Serratia marcescens biocontrol bacterium for efficiently inhibiting aspergillus flavus compounded aflatoxin and its application
A technology of Serratia marcescens and aflatoxin, applied in the field of microbiology, can solve the problem that Serratia marcescens inhibits Aspergillus aflatoxin, etc.
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Embodiment 1
[0017] 1) Activate Serratia marcescens 3J4SM on LB plates, culture them in a 37°C incubator for 24 hours, pick a single colony of the activated Serratia marcescens with a pick, and transfer to a plate containing 15 mL of LB liquid In the Erlenmeyer flask of the culture medium, shake culture at 28°C, 200r·min-1 for 12h. Aspirate 1% of the culture solution and transfer it to a Erlenmeyer flask filled with 15mL LB liquid medium for culture, and culture with shaking at 28°C and 200r·min-1 for 12h to obtain the fermentation broth of the antagonistic strain.
[0018] 2) Serratia marcescens fermentation broth (final concentration of 1 × 107 CFU / mL) was cultured for 7 days and vigorously growing Aspergillus flavus suspension (final concentration of spores was 5.0 × 10 5 Spores mL-1) were cultured together in Sabouraud liquid medium at 28°C and 200 rpm for 5 days, and each treatment was repeated 3 times.
[0019] 3) Measure the content of aflatoxin B1 in its culture solution (Table 2)...
Embodiment 2
[0024] 1) Take Zhonghua No. 6 peanut grains from the peanut field in Hubei and grind them into powder, weigh 1g of peanut powder in a petri dish, and add 1ml of Aspergillus flavus spore liquid (5×10 5 cells / mL) and 1ml CCTCC M 2017328 bacterial fluid (1×107 CFU / mL), with Sabouraud medium instead of CCTCC M 2017328 bacterial fluid as the control;
[0025] 2) Incubate the inoculated peanut powder in an incubator at 28° C. for 9 days, add 15 mL of 70% methanol water, vortex and put it on a shaker for 30 minutes. Take 3mL supernatant, add 8mL ultrapure water and vortex centrifuge;
[0026] 3) Take 8 mL of the supernatant and use the immunoaffinity column-HPLC method to determine the content of aflatoxin B1 (Table 3), and there are 3 repetitions in the experiment.
[0027] Table 3 Control effect of biocontrol bacteria on peanut aflatoxin
[0028]
[0029] It can be seen from the experimental results that the inhibition rate of CCTCC M 2017328 strain on the toxin production of ...
Embodiment 3
[0031] 1) Take 10 grains of Luhua No. 8 peanuts from Anhui peanut field, coat the surface of the peanuts with Serratia marcescens 3J4SM fermentation liquid, and add 1ml of Aspergillus flavus spore liquid (5×10 5 Individual / mL), with Sabouraud medium instead of CCTCC M 2017328 fermentation broth as a control;
[0032] 2) Cultivate the inoculated peanut grains in an incubator at 28°C for 9 days, then grind the peanut grains into peanut powder, add 15mL of 70% methanol water, vortex and place on a shaker for 30min. Take 3 mL supernatant, add 8 mL ultrapure water and vortex centrifuge;
[0033] 3) Take 8 mL of the supernatant and use the immunoaffinity column-HPLC method to determine the content of aflatoxin B1 (Table 4), and there are 3 repetitions in the experiment.
[0034] Table 4 Control effect of biocontrol bacteria on peanut Aspergillus flavus
[0035]
[0036] From the experimental results, it can be seen that the inhibition rate of CCTCC M 2017328 strain on Aspergill...
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