated Xcc jx-6 bacterial suspension to Benshi tobacco cultured in a greenhouse for two months by using a needleless injector, and continuously culturing the tobacco in the greenhouse after inoculation, and photographing and observing after 4-6 days.
2. Experimental results:
as a result of the experiment, referring to FIG. 2, it can be seen that compound Z-10 has a significant inhibitory effect on the HR of Xcc jx-6 on tobacco, a non-host plant.
Example 5 Effect of Compound Z-10 on the pathogenicity of Xcc Strain on citrus
1. The experimental method comprises the following steps:
the citrus is Wo-gan, and is cultivated in a greenhouse. Xcc jx-6 Strain was cultured in NB Medium to OD 600 About 0.6 to 0.8, centrifugally collecting the thalli, re-suspending the thalli with sterile water, and adjusting the OD 600 To 0.2 (1.0X10) 8 CFU/mL), compound Z-10 at final concentrations of 100. Mu.M, 150. Mu.M, 200. Mu.M, and kasugamycin at final concentrations of 200. Mu.M were added as positive reagent controls, equal volumes of DMSO as solvent controls, sterile water and sterile water containing 200. Mu.M compound Z-10 as blank controls. The compound and the bacterial liquid are fully and evenly mixed and treated for 2 hours at the temperature of 28 ℃. Taking 5-10 mu L of the treated bacterial suspension or sterile water, and extruding the bacterial suspension or sterile water into the back surface of the citrus leaves by using a 1mL needleless injector. And (5) continuously placing the strain in a greenhouse for culturing for 3-7 days after inoculation, and observing the disease condition.
2. Experimental results:
the results of inoculation are shown in FIG. 3. The right side of the veins are respectively from top to bottom: sterile water control, sterile water control for compound Z-10, dmso+wt (wild type Xcc jx-6 strain) solvent control, kasugamycin+wt positive agent control; two of the blank controls had no sign of onset at all, dmso+wt solvent control was normal onset, and a veramycin+wt positive agent control also showed significant plaque. The left side of the veins is respectively from top to bottom: compound Z-10+wt experiments at concentrations of 100 μm, 150 μm, 200 μm; it can be seen that compound Z-10 had significant lesions at a concentration of 100. Mu.M, significantly smaller lesions at a concentration of 150. Mu.M, and were completely non-pathogenic at a concentration of 200. Mu.M.
In order to more accurately count the control effect of the compound, the inventors cut out leaves with the diameter of 5mm by using a puncher with the diameter of 5mm and taking an inoculation point as a circle center, grind the leaves into homogenate by using sterile water in a mortar after sterilization, take 100 mu L to coat on an NA flat plate after gradient dilution to proper concentration, and culture the flat plate for 2-3 days at the temperature of 28 ℃ for colony counting. The coating results are shown in fig. 4, and the counting results are shown in fig. 5. The result shows that the compound Z-10 has very remarkable control effect on citrus canker at the concentration of 200 mu M compared with the control, and has better effect than the kasugamycin positive medicament at the same concentration.
EXAMPLE 6 Compound Z-10 and biocontrol Strain HN-8 have effect of controlling citrus canker
1. The experimental method comprises the following steps:
citrus variety: wo citrus reticulata (R) is used. Xcc jx-6 strain and biocontrol strain HN-8 (Burkholderia anthina HN-8, patent No. 201711405448.1) were cultured in NB medium to OD 600 About 0.6 to 0.8, collecting the cells by centrifugation, re-suspending the cells with sterile water, and subjecting the Xcc jx-6 to OD 600 Adjusted to 0.2 (1.0X10) 8 CFU/mL), the concentration of the compound Z-10 was selected to be 100. Mu.M with substantially no control effect, and the concentration of biocontrol strain HN-8 was selected to be pathogen Xcc jx-6OD 600 2 times the value, i.e. to OD 600 =0.4. The following experimental groups were set up: Z-10+WT, HN-8+WT, Z-10+HN-8+WT, DMSO+WT solvent control, sterile water as a blank, and treatment at 28℃for 2h. Taking 5-10 mu L of the treated bacterial suspension and sterile water, extruding the bacterial suspension and sterile water into the back of citrus leaves by a 1mL syringe without a needle, inoculating, continuously placing the citrus leaves in a greenhouse for culturing for 3-7 days, and observing the disease condition. And colony counting was performed on the inoculated portions with reference to the plating counting method in example 4.
2. Experimental results:
the coating results are shown in fig. 6, and the counting statistics are shown in fig. 7. Therefore, after 100 mu M of compound Z-10 without control effect is compounded with biocontrol strain HN-8, the effect is better than that of the compound Z-10 and biocontrol strain HN-8 which are singly used, and obvious synergistic effect is achieved, so that the compound Z-10 has good compounding potential.
In summary, the above examples all show that the compound Z-10, namely the compound 1-methyl-5-mercapto-1H-tetrazole, can strongly inhibit the expression of the hpa1 gene promoter of the T3SS system of the compound while not affecting the growth of pathogenic bacteria Xcc jx-6, thereby obviously reducing the pathogenicity of the pathogenic bacteria, and can achieve the effect of preventing and/or treating plant diseases caused by the pathogenic bacteria while not affecting the growth of the pathogenic bacteria, thereby avoiding the generation of drug resistance of the pathogenic bacteria and prolonging the effective service life of the compound.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.

Claims (10)

  1. The application of 1-methyl-5-mercapto-1H-tetrazole in preventing and treating citrus canker is characterized in that the 1-methyl-5-mercapto-1H-tetrazole is simply called a compound Z-10, and has the following structure:
  2. 2. use according to claim 1, wherein the pathogenic bacteria of citrus canker is citrus canker Xanthomonas citri subsp.
  3. 3. Use according to claim 2, characterized in that the compound Z-10 inhibits the plant pathogenic bacterial type III secretion system.
  4. 4. Use according to claim 3, characterized in that said compound Z-10 inhibits the expression of the hpa1 gene promoter in the secretory system of plant pathogenic bacteria type III.
  5. 5. The use according to any one of claims 1 to 4, wherein the 1-methyl-5-mercapto-1H-tetrazole is also a pharmaceutically acceptable salt or hydrate thereof.
  6. The application of 1-methyl-5-mercapto-1H-tetrazole as a plant pathogenic bacteria type III secretion system inhibitor is characterized in that the 1-methyl-5-mercapto-1H-tetrazole is simply called a compound Z-10, and has the following structure:
  7. 7. the use according to claim 6, wherein the 1-methyl-5-mercapto-1H-tetrazole is also in the form of a pharmaceutically acceptable salt or hydrate thereof.
  8. 8. The use according to claim 6 or 7, wherein the inhibitor is in the form of a solid or liquid formulation.
  9. 9. A composition for controlling plant diseases, which is characterized in that the composition contains 1-methyl-5-mercapto-1H-tetrazole or pharmaceutically acceptable salts or hydrates thereof and one or more of bactericides, biocontrol bactericides, plant growth regulators and pesticides.
  10. 10. The composition of claim 9, wherein the plant disease is citrus canker.
CN202310746981.3A 2023-06-21 2023-06-21 Application of 1-methyl-5-mercapto-1H-tetrazole in preventing and treating citrus canker Pending CN116831130A (en)

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