A kind of plant endophytic fungus Penicillium breenensis f4a and its application
A technology of plant endophytic fungi and Penicillium, applied in the field of microorganisms, can solve the problems of low BFA production, achieve reduced treatment and pollution, good industrialization prospects, strong antibacterial and insecticidal effects
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Embodiment 1
[0029] The identification of embodiment 1 plant endophyte Penicillium brescens F4a
[0030] The plant endophytic fungus F4a was preserved in the China Center for Type Culture Collection on December 18, 2012. The address of the preservation unit is China. Wuhan. Wuhan University, the preservation number: CCTCC M 2012531, and the taxonomic name is Penicillium brezheng F4a ( Eupenicillium brefeldianum F4a).
[0031] Strain F4a grows well on PDA and Martin's medium; the early colonies are pure white, and gradually the center of the colony turns yellow-green; the surface of the colony is fluffy, without exudate and soluble pigment; after maturity, the colony has radial folds, and the center Slightly raised, forming a round plaque with a distinct color, yellowish on the back. Observed under the microscope, there is no enlarged apical capsule at the top, and the conidia structure is scarce, and the spores are born on the conidiophores in a broom shape. Size 2.5-2.0×2.2-1.6μm ( fig...
Embodiment 2
[0035] Example 2 Fermentation of strain F4a to prepare BFA
[0036] 2.1 Preparation of seed solution: first inoculate the strain F4a on the prepared PSA medium plate by streaking method, and culture at a constant temperature of 28°C for 5 days; In a 250mL Erlenmeyer flask filled with 50mL PSB medium, culture on a constant temperature shaker at 180rpm at 28°C for 72h;
[0037] The PSB medium is: 20.0g sucrose, 200g potato extract, balanced to 1L with distilled water, pH 5.0-8.0, sterilized at 115°C for 30min.
[0038] PSA is a solid medium with 18 g of agar added per liter of PSB.
[0039] 2.2 Fermentation in large bottles: use a 3L Erlenmeyer flask, fill each bottle with 0.70L of PSB medium, and sterilize at 115°C for 30 minutes before use. 50mL of inoculated seed solution per bottle, cultivated in a constant temperature shaker at 28°C and 180rpm for 7-13d, and fermented 10L in total;
[0040] 2.3 Obtaining crude extract: filter the obtained fermentation broth with 4-6 laye...
Embodiment 3
[0043] The structural analysis of embodiment 3 BFA
[0044] BFA is a colorless crystal, ESI-MS gives ( image 3 )m / z 303.2[M+Na] + (Calcd. for C 16 h 24 o 4 Na, 303.1572) infer that its molecular formula is C 16 h 24 o 4 . exist 1 H-NMR (600 MHz, CD 3 OD) spectrum (Table 1 and Figure 4 ), the low field region δ H 7.46 (1H, dd, J = 15.5, 2.2 Hz) and 5.82 (1H, d, J = 15.5 Hz) form a trans double bond, and one end is connected to the quaternary carbon; δ H 5.27 (H, dd, J = 15.1, 9.7 Hz) and 5.77 (1H, ddd, J = 15.1, 9.7, 4.4 Hz) form another trans double bond, and the two ends are respectively connected to CH and CH 2 group. Hydrogen δ on 3 oxygenated carbons H 4.04 (1H, brd, J = 9.1 Hz), 4.80 (1H, m) and 4.21 (1H, m) and the signal δ of a methyl proton attached to CH H 1.24 (3H, d, J = 6.2 Hz). 13 C-NMR (150 MHz, CD 3 OD) gives 16 carbon signals (Table 1 and Figure 5 ), where one ester carbonyl carbon signal δ C 166.96; 4 unsaturated carbon signals δ C 1...
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