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
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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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