A strain of Bacillus amyloliquefaciens b4216 and its application
A technology of amylolytic spores, B4216CGMCCNO.18425, applied in the fields of environmental biotechnology and plant disease biological control, which can solve the problems of shortening the fermentation cycle, less research, and accelerating the decomposition of compost substrates, and achieves broad antibacterial spectrum, simple cultivation, and good Antagonistic effect
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Embodiment 1
[0029] Example 1 Isolation and identification of Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) strain B4216
[0030] Secondary fermentation samples of cow dung compost were collected, and the bacteria were isolated by beef extract peptone medium, and the strains that had inhibitory effect on Fusarium oxysporum f.sp.cubense were initially isolated by confrontation culture.
[0031] The preparation method of the above-mentioned beef extract peptone medium is as follows (take the preparation of 1L medium as an example): 3g beef extract, 10g peptone, 5g NaCl, 20g agar, 1000ml water, pH: 7.4-7.6, sterilized at 121°C for 30 minutes.
[0032] The isolated strain was named strain B4216, and its morphology was as follows: figure 1As shown, it is a rod-shaped Gram-negative bacteria that can form spores. The optimum growth temperature of the strain is about 37°C, and the optimum growth pH range is 5.0-8.8. Extract the genomic DNA of the strain, and use gyrA primers to carry o...
Embodiment 2
[0038] Antagonism of Example 2 bacterial strain B4216 to phytopathogenic fungi
[0039] Botrytis cinereal, Fusarium oxysporum f.sp.cubense, Fusarium oxysporum f.sp.cucumerinum, Phytophthora capsici, Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) strain B4216 was inoculated at a distance of 1.5 cm from the center of the wheat root rot fungus (Bipolarissorokiniana) bacteria cake disc, inoculated in the center of the PDA medium plate, and the control group was not inoculated with B4216, and placed in a dark place at 28 ° C. Photographs were taken after 120 h of cultivation. By observing the inhibition zone to determine whether the strain B4216 has an antagonistic effect on the phytopathogenic fungi, it was found that the bacterial strain B4216 has a good antagonism effect on the above-mentioned phytopathogenic fungi. ( figure 2 )
[0040] The experimental results show that: Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) B4216 has a good antagonistic effect o...
Embodiment 3
[0042] Application of embodiment 3 bacterial strain B4216 in secondary composting
[0043] 1) The bacterial strain B4216 was inoculated in LB liquid medium, and after 48 hours of liquid shake flask culture, the bacteria or spore count was greater than 1 × 10 8 cfu / ml culture solution.
[0044] The culture conditions are: initial natural pH, culture temperature 30° C., shaker 200 rpm for 48 hours. At this time, the bacterial solution contains more than 1 bacterium or spores × 10 8 cfu / ml;
[0045] The preparation method of described LB liquid culture medium is (take configuration 1L culture medium as example):
[0046] Add 10 g of tryptone, 5 g of yeast extract, and 10 g of sodium chloride into 950 ml of deionized water, dilute to 1000 ml with deionized water, and sterilize at 121°C for 20 minutes at natural pH.
[0047] 2) Prepare a mixed compost of chicken manure, cow dung and mushroom dregs (mass ratio 6:5:5) after primary fermentation.
[0048] Prepare the culture br...
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