Coral-associated marine bacillus amyloliquefaciens strain CoMb-9 and application thereof
A technology of amylolytic spores and comb-9, applied in the field of agricultural biology, can solve problems such as structural damage, soil compaction, and deterioration of agricultural product quality, and achieve good growth promotion, plant growth, and good application prospects
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Embodiment 1
[0039] Example 1 Isolation and identification of coral symbiotic marine bacillus amyloliquefaciens strain CoMb-9
[0040] 1. Isolation of strains
[0041] Take healthy living coral, wash the surface with sterile distilled water, add sterile seawater to crush, and after standing still, take the leaching solution and spread it on a seawater nutrient agar medium plate, and after cultivating in the dark at 25°C for 48 to 72 hours, select round, Pale milky yellow, raised in the middle, opaque, rough and dry surface, and a single colony with serrated edges. After purification, it is temporarily stored in a 4°C refrigerator with seawater NA medium.
[0042] Wherein, the formula of the seawater nutrient agar medium: 3.0g of beef extract, 1.0g of yeast extract, 5.0g of peptone, 10.0g of glucose, 20.0g of agar, 1000mL of aged seawater, pH 7.0-7.5; referred to as seawater NA.
[0043]2. Antibacterial experiment
[0044] The antagonism of the above-mentioned isolated bacterial strains t...
Embodiment 2
[0056] The bacteriostatic active substance that embodiment 2 bacterial strain CoMb-9 secretes produces and the optimization of culture condition
[0057] 1. In order to make the strain CoMb-9 better secrete and produce antibacterial active substances, the culture conditions of the optimized strain were determined by single factor and response surface methods, including culture medium, culture temperature, initial pH of the culture medium, culture time, inoculum size , speed, liquid volume, etc.
[0058] 2. Results
[0059] The results show that the best culture medium for the secretion of coral symbiotic marine bacillus amyloliquefaciens strain CoMb-9 to produce active substances that inhibit Phytophthora soybean and other phytopathogens is GBE medium, and its formula is: beef extract 10.0 g, glucose 25.0g, MgSO 4 0.5g, KH 2 PO 4 0.5g, distilled water 1000mL.
[0060] The optimal culture conditions are: culture temperature 18℃, culture medium initial pH 7.3, culture time ...
Embodiment 3
[0063] Colonization of Example 3 bacterial strain CoMb-9 in soybean
[0064] 1. The coral symbiotic marine Bacillus amyloliquefaciens strain CoMb-9 of the present invention can grow in a rifampicin concentration of 500 μg / mL by inducing resistance to rifampicin and at the same time can antagonize Phytophthora sojae Double antibody marker CoMb-9 for Phytophthorasojae Rif Strain, take double antibody labeled CoMb-9 Rif Strain fermentation broth (concentration is 1.0×10 7 cfu / mL) to evenly water the rhizosphere soil of soybean plants, use sterile water as a control, use rifampicin-containing medium to recover the marker strains in soybeans after treatment, and determine the antagonistic effect of the recovered strains on Phytophthora soybean.
[0065] 2. The results showed (Table 2) that the marker strain CoMb-9 Rif The marked strains could be isolated from soybean roots, stems and leaves after root irrigation treatment, which indicated that the strains could enter soybean pla...
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