Compound with plant pathogenic fungus resistant activity and preparation method of compound
A technology of phytopathogenic fungi and compounds, applied in the field of compounds with activity against phytopathogenic fungi and their preparation, can solve problems such as negative impacts on the environment and human health, and achieve industrialized large-scale production, easy operation and implementation, and good practicality value effect
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Embodiment 1
[0035] Compound Fermentation:
[0036]1) Inoculate Bacillus subtilis (Bacillus subtilis) Pc3 in 5mL LB medium (in g / L: yeast extract 5, peptone 10, sodium chloride 10) for activation culture, at 28°C, 180r / min Cultivate on a shaking table for 12 hours;
[0037] 2) Inoculate the activated culture solution of Bacillus subtilis (Bacillus subtilis) Pc3 in step 1), the inoculation volume is 5mL, inoculate in 1L Erlenmeyer flask, each bottle contains 500mL fermentation medium (in g / L: Glucose 20, KH 2 PO 4 6. K 2 HPO 4 14. MgSO 4 0.2, (NH4) 2 SO 4 4. Trace elements 2mL (FeSO 4 4H 2 O, CaCl 2 2H 2 O, MnSO 4 4H 2 O and ZnCl 2 1 mM each), cultured on a shaking table at 28° C. and 180 r / min for 48 hours to obtain a fermentation broth containing the compound having anti-phytopathogenic fungal activity of the present invention.
Embodiment 2
[0039] Compound preparation:
[0040] Centrifuge the fermentation culture liquid for 10 min under the condition of 8000r / min to obtain the fermentation supernatant, and precipitate the fermentation supernatant containing the compound having anti-phytopathogenic fungal activity of the present invention with 3 times the volume of absolute ethanol, and use an equal volume of Ethyl acetate was extracted 3 times, and the obtained extract was concentrated. The obtained ethyl acetate part was separated by reversed-phase high-performance liquid chromatography. The chromatographic column was a Shim-Pack RPC-ODS column (20mm×25cm; No.2282346493), and the detection wavelength was 214nm. The mobile phase used was methanol-water system at 8.0 mL / min flow rate for elution, the elution program is methanol 0 ~ 20min, 5% ~ 70%; 20 ~ 30min, 70% ~ 100%; 30 ~ 35min, 100% ~ 5% Chromatographic peak; then through reverse high performance liquid chromatography with 8.0mL / min 55% methanol isocratic e...
Embodiment 3
[0042] Compound Identification:
[0043] The present invention has anti-phytopathogenic fungal compound molecular formula according to mass spectrometry HR-ESI-MS + presumed to be C 11 h 12 N 2 o 2 ([M+Na] + 227.0792) such as figure 1 shown.
[0044] The NMR data and signal assignments are shown in Table 1.
[0045] Table 1 is the nuclear magnetic resonance data of the compound with anti-phytopathogenic fungus activity of the present invention
[0046]
[0047] according to 13 C-NMR and DEPT spectra show that its structure has 11 carbons, 1 methylene, 6 methines, and 4 quaternary carbons. According to the peak type of proton and H-C(7) and C(9), C(10); H-C(8) and C(9); H-C(9) and C(7), C(10); H-C(10) Long range correlation with C(4); H-C(6) and C(7), C(8); H-C(5) and C(1), C(2), C(8), C(9) , and H-C(7) and H-C(8); H-C(8) and H-C(7), H-C(10); H-C(9) and H-C(4); H-C(10) and H-C(8) have 1 H- 1 The relationship of H COZY, it can be speculated that the indole compos...
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