Bacillus velezensis, microbial agent thereof, microbial agent preparation method and application
A microbial agent, bacillus technology, applied in the field of biological control, to achieve the effect of reducing the occurrence of diseases, ensuring the number of antagonistic bacteria, and strengthening the inhibitory effect of the plate
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[0048] The preparation method of microbial agents, including the following steps,
[0049] (A) Inoculation of Belles Bacillus to LB ramp medium;
[0050] (B) Protecting the activated Belles branch bacterium to the LB liquid medium to obtain a culture medium;
[0051] (C) Placing the culture solution in the shaker, at a temperature of 120 rpm / min ~ 180 rpm / min and 10 ° C to 50 ° C, 10 h ~ 84h, i.e., a liquid fungicide was prepared. More preferably, the culturing temperature of the Belets Bacillus is 25 ° C to 35 ° C.
[0052] The NaCl concentration in the culture solution was 0.5% to 4%; the pH of the culture solution was 4.0 to 8.0.
[0053] The growth of Belles Bacillus in the culture solution is based on β-formyl-D-glucoside, α-D-glucose, D-mannose, D-fructose or D-galactose or A variety of monosaccharide compounds as a carbon source; the growth of Belets Bacillus, one or more of D-maltose, D-trehalose, α-D-lactose or honeydose compounds As a carbon source; the growth of Bel...
Embodiment 1
[0063] Example 1: Separation, screening and identification of Bacilli Bacillus Velezensis CGMCNS VELEZENSIS CGMCNS19860
[0064] (1) Separation and screening of antagonistic bacteria
[0065] Gradient dilution separation method: Choose a robust tomato plant in the pathogenesis in the Wenzhou area, gently bring the soil and discharge plant, gently shake the loose attachment of the replacement plant, cut off the roots, weigh 10g plant roots and Root soil, placed in conical bottles with sterilized glass beads and 90 ml sterile water, temperature and 150R · min -1 The stirring speed is oscillated for 30 min, resulting in a concentration of 10 -1 Root soil suspension, then dilute into 10 using gradient concentration dilution -4 10 -5 And 10 -6 Diluent; each take 0.1ml 10 -4 10 -5 And 10 -6 The dilutions of the three gradients were coated on the LB medium, respectively, each gradient repeated 3 times, inverted culture 18h under temperature conditions at 30 ° C, and picked a single typic...
Embodiment 2
[0079] Example 2: Growth conditions of Bacillus Bacillus Bacilli Bacillus Velezensis Cgmcc No.19860
[0080] (1) Effect of temperature on the growth of strains WSW2
[0081] The medium was produced by LB medium, the initial pH was 7.0, and the shake bottle was 50 ml / 250 ml, the inoculation amount was 1% (V / V), and the culture temperature was set to 10 ° C, 20 ° C, 30 ° C, 40 ° C and 50 ° C, shaker rotation speed 150 rpm, culture time 24 h, the end of different temperatures at different temperatures were measured after the end of culture. 600 Value, each temperature is repeated 3 times.
[0082] RESULTS: The strain WSW2 cultured at different temperatures for 24h bacterial liquid OD 600 Such as Figure 5 As shown, from Figure 5 It can be seen in that, when the culture temperature is 10 ° C, the concentration of strain WSW2 is low, and the average OD is. 600 0.085, strain WSW2 is basically not yet grown; the growth amount of strains at 30 ° C is the highest, OD 600 It reached 2.50...
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