Bacillus megaterium and application thereof
A technology of Bacillus megaterium and plant growth-promoting bacteria, which is applied to Bacillus megaterium and its application field, can solve problems such as soil environment that may not be adapted to fluvo-aquic soil, and achieve good growth-promoting effect, improving utilization rate and high utilization rate.
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Embodiment 1
[0039] First prepare the following three media.
[0040] LB medium: peptone 10g, yeast extract 5g, sodium chloride 10g, agar 20g, distilled water 1000ml, pH7.0-7.2, sterilized at 121°C for 20min.
[0041] LB liquid medium: without agar, other conditions are the same as above.
[0042] Inorganic phosphorus bacteria medium (PKO medium): 5g tricalcium phosphate, 10g glucose, 0.5g ammonium sulfate, 0.3g sodium chloride, 0.3g magnesium sulfate heptahydrate, 0.3g potassium chloride, 0.03g manganese sulfate, heptahydrate Ferrous sulfate 0.03g, pH7.0 agar 20g, distilled water 1000ml, pH7.0~7.2. Sterilize at 121°C for 20min.
[0043] Inorganic salt medium: ammonium sulfate 2.0g; sodium dihydrogen phosphate 0.5g; dipotassium hydrogen phosphate 0.5g; magnesium sulfate heptahydrate 0.2g; calcium chloride dihydrate 0.1g, distilled water 1000mL, pH7.0, sterilized at 121℃ , 20min.
[0044] Weigh 10g of the fluvo-aquic soil collected from Banqiao, Nanjing, and put it in a 250ml Erlenmeyer ...
Embodiment 2
[0052] Aerobic test
[0053] Pour the sterilized LB culture medium into 3 sterilized test tubes, at about 2 / 3, on the aseptic operating table, pick up the JX15 (CGMCC No.5622) cultured on the slant with an inoculation needle, and puncture Inoculate into the above medium (must be pierced to the bottom of the tube). Cultivate at 30°C, and observe the results in 3 days to 7 days respectively. Those that grow on the surface of the agar column are aerobic bacteria, and those that grow along the puncture line are anaerobic or facultative anaerobic bacteria. The test results showed that JX15 (CGMCC No.5622) colonies grew along the surface of the agar column, and no colonies grew in the puncture line, which was strictly aerobic.
[0054] Determination of catalase
[0055] Put 1 drop of 3% H on a clean slide 2 o 2 , take 1 ring of 18~24h LB slant culture, in H 2 o 2 Smear in the middle, if bubbles are produced, it is positive, otherwise it is negative. The test results showed t...
Embodiment 3
[0081] In order to further verify the ability and optimal conditions of the plant growth-promoting bacteria JX15 (CGMCC No.5622) obtained in Example 1 to produce indole acetic acid, the following is to explore the indole acetic acid for different pH, liquid volume, different carbon sources, and different nitrogen sources. Effect on acetic acid production.
[0082] Put 25ml, 50ml, 75ml, 100ml, 150ml of LB liquid medium containing L-tryptophan (100mg / L) into a 250mL Erlenmeyer flask, and inoculate at 1% (v / v) inoculum in logarithmic growth phase After JX15 (CGMCC No.5622), put it at 30℃, 180r·min -1 Cultivate on a shaking table for 24 hours, and measure the amount of IAA produced by a quantitative method. The result is as figure 2 As shown, because the strain JX15 (CGMCC No.5622) has a good metabolism, the aeration rate affects the efficiency of the strain to produce IAA. When the liquid volume is 50mL, the strain produces the most IAA, and then as the liquid volume increases...
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