Method for researching action mechanism of bactericide based on metabolomics

A mechanism of action and metabolomics technology, applied in scientific instruments, measuring devices, instruments, etc., to achieve rapid identification and high-throughput analysis, rapid method, and strong specificity

Inactive Publication Date: 2018-03-13
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports at home and abroad on the biomarkers of the mechanism of action of fungicides and their research methods.

Method used

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  • Method for researching action mechanism of bactericide based on metabolomics
  • Method for researching action mechanism of bactericide based on metabolomics
  • Method for researching action mechanism of bactericide based on metabolomics

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Screening of Mechanism-specific Biomarkers of Succinate Dehydrogenase Inhibitors

[0051] The process used is as figure 1 As shown, the specific implementation method is as follows:

[0052] 1. Pretreatment of Botrytis cinerea mycelium samples

[0053] 1) Cultivation of Botrytis cinerea

[0054] Take the PDA medium plate of Botrytis cinerea SP2-6 that has grown for 3 days, and use a 0.5cm puncher to punch out the bacterial cake along the outer edge of the colony in the ultra-clean workbench, pick the bacterial cake with a pick needle and connect it to the cellophane-covered Contains EC 50 Drug concentration on solid PDA medium plate, i.e. 0.04 μg mL -1 Carbendazim, 0.4 μg mL -1 Thiophanate-methyl, 0.08μg mL -1 Boscalid, 0.1 μg mL -1 Kresoxim-methyl, 9.8μg mL -1 Cyazofamid, 0.7μg mL -1 Pyrimethanil, 0.5 μg mL -1 Cyprodinil, 0.01 μg mL -1 Fludioxonil or 4.6 μg mL -1 For imazalil, the treatment containing the same amount of DMSO and no drug was used a...

Embodiment 2

[0080] Example 2 Application of succinic acid in the recognition of the mechanism of action of wiltapyl and fluopyram on Botrytis cinerea

[0081]1. Botrytis cinerea mycelium culture and GC-MS detection of succinic acid in mycelium

[0082] The pretreatment method that adopts and the detection method of succinic acid see embodiment 1 for details, wherein the cultivation of Botrytis cinerea Bc52 is specifically implemented as follows:

[0083] Take the PDA medium plate of Botrytis cinerea Bc52 that has grown for 6 days, and use a 0.5cm hole puncher to take the bacterial cake along the outer edge of the colony in the ultra-clean workbench, pick the bacterial cake with a pick needle and connect it to a cellophane-lined With drug (10μg mL -1 Carboxyl or 1 μg mL -1 Fluopyram, which inhibits the growth of mycelium by 50%), was cultured on a solid PDA medium plate for 6 days at 17.5-18.5° C. to obtain a mycelium culture.

[0084] 2. Changes of succinic acid content in Botrytis cin...

Embodiment 3

[0086] Example 3 Application of succinic acid in the identification of the mechanism of action of boscalid against Rhizoctonia solani

[0087] 1. Mycelia culture of Rhizoctonia solani and GC-MS detection of succinic acid in mycelia

[0088] The pretreatment method that adopts and the detection method of succinic acid see embodiment 1 for details, wherein the cultivation of Rhizoctonia solani D179 is specifically implemented as follows:

[0089] Take the PDA medium plate of Rhizoctonia solani D179 that has grown for 1 day, and use a 0.5cm puncher to punch out the bacterial cake along the outer edge of the colony in the ultra-clean workbench, pick the bacterial cake with a pick needle and connect it to the Drug-carrying cellophane (0.93μg mL -1 Boscalid, which inhibits the growth of mycelium by 50%) is cultured on a solid PDA medium plate at 24.5-25.5° C. for 1 day to obtain a mycelium culture.

[0090] 2. Changes of succinic acid content in Rhizoctonia solani D179 under the t...

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Abstract

The invention discloses a method for researching the action mechanism of a bactericide based on metabolomics. According to the method, pathogen hyphae respectively cultured by a bactericide-containingmedium and a bactericide-free control medium are subjected to metabolomics detection, differential metabolites are found through comparing the metabolomics research methods, common metabolites with different mechanisms of action are eliminated, a specific biomarker with a target bactericide action mechanism is acquired, and the bactericide action mechanism is identified and is subjected to high-throughput analysis through biomarker content change. The invention also discloses a use of succinic acid as a biomarker of a succinate dehydrogenase inhibitor. The method is fast and reliable and canproduce a biomarker with strong specificity. Through biomarker specificity change, the rapid identification and high-throughput analysis of the active substance action mechanism are realized.

Description

technical field [0001] The invention relates to the technical field of fungicide pharmacology and metabolome analysis, in particular to a method for researching the action mechanism of fungicides based on metabolomics. Background technique [0002] Fungicides are an important means of production to protect crops from disease losses in agricultural production. With the emergence of pathogen resistance to common fungicides and people's concerns about food safety and ecological health, researchers are required to continuously search for new biological target proteins Compounds with new active structures have discovered a large number of new low-toxicity, high-efficiency, and low-residue antibacterial compounds, which provide excellent fungicide potential resources for the effective control of field diseases. However, the successful development of a drug requires not only attention to its biological activity, but also the mechanism of action of the compound on harmful organisms,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/72
CPCG01N30/02G01N30/06G01N30/72G01N2030/067
Inventor 刘鹏飞胡志宏母丹任纪龙代探梁莉刘盼晴刘西莉
Owner CHINA AGRI UNIV
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