Agrobacterium sp. DP3, microbial agent comprising Agrobacterium sp. DP3 and application in field of biological fertilizer preparation
A technology of Agrobacterium and bio-fertilizer, applied in soil conditioning materials, microbial-based methods, applications, etc., can solve the problems that plants cannot directly absorb and utilize
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Embodiment 1
[0051] Example 1 Screening and Identification of Salt-tolerant Phosphate-Solubilizing Bacteria
[0052] 1. Configuration of culture medium
[0053] LB liquid medium: tryptone 5g / L, yeast extract 2.5g / L, sodium chloride 5g / L, deionized water 1000mL, pH=7.4.
[0054] LB solid medium: tryptone 5g / L, yeast extract 2.5g / L, sodium chloride 5g / L, agar 20g / L, deionized water 1000mL, pH=7.4.
[0055] PKO liquid medium: glucose 10g / L, tricalcium phosphate 5g / L, ammonium sulfate 0.5g / L, sodium chloride 0.2g / L, potassium chloride 0.2g / L, magnesium sulfate heptahydrate 0.3g / L, sulfuric acid Manganese 0.03g / L, ferrous sulfate 0.003g / L, yeast powder 0.5g / L, distilled water 1000mL, pH=7.0±0.2, high temperature and high pressure sterilization at 121℃ for 30min.
[0056] PKO liquid medium: glucose 10g / L, tricalcium phosphate 5g / L, ammonium sulfate 0.5g / L, sodium chloride 0.2g / L, potassium chloride 0.2g / L, magnesium sulfate heptahydrate 0.3g / L, sulfuric acid Manganese 0.03g / L, ferrous sulfate...
Embodiment 2
[0077] The saline-alkali tolerance test of embodiment 2 phosphate-dissolving bacteria
[0078] 1. Solution configuration
[0079] Configuration of different gradient salinity solutions: set the salinity to 0.1g / L, 0.5g / L, 1.0g / L, 1.5g / L, 3.0g / L, set the alkalinity to 7.2, 7.6, 8.0, 8.4, A total of 20 treatments. The salinity is prepared with sodium chloride, and the alkalinity is adjusted with hydrochloric acid and sodium hydroxide.
[0080] The configuration of the non-salinity LB liquid medium: tryptone 5g / L, yeast extract 2.5g / L, deionized water 1000mL, pH natural.
[0081] 2. Determination of the growth curve of DP3 under different salinity
[0082] Inoculate strain DP3 into liquid LB medium and cultivate to OD=0.1 (bacteria density is about 10 8 cells / mL) and stored in a 4°C refrigerator for later use.
[0083] Inoculate the bacterium liquid with OD=1.0 with 5% inoculation amount into the culture medium with different salinity and alkalinity containing 10% non-salini...
Embodiment 3
[0085] Corn germination experiment under the inoculation of embodiment 3 phosphorus-solubilizing bacteria DP3
[0086] Select large, round and plump corn seeds, soak them in 70% absolute ethanol for 2 minutes for disinfection, and rinse them with tap water. Cover each petri dish with filter paper, sow 10 corn seeds, inoculate 10 9 DP3 bacteria and irrigated with water with different saline-alkaline gradients. The salinity gradient is 0.1g / L, 0.5g / L, 1.0g / L, 1.5g / L, 3.0g / L, the alkalinity gradient is 7.2, 7.6, 8.0, 8.4, a total of 20 treatments. Two parallels were set up for each saline-alkaline gradient. Bacillus megaterium, which also has the ability to dissolve phosphorus, was purchased from the China General Microorganism Culture Collection Center (CGMCC) as a control. Observe how the corn seeds germinate after a week.
[0087] The results showed that the germination rate of corn seeds inoculated with DP3 could reach more than 70% at any alkalinity and salinity of 3.0g / L,...
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