Small molecular inhibitor pyrazinamide used in plant ethylene synthesis and application thereof
A pyrazinamide agent and a synthesis route technology, which is applied in the field of small molecule inhibitor pyrazinamide, can solve the problems of high price, environmental hazards, non-specific action sites, etc., and achieves easy popularization and application, reduction of synthesis amount, and inhibition effect. strong effect
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Embodiment 1
[0017] Example 1 Screening of Small Molecule Inhibitors that Inhibit Plant Ethylene Synthesis Pathway
[0018] Using chemical genetics, the small molecule inhibitor screening method, to screen 2,000 small chemical molecules selected from the microsource spectrum database (SP2000, http: / / www.msdiscovery.com) provided by the University of California, Riverside. All the small molecules in the library were made into a stock solution with a concentration of 0.5 mg / 100 μL in DMSO. Related small molecule information can be downloaded from (http: / / www.msdiscovery.com / downloads.html). The structures, molecular formulas, molecular weights, names and functions of the small molecules in the screening library are known, and the concentration of the small molecules in the screening test is 50-100 μM. By screening the library containing 2000 small molecules (gifted by Yang Zhenbiao, University of California, Riverside) (reference: Girke T, Cheng LC, et al. (2005). "ChemMine. A compound mini...
Embodiment 2
[0019] Embodiment 2 The present invention inhibits the medium of plant ethylene synthesis pathway
[0020] The MS medium for inhibiting plant ethylene synthesis pathway of the present invention is prepared by using the small molecule inhibitor pyrazinamide (PZA for short), that is, the MS medium with a concentration of pyrazinamide of 30-50 μM (micromoles per liter) is prepared. The control is commonly used MS medium.
[0021] The experimental materials were Arabidopsis thaliana ethylene-oversynthesis mutant strain eto1-2, and Arabidopsis wild-type Col-0 with normal expression of ethylene.
[0022] Culture eto1-2 and Col-0 with MS medium that inhibits plant ethylene synthesis pathway (that is, MS medium with pyrazinamide added) and control MS medium, and culture them in a 22° incubator for 3 days in the dark and dark .
[0023] Observation after 3 days of cultivation, the results are shown in figure 2 ( figure 2 The effect of adding the small molecule inhibitor pyrazinam...
Embodiment 3
[0024] Embodiment 3 Determination of pyrazinamide action target
[0025] In order to determine the action target of the small-molecule inhibitor pyrazinamide of the present invention, the action of the small-molecule inhibitor was observed after applying excess ethylene synthesis precursor ACC. If the inhibitor acts on the ethylene synthesis pathway, in the presence of excess ethylene synthesis precursor ACC, the small molecule inhibitor pyrazinamide will no longer have a significant inhibitory effect on ethylene synthesis; but if the inhibitor target is located downstream of the ethylene synthesis pathway ethylene signal transduction pathway, in the presence of excess ethylene synthesis precursor ACC, the inhibitory effect of pyrazinamide on ethylene will not be affected, that is, excess ethylene synthesis precursor ACC cannot affect the downstream ethylene signal pathway inhibitors on ethylene Inhibition of the signal.
[0026] The experimental method was referred to Exampl...
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